Combined point mutations in codon 12 and 13 of KRAS oncogene in prostate carcinomas

Prostate cancer is a common malignancy that develops by structural mutation(s) and/or other genetic alterations in specific genes.The G to T transversions in codon 12 and C to T transitions in codon 13 of KRAS proto-oncogene are predominant point mutations that occur in about 20% of different cancers in human. In the current study it was aimed to investigate the prevalence and predictive significance of KRAS mutations in patients with prostate carcinomas. In a total of 30 fresh tumoural tissue specimens were investigated in patients with prostate carcinoma. All tumoural specimens were histo-pathologically diagnosed and genotyped for codon 12, 13 KRAS point mutations by reverse hybridisation and direct sequencing methods. KRAS mutations were found in 12 (40%) samples with 29 samples deriving from adenocarcinomas and 1 sample was small cell prostate carcinoma. In 1 (3.44%) sample codon 12 was found to be mutated and in 2 (6.8%) samples codon 13 and in 9 (31%) samples combined codon 12 and 13 were found to be mutated particularly in higher grade of tumoural tissues. Our study, based on representative collection of human prostate tumours, indicates that combined mutations in codons 12 and 13 KRAS are relatively infrequent and most commonly occur in prostate carcinomas.     

Prevalence of known mutations in the MEFV gene in a population screening with high rate of carriers

The Familial Mediterranean Fever (FMF) shows an autosomal recessive pattern of inheritance and affects certain ethnic groups. Disease is caused by mutations in MEFV gene and more than 180 mutations have been defined in affected individuals. Current study aimed to determine the frequency-type of the mutations for MEFV gene in Sivas??middle Anatolian city. The cohort was composed of 3340 patients. MEFV gene mutations were studied by multiplex PCR based reverse hybridization stripAssay method. Patients?? clinical features were; family history: 68%, erysipelas-like erythema: 17.6%, fever: 89.9%, abdominal pain: 84.2%, peritonitis: 90.2%, arthritis: 33%, pleuritis: 14.2%, parental consanguinity: 21.2%. Current results revealed that M694V is the most frequent mutation (43.12%), followed by E148Q (20.18), M680I(G/C) (15.00%) and V726A (11.32%). The study population has a high rate of carriers and the E148Q mutation frequency was found to be highest when compared to the other regions of Turkey and other Mediterranean groups.     

Micronucleus frequency and lipid peroxidation in <Emphasis Type="Italic">Allium sativum</Emphasis> root tip cells treated with gibberellic acid and cadmium

Gibberellic acid (GA₃) is a very potent hormone whose natural occurrence in plants controls their development. Cadmium is a particularly dangerous pollutant due to its high toxicity and great solubility in water. In this study, the effect of GA₃ on Allium sativum root tip cells was investigated in the presence of cadmium. A. sativum root tip cells were exposed to CdNO₃ (50, 100, 200 μM), GA₃ (10-3 M), both CdNO₃ and GA₃. Cytogenetic analyses were performed as micronucleus (MN) assay and mitotic index (MI). Lipid peroxidation analysis was also performed in A. sativum root tip cells for determination of membrane damage. MN exhibited a dose-dependent increase in Cd treatments in A. sativum. GA₃ significantly reduced the effect of Cd on the MN frequency. MN was observed in GA₃ and GA₃ + 50 μm Cd treatments at very low frequency. MI slightly decreased in GA₃ and GA₃ + Cd treatments. MI decreased more in high concentrations of Cd than combined GA₃ + Cd treatments. The high concentrations of cadmium induce MN, lipid peroxidation and lead to genotoxicity in A. sativum. Current work reveals that the effect of Cd on genotoxicity can be partially restored with GA₃ application.     

Enzymatic hydrolysis of complex substrates following thermal pretreatment operations

Summary Most agroindustrial wastes generally undergo various thermal treatments during processing stages. The aim of this study was to determine the effect of different heat treatment methods on hydrolysis efficiency of such natural substrates. It was found that a wet thermal treatment generally increases the rate and degree of hydrolysis, while a dry one has a detrimental effect.     

Determination of saturated and unsaturated Fatty acids amount in leukocyte membranes from subjects fed with solid and fluid oils

Modifications in dietary fatty acid intake might lead to a modification in membrane phospholipid fatty acid composition. The purpose of this study was to investigate relationship between different type of oil consumption and leukocyte membrane phospholipid composition. This study was carried out in subjects utilizing butter (n = 15), margarine (n = 15), fluid oil (n = 15) and mixed types of oils (n = 15) in total 60 subjects. Leukocytes were separated from total blood by dextran sedimentation method. Membrane lipids and proteins were isolated following the cell disruption. Fatty acids of membrane phospholipids were isolated by hydrolysation with phospholipase B under ultrasonic dismembranator. Free fatty acids were identified with gas chromatography at chloroform phase. The results obtained were compared with data obtained by chromatograms of the standards. Results more prominent values of arachidic, dihomo-gamma-linolenic and palmitoleic acids were found in butter-or mixed oil-user groups; eicosadienoic, eicosamonoenoic, dihomo-gamma-linolenic and behenic acids in fluid oil heptanoic, valeric, eicosadienoic and linolenic acids in margarine groups. The fatty acid composition of mixed oil; was similar to butter, while other two oils were so different. From this study, it was concluded that the type of oil consumption might have an influence on phospholipid components of plasma membranes.     

