Flask and reactor scale production of plant growth regulators by Inonotus hispidus: optimization, immobilization and kinetic parameters.

The aims of the presented study are to compare submerged, static, and solid-state fermentation in the production of gibberellic acid (GA3), indole acetic acid (IAA), and abscisic acid (ABA) by Inonotus hispidus, to optimize with a statistical approach, and to determine the kinetic parameters under flask and reactor conditions. The maximum concentrations of GA3, (2478.85 ± 68.53 mg/L), ABA, (273.26 ± 6.17 mg/L) and IAA (30.67 ± 0.19 mg/L) were obtained in submerged conditions. After optimization, these values reached 2998.85 ± 28.85, 339.47 ± 5.50, and 34.56 ± 0.25 mg/L, respectively. Immobilization of fungal cells on synthetic fiber, polyurethane foam, and alginate beads resulted in an increase in plant growth regulators (PGR) production by 5.53%- 5.79% under optimized conditions. At the reactor scale, a significant increase was observed for GA3 concentration, 5441.54 mg/L, which was 2.14 and 1.45 times higher than non-optimized and optimized conditions in the flask scale, respectively. The maximum values for ABA and IAA were 390.39 and 44.79 mg/L, respectively. Although the specific growth rate (µ) decreases relatively from non-optimized flask conditions to optimized reactor conditions, it was observed that the PGR amounts produced per liter medium (rp) and per gram biomass (Qp) increased significantly. This is the first report on the synthesis of PGR by Inonotus hispidus which could be crucial for sustainable agriculture.

Anticancer and antimetastatic activity of Hypomyces chrysospermus, a cosmopolitan parasite in different human cancer cells.

The importance of microbial natural compounds in drug research is increasing every year and they are used to prevent or treat a variety of diseases. Hypomyces chrysospermus is a cosmopolitan parasite of many Boletaceae members. Since not much work has been conducted to date, this study is undertaken to explore the anticancer effect, including the antiproliferative and antimetastatic activity of Hypomyces chrysospermus. The aim of this study is to determine the antiproliferative and antimetastatic activity of Hypomyces chrysospermus ethyl acetate extract, having antioxidant activity, against A549, Caco2, MCF-7 human cancer and CCD-19 Lu and CCD 841 CoN healthy human cell lines. Firstly, cytotoxic activity was determined by the WST-1 assay. After cell proliferations and anti-metastatic effects were investigated by a real-time cell analysis system (RTCA-DP) and IC50 concentrations were calculated for each cell line. In addition, the expression levels of Apaf-1, TNF and NF-kB mRNA in cancer cells were investigated with RealTime-PCR. The ethyl acetate extract of Hypomyces chrysospermus presented anticancer activities including antiproliferative and antimetastatic effects. Hypomyces chrysospermus as a source of biologically active metabolites can be used as an important resource in the development of new anticancer effective agents.

Relationship of spontaneous pneumothorax cases seen in Eastern Black Sea region with meteorological changes.

The relationship of climate changes or weather conditions with the incidence of pneumothorax has been explored for many years. We aimed at revealing the effects of meteorological changes on the incidence of pneumothorax in the Eastern Black Sea region where spontaneous pneumothorax cases are seen relatively more frequently. The records of 195 subjects (179 males and 16 females) who had been monitored and treated due to spontaneous pneumothorax between January 2006 and December 2012 at our clinic were reviewed retrospectively, and their relationship was investigated with the meteorological data obtained by going through the database archive records of the 11th Regional Meteorology Directorate for the years between 2006 and 2012. Wind velocity was observed to be less in the days of having spontaneous pneumothorax than in the days of having no spontaneous pneumothorax, and the difference was found statistically significant (P = 0.026). The people of our region whose active lifestyle is reflected in their working life, social life, and even in their folk dances usually take a rest in the days of slower wind speed. We think that this state of resting leads to an increase in the frequency of spontaneous pneumothorax.

Statistical optimization of lovastatin production by Omphalotus olearius (DC.) singer in submerged fermentation.

In this study, culture conditions were optimized to improve lovastatin production by Omphalotus olearius, isolate OBCC 2002, using statistical experimental designs. The Plackett-Burman design was used to select important variables affecting lovastatin production. Accordingly, glucose, peptone, and agitation speed were determined as the variables that have influence on lovastatin production. In a further experiment, these variables were optimized with a Box-Behnken design and applied in a submerged process; this resulted in 12.51 mg/L lovastatin production on a medium containing glucose (10 g/L), peptone (5 g/L), thiamine (1 mg/L), and NaCl (0.4 g/L) under static conditions. This level of lovastatin production is eight times higher than that produced under unoptimized media and growth conditions by Omphalotus olearius. To the best of our knowledge, this is the first attempt to optimize submerged fermentation process for lovastatin production by Omphalotus olearius.

Effects of polysaccharide from fruiting bodies of Agaricus bisporus, Agaricus brasiliensis, and Phellinus linteus on alcoholic liver injury.

In the present study, the curative effects of crude polysaccharides (PSs) from mushrooms on the symptoms of alcoholic liver injury were investigated. PSs from Agaricus bisporus, Agaricus brasiliensis, and Phellinus linteus fruiting bodies were administered by gavage at levels of 100 mg per kg body weight per day for 7 d after the onset of the disease. The caspase-3 activity, mitochondrial membrane potential, mitochondrial outer membrane integrity of the liver tissues of sacrificed rats, and the serum alanine aminotransferase (ALT) levels were determined. In addition, light and transmission electron microscope (TEM) studies were performed for histopathological and cytological evaluations on liver sections. PSs from A. brasiliensis decreased ALT level and mitochondrial membrane potential and increased the outer membrane integrity; microscopic examinations also revealed normal hepatocytes and tissue. On the basis of our data, it can be argued that crude PSs from Agaricus brasiliensis have therapeutic potential for alcoholic liver injury.

Consumption of Coprinus comatus polysaccharide extract causes recovery of alcoholic liver damage in rats.

CONTEXT: Excess use of alcohol is known to be associated with liver diseases such as fatty liver, alcoholic hepatitis, and cirrhosis. Various practices may be applied to prevent or treat the damage caused by chronic alcoholism. Coprinus comatus (O.F. Müll.) Pers. (Agaricaceae) is a macrofungus that has been reported to aid the recovery of murine livers damaged by benzopyrene. OBJECTIVE: In this study, the possible therapeutic effects of three different doses (50, 100, and 150 mg/kg) of C. comatus polysaccharide (PS) extract were studied in rats subjected to an alcoholic diet. The histological and biochemical results were compared between the control and experimental groups. MATERIALS AND METHODS: Modified Lieber-Decarli's calorie-adjusted liquid alcohol diet was given orally for 60 d. In addition to histopathology, alanine transaminase (ALT), aspartate transaminase (AST), mitochondrial membrane integrity, total cytochrome-c oxidase activity (TotalStCox), total mitochondrial cytochrome-c oxidase activity (TotalMtStCox), and caspase-3 values were used as liver parameters, and liver sections from all experimental groups were examined by electron microscopy. RESULTS: Using histopathological assessment, it was observed that there was a decline in liver hepatocyte vacuolization in the treatment group fed 50 mg PS/kg. The TotalStCox and TotalMtStCox values of this group differed from the EtOH control group (p < 0.05). DISCUSSION AND CONCLUSION: Daily administration of 50 mg/kg of C. comatus PS extract considerably reduced the negative effects of alcohol on liver structure and function.

Ochratoxin A biodegradation by Agaricus campestris and statistical optimization of cultural variables.

The goal of this study is to identify the optimum conditions for ochratoxin A (OTA) biodegradation by the supernatant of Agaricus campestris strain. The Plackett-Burman and Box-Behnken methods were used to determine optimum OTA degradation conditions of Agaricus campestris under various incubation conditions. The Plackett-Burman method was planned through 16 varied experiments with 15 variants. The three most potent variants, sucrose, yeast extract and wheat bran, were selected using the Box-Behnken methodology. Ochratoxin A biodegradation ratio of 46.67% has been specified in only 1 h under ideal growing conditions. This is the first report on the optimization of OTA biodegradation by Agaricus campestris. When compared to previously published articles, it can be asserted that Agaricus campestris has promise based on its OTA biodegradation ratio in only 1 h of reaction time.

Curative effect of crude exopolysaccharides of some macrofungi on alcohol-induced liver damage.

Abstract The authors investigate the curative effects of crude exopolysaccharides (EPS) produced by four Basidomycetes strains on the symptoms of alcoholic liver injury. EPSs were administered to experimental groups at levels of 100 mg per kg body weight per day for 7 days using an oral zonde needle after the onset of the disease. Serum levels of alanine transaminase (ALT), lactate dehydrogenase (LDH), DNA fragmentation, caspase-3 activities, and mitochondrial outer membrane integrity were determined following sacrifice of the rats. Light and transmission electron microscope (TEM) studies were performed on liver sections for histopathological and cytological evaluations. EPS that was obtained from Coprinus comatus OBCC 1014 decreased serum ALT level and increased outer membrane integrity, and allowed for the regaining of histologically and cytologically normal hepatocyte and tissue views. As a result, based on the obtained data, it can be argued that among all studied mushroom strains crude exopolysaccharides from Coprinus comatus OBCC 1014 strain have therapeutic potential for alcoholic liver injury according to control groups.

Optimization and reactor-scale production of plant growth regulators by Pleurotus eryngii.

The aims of the this study are to select the best cultivation type for plant growth regulator (PGR) production, to optimize PGR production with statistical experimental design, and to calculate bioprocess parameters and yield factors during PGR production by P. eryngii in flask and reactor scales. Submerged fermentation was the best cultivation type with 4438.67 ± 37.14, 436.95 ± 27.31, and 54.32 ± 3.21 mg/L of GA3, ABA, and IAA production values, respectively. The Plackett-Burman and Box-Behnken designs were used to determine effective culture parameters and interactive effects of the selected culture parameters on PGR production by Pleurotus eryngii under submerged fermentation. The statistical model is valid for predicting PGR production by P. eryngii. After these studies, maximum PGR production (7926.17 ± 334.09, 634.92 ± 12.15, and 55.41 ± 4.38 mg/L for GA3, ABA, and IAA, respectively) was reached on the 18th day of fermentation under optimized conditions. The optimum formula was 50 g/L fructose, 3 g/L NaNO3, and 1.5 g/L KH2PO4, 1 mg/L thiamine, incubation temperature 25 °C, initial medium pH 7.0, and an agitation speed of 150 rpm. The kinetics of PGR production was investigated in batch cultivation under 3-L stirred tank reactor conditions. Concentrations of GA3, ABA, and IAA of 10,545.00 ± 527.25, 872.32 ± 21.81, and 60.48 ± 3.48 mg/L were obtained at the reactor scale which were 4.1, 3.4, and 2.3 times higher than the initial screening values. The specific growth rate (µ), the volumetric (rp) and specific (Qp) PGR production rates, 486.11 mg/L/day and 107.43 mg/g biomass/day for GA3, confirmed the successful transfer of optimized conditions to the reactor scale. In the presented study, PGR production of P. eryngii is reported for the first time.

Screening of medicinal higher Basidiomycetes mushrooms from Turkey for lovastatin production.

As a first attempt, a study was carried out to test for lovastatin production ability in local higher Basidiomycetes mushroom isolates from Turkey. An extended screening was performed for lovastatin production in yeast lactose agar medium, among a total of 136 macrofungi isolates from the Basidiomycetes Culture Collection of Eskisehir Osmangazi University. Lovastatin production was evaluated by disc diffusion method and was also confirmed by TLC and HPLC. Only six isolates were found to be lovastatin producers. The highest production of lovastatin was obtained from the extracts from Omphalotus olearius OBCC 2002 and Pleurotus ostreatus OBCC 1031. The lovastatin amount produced by commercial strains, Aspergillus terreus NRRL 255 (7.0 mg/L) and Penicillium citrinum NRRL 1841 (7.0 mg/L), was nearly comparable to the amount produced by Pleurotus ostreatus OBCC 1031 (5.8 mg/L) and Omphalotus olearius OBCC 2002 (4 mg/L).

Comparative Analysis of Antioxidant and Anticancer Activities of Fruiting Bodies, Mycelial Biomass and Culture Liquid of Omphalotus olearius OBCC 2503 (Agaricomycetes) from Turkey.

Up to now, the comparative analysis of the antioxidant and anticancer activities of the ethyl acetate extracts from fruiting body, mycelial biomass, and culture liquid of Omphalotus olearius are not reported. In this study, antioxidant activity of different samples was compared by six different methods: DPPH and ABTS radical scavenging activities, metal chelating activity, reducing power, inhibition of ß-carotene bleaching, and lipid peroxidation through ferric thiocyanate assay. Total phenolic contents of the samples were also determined. The mycelial biomass produced in vitro conditions presented the highest activity by five of them which was following by culture liquid. The potential antiproliferative effects of the extracts in cell culture, human cancer (A549, Caco2 and MCF-7) and healthy (CCD-19LU and CCD 841 CoN) cell lines by WST-1 assay was also evaluated. The highest anticancer effects were determined on A549 lung cancer cells treated with mycelial biomass extract and Caco2 colon cancer cells treated with culture liquid extract. The culture liquid was more effective on the cells than mycelial biomass according to IC50 values. All tested extracts were also evaluated for their toxicity against brine shrimp larvae (Artemia salina). O. olearius culture liquid and mycelial biomass extracts showed highest and lowest toxicity against A. salina, respectively.

