Metabolomic profiling of liver tissues after acute administration of vardenafil in rats.

Erectile dysfunction (ED) diseases have almost affected 100 million men all over the world. Orally administered phosphodiesterase 5 (PDE 5) inhibitors are the most used pharmaceutical formulations for the treatment of ED. In this study, it is aimed to investigate the metabolomics feature of orally administered vardenafil in rats. To carry out the experimental procedure eight male Wistar albino rats were used. Their livers were gently removed and metabolomics profiles of each sample were determined by UPLC Q-TOF MS. Identification of metabolites was achieved by the METLIN database. Cluster analysis was also performed via Principle Component Analysis. Several metabolites were identified and results were evaluated by XCMS software. UPLC Q-TOF MS could be successfully applied to profile biomarkers and help us understand the molecular mechanisms of vardenafil usage. It was concluded that the level of some metabolites, responsible for the collagen synthesis and Kreb's cycle, has been statistically significant after the vardenafil administration.

Nitrite, neopterin levels and tryptophan degradation in allergic conjunctivitis.

PURPOSE: The study aims to evaluate changes in neopterin levels and tryptophan degradation which are induced by Th1-type immune response and nitric oxide metabolism which may be involved in allergic inflammation. METHODS: Serum nitrite, kynurenine, tryptophan and neopterin levels were evaluated in 36 patients with seasonal allergic conjunctivitis, along with these values in 41 healthy subjects. All these parameters have been compared with symptom and sign scores. RESULTS: Tryptophan and kynurenine concentrations were not significantly changed, while serum nitrite concentrations were significantly low, and neopterin levels were significantly increased in patients compared to healthy subjects (p < 0.05). There was a significant relationship between symptom scores and serum nitrite levels in patients. CONCLUSIONS: This preliminary study demonstrates that serum nitric oxide metabolism might have a role in allergic conjunctivitis. Serum neopterin levels but not tryptophan metabolism could serve as a biomarker in patients with seasonal allergic conjunctivitis.

Lycopene restores trace element levels in ochratoxin A-treated rats.

This study was designed to investigate the in vivo effects of ochratoxin A (OTA) and/or lycopene on the levels of selenium, zinc, and copper in the liver, kidneys, and testes of male Sprague-Dawley rats. The rats were treated with OTA (0.5 mg kg-1 day-1) and/or lycopene (5 mg kg-1 day-1) by gavage for 7 or 14 days. Trace element levels were measured by atomic absorption spectrometry. OTA significantly lowered selenium (20 % in the liver, 17 % in the kidney, and 40 % in the testis), zinc (24 % in the liver, 23 % in the kidney, and 26 % in the testis), and copper levels (40 % in the liver and 10 % in the kidney). Lycopene alone did not affect the trace element levels in any of the organs. In combination with OTA, however, it significantly restored liver, kidney, and testis selenium and zinc levels compared to the group treated with OTA alone. Our results have confirmed that depletion of trace elements in different organs is one of the mechanisms of action of OTA. They also suggest that lycopene interferes with this depleting effect and restores trace element levels, the implications of which need to be further investigated.

Possible changes of New-Generation inflammation markers with occupational lead exposure.

OBJECTIVES: Occupational lead (Pb) exposure is still an important health problem in the world. Long-term Pb exposure causes several adverse effects. The aim of this study was to investigate the changes of inflammation markers with chronic Pb exposure by analyzing neopterin levels and kynurenine (Kyn) to tryptophan (Trp) ratio that reflects indolamine 2,3-dioxygenase activity and to compare with healthy volunteers' parameters. METHODS: Blood lead levels (BLLs) were analyzed by atomic absorption spectrometry. Urinary neopterin and serum Kyn and Trp levels were analyzed by high-performance liquid chromatography. RESULTS: According to our results, mean BLL of the 29 workers was 20.4±9.6 µg/dl. Urinary neopterin levels, serum Kyn levels, and Kyn/Trp of Pb workers (188±52 µmol/mol creatinine, 2.70±0.66 µM, and 43.19±10.38 µmol/mmol, respectively) were significantly higher than controls (144±35 µmol/mol creatinine, 2.08±0.34 µM, and 32.24±7.69 µmol/mmol, respectively). Pb-exposed workers were divided into further three groups according to their BLLs: as 10-19 µg/dl (n=18), 20-29 µg/dl (n=8), and 30-49 µg/dl (n=3). Neopterin levels of the workers with BLL of 30-49 µg/dl were significantly higher than those of BLL with 10-29 µg/dl, while Trp levels decreased. Kyn/Trp of workers with BLL of 30-49 µg/dl were elevated significantly compared with the workers with BLL<30 µg/dl. In addition to neopterin, Kyn and Kyn/Trp levels were positively influenced by Pb exposure. CONCLUSIONS: Increased level of inflammation markers confirms the adverse effects of Pb even low BLLs, and we suggest that monitoring BLLs with inflammation markers could help to prevent serious occupational health problems.

