Effects of thymoquinone on alpha-amanitin induced hepatotoxicity in human C3A hepatocytes

Abstract Thymoquinone (TQ) has shown hepatoprotective effects in various experimental studies. We aimed to investigate the possible beneficial effects of TQ regarding its prevention of alpha-amanitin induced hepatotoxicity in human C3A hepatocytes. After administering alpha-amanitin in a concentrations of 1 and 10µg/mL on the cells in a hepatocyte cell line, TQ was administered in various concentrations (10, 5, 1, 0.5, 0.1, 0.05, 0.01, 0.005 µg/mL). The MTT test was used to determine cell viability. For the groups given only TQ at various concentrations, the cell viability rates at 48 hours post-administration were found at 82.6, 98.3, 102.1, 102.5, 99.4, 99.4, 101.9 and 106.3%, respectively. For the group with 1μg/mL alpha-amanitin and various TQ concentrations, the cell viability rates were found at 74.6, 88.5, 87.4, 88.7, 85.7, 86.8, 88.4, and 92.9%, respectively. For the group with 10μg/mL alpha-amanitin and various TQ concentrations, the cell viability rates for each TQ subgroup were found at 65.2, 79.2, 81.4, 81.1, 81.8, 81.8, 82.2 and 91.9%, respectively. Our study is the first in vitro study that investigates TQ's effects on alpha-amanitin induced hepatotoxicity. Although TQ had beneficial effect in low doses did not significantly increase cell viability in liver damage due to alpha-amanitin toxicity.

Determination of histological, immunohistochemical and biochemical effects of acute and chronic grayanotoxin III administration in different doses in rats.

Grayanotoxin (GTX)-III is a Na-channel neurotoxin. Grayanotoxins can be found in the nectar, pollen, and other plant parts of the Rhododendron genus plants from the Ericaceae family. It is widely believed that honey produced from these plants, which are concentrated in the Black Sea region, is traditionally characterized as enhancing sexual performance. It is thought that the effective factor is dose for this compound, which has both beneficial and toxic effects reported. Therefore, it is aimed to evaluate the histological, immunohistochemical, and biochemical effects of acute and chronic impact of GTX-III in different doses on testes tissue in this study. For this purpose, 100 Sprague-Dawley male rats were divided into 5 separate groups for acute and chronic research. While dose groups were (control, 0.1, 0.2, 0.4, ve 0.8 µg/kg/bw) for experimental groups, a single dose (i.p.) was administered for acute impact whereas the same doses were administered daily for 3 weeks to assess chronic effect. At the end of the experiment, Johnsen testicular biopsy scoring was performed on testicular tissue samples, seminiferous tubule diameters were measured, and apoptotic cells were evaluated by TUNEL method. Testosterone, LH, and FSH levels were measured by ELISA method in serum and tissue specimens. It was found that Johnsen score of acute doses was significantly lower than the control group, and the diameter of the seminiferous tubules decreased significantly in acute and chronic dose-administered groups compared to the control. Hemorrhage, epithelial shedding, irregularity in seminiferous epithelium, and vacuolization were observed in acute and chronic dose-administered groups, and increase in apoptotic cells was determined. Hormone levels varied depending on the dose. In conclusion, it was found that dose-dependent acute and chronic effects of GTX-III are different, and this factor should be taken into account in studies to be carried out due to the adverse effects of high doses.

Omentin Val/Val genotype increases predisposition to acne vulgaris without changing omentin serum level.

Acne vulgaris is the most frequent and multifactorial inflammatory skin disorder in all races. Obesity is considered to be a risk factor for acne due to its contribution to inflammation. The involvements of inflammatory (leptin and resistin) and anti-inflammatory (adiponectin) adipokines in the pathogenesis of acne were reported. Omentin resembles adiponectin in terms of having inhibitory effect on tumor necrosis factor-α (TNF-α) induced inflammation, a vital process in the acne formation. This study was designed to investigate the putative involvement of omentin in acne formation. The genotyping was performed by restriction fragment length polymorphism (RFLP) method. Serum omentin protein levels were analyzed by enzyme-linked immunosorbent assay (ELISA). Serum omentin level was not significantly changed between groups. However, the decreased serum omentin level was observed as the mean value of BMI increased. The Asp/Asp, Val/Asp and Val/Val genotypes distributions for control and patient groups (19[17.4%], 22[20.2%], and 3[2.8%] respectively, vs. 31[28.4%], 25[22.9%], and 9[8.3%], respectively) were obtained. The Val/Val (mutant homozygote) genotype was found nearly 1.8 times more in the patient group (p=0.403, OR=1.839 (0.442-7.653)). This is the first time to clarify a linkage between anti-inflammatory omentin and acne vulgaris. Omentin Val109Asp polymorphism affects the overall function of the protein. In conclusion, omentin Val/Val (mutant homozygote) genotype increases predisposition to acne vulgaris by probably disrupting overall protein function of omentin.

