Oksidacijski i apoptotski učinci fluoksetina i njegova metabolita norfluoksetina u vodenbuhe Daphnia magna

The aim of this study was to investigate the oxidative and apoptotic potential of fluoxetine, a widely used antidepressant in Turkey and the world, and of its metabolite norfluoxetine on a model non-target organism, Daphnia magna to see how exposure to this group of antidepressants (specific serotonin reuptake inhibitors) could affect the aquatic environment in which they end up. Juvenile D. magna specimens were chronically exposed to fluoxetine and norfluoxetine alone and in combination at concentrations found in the aquatic environment (0.091 and 0.011 μg/L, respectively) and to their 10-fold environmental concentrations for 21 days. Another group of 17-day-old animals were subacutely exposed to 100-fold environmental concentrations for four days. After exposure, we measured their glutathione peroxidase (GPx) and cholinesterase (ChE) activities, thiobarbituric acid-reactive substances (TBARS), and total protein content spectrophotometrically, while mitochondrial membrane potential (MMP) was analysed by fluorescence staining, and cytochrome c and ERK1/2 protein content by Western blotting. This is the first-time cytochrome c and ERK1/2 were determined at the protein level in D. magna. We also measured their carapace length, width, and caudal spine length microscopically. At environmental concentrations fluoxetine and norfluoxetine caused an increase in ChE activity and brood production. They also caused a decrease in juvenile carapace length, width, and caudal spine length and depolarized the mitochondrial membrane. At 10-fold environmental concentrations, GPx activity, lipid peroxidation levels, cytochrome c, and ERK1/2 protein levels rose. The most pronounced effect was observed in D. magna exposed to norfluoxetine. Norfluoxetine also decreased brood production. Similar effects were observed with subacute exposure to 100-fold environmental concentrations. However, total protein content decreased. All this confirms that fluoxetine and norfluoxetine have oxidative and apoptotic potential in D. magna. Daphnia spp. have a great potential to give us precious insight into the mechanisms of environmental toxicants, but there is still a long way to go before they are clarified in these organisms.Cilj je ovoga istraživanja bio utvrditi oksidacijski i apoptotski potencijal fluoksetina, antidepresiva raširenoga u Turskoj i svijetu, i njegova metabolita norfluoksetina na modelu vodenbuhe Daphnia magna koji nije ciljani organizam djelovanja spojeva. Također smo željeli vidjeti kako izloženost toj skupini antidepresiva (specifičnih inhibitora ponovne pohrane serotonina) može utjecati na vodeni okoliš u kojem oni završe. Mlade jedinke vodenbuhe bile su izložene fluoksetinu (0,091 μg/L) i norfluoksetinu (0,011 μg/L), odvojeno i u kombinaciji, pri koncentracijama zamijećenima u okolišu I deseterostrukim okolišnim koncentracijama u trajanju od 21 dan (kronična izloženost). Jedna je skupina 17 dana starih vodenbuha također bila izložena stostrukoj okolišnoj koncentraciji u trajanju od četiri dana (subakutna izloženost). Potom su u životinja spektrofotometrijom izmjerene aktivnosti enzima glutation peroksidaze (GPx) i kolinesteraza (ChE) te razine reaktivnih tvari tiobarbituratne kiseline (TBARS) i ukupnih proteina. Potencijal mitohondrijske membrane (MMP) utvrđen je fluorescencijom, a proteini citokrom c i ERK1/2 Western blot metodom. Ovo je prvi put da su se u vodenbuhi citokrom c i ERK1/2 utvrđivali na razini proteina. Također je mikroskopski izmjerena dužina i širina oklopa i dužina repne bodlje vodenbuha. Pri okolišnim koncentracijama fluoksetin i norfluoksetin doveli su do povišene aktivnost ChE i većeg razmnožavanja te smanjenja (dužine i širine) karapaksa i repne bodlje u podmlatka i depolarizacije mitohondrijske membrane. Pri deseterostrukim okolišnim koncentracijama porasle su razine aktivnosti GPx, lipidne peroksidacije, citokroma c i ERK1/2 proteina. Norfluoksetin je pritom iskazao najsnažnije djelovanje te doveo do pada razmnožavanja. Slični su učinci primijećeni kod subakutne izloženosti stostrukim okolišnim koncentracijama fluoksetina i norfluoksetina, osim što je ona dovela i do pada ukupnih proteina. Naši rezultati potvrđuju da fluoksetin i norfluoksetin imaju oksidacijski i apoptotski učinak u vodenbuhe. Taj životinjski model može odlično poslužiti za stjecanje uvida u mehanizme toksičnoga djelovanja tvari u okolišu, no potrebna su daljnja istraživanja prije nego što ti mehanizmi postanu potpuno jasni