A new update for radiocontrast-induced nephropathy aggravated with glycerol in rats: the protective potential of epigallocatechin-3-gallate.

Contrast media (CM) is known to have nephrotoxic adverse effects. Epigallocatechin-3-gallate (EGCG) is the most abundant and active catechin in green tea, and has strong antioxidant and anti-inflammatory properties. This study investigated whether EGCG can reduce contrast-induced nephrotoxicity (CIN), alone or with glycerol (GLY)-induced renal damage, and to understand its mechanisms of protection against toxicity, using models of GLY and CIN in rats. The rats were separated into eight groups (n = 6 in each), as follows: Healthy, GLY, CM, GLY + CM, CM + EGCG 50 mg/kg (po), GLY + CM + EGCG 50 mg/kg (po), CM + EGCG 100 mg/kg (po), and GLY + CM + EGCG 100 mg/kg (po). Both doses of EGCG protected against CM-induced renal dysfunction, as measured by serum creatinine and blood urea nitrogen (BUN). In addition, EGCG treatment markedly improved CIN-induced oxidative stress, and resulted in a significant down-regulatory effect on tumor necrosis factor (TNF)-α and nuclear factor (NF)-κB mRNA expression. Moreover, histopathological analysis showed that EGCG also attenuated CM-induced kidney damage. Considering the potential clinical use of CM and the numerous health benefits of EGCG, this study showed the protective role of multi-dose EGCG treatment on CIN and GLY-aggravated CIN through different mechanisms.

The Use of Breast Magnetic Resonance Imaging Parameters to Identify Possible Signaling Pathways of a Serum Biomarker, HE4.

OBJECTIVES: This study aimed to investigate the relationship between breast magnetic resonance imaging (MRI) parameters; clinical features such as age, tumor diameter, N, T, and TNM stages; and serum human epididymis protein 4 (HE4) levels in patients with breast carcinoma and use this as a means of estimating possible signaling pathways of the biomarker, HE4. METHODS: Thirty-seven patients with breast cancer were evaluated by breast MRI and serum HE4 levels before therapy. Correlations between parameters including age, tumor diameter T and N, dynamic curve type, enhancement ratio (ER), slope washin (S-WI), time to peak (TTP), slope washout (S-WO), and the serum level of HE4 were investigated statistically. Human epididymis protein 4 levels of early and advanced stage of disease were also compared statistically. RESULTS: Breast MRI parameters showed correlation to serum HE4 levels and correlations were statistically significant. Of these MRI parameters, S-WI had higher correlation coefficient than the others. Human epididymis protein 4 levels were not statistically different in early and advanced stage of disease. CONCLUSIONS: High correlation with MRI parameters related to neoangiogenesis may indicate signaling pathway of HE4.

Neopterin Release and Tryptophan Degradation in Patients with Uveitis.

PURPOSE: Uveitis is an intraocular inflammation affecting the highly vascularized uveal tissue. Early recognition and appropriate therapeutic intervention of uveitis are important since the condition may be associated with systemic disease and untreated uveitis may lead to blindness. Neopterin, an unconjugated pteridine, is an important biomarker of cell-mediated immunity and has a potential function in the process of inflammation. In addition to neopterin release, cellular immune activation also induces indoleamine 2,3-dioxygenase (IDO). In this study, the aim was to investigate possible immune changes in uveitis by determination of neopterin concentrations and tryptophan (Trp) degradation. MATERIALS AND METHODS: The participants who attended to the ophthalmology clinic with uveitis were divided into two groups: active (n = 63) and remission (n = 41). Additionally 30 healthy subjects were recruited as a control group. RESULTS: In total, in 104 uveitis patients, urinary and serum neopterin, kynurenine (Kyn), and Kyn/Trp were found to be statistically higher than the 30 controls (all, p < 0.05). It was observed that all of the measured parameters did not differ between active and remission uveitis groups (all, p > 0.05), except for the Kyn/Trp ratio (p < 0.05). Urinary and serum neopterin levels were positively correlated with Kyn/Trp in the uveitis patients (both p < 0.05). CONCLUSIONS: From these results, it can be concluded that uveitis can cause alterations in neopterin levels and the Kyn pathway. It seems that the measured parameters can be useful markers of cellular immune response in uveitis, although they might not be used to differentiate active or remission uveitis.

Endothelial dysfunction biomarker, endothelial cell-specific molecule-1, and pediatric metabolic syndrome.

BACKGROUND: The aim of this study was to compare serum endothelial cell-specific molecule-1 (endocan) in pediatric patients with metabolic syndrome (MetS) and in healthy children, and to determine whether it can be used as an indicator of endothelium damage-induced complications in pediatric MetS patients. METHODS: The study included 30 patients, aged 6-16 years, who were diagnosed with MetS. Another 30 children with no diseases were recruited as healthy controls. Endocan concentration was measured using enzyme-linked immunosorbent assay. RESULTS: Endocan was increased almost threefold in the MetS group compared with the healthy group. Systolic arterial tension and diastolic arterial tension, serum triglyceride, total cholesterol, and low-density lipoprotein cholesterol were higher, and high-density lipoprotein cholesterol was lower, in the MetS children than in the healthy group. Fasting blood glucose (FBG), hemoglobin A1c (HBA1C), and homeostasis model assessment insulin resistance (HOMA-IR) were also significantly increased in the children with MetS compared with the healthy group. CONCLUSIONS: Serum endocan level in pediatric MetS patients could be an important indicator of cardiovascular risk in adulthood.

Levels of endothelial cell-specific molecule-1 (ESM-1) in overt hypothyroidisim.

PURPOSE: Endothelial cell-specific molecule-1, endocan, is a proteoglycan that is expressed by the vascular endothelium. Endocan can be a biomarker of endothelial dysfunction caused by endothelial cell-dependent disorders. Endothelial dysfunction is an early step of atherosclerosis and is developed in hypothyroid patients, which indicates an association between hypothyroidism and atherosclerosis. Therefore, we aimed to investigate whether circulating endocan levels are associated with endothelial dysfunction in overt hypothyroid patients. MATERIALS AND METHODS: Forty patients with hypothyroidism diagnosed in the last 5 years and 30 healthy subjects were recruited. RESULTS: The mean endocan value in all patients was 0.63 ± 0.26 pg/ml, which was higher than that in controls (0.36 ± 0.10 pg/ml, p < 0.05). When we subgrouped the patients as hypothyroid and euthyroid, all groups demonstrated significantly different endocan levels, and hypothyroid patients had the highest endocan levels. A correlation analysis demonstrated that endocan levels were positively correlated with body mass index (BMI), thyroid-stimulating hormone (TSH), anti-thyroid peroxidase, and anti-thyroglobulin and negatively correlated with free thyroid hormone 4 (FT4) and vitamin D levels. In addition, in the patient group, endocan levels were correlated with FT4 levels independently in a covariance analysis. CONCLUSIONS: The circulating endocan level increased in hypothyroid patients, suggesting that endocan levels may be an early biomarker of the development of endothelial dysfunction in patients with hypothyroidism. They may also prove useful in the prediction of cardiovascular diseases after further studies using cardiovascular disease biomarkers. In addition, targeting endocan levels to decrease cardiovascular risk may be a new treatment strategy in these patients.

The Role of RAAS Inhibition by Aliskiren on Paracetamol-Induced Hepatotoxicity Model in Rats.