Evaluation of the corrosion inhibiting efficacy of a newly synthesized nitrone against St37 steel corrosion in acidic medium: Experimental and theoretical approaches.

A novel amphiphilic nitrone, N-phenyl-1-(4-((11-(pyridin-1-ium-1yl) undecanoyl) oxy)phenyl)methanimine oxide bromide (NP-1-4-11-PUOPMOB) has been synthesized from a fatty acid derivative as a starting material. Structural characterization of the new compound has been realized by spectroscopic techniques (FTIR, 1H NMR, and 13C NMR). The corrosion inhibition effect of the compound for St37 steel corrosion in 1 M HCl medium has been investigated using experimental (weight loss, electrochemical impedance spectroscopy, potentiodynamic polarization, dynamic electrochemical impedance spectroscopy) and theoretical approaches complemented by surface morphological examination using energy dispersive X-ray spectroscopy, scanning electron microscope, and atomic force spectroscopy. Results from both chemical and electrochemical techniques reveal that the presence of the nitrone in the acid solution impedes St37 steel corrosion. The inhibition efficiency obtained at 125 ppm and 150 ppm concentrations for all methods is found to be over 90%. NP-1-4-11-PUOPMOB behaves as a mixed type corrosion inhibitor according to the potentiodynamic polarization studies. The adsorption of NP-1-4-11-PUOPMOB molecules onto the metal surface follows Langmuir adsorption isotherm and the calculated Kads (equilibrium constant of the adsorption process) value reflects strong interaction. There is evidence of NP-1-4-11-PUOPMOB adsorption on the metal surface from SEM, EDAX, and AFM studies. Experimental and theoretical results are in good agreement.

Gum Arabic-silver nanoparticles composite as a green anticorrosive formulation for steel corrosion in strong acid media.

A green anticorrosive composite (GA-AgNPs) has been formulated for steel in 15% HCl and 15% H2SO4 media. Characterization of GA-AgNPs is achieved via FTIR, UV-vis, EDAX, and SEM. Gravimetric, electrochemical (EIS, EFM, DEIS, & TP), and surface assessment (SEM, EDAX, AFM, & XPS) techniques have been deployed in the anticorrosion studies. Results from all applied methods potray GA-AgNPs as effective anticorrosive agent. Inhibition is by adsorption mechanism and follows Langmuir isotherm. GA-AgNPs acts as mixed type inhibitor in 15% H2SO4 solution but as anodic type in 15% HCl solution. Results from surface techniques confirm adsorption of GA-AgNPs molecules on specimen surface. Oxides, hydroxides, carbonates, and sulphates (H2SO4 medium) or chlorides (HCl medium) are the corrosion products in the free corrodent according to XPS results. In the presence of composite, both ionic and neutral forms of GA-AgNPS are adsorbed. AgNPs are present on the surface in the form: Ag°, Ag2O, and AgO.

Comparison of Changes in Anxiety and Depression Level Between Dabigatran and Warfarin Use in Patients With Atrial Fibrillation.

We hypothesized that patients taking warfarin require frequent hospital follow-up and they are at higher risk for complications, so the incidence of depression and anxiety is higher in patients with atrial fibrillation (AF) in the period of taking warfarin compared to the period of taking dabigatran. Fifty patients having AF without valvular diseases under treatment of warfarin in whom a transition to dabigatran was planned were consecutively enrolled in this study and followed up prospectively between July 2013 and July 2014. All patients completed Beck Depression Inventory and Hamilton Anxiety Scale (HAS) at the initiation of study and 6 months after initiation of study. Of the patients enrolled in the study, age, gender, smoking status, and comorbidities were questioned. A total of 50 patients (28 women; mean age 74.6 ± 8.7 years) treated with warfarin in whom a transition to dabigatran was planned were included. Basal mean value of BDS (15.6 ± 7.8 vs 11.5 ± 4.8, P < .001) and HAS (16.8 ± 10.4 vs 12.6 ± 8.1, P < 0.001) was significantly higher in patients when they used warfarin than when they switched to dabigatran. In categorical analysis, frequency of patients with depression (mild, moderate, and severe) was significantly higher in period of warfarin use than after dabigatran transition (n = 24, 48% vs n = 14, 28%, P = .039). Our study demonstrates that patients with nonvalvular AF under treatment of dabigatran had lower BDS and HAS scores compared to warfarin. These findings suggest that dabigatran may increase quality of life and decrease morbidity and mortality due to reduction in anxiety and depression.

Effect of Montelukast on Spinal Cord Ischemia- Reperfusion Injury.

AIM: Paraplegia due to ischemia-reperfusion (I/R) injury of the spinal cord is a devastating complication of thoracoabdominal aortic surgery. Cysteinyl leukotrienes are potent mediators of inflammation that are associated with I/R injury. The present study was designed to investigate the role of montelukast, a selective reversible CysLT1 receptor antagonist, on spinal cord I/R injury in an experimental model. MATERIAL AND METHODS: Twenty-one male Sprague-Dawley rats were randomly assigned to three groups (n=7 per group) as G1 (no aortic occlusion and montelukast administration), G2 (45 min. aortic occlusion; no montelukast administration) and G3 (45 min. aortic occlusion, 10 mg/kg montelukast administration). After neurologic evaluation using the Motor Deficit Index (MDI) score at the 48th hour of reperfusion, lumbar spinal cords were removed for histopathological evaluation and immunohistochemical staining for HSP70, interleukin-6 and myeloperoxidase (MPO). RESULTS: All rats in the G1 group had a normal neurological status and their MDI score was 0 (p < 0.05). The MDI score of G3 was significantly lower than G2 group (2.8 vs. 5.5; p < 0.05). Vacuolar congestion was found to be significantly lower in G1 than the other groups (p=0.0001). The interleukin-6 receptor level was found to be significantly lower in G3 group than the control group (p=0.013). There was no statistically significant difference found among the groups in terms of the degree of HSP70 and MPO staining. CONCLUSION: Increased generation of leukotrienes in postischemic organs play an important role in I/R injury. The findings of the current study demonstrated that montelukast improved motor recovery and decreased IL-6 levels in spinal cord I/R injury.

Erythropoietin stimulates patellar tendon healing in rats.

BACKGROUND: Erythropoietin (EPO), regulating erythropoiesis, is used to provide protective and regenerative activity in non-haematopoietic tissues. There is insufficient knowledge about the role of EPO activity in tendon healing. Therefore, we investigated the effect of EPO treatment on healing in rat patellar tendons. METHODS: One hundred and twenty-six, four-month-old male Sprague-Dawley rats were randomly assigned to three experimental groups: 1, no treatment; 2, treatment with isotonic saline (NaCl) and 3, treatment with EPO. Each group was randomly subdivided into two groups for sacrifice at three (1a, 2a, 3a) or six weeks (1b, 2b, 3b). Complete incision of the left patellar tendon from the distal patellar pole was performed. We applied body casts for 20 days after the incised edges of the patellar tendon were brought together with a surgical technique. Both legs were harvested and specimens from each group underwent histological, biomechanical, and protein mRNA expression analyses. RESULTS: There were statistically significant differences in the ultimate breaking force between the EPO group and others at both weeks three and six (p<0.05); significant differences in fibroblast proliferation, capillary vessel formation, and local inflammation were found between groups 1a and 3a, and 2a and 3a (p<0.05). There were statistical differences between 1a, 3a and 2a, 3a for Col III, TGF-ß1, and VEGF and between 1b, 3b and 2b, 3b for Col I, Col III, TGF-ß1, and VEGF mRNA expressions. CONCLUSION: EPO had an additive effect with surgery on the injured tendon healing process in rats compared to the control groups biomechanically, histopathologically and with tissue protein mRNA expression. CLINICAL RELEVANCE: This is the first experimental study to analyze the relationship between EPO treatment and the patellar tendon repair process by biomechanical, histopathological, and tendon tissue mRNA expression methodologies.

Evaluation and comparison of alpha- and beta-amanitin toxicity on MCF-7 cell line.

BACKGROUND/AIM: Alpha- and beta-amanitins are the main toxins of the poisonous Amanita phalloides mushroom. Although there are many studies available concerning alpha-amanitin, there are limited data about beta-amanitin in the literature. Therefore, this study is aimed at comparing the toxic effects of alpha- and beta-amanitin on the MCF-7 cell line. MATERIALS AND METHODS: The alpha- and beta-amanitins used for this research were purified from Amanita phalloides by preparative high-performance liquid chromatography. The MCF-7 breast cancer cell line was used, and specific concentrations of the toxins (100, 10, 1, 0.1, and 0.01 µg/mL) were applied to the cells. The MTT test was performed to determine the level of toxicity, and the quantity of protein in the cell was measured using the biuret test. RESULTS: The aLpha-amanitin showed a higher toxicity at 36 h, while the highest inhibition of protein synthesis by the beta-amanitin was observed at 24 h. CONCLUSION: It was shown that the beta-amanitin may be responsible for toxicity, like alpha-amanitin, in Amanita phalloides mushroom poisoning. The early inhibition of protein synthesis for beta-amanitin might be useful for future experiments and research.

Polymorphisms in MMP-2 and TIMP-2 in Turkish patients with prostate cancer.

AIM: Prostate cancer is the most commonly diagnosed malignancy and the second most common cause of cancer deaths in the Western male population. Matrix metalloproteinases (MMPs) and tissue inhibitors of MMPs (TIMPs) modulate the remodeling of the extracellular matrix (ECM). The imbalance between MMPs and TIMPs may lead to an emergence of pathological processes such as cancer. In this study, the association between TIMP-2 (-418 G/C) and MMP-2 (-1306 C/T) polymorphisms and prostate cancer in the Turkish population was investigated. MATERIALS AND METHODS: Sixty-one prostate cancer patients and 46 healthy subjects were included in the study. DNA was isolated from 2 mL of peripheral blood taken from subjects, and genotypes were analyzed by the polymerase chain reaction-restriction fragment length polymorphism method. RESULTS: The TIMP-2 -418 (GC) genotype was found in 15 cases (32.6%) in the control group and in 9 cases (14.8%) in the patients group, and statistical significance was determined (P = 0.037, OR = 0.346). The MMP-2 -1306 (CT) genotype was found 2.17 times more in the patient group than in the control group (P = 0.149, OR = 2.17). CONCLUSION: Our results show that the TIMP-2 -418 (GC) genotype had a putative protective effect against prostate cancer.

Extracorporeal shockwave increases the effectiveness of systemic antibiotic treatment in implant-related chronic osteomyelitis: experimental study in a rat model.

Implant-related chronic osteomyelitis is a serious complication of orthopedic surgery requiring implant removal and radical debridement. Extracorporeal shockwave (ESW) have demonstrated significant bactericidal effectiveness in vitro and effectiveness and safety were evaluated in an animal model of osteomyelitis. In this experimental study, we aimed to test our hypothesis that the use of ESW together with systemic antibiotic treatment will provide synergy for the treatment of implant-related chronic osteomyelitis caused by methicillin-susceptible Staphylococcus aureus (MSSA). The proximal tibia of 32 rats was contaminated with (10) 8 CFU/ml methicillin-sensitive S. aureus (MSSA-ATCC 29213) and Kirschner-wires were placed into the medulla of the tibia. After 4 weeks, Kirschner-wires were removed and the rats were randomly divided into four groups: group I, untreated contaminated control group; group II, receiving only ESW therapy; group III, receiving only systemic teicoplanin; group IV, treated with a combination of ESW and systemic teicoplanin. ESW was applied twice to the infected limbs and all rats were sacrificed at the end of 8th week. The degree of tibial osteomyelitis was assessed by quantitative culture analysis. Bacterial counts in groups III and IV were significantly reduced relative to the control (p=0.002 and 0.001, respectively). The decrease in bacterial counts was more pronounced and significant in group IV compared to group III (p=0.024). In group II, bacterial counts also decreased, but the differences were in significant (p=0.068). Our experimental model suggests that ESW provides significant synergy for systemic antibiotic treatment. However, further clinical trials are required in order to use this treatment modality safely in patients, even though our study demonstrated successful results in the treatment of implant-related chronic osteomyelitis in rats.

Sedoanalgesia in pediatric daily surgery.

PURPOSE: The present report was focused on clinical advantages of sedoanalgesia in the pediatric outpatient surgical cases. METHOD: Sedoanalgesia has been used to sedate patients for a variety of pediatric procedures in our department between 2007 and 2010. This is a retrospective review of 2720 pediatric patients given ketamine for sedation with midazolam premedication. Ketamine was given intravenously (1-2 mg/kg) together with atropine (0.02 mg/kg) and midazolam (0.1 mg/kg) + a local infiltration anesthetic 2 mg/kg 0.5% bupivacaine hydrochloride. RESULT: Median age of the patients included in the study was 5.76 ± 2.12 (0-16 years). The main indications for ketamine include circumcision (69%), inguinal pathologies (inguinal hernia (17%), orchidopexy (2.68%), hydrocele (3.38%), hypospadias (1.94%), urethral fistula repair (0.33%), urethral dilatation (0.25%), and other conditions. All of our patients were discharged home well. In this regard, we have the largest group of patients ever given ketamine. CONCLUSION: Sedoanalgesia might be used as a quite effective method for daily surgical procedures in children.

Efficacy of iloprost and montelukast combination on spinal cord ischemia/reperfusion injury in a rat model.

BACKGROUND: The thoracic or thoracoabdominal aortic aneurysm surgery may cause spinal cord ischemia because of aortic cross-clamping and may result in severe postoperative complications caused by spinal cord injury. Ischemia/reperfusion injury may directly or indirectly be responsible for these complications. In this study we sought to determine whether combination of iloprost and montelukast can reduce the ischemia/reperfusion injury of spinal cord in a rat model. METHODS: Medulla spinalis tissue concentrations of interleukin-6 (IL-6), myeloperoxidase (MPO) and heat shock protein 70 (HSP-70) were determined in 3 groups of Spraque Dawley rats: control group (operation with cross clamping and intraperitoneal administration of 0.9% saline, n = 7), sham group (operation without cross clamping, n = 7), and study group (operation with cross-clamping and intraperitoneal administration of iloprost (25 ng/kg) and montelukast (1 mg/kg), n = 7). The abdominal aorta was clamped for 45 minutes, with a proximal (just below the left renal artery) and a distal (just above the aortic bifurcation) clip in control and study groups. Hindlimb motor functions were evaluated at 6, 12, 24, and 48 hours using the Motor Deficit Index score. All rats were sacrificed 48 hours after the procedure and spinal cord tissue levels of myeloperoxidase, interleukin-6, and heat shock protein (HSP-70) were evaluated as markers of oxidative stress and inflammation. Histopathological analyses of spinal cord were also performed. RESULTS: The tissue level of HSP-70 was found to be similar among the 3 groups, however, MPO was highest and IL-6 receptor level was lowest in the control group (p = 0.007 and p = 0.005; respectively). In histopathological examination, there was no significant difference among the groups with respect to the neuronal cell degeneration, edema, or inflammation, but vascular congestion was found to be significantly more prominent in the control group than in the sham or in the study group (p = 0.05). Motor deficit index scores at 24 and 48 hours after ischemia were significantly lower in the study group than in the control group. CONCLUSION: This study suggests that combined use of iloprost and montelukast may reduce ischemic damage in transient spinal cord ischemia and may provide better neurological outcome.

Branch retinal vein occlusion associated with tamoxifen use.

Tamoxifen is a selective estrogen receptor modulator widely used in the treatment of hormone-responsive breast cancer. Tamoxifen-induced ocular complications are very rare. A post-menopausal woman with carcinoma of the left breast had presented with sudden loss of vision. The patient had been on tamoxifen therapy 20 mg daily for the last three years. Fundus examination showed left branch retinal vein occlusion. Fluorescein angiography and optical coherence tomography confirmed the diagnosis. Tamoxifen therapy was discontinued. Although branch retinal vein occlusion is rare, careful evaluation of patients on tamoxifen therapy with visual symptoms is required.

Neutrophilic eccrine hidradenitis induced by cetuximab.

Cetuximab is an epidermal growth factor receptor inhibitor used in metastatic colorectal cancer, and head and neck cancers. Several cutaneous side effects due to cetuximab such as acne-like rash, pruritus, dry skin, desquamation, hypertrichosis, and paronychia have been reported so far. A 59-year-old male patient with metastatic colon cancer referred to our outpatient clinic for his lesions on the dorsal surfaces of his hands and wrists, and on thighs developing after the chemotherapy. He was diagnosed as neutrophilic eccrine hydradenitis related to cetuximab in the light of clinical and histopathological findings. According to our knowledge, this is the first reported case of neutrophilic ecrine hydradenitis due to cetuximab.