A comparison of leptin and ghrelin levels in plasma and saliva of young healthy subjects

In the last 10 years, saliva has been increasingly used as a diagnostic fluid and in predictions of disease progression. Leptin and ghrelin are synthesized in several tissues including the salivary glands. The action of ghrelin is antagonistic to that of leptin. This study was undertaken to measure and compare the saliva ghrelin-leptin and plasma ghrelin-leptin levels in healthy young subjects. In 30 healthy subjects, after an overnight fast, saliva and plasma leptin levels were measured using the ELISA method while saliva and plasma immunoreactive ghrelin levels were measured using a commercial radioimmunoassay (RIA). The latter uses 125I-labeled bioactive ghrelin as a tracer and a rabbit polyclonal antibody raised against full-length octanoylated human ghrelin (Phoenix, Europe, Karlsruhe, Germany). The results of this investigation revealed that saliva leptin levels (6.19+/-2.10 microg/l) were lower than plasma levels (7.39+/-3.23 microg/l) while saliva ghrelin levels (188.5+/-84.7 pg/ml) were higher than plasma levels (126.4+/-38.5 pg/ml), when male and female subjects were considered together. Saliva leptin levels (5.93+/-1.94 microg/l) were lower than plasma levels (6.22+/-2.92 pg/ml) while saliva ghrelin levels (190.3+/-80.2 pg/ml) were higher than plasma levels (120.4+/-35.7 pg/ml) in young males. Saliva leptin levels (6.47+/-2.29 microg/l) were lower than plasma levels (8.73+/-3.14 microg/l) while saliva ghrelin levels (183.2+/-90.2 pg/ml) were higher than plasma levels (129.3+/-42.8 pg/ml) in young females, and both saliva and plasma leptin levels were slightly lower in male subjects in comparison with female subjects. Also, Immunohistochemistry study indicated that ghrelin positivity was found in ductus epithelium of salivary gland. We have demonstrated for the first time that saliva ghrelin levels were higher than in plasma while saliva leptin levels were almost the same as in plasma. Measurements of ghrelin and leptin in saliva is non-invasive, simple, and generally much preferred by patients and thus may be an acceptable alternative to plasma sampling.     

RAB3GAP1 and RAB3GAP2 modulate basal and rapamycin-induced autophagy

Macroautophagy is a degradative pathway that sequesters and transports cytosolic cargo in autophagosomes to lysosomes, and its deterioration affects intracellular proteostasis. Membrane dynamics accompanying autophagy are mostly elusive and depend on trafficking processes. RAB GTPase activating proteins (RABGAPs) are important factors for the coordination of cellular vesicle transport systems, and several TBC (TRE2-BUB2-CDC16) domain-containing RABGAPs are associated with autophagy. Employing C. elegans and human primary fibroblasts, we show that RAB3GAP1 and RAB3GAP2, which are components of the TBC domain-free RAB3GAP complex, influence protein aggregation and affect autophagy at basal and rapamycin-induced conditions. Correlating the activity of RAB3GAP1/2 with ATG3 and ATG16L1 and analyzing ATG5 punctate structures, we illustrate that the RAB3GAPs modulate autophagosomal biogenesis. Significant levels of RAB3GAP1/2 colocalize with members of the Atg8 family at lipid droplets, and their autophagy modulatory activity depends on the GTPase-activating activity of RAB3GAP1 but is independent of the RAB GTPase RAB3. Moreover, we analyzed RAB3GAP1/2 in relation to the previously reported suppressive autophagy modulators FEZ1 and FEZ2 and demonstrate that both reciprocally regulate autophagy. In conclusion, we identify RAB3GAP1/2 as novel conserved factors of the autophagy and proteostasis network.     

Dephenolization and decolorization of olive mill wastewater through sequential batch and co-culture applications

In a previous work it was reported adapted Trametes versicolor FPRL 28A INI culture was used to treat undiluted olive mill wastewater (OMW) without addition of any nutrients with significant amount of total phenolics were removed. However, decolorization was not so pronounced. Therefore, the aim of this study is to enhance the efficiency of dephenolization and decolorization of the primary treatment with adapted Trametes versicolor, incorporating a secondary biological treatment step using different microorganisms with sequential batch and co-culture applications. Through sequential batch applications Funalia trogii ATCC 20080 was found to have a higher potential in terms of total phenolics removal and decolorization amongst the tested organisms and better results were obtained from sequential batch applications as compared to co-culture experiments. In sequential batch applications, up to 91% total phenolics were removed and 64% decolorization was achieved after 24 days with 20% (v/v) inoculation rate of F. trogii when malt extract broth was used in inoculum preparation. In addition, significant accumulation of laccase (2019 ± 121.13Ul⁻¹) and manganese peroxidase (463 ± 33.89 Ul⁻¹) activities were attained. In co-culture applications highest total phenolics removal and decolorization were 78 and 39%, respectively, with non-adapted T. versicolor, whereas highest laccase and manganese peroxidase acitivