Recep Bayram

Apoptotic, antioxidant and antiradical effects of majdine and isomajdine from Vinca herbacea Waldst. and kit

In the present study, apoptotic, antioxidant and antiradical effects of majdine and isomajdine from Vinca herbacea Waldst. and Kit were studied. For testing the possible apoptotic effects of majdine and isomajdine from V. herbacea, DNA fragmentation assay was conducted on the rat brain cortical tissue homogenates, in vitro. Also their possible effects on mitochondrial activity were tested by using the same tissue samples of rats. In addition, the antioxidant activity of isomajdine and majdine was determined using various in vitro antioxidant assays, including 2,2′-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS•+) radical scavenging and N,N-dimethyl-p-phenylenediamine (DMPD•+) radical scavenging, ferric ions (Fe3+) and cupric ions (Cu2+) reducing abilities and ferrous ions (Fe2+) chelating activity. On the other hand, butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), α-tocopherol and trolox (6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid) were used as reference antioxidants.

Amanitin and phallotoxin concentration in Amanita phalloides var. alba mushroom

Although rarely seen, Amanita phalloides var. alba, a variety of A. phalloides type mushrooms, causes mushroom poisoning resulting in death. Since it is frequently confused with some edible mushrooms due to its white colored cap and macroscopic appearance, it becomes important in toxicological terms. Knowledge of the toxin amount contained in this mushroom type is invaluable in the treatment of cases involving poisoning. In this study, we examined the toxin levels of various parts of the A. phalloides var. alba mushroom growing Duzce region of Turkey. Toxin analyses were carried out for A. phalloides var. alba, which were collected from the forests Duzce region of Turkey in 2011, as a whole and also separately in its spore, pileus, gills, stipe and volva parts. The alpha amanitin, beta amanitin, gamma amanitin, phalloidin and phallacidine analyses of the mushrooms were carried out using the RP-HPLC method. A genetic analysis of the mushroom showed that it had similar genetic characteristics as A. phalloides and was a variety of it. The lowest toxins quantity was detected in spores, volva and stipe among all parts of the mushroom. The maximum amount of amatoxins was measured in the gills. The pileus also contained a high amount of amatoxins. Generally, amatoxins and phallotoxin concentrations were lower as compared to A. phalloides, but interestingly all toxins other than gamma toxin were higher in the spores of A. phalloides var. alba. The amount of toxin in all of its parts had sufficient concentrations to cause death. With this study, the amatoxin and phallotoxin concentrations in A. phalloides var. alba mushroom and in its parts have been revealed in detail for the first time.

Amatoxin and phallotoxin concentration in Amanita phalloides spores and tissues.

Most of the fatal cases of mushroom poisoning are caused by Amanita phalloides. The amount of toxin in mushroom varies according to climate and environmental conditions. The aim of this study is to measure α-, β-, and γ-amanitin with phalloidin and phallacidin toxin concentrations. Six pieces of A. phalloides mushrooms were gathered from a wooded area of Düzce, Turkey, on November 23, 2011. The mushrooms were broken into pieces as spores, mycelium, pileus, gills, stipe, and volva. α-, β-, and γ-Amanitin with phalloidin and phallacidin were analyzed using reversed-phase high-performance liquid chromatography. As a mobile phase, 50 mM ammonium acetate + acetonitrile (90 + 10, v/v) was used with a flow rate of 1 mL/min. C18 reverse phase column (150 × 4.6 mm; 5 µm particle) was used. The least amount of γ-amanitin toxins was found at the mycelium. The other toxins found to be in the least amount turned out to be the ones at the spores. The maximum amounts of amatoxins and phallotoxin were found at gills and pileus, respectively. In this study, the amount of toxin in the spores of A. phalloides was published for the first time, and this study is pioneering to deal with the amount of toxin in mushrooms grown in Turkey.

Clinical importance of toxin concentration in Amanita verna mushroom.