The Anticancer Activity of Cetraria Islandica (L.) Ach in Breast Cancer Cells Through Crosstalk of Ampk-α1 and Erk1/2 Signalling

In the present study, we aimed to evaluate the anticancer activities of Cetraria islandica (C.islandica) extracts on MCF-7 breast cancer cell lines. Cell viability, protein levels, apoptotic cells number, F-actin distribution were measured. Cell viability of MCF-7 breast cancer cells was found to be reduced in a dose-dependent manner.EC50 values of C.islandica on MCF-7 cells were found to be 9.2047 E-5 g/ml (cell amount) by using intelligence system. Expressions of p53, caspase 3 and Bcl-2, were shown to be elevated after low doses of extract and diminished after high dose treatments. PPAR- protein level was decreased, although AMP-activated kinases-α1 (AMPK-α1) protein level was increasedin its extract groups. ERK1/2 protein level was also elevated in its extract groups. 125 mg/ml of extract treated cells show a low decrease in actin filament density. MCF-7 cells with C.islandica treatment for 24 h increased the apoptotic cell percentage, though the cells-treated with C.islandica for 48 was high necrotic cells percentage. Consequently, the C.islandica extract treatment causes to elevate ERK1/2 and AMPK-α1 protein levels, resulting in PPAR- and then triggers the apoptosis by modulation caspase-3 and P53 protein levels. Therefore, C.islandica might be a good candidate for anticancer tissue, especially soft tissue tumours

The Investigation of Antidiabetic Effects of Leontice leontopetalum Extract on Human Pancreatic β Cell Lines (1.1B4) Treated with Streptozotocin

One of the alternative therapeutic methods is herbal medicine. Leontice leontopetalum belongs to Berberidaceae family. The aim of study was investigated the extract of LL on human pancreatic beta cell-treated with STZ. Materials and methods: The human pancreatic beta cell (1.1B4) line was used the current study. LL’s extracts (1, 10, 100, and 1000 ug/ml) were supplemented in media for twenty-four hours and/or after STZ treatment (10 and 20 mM). Cells survivals (MTT), cells proliferation were shown by using xCelligence. Insulin content and releasing were measured at 1.1, 8.4 and 16.7 mM glucose concentrations. Results: The result of MTT was shown that cell survival was decreased, based on dose-dependent. When looked at xCelligence results, cell proliferation in STZ groups and the lowest and highest concentrations of LL were attenuated in a dose-dependent manner. Also, cotreatments of LL and STZ were decreased as well. The result of insulin-releasing on glucose induction was shown that STZ concentration gave rise to reduce insulin content at low and high glucose levels. Also, co-treatment of LL and STZ attenuated insulin content based on dose. Conclusion: It was considered that LL treatment led to increased insulin realizing, resulting from decreasing insulin content in diabetic beta cells, but decrease cell survival

The frequency of Duchenne muscular dystrophy/Becker muscular dystrophy and Pompe disease in children with isolated transaminase elevation: results from the observational VICTORIA study

IntroductionElevated transaminases and/or creatine phosphokinase can indicate underlying muscle disease. Therefore, this study aims to determine the frequency of Duchenne muscular dystrophy/Becker muscular dystrophy (DMD/BMD) in male children and Pompe disease (PD) in male and female children with isolated hypertransaminasemia.MethodsThis multi-center, prospective study enrolled patients aged 3–216 months with serum alanine transaminase (ALT) and/or aspartate transaminase (AST) levels >2× the upper limit of normal (ULN) for ≥3 months. Patients with a known history of liver or muscle disease or physical examination findings suggestive of liver disease were excluded. Patients were screened for creatinine phosphokinase (CPK) levels, and molecular genetic tests for DMD/BMD in male patients and enzyme analysis for PD in male and female patients with elevated CPK levels were performed. Genetic analyses confirmed PD. Demographic, clinical, and laboratory characteristics of the patients were analyzed.ResultsOverall, 589 patients [66.8% male, mean age of 63.4 months (standard deviation: 60.5)] were included. In total, 251 patients (188 male and 63 female) had CPK levels above the ULN. Of the patients assessed, 47% (85/182) of male patients were diagnosed with DMD/BMD and 1% (3/228) of male and female patients were diagnosed with PD. The median ALT, AST, and CPK levels were statistically significantly higher, and the questioned neurological symptoms and previously unnoticed examination findings were more common in DMD/BMD patients than those without DMD/BMD or PD (p < 0.001).DiscussionQuestioning neurological symptoms, conducting a complete physical examination, and testing for CPK levels in patients with isolated hypertransaminasemia will prevent costly and time-consuming investigations for liver diseases and will lead to the diagnosis of occult neuromuscular diseases. Trial RegistrationClinicaltrials.gov NCT04120168

Melatonin prevents mitochondrial damage induced by doxorubicin in mouse fibroblasts through Ampk-Ppar gamma-dependent mechanisms.

İstanbul Bilim Üniversitesi, Sağlık Yüksekokulu.Background: Doxorubicin (brand name: Adriamycin®) is used to treat solid tissue cancer but it also affects noncancerous tissues. Its mechanism of cytotoxicity is probably related to increased oxidation, mitochondrial dysfunction, and apoptosis. Melatonin is reported to have antiapoptotic and antioxidative effects. The aim of this study was to determine whether melatonin would counteract in vitro cytotoxicity of doxorubicin in mouse fibroblasts and determine the pathway of its action against doxorubicin-induced apoptosis. Material/Methods: We measured markers of apoptosis (cytochrome-c, mitochondrial membrane potential, and apoptotic cell number) and oxidative stress (total oxidant and antioxidant status) and calculated oxidant stress index in 4 groups of fibroblasts: controls, melatonin-treated, doxorubicin-treated, and fibroblasts concomittantly treated with a combination of melatonin and doxorubicin

Preparation and characterization of papaverine-loaded poly[(R)-3- hydroxybutyrate] membranes to be used in the prevention of vasospasm

PubMed ID: 21502032The objective of this study is the preparation and characterization of poly[(R)-3-hydroxybutyrate] (PHB) membrane loaded with a vasodialative agent (i.e., papaverine hydrochloride) as a blood vessel coverage strip to be used in the prevention of undesired vascular vasospasm. Papaverine-loaded PHB membranes were especially designed to act locally and provide an efficient, long term, and sustained prevention of vasospasm at the site of the newly created vascular anastomosis without any systemic vasodilation effect of the papaverine, which may be life-threatening for the patient. The membranes were prepared by gelation of PHB followed by solvent casting. PHB membranes were characterized in terms of morphology, chemical structure, swelling behavior, in vitro drug release, degradation, and blood compatibility studies as in vitro coagulation tests. Activated partial thromboplastin time, prothrombin time, and fibrinogen concentration were measured by blood coagulation assays. Investigated and evaluated parameters for in vitro drug release, degradation, and in vitro coagulation studies were the concentration of PHB and drug content. Similar effective parameters were used for swelling behavior studies (i.e., concentration of PHB and drug content). Drug release, swelling behavior, degradation, and in vitro coagulation of the membranes were found to be influenced by these parameters. Obtained results showed that papaverine-loaded PHB membranes provide an efficient and advantegous means for the prevention of vasospasm during vascular anastomosis as a local application. ©PDA, Inc. 2010

Role of Vitamin D in Intima Media Thickness in Patients with Type 1 Diabetes Mellitus

Increased carotid intima media thickness indicates subclinical atherosclerosis. We evaluated the relation between vitamin D level and intima media thickness in patients with type 1 DM. 93 patients (female/male: 48/45, aged 31.5 +/- 11.9 years, A1c 9.48 +/- 2.43, vitamin D [15.9 (12.1-19.2)]) with type 1 DM were included into the study. Common carotid artery IMT was measured by real time B mode ultrasonography (MyLab 70 XVG, Esaote SpA, Genoa, Italy). Vitamin D was measured using radioimmunassay. Male and female patients (n = 14, 15%) had similar rates of plaque presence (p = 0.377). IMT was similar according to gender. IMT [0.45 (0.40-0.50)] was positively correlated with age, duration of diabetes, creatinine, LDL/HDL ratio, and ALP. Median IMT was higher in current smokers, patients with retinopathy, and nephropathy, and overweight/obese patients. IMT was not different according to vitamin D status. However calcium level corrected for albumin was in positive correlation with mean IMT (r = 0.221, p = 0.033). We detected high frequency of vitamin D deficiency (78%) defined as less than 20 ng/ml. Vitamin D and diabetes control defined as A1c have no effect on intima media thickness in type 1 DM. Traditional cardiovascular risk factors including age, duration of DM, smoking, and BMI adversely affect intima media thickness