Downregulation of Yap1 during limb regeneration results in defective bone formation in axolotl.

The Hippo pathway plays an imperative role in cellular processes such as differentiation, regeneration, cell migration, organ growth, apoptosis, and cell cycle. Transcription coregulator component of Hippo pathway, YAP1, promotes transcription of genes involved in cell proliferation, migration, differentiation, and suppressing apoptosis. However, its role in epimorphic regeneration has not been fully explored. The axolotl is a well-established model organism for developmental biology and regeneration studies. By exploiting its remarkable regenerative capacity, we investigated the role of Yap1 in the early blastema stage of limb regeneration. Depleting Yap1 using gene-specific morpholinos attenuated the competence of axolotl limb regeneration evident in bone formation defects. To explore the affected downstream pathways from Yap1 down-regulation, the gene expression profile was examined by employing LC-MS/MS technology. Based on the generated data, we provided a new layer of evidence on the putative roles of increased protease inhibition and immune system activities and altered ECM composition in diminished bone formation capacity during axolotl limb regeneration upon Yap1 deficiency. We believe that new insights into the roles of the Hippo pathway in complex structure regeneration were granted in this study.

Tissue Factor and Vascular Endothelial Growth Factor in Detecting Thromboembolic Complications in Diabetic Atherosclerotic Patients.

Background: Atherosclerosis, which is one of the leading causes of death all over the world, can create major or minor thromboembolic complications with the exponentially increasing diabetic status. Despite all the studies, the mechanism by which endothelial damage in atherosclerosis is triggered in diabetic setting is still not fully understood. Methods: In this study, tissue factor (TF), which is thought to act together in the formation of vasular endothelial growth factor (VEGF-A) and coagulopathy in diabetic atherosclerotic patients, may be an important indicator in this regard, a total of 100 cases who were undergone off-pump coronary artery bypass (OPCAB) which were at same risk group examined by dividing into diabetic status. Early postoperative process and biochemical parameters analyzed in terms of TF and VEGF-A levels measured before and after the operation. Results: TF and VEGF-A expression of the T1DM group were statistically high compared to non-diabetics. Significantly longer hospital stays with changes in TF and VEGF-A were found in patients in the diabetic group compared to pre- and postoperatively, respectively; TF (95% CI: 0.879-0.992; p = 0.025), VEGF-A (95% CI: 0.964-0.991; p = 0.001) and hospital stay (95% CI: 1.96-7.49; p = 0.0001). Preoperatively measured carotid intima-media thickness (CT) was higher in diabetics and was significantly associated with atrial fibrillation (AF), (r = 0.873). Surgical team and protocols were same and OPCAB procedures were routinely applied to all patients in our clinic. No minor or major events were observed in any of the cases. Conclusion: TF and VEGF-A values in patients with diabetic atherosclerosis may be important in the early detection of thromboembolic complications.

Phosphoproteomic strategies in cancer research: a minireview.

Understanding the cellular processes is central to comprehend disease conditions and is also true for cancer research. Proteomic studies provide significant insight into cancer mechanisms and aid in the diagnosis and prognosis of the disease. Phosphoproteome is one of the most studied complements of the whole proteome given its importance in the understanding of cellular processes such as signaling and regulations. Over the last decade, several new methods have been developed for phosphoproteome analysis. A significant amount of these efforts pertains to cancer research. The current use of powerful analytical instruments in phosphoproteomic approaches has paved the way for deeper and sensitive investigations. However, these methods and techniques need further improvements to deal with challenges posed by the complexity of samples and scarcity of phosphoproteins in the whole proteome, throughput and reproducibility. This review aims to provide a comprehensive summary of the variety of steps used in phosphoproteomic methods applied in cancer research including the enrichment and fractionation strategies. This will allow researchers to evaluate and choose a better combination of steps for their phosphoproteome studies.

Dose-dependent effects of vitamin 1,25(OH)2D3 on oxidative stress and apoptosis.