Poisoning from Amanita group of mushrooms comprises approximately 3% of all poisonings in our country and their being responsible for nearly the entire fatal mushroom poisonings makes them important. These mushrooms contain primarily two types of toxins, amatoxins and phallotoxins. Phallotoxins have a more limited toxicity potential and they primarily consist of phalloidin (PHN) and phallacidin (PCN). Amatoxins, on the other hand, are very toxic and they primarily consist of alpha-amanitin (AA), beta-amanitin (BA) and gamma-amanitin (GA). Toxin levels can vary among various species, even among varieties of the same species, of Amanita mushroom family. Revealing the differences between the toxin compositions of the Amanita species that grow in our region may contribute to the clinics of poisonings. Our study aims at showing in detail the toxin levels in various parts of Amanita verna mushroom. A. verna mushrooms needed for toxin analysis were collected from Kozak Plateau near Ayvalik county of Balıkesir, Turkey in April 2013. The mushrooms were divided into their parts as pileus, gills, stripe and volva. Following the procedures required before the analysis, the AA, BA, GA, PHN and PCN levels were measured using the RP-HPLC method. While the lowest level of amatoxin was in the volva of the mushroom, the highest was measured in the gills. This was followed by pileus and stripe where the levels were close to each other. Similarly, the highest level of phallotoxin was measured in the gills. Gamma toxin and phalloidin were at lower amounts than the other toxins. A. verna is frequently confused with edible mushrooms with white caps due to its macroscopic similarity. If just one of them is eaten by mistake by an adult person with no mushroom experience, it can easily poison them. The amount of amatoxin is more as compared to Amanita phalloides and A. phalloides var. alba. Particularly, the AA and BA levels are approximately three times higher, whereas GA levels are lower. Similarly, the level of PCN is approximately four times higher as compared to A. phalloides and A. phalloides var. alba; by contrast, the level of PNH is about a half of theirs. In summary, it can be said that A. verna is a more toxic mushroom than A. phalloides and has a higher rate of mortality. With our study, the amatoxin and phallotoxin concentrations and distribution in A. verna mushrooms were shown in detail for the first time and it would be useful to carry out more similar studies with other members of Amanita family growing in various parts of the world.

Purification and in vitro toxicity of gamma amanitin

Abstract We aimed to obtain gamma amanitin with high purity through a purification process and compare toxic effects of alpha, beta, and gamma amanitin. Specific concentrations of the toxins (25, 10, 1, and 0.1 μg/mL) were applied to the C3A human hepatocytes. A MTT test was performed to determine the level of toxicity. Alpha amanitin showed a higher toxicity in 48 h while the lowest toxicity was observed in beta amanitin. The toxicity level of gamma amanitin was found between the alpha and beta amanitin toxicity. Our method is suitable for obtaining gamma amanitin with high purity (>99%) as well as for obtaining alpha amanitin and beta amanitin. Gamma amanitin has been shown to have equal responsibility for toxicity as alpha amanitin in amanita poisoning.

Toxin content and toxicological significance in different tissues and development stages of Lepiota brunneoincarnata mushroom

Abstract We investigated the quantity and concentration of toxins in different parts and in different growth phases of Lepiota brunneoincarnata mushroom. The amatoxins and phallotoxin levels were measured using the reversed phase high-performance liquid chromatography (RP-HPLC) method. Alpha amanitin (2.38 ± 0.70 mg/g) was followed by beta amanitin (1.97 ± 0.52 mg/g) and gamma amanitin (0.04 ± 0.01 mg/g) in trace amounts; it did not contain any phallotoxin. The cap part is richer in amatoxins than the stipe part. While medium mushrooms were quite rich in amatoxins, less levels of toxin were measured in fully developed mushrooms. The study showed in detail the toxin concentrations of L. brunneoincarnata with regard to different developmental stages and different segments.

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 108 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.

Trimetazidine increases cell survival and inhibits the activation of inflammatory response in sodium taurocholate-induced acute pancreatitis

Objective: To evaluate the therapeutic effects of trimetazidine (TMZ) in an experimental acute pancreatitis (AP) model induced with sodium taurocholate (STC). Summary of Background Data: At present...

Effects of erdosteine on alpha amanitin-induced hepatotoxicity in mice

Abstract The aim of this study was to investigate beneficial effects of erdosteine in the alpha amanitine-induced hepatotoxicity in mice. Three hours after giving alpha amanitin (0.5 mg/kg, i.p.) to the mice, they were administered silibinin (50 mg/kg/d, i.p.) or erdosteine (100 mg/kg/d, oral) therapies once a day for 3 d. A histopathological examination of their liver tissues was carried out 24 h after the last treatment; transaminase levels, blood urea nitrogen, urea, and creatinine were analyzed in serum. Erdosteine showed a beneficial effect by significantly improving the functional parameters particularly in alpha amanitin-induced hepatotoxicity and partially in renal toxicity. In the histopathological evaluation, the toxicity that was generated with alpha amanitin was significantly reduced by erdosteine, showing a possible hepatoprotective effect.