Paracetamol is one of the most popular and widely used analgesic and antipyretic agents, but an overdose can cause hepatotoxicity and lead to acute liver failure. Aliskiren directly inhibits renin which downregulates the renin-angiotensin-aldosterone system (RAAS). Recent findings suggest that RAAS system takes part in the pathogenesis of liver fibrosis. We aimed to reveal the relationship between hepatotoxicity and the RAAS by examining paracetamol induced hepatotoxicity. Rats were separated into five groups as follows: control, 100 mg/kg aliskiren (p.o.), 2 g/kg paracetamol (per os (p.o.)), 2 g/kg paracetamol + 50mg/kg aliskiren (p.o.), and 2 g/kg paracetamol + 100 mg/kg aliskiren(p.o.). Samples were analyzed at the biochemical, molecular, and histopathological levels. Paracetamol toxicity increased alanine aminotransferases (ALT), aspartate aminotransferases (AST), renin, and angiotensin II levels in the serum samples. In addition, the SOD activity and glutathione (GSH) levels decreased while Lipid Peroxidation (MDA) levels increased in the livers of the rats treated with paracetamol. Paracetamol toxicity caused a significant increase in TNF-α and TGF-ß. Both aliskiren doses showed an improvement in ALT, AST, oxidative parameters, angiotensin II, and inflammatory cytokines. Only renin levels increased in aliskiren treatment groups due to its pharmacological effect. A histopathological examination of the liver showed that aliskiren administration ameliorated the paracetamol-induced liver damage. In immunohistochemical staining, the expression of TNF-α in the cytoplasm of the hepatocytes was increased in the paracetamol group but not in other treatment groups when compared to the control group. In light of these observations, we suggest that the therapeutic administration of aliskiren prevented oxidative stress and cytokine changes and also protected liver tissues during paracetamol toxicity by inhibiting the RAAS.

Study of the boron levels in serum after implantation of different ratios nano-hexagonal boron nitride-hydroxy apatite in rat femurs.

Boron and its derivatives are effective in bone recovery and osteointegration. However, increasing the boron levels in body liquids may cause toxicity. The aim of our study is to investigate serum boron levels using ICP-MS after implantation of different ratios of nano-hBN-HA composites in rat femurs. All rats were (n=126) divided into five experimental groups (n=24) and one healthy group (6 rats); healthy (Group1), femoral defect + %100 HA (Group2), femoral defect + %2.5 hBN + %97.5 HA (Group3), femoral defect + %5 hBN + %95 HA (Group4), femoral defect + %10 hBN + %90 HA (Group5), femoral defect + %100 hBN (Group6). The femoral defect was created in the distal femur (3mm drill-bit). Each implant group was divided into four different groups (n=24) also 6 rats sacrificed for each groups in one week intervals during four weeks. In our results; at 1, 2, 3, and 4 weeks after implantation near bone tissue, serum levels of boron were evaluated using ICP-MS. We demonstrated that neither short-term nor long-term implantation of hBN-HA composite resulted in statistically increased serum boron levels in experimental groups compared to healthy group. In conclusion, this study investigated the implant material produced form hBN-HA for the first time. Our data suggest that hBN is a new promising target for biomaterial and implant bioengineers.

Erythrocyte folate status and serum iron levels in patients undergoing hemodialysis.

The present study was aimed to evaluate erythrocyte folate and the iron levels in diabetes and hypertension patients treated with/without hemodialysis. The effects of erythropoietin and iron treatment as well as vitamin supplementation on measured parameters were considered. The 67 controls consisted of healthy subjects (n = 22), hypertensive subjects (n = 22), and diabetic subjects (n = 23) without any renal disorder. According to primary renal disorders, the patients undergoing hemodialysis (n = 68) were classified into four groups as diabetic nephropathy, hypertensive nephropathy, reflux nephropathy or interstitial nephritis, and renal insufficiency depending on other causative factors. The mean value of erythrocyte folate levels of all patients undergoing hemodialysis was higher than the healthy control group (P < 0.05). Erythrocyte folate levels in hypertensive and diabetic nephropathy patients were higher than their own hypertensive or diabetic controls and also healthy controls (both, P < 0.05). Serum iron levels of all subgroups in hemodialysis patients were found to be similar with healthy controls (all, P > 0.05). The only significance observed within the subgroups was between diabetic controls and diabetic nephropathy patients (P < 0.05). None of the treatment or supplementation of erythropoietin, iron and vitamin affected erythrocyte folate levels (all, P > 0.05). The increase in erythrocyte folate status of patients with end stage renal diseases might be the result of sum or individual effects of causative factors such as renal pathology, compensation mechanism against renal anemia, or routine folate supplementation.

Immunomodulation and oxidative stress in denim sandblasting workers: changes caused by silica exposure.

Workers in denim sandblasting are at a high risk of developing silicosis, an occupational lung disease caused by inhaling crystalline silica dust. The development and progress of silicosis is associated with the activation of the immune system and oxidative stress. In the former, interferon-gamma induces both neopterin release and the enzyme indoleamine [2, 3]-dioxygenase (IDO) in various cells. The determination of the kynurenine-to-tryptophan ratio and neopterin concentration has proven to be an efficient method to monitor the activation status of IDO and cellular immunity. The present study aimed to investigate whether occupational silica exposure leads to any alterations in neopterin levels, tryptophan degradation, and activities of superoxide dismutase (SOD) and catalase (CAT), agents in the antioxidant defense system. Fifty-five male denim sandblasting workers and twenty-two healthy men as controls were included. Mean neopterin and kynurenine levels, kynurenine-to-tryptophan ratio, and SOD activity were higher in subjects with silicosis compared to non-exposed controls (all, p<0.05). Neopterin levels and kynurenine-totryptophan ratios were positively correlated (p<0.05); however, no correlation was observed between length of employment and the measured parameters. Some of the measured parameters were significantly affected by the severity of the pathology. Our results suggest that silica exposure activates the cellular immune response. The increased neopterin levels and tryptophan degradation confirm the possibility of their use as an indicator of cellular immune response.

The carotenoid lycopene protects rats against DNA damage induced by Ochratoxin A.

Ochratoxin A (OTA), one of the most prevalent mycotoxins in the world, has nephrotoxic and hepatotoxic properties. Lycopene is an important carotenoid and has a high singlet-oxygen and free-radical scavenging capacity. This study was designed to investigate the possible protective effects of lycopene against the genotoxicity of OTA in rat tissues using the alkaline comet assay. Male Sprague-Dawley rats were used in the experiments. OTA (0.5 mg/kg b.w./day) was administered by gavage for 14 days, whereas lycopene was applied on the last 7 days or for 14 days of the feeding period, with OTA treatment. OTA caused marked increases in tail length, tail moment, and tail intensity vs. control both in the kidney and liver cells, but not in the lymphocytes. Lycopene administration alone for 7 and 14 days did not provide any significant change in DNA damage of the lymphocytes, renal and hepatic cells vs. controls. However, lycopene for both 7 and 14 days, with OTA exposure in renal and hepatic cells, supplied significant decreases in tail length, tail moment, and tail intensity vs. OTA-exposed rats. The effect of 14 days supplementation seemed to be more protective, particularly against hepatic cells. These results suggest that lycopene may protect hepatic and renal tissue from OTA-induced DNA damage.

HPLC method for determination of diclofenac in human plasma and its application to a pharmacokinetic study in Turkey.

A simple high-performance liquid chromatography (HPLC) method has been developed for determination of diclofenac in human plasma. The method was validated on Ace C(18) column using UV detection. The mobile phase consisted of 20 mM phosphate buffer (pH 7) containing 0.1% trifluoroacetic acid-acetonitrile (65:35, v/v). Calibration curve was linear between the concentration range of 75-4000 ng/mL. Intra- and inter-day precision values for diclofenac in plasma were less than 3.6, and accuracy (relative error) was better than 5.3%. The limits of detection and quantification of diclofenac were 25 and 75 ng/mL, respectively. Also, this assay was applied to determine the pharmacokinetic parameters of diclofenac in healthy Turkish volunteers who had been given 50 mg diclofenac.