Background The purpose of this study is to examine the dose-dependent effects of vitamin 1,25(OH)2D3 on apoptosis and oxidative stress. Methods In this study, 50 male Balb/c mice were used as control and experiment groups. The mice were divided into 5 groups each consisting of 10 mice. Calcitriol was intraperitoneally administered as low dose, medium dose, medium-high dose and high dose vitamin D groups (at 0.5, 1, 5 and 10 µg/kg, respectively), for three times a week during 14 days. At the end of the study, annexin V was measured by enzyme-linked immunosorbent assay method, and total antioxidant capacity and total oxidant status values were measured by colorimetric method in serum. Hematoxylin eosin staining was performed in liver tissues and periodic acid schiff staining was performed in kidney tissues. Results While comparing the results of medium-high dose (5 µg/kg) and high dose (10 µg/kg) vitamin D administration to that of the control group, it was observed that serum antioxidant status and annexin V levels decreased and glomerular mesenchial matrix ratio increased in kidney (p<0.05). In addition to these findings, in the group receiving high dose vitamin D (10 µg/kg), it was observed that the damage to the liver increased together with the the oxidative stress index values (p<0.05). Conclusions As a result, this study was the first in the literature to report that use of high-dose vitamin D (10 µg/kg) results in oxidant effect, rather than being an antioxidant, and causes severe histopathological toxicity in the liver and kidney.

Effects of altered thyroid states on oxidative stress parameters in rats.

BACKGROUND: Thyroid hormones are effective on oxidant-antioxidant balance by leading basal metabolic rate. In this study, the effects of altered thyroid states on low density lipoprotein (LDL) oxidation and oxidative stress parameters were investigated in an experimental animal model. METHODS: Thirty female Wistar Albino rats were equally divided into 3 groups as follows: control group; hypothyroid group (methimazole (75 mg/100 g was added to diet); hyperthyroid group [L-thyroxine (0.4 mg/100 g was added to diet)]. Oxidized LDL (ox-LDL) levels, thyroid, and lipid parameters were determined in serum. Also lipid peroxidation (LPO), sialic acid (SA) and glutathione levels (GSH), as well as superoxide dismutase (SOD) and catalase (CAT) activities were determined in tissue samples. RESULTS: A significant increase in lipid parameters was observed in hypothyroid group, whereas these parameters were decreased in hyperthyroid group compared to control group. For ox-LDL levels, a significant increase was observed both in hypothyroid and hyperthyroid groups. In brain, liver and kidney tissues, LPO and SA levels were increased, whereas GSH levels were decreased both in hypothyroid and hyperthyroid groups. The SOD and CAT activities were significantly decreased in hypothyroid group, however, they were increased in hyperthyroid group compared to control group. Both hypothyroid and hyperthyroid conditions modify the oxidant-antioxidant state in serum and tissues. CONCLUSIONS: Increased SOD and CAT activities in hyperthyroid group suggest that elevated thyroid hormones can reduce oxidative stress by maintaining antioxidant defense and they might have a protective effect on some tissues against oxidants.

Galectin-3 and plasma cytokines in patients with acute myocardial infarction.

OBJECTIVE: To investigate the concentrations of plasma cytokines and Galectin-3 (Gal-3) as inflammatory markers in patients with acute myocardial infarction (AMI). METHODS: The study population consisted of 29 patients with AMI and 29 healthy control subjects. We measured Gal-3, tumor necrosis factor-alpha (TNF-α), and interleukin-6 (IL-6) levels in plasma using enzyme-linked immunosorbent assays (ELISAs). We measured levels of C-reactive protein (CRP) via the nephelometric method. RESULTS: Patients with AMI showed significantly higher plasma Gal-3, TNF-α, and IL-6 levels compared with controls. Gal-3 levels were positively and significantly correlated with plasma IL-6, TNF-α, and CRP levels in the control and patient groups. CONCLUSION: Our findings suggest that Gal-3 can be a new circulating biomarker of inflammation associated with AMI.

Levels of C-reactive protein and protein C in periodontitis patients with and without cardiovascular disease.

Since periodontitis is a chronic and inflammatory disease, a number of hypotheses have proposed that it has an etiological or modulating role in cardiovascular disease (CVD). This study aimed to ascertain the changes in the plasma levels of C-reactive protein (CRP) and protein C (PC), a natural anticoagulant also having an anti-inflammatory effect, in patients who have mild-to-severe periodontitis with or without CVD. The test group consisted of 26 patients with CVD and chronic periodontitis and the control group consisted of 26 patients with chronic periodontitis and no systemic disease. In both groups Community Periodontal Index of Treatment Needs scores were recorded and blood samples were collected. CRP levels were significantly high and PC activity was significantly low in the test group compared to the control group (p < 0.001). There was a negative correlation between tooth loss and PC and between CRP and PC. How PC is affected by the inflammatory events and its association with CRP is an active area of investigation.

Serum and saliva sialic acid in periodontitis patients with and without cardiovascular disease.