Synthesis, antimicrobial activity and modeling studies of thiazoles bearing pyridyl and triazolyl scaffolds.

In this study, novel 4-(5-((2/3/4-substituted benzyl)thio)-4-(4-substituted phenyl)-4H-1,2,4-triazol-3-yl)-2-(pyridin-3/4-yl)thiazoles were synthesized following a multi-step synthetic procedure. All the compounds were screened with a panel of gram positive/negative bacteria, yeasts, and molds for antimicrobial activity using the disc diffusion method. Then, the minimum inhibitor concentration (MIC) and the minimum bactericidal concentration (MBC) values of active compounds were determined against Micrococcus luteus, Bacillus cereus, Listeria monocytogenes, and Staphylococcus aureus using the broth microdilution technique. These compounds were also screened for their inhibitory activities against S. aureus DNA gyrase by supercoiling assay. Furthermore, the crystal structure of S. aureus DNA gyrase B ATPase was subjected to a docking experiment to identify the possible interactions between the most active ligand and the active site. Lastly, the in silico technique was performed to analyze and predict the drug-likeness, molecular and ADME properties of the synthesized molecules.

First Report on the In Vivo Prebiotic, Biochemical, and Histological Effects of Crude Polysaccharide Fraction of Golden Chantharelle Mushroom, Cantharellus cibarius (Agaricomycetes).

In this study, we hypothesize that Cantharellus cibarius polysaccharide (CCP) could contribute to the growth performance, specific growth rate, and bacteriological, biochemical, and histological aspects of the gut in rats. Three groups of seven rats received one of three diets; negative control, treatment, and positive control groups were supplemented with commercial feed, + 0.25% CCP, and + 0.25% inulin, respectively. The highest Lactobacillus and the lowest Clostridium counts (1.84 × 109 and 0.03 × 109 CFU/ml, respectively) were obtained in the colon content of rats fed with CCP. The prebiotic index value of the treatment group (9.39) was higher than that of the positive control, inulin (7.74). There was a slight increase in total cholesterol and low-density lipoprotein and a slight decrease in high-density lipoprotein and triglyceride levels in the treatment group. Histologically, any important increase in mesenteric mast cell counts and degranulation has not been determined. The results indicate that CCP displays a prebiotic effect through the change of the intestinal microbiota. To our knowledge, the prebiotic effect of polysaccharide of C. cibarius fruiting body under in vivo conditions is reported here for the first time.

Synergistic activities of three triazoles with caspofungin against Candida glabrata isolates determined by time-kill, Etest, and disk diffusion methods.

Combinations of voriconazole, fluconazole, and itraconazole with caspofungin were evaluated against 50 Candida glabrata isolates by the time-kill, disk diffusion, and Etest methods. The majority of antifungal combinations were indifferent. By the time-kill method, synergistic activity was detected with eight (16%) of the caspofungin-voriconazole and seven (14%) of the caspofungin-fluconazole combinations, but synergy was not seen with the caspofungin-itraconazole combination. Further comparisons of the Etest and disk diffusion synergy techniques with the time-kill method are warranted.

Selection of Streptomyces isolates from Turkish karstic caves against antibiotic resistant microorganisms.

In this work, actinomycetes isolates were isolated from rock wall and speleothem surfaces and soil samples of 19 karstic caves in Turkey. Out of 290 isolates isolated, 180 isolates (62%) exhibited antimicrobial activity against a panel of four bacteria, two yeasts and four filamentous fungi in the screening program. One of them, Streptomyces sp. 1492, was examined for antibiotic production in batch culture. The maximum of antimicrobial activity was shown at 5th day. Antimicrobial activity of the extracted active compound was recorded as dose dependent bacteriostatic or bactericidal against antibiotic resistant clinical bacteria strains; methicillin-resistant Staphylococcus aureus (MRSA), vancomycin resistant Enterobacter faecium (VRE), and Acinetobacter baumanii. Minimum inhibitor concentration and minimum bactericidal concentrations were determined as lower than standard antibiotic streptomycin; 125 microg/ml and 250-1000 microg/ml, respectively. Active component was found as heat-stable.

Statistical optimization of cultural variables for enzymatic degradation of aflatoxin B1 by Panus neostrigosus.

The aim of this study is to determine the best aflatoxin B1 degradation conditions which was optimized using a combination of the Plackett-Burman and Box-Behnken methods with Panus neostrigosus culture filtrate. Panus neostrigosus was grown in a modified Kirk Broth medium to determine optimal degradation conditions. As a result, aflatoxin B1 was degraded under varying culture conditions. The Plackett-Burman method was designed after sixteen different experiments with fifteen variables. The three most effective variables (Sucrose, yeast extract, wheat bran) were chosen for the Box-Behnken methodology. The aflatoxin B1 degradation rate was 49% in just 1 h exposure to culture filtrate which was obtained under optimal growth conditions; (g-ml/L) sucrose 10, yeast extract 3, wheat bran 3, soytone 5, KH2PO4 2, MgSO4.7H2O 0.5, CaCl2.H2O 0.1, ammonium tartrate 2, trace element solution 10; 28 °C of incubation temperature, medium pH 5, 7.5% inoculum rate, 125 rpm of agitation speed, and a twelve-day incubation period. The SDS-PAGE studies show that the enzyme responsible for AFB1 degradation has 38 kDa molecular weight and has no laccase or MnP activity. To the best of our knowledge, this is the first report for AFB1 degradation by Panus neostrigosus.

Antifungal activity of caspofungin in combination with amphotericin B against Candida glabrata: comparison of disk diffusion, Etest, and time-kill methods.

The in vitro activities of caspofungin plus amphotericin B against 50 Candida glabrata isolates were evaluated by the time-kill, disk diffusion, and Etest methods. In vitro experiments showed a positive interaction. Even though each of these methods uses different conditions and endpoints, the results of the different methods frequently agreed.

Evaluation of the antioxidant activity of four edible mushrooms from the Central Anatolia, Eskisehir - Turkey: Lactarius deterrimus, Suillus collitinus, Boletus edulis, Xerocomus chrysenteron.

The methanolic extracts of Lactarius deterrimus, Suillus collitinus, Boletus edulis, Xerocomus chrysenteron were analyzed for their antioxidant activities in different test systems namely beta-carotene/linoleic acid, DPPH free radical scavenging, reducing power and metal chelating activities in addition to their total phenolic and flavonoid contents. In beta-carotene/linoleic acid and DPPH systems, L. deterrimus and B. edulis showed the strongest activity patterns. Their activities were as strong as the positive controls. The reducing power of the species was excellent. Chelating capacity of the extracts was increased with the increasing concentration. On the other hand, B. edulis found to have the highest phenolic content. Total flavonoid content of S. collitinus found the superior to the other mushrooms.

Effects of Black Hoof Medicinal Mushroom, Phellinus linteus (Agaricomycetes), Polysaccharide Extract in Streptozotocin-Induced Diabetic Rats.

In this article we report the healing effects of a Phellinus linteus fruiting body hot water extract (PLE) in streptozotocin (STZ)-induced diabetic rats. PLE was given before and after STZ. The preprotective, protective, and postprotective effects of PLE on STZ-induced oxidative stress were studied using biochemical (caspase 3 activity, cytosolic-to-lysosomal ratio of cathepsin B and L, DNA fragmentation levels), ordinary histological and immuno-histochemical investigation parameters. Following oral administration of PLE after STZ application, the serum glucose concentration significantly decreased up to 41.13% compared with the control group (P < 0.05). The hypoglycemic potential of the PLE was further supported by an increase of insulin secretion in the islets of Langerhans. In addition, the number of cells in Langerhans islets increased by 45.89% when PLE was given after STZ application. On the other hand, the use of PLE before oxidative stress could not prevent the onset of diabetes. This is, to our knowledge, the first study of the effect of application time of orally administered Ph. Linteus hot water extract on STZ-induced diabetes.

Enhanced Bioactive Exopolysaccharide Production by Mossy Maze Polypore, Cerrena unicolor (Higher Basidiomycetes) in Submerged Culture Conditions.

In this study, the culture requirements of the Cerrena unicolor OBCC 5005 strain were determined to optimize bioactive exopolysaccharide production in submerged culture. The effects of initial medium pH, carbon and nitrogen sources, inoculum age and amount, and mineral source on exopolysaccharide and mycelial biomass production by the C. unicolor OBCC 5005 strain were studied using a one-factor-at-a-time method. The highest exopolysaccharide production was obtained when culture parameters were used as initial medium pH: 5.5, 5% sucrose, 5% mycological peptone, and 5% of 4-day inoculants in the presence of 5 mM Fe2+. Optimized culture conditions at a flask scale were applied to a 3-L stirred tank reactor. As a result, 7.92 g/L and 7.34 g/L maximum exopolysaccharide production in optimized conditions at flask and stirred-tank reactor scales were achieved, respectively. The present study is the first to prove that C. unicolor can yield high bioactive exopolysaccharide production at flask and stirred-tank reactor scales.