Serum total sialic acid (sTSA) has recently been shown to be a cardiovascular risk factor. However, there is little information about the role of sTSA and TSA in saliva in periodontitis, a chronic and inflammatory disease known to be a risk factor for cardiovascular disease (CVD). We aimed to investigate the changes in sTSA and TSA levels in saliva in patients having both periodontitis and CVD versus periodontitis patients without diagnosed CVD. The study group consisted of 26 patients with proven periodontitis and 26 controls with no diagnosed systemic disease but periodontitis. sTSA and saliva TSA levels were determined by the thiobarbituric acid method, and C-reactive protein (CRP) was evaluated by the nephelometric method. The severity of periodontitis has been determined by the community periodontal index of treatment needs (CPITN). TSA in blood and saliva and CRP levels in blood were significantly increased in CVD patients compared with the control group. CPITN ranged from 2 to 4 in both groups. Significant and positive correlations were found between sTSA and saliva SA levels in patients and controls and between tooth loss and TSA both in blood and saliva. Therefore, TSA in saliva may be a useful marker similar to sTSA in CVD patients.

Effects of short-term streptozotocin-induced diabetes and vitamin C on platelet non-enzymatic glycation.

Diabetes mellitus is one of the most prevalent metabolic syndromes worldwide. Glycation, a chemical modification of proteins with reducing sugars, indicates a possible explanation for the association between hyperglycemia and the wide variety of tissue pathologies. Non-enzymatic glycation (NEG) of platelet proteins is one of the key mechanisms in the pathogenesis of diabetic complications and may be significant in diabetic atherothrombosis. The aim of this study was to investigate the effects of streptozotocin (STZ)-induced short-term experimental diabetes on the glycation of platelets and to find out if vitamin C affected this glycation. A total of 40 male Wistar albino rats, 200-250 g, were randomly divided into 4 groups (2 diabetic and 2 control groups). The diabetic groups were made diabetic by intraperitoneal injection of STZ (65 mg/kg, citrate buffer pH 4.5). By daily intraperitoneal injection, 80 mg/kg vitamin C (Roche, Turkey) was administered until the end of the experiment. Blood glucose levels of the diabetic groups were significantly higher than those at day 0 and also higher than those of the non-diabetic control groups. The changes in total protein, NEG and vitamin C levels were not statistically significant. Although the differences among the groups were not statistically significant, vitamin C administration increased NEG levels in the diabetic group. The results of this study demonstrate that 8 days of STZ-induced short-term diabetes did not cause a significant increase in NEG of platelets. However, the effect of vitamin C on platelet NEG needs to be further investigated.

Investigation of inflammatory and hemostatic parameters in female patients undergoing total knee arthroplasty surgery.

Tendency to hypercoagulation is a common phenomenon in primary osteoarthritis patients (POA) undergoing total knee arthroplasty (TKA) surgery, but the clinical implications of this condition are not clear. Therefore we aimed to evaluate the inflammatory and coagulation parameters in the patient group and find a possible explanation for the tendency to hypercoagulation occurring in plasma and synovia of inflamed joints. Of the evaluated factors involved in inflammation and coagulation, galectin-3, C reactive protein (CRP), erythrocyte sedimentation rate (ESR), fibrinogen, FVIIa:C, FXII:C, and platelet count increased, whereas tissue factor (TF) activity in synovia, PT, APTT and FVII:C in plasma and synovia were decreased. In conclusion, activation of inflammation and tendency to hypercoagulation is observed in preoperative plasma and synovia of patients undergoing TKA surgery.

[Thromboelastography and its use in cardiac surgery]. / Tromboelastografi ve kalp cerrahisinde kullanimi.

Thromboelastography is an alternative method to conventional coagulation tests for the general evaluation of hemostatic system. Cardiac surgery with cardiopulmonary bypass is accomplished by complex alterations of hemostasis, including acquired dysfunction of platelets, consumption coagulopathy and increased fibrinolysis. Despite major advances in blood conservation methods and perioperative care of the patients, transfusion rates in cardiac surgery remain high. Thromboelastography has an ability to assess almost all components of haemostatic system globally. Currently, thromboelastography is used with standard coagulation tests to decrease the microvascular bleeding and homologous blood transfusion in cardiac surgery with cardiopulmonary bypass. In this review, we aimed to discuss thromboelastography technology and its usage in cardiac surgery.

The relation between plasma tissue factor and oxidized LDL levels in acute coronary syndromes.

AIM: Tissue factor (TF) is a low-molecular-weight glycoprotein responsible for the initiation of the coagulation cascade. The relation between oxidized low-density lipoprotein (Ox-LDL), that has been shown to be involved in atherogenesis, and TF has not been evaluated before in circulating plasma. The aim of this study was to determine plasma levels of TF and Ox-LDL in acute coronary syndrome (ACS) and stable coronary artery disease (SCAD). METHODS: The study group consisted of 41 patients with ACS and 26 patients with SCAD. Among the ACS patients, 12 were diagnosed with unstable angina pectoris (UAP) and 29 were diagnosed with acute myocardial infarction (AMI). The control group consisted of 30 healthy volunteers. TF and Ox-LDL levels were evaluated by ELISA kits. RESULTS: Ox-LDL levels were significantly higher in UAP and AMI patients compared with the control (p < 0.001) and SCAD (p < 0.01 and p < 0.001, respectively) groups. TF levels were significantly higher in the UAP, AMI and SCAD groups compared with the control group (p < 0.001, p < 0.001 and p < 0.01, respectively). In the AMI group a significant increase was observed in TF levels when compared with the SCAD group (p < 0.01). Plasma Ox-LDL levels were significantly and positively correlated with TF levels in the UAP and AMI groups (p < 0.05, r = 702, and p < 0.0001, r = 0.679, respectively). CONCLUSION: The potential link between Ox-LDL and TF in circulating blood in ACS may strengthen the evidence supporting a relationship between oxidant stress, lipids and thrombosis and consequently may contribute to understanding the mechanism through which Ox-LDL and TF may mediate the pathogenesis of CAD.

Effect of fenofibrate on serum and tissue sialic acid levels in short-term experimental hypercholesterolemia.

In recent years, sialic acid is considered to be a possible marker for cardiovascular diseases. The aim of this study was to investigate the effect of two different treatment periods of fenofibrate (CAS 49562-28-9) on serum, heart and liver sialic acid levels in experimental hypercholesterolemia. Serum, heart and liver total sialic acid levels were determined by Warren's thiobarbituric acid (TBA) method and serum lipid levels by commercial kits at the end of the fenofibrate treatment for 3 and 6 weeks. Fenofibrate treatment reduced serum total sialic acid levels significantly in the control and hypercholesterolemic groups at the end of the 3rd week and only in the control group at the end of the 6th week. Serum sialic acid levels of fenofibrate-treated hypercholesterolemic rats in the 6-week period were significantly higher than those in the 3-week period. Neither the hypercholesterolemic diet nor fenofibrate had any significant effect on heart and liver sialic acid levels. In conclusion, decreased serum sialic acid levels in control and hypercholesterolemic groups by fenofibrate short-term treatment may contribute to the decreased risk of cardiovascular diseases that accompanies the hypercholesterolemic complications. The decreased serum sialic acid levels further indicate the clinical efficacy of fenofibrate as an anti-inflammatory agent.

Effects of glibornuride versus metformin on eye lenses and skin in experimental diabetes.

The aim of the study was to investigate the effect of glibornuride (CAS 26944-48-9) and metformin (CAS 657-24-9) on eye lenses and skin of streptozotocin (STZ)-induced diabetic rats. The drugs were administered daily to one diabetic and one control group separately from day 14 to day 42. After 42 days, diabetes caused significant increases in blood glucose levels, non-enzymatic glycosylation (NEG) of skin and lens proteins and skin lipid peroxidation (LPO) levels as well as decreases in body weights and lens glutathione (GSH) levels. Metformin administration to the diabetic rats produced more significant reduction in blood glucose than glibornuride. Metformin produced non-significant increase in NEG levels in lenses and skin. Unlike metformin, glibornuride increased NEG levels significantly in lenses. Both drugs produced non-significant increase in lens GSH levels and decreases in skin LPO levels in diabetic rats. Sodium dodecyl sulphate (SDS) polyacrylamid gel electrophoresis revealed no significant difference in any of the protein bands between any of the groups. These observations suggest that metformin and glibornuride as oral antidiabetics have similar protective effects on tissues in STZ induced diabetic rats.

Melatonin reduces oxidative damage to skin and normalizes blood coagulation in a rat model of thermal injury.

This study was designed to determine the effect of melatonin treatment on the glutathione (GSH) and lipid peroxidation (LPO) levels in the skin as well as prothrombin time (PT) and fibrin degradation products (FDPs) in the blood of rats with thermal injury. Under ether anaesthesia, the shaved dorsum of the rats was exposed to 90 degrees C bath for 10 s to induce burn injury. Rats were decapitated either 3 or 24 hours after burn injury. Melatonin (10 mg/kg) was administered i.p. immediately after burn injury to same animals. In the 24 hour burn group, melatonin injections were repeated for two more occasions 8 and 16 h after burn injury. In the control group the same protocol was applied except that the dorsum was exposed to a 25 degrees C water bath for 10 s. Severe skin scald injury (30% of total body surface area) caused a significant decrease in PT at post burn 3 and 24 hours. FDPs was not increased at post burn 3 hour but was significantly increased at post burn 24 hour. GSH levels were significantly depressed at post burn 3 hour but were not changed at post burn 24 hour. LPO levels were significantly increased both at post burn 3 and 24 hours. Skin protein levels were significantly reduced at post burn 24 hour as evidenced by electrophoresis. Treatment of rats with melatonin normalized PT levels both at post burn 3 and 24 hours. FDP decreased at post burn 24 hour due to melatonin treatment. GSH levels significantly increased as a result of melatonin treatment both at post burn 3 and 24 hours melatonin treatment. LPO levels were not changed by melatonin at post burn 3 hour; however, the melatonin significantly decreased LPO values at post burn 24 hours. In conclusion, exogenously administered melatonin reduced skin oxidant damage and normalized the activated blood coagulation induced by thermal trauma.

6,7-Dihydroxy-3-phenylcoumarin inhibits thromboplastin induced disseminated intravascular coagulation.

6,7-Dihydroxy-3-phenylcoumarin (DHPC) was tested to determine whether it had any effect on vitamin K inhibition, by investigating the prothrombin time (PT), activated partial thromboplastin time (APTT), fibrinogen level and platelet count. The anticoagulant and antithrombotic effects of DHPC were compared with those of warfarin by conducting a 4 h acute trial on thromboplastin-induced disseminated intravascular coagulation (DIC), investigating various haemostatic and antioxidant system parameters and performing a haemogram. Of most significance was that in the 5-d DHPC trial on healthy controls, PT, APTT, fibrinogen, platelet count remained within normal levels. In the 4-h DIC trial, both DHPC (0.025 mg/kg, i.v.) and warfarin (0.25 mg/kg, i.v.) significantly inhibited DIC, by reducing the PT, APTT, and fibrin degradation products and increasing fibrinogen levels and platelet count. In the DIC drug groups, lipid peroxidation significantly increased only in the warfarin group and glutathione significantly increased only in the DHPC group. However leucocyte count was significantly higher in the DHPC than the warfarin group. Further investigation is required for why DHPC is effective on the parameters investigated, at doses one-tenth of those of warfarin.

Biochemical and morphological alterations in lungs induced by experimental inhibition of fibrinolytic activity.

The fibrinolytic system is known to play an important role in the protection of lung architecture and function. This study investigated the effects on lungs of inhibiting the fibrinolytic system using tranexamic acid (TXA). Thirty cats were used, 15 experimental and 15 control. TXA was administered intravenously to the experimental animals for 3 h at 200 mg/kg (acute) and 7 days at 100 mg/kg (chronic). Blood samples were obtained from the carotid artery. The acute dose cats were sacrificed at 3 h and 24 h and the chronic dose cats at 8 days. Samples of inflated and fixed lung were examined morphologically and their collagen contents were determined. Fibrinolytic activity in blood samples was determined by fibrinogen degradation products levels, fibrin plate lytic area diameter, and the euglobulin lysis time. Hyperemia, lung interstitial oedema, haemorrhaging, inflammatory cell infiltration, pneumocyte type II cell proliferation, thrombosis and emphysema-related changes, characterized by enlargement of air spaces accompanied by destruction of alveolar walls, were observed in experimental cats group. None of these alterations except hyperemia and lung interstitial oedema were observed in two control animals. Electron microscopy results revealed oedema fluid in the interstitium, proliferation of pneumocyte type II cells, thickening of the alveolar septa and presence of marked amounts of collagen. Vacuoles were seen in the capillary endothelial cells. Elastic tissue was observed as elastic masses and partly disrupted, although elastic fibers were not prominent in all parts of the interstitium. Collagen content in the chronic dose experimental group was significantly higher than in all control and acute dose experimental groups. The inhibition of fibrinolytic system appears to have caused the emphysematous alterations, alveolar wall destruction and collagen accumulation possibly by causing microthromboses leading to mechanical blockage-ischemic changes, or by causing secondary fibrinolysis as a result of fibrin degradation products affecting local plasminogen activators and proteases. An injury-repair process also appears to have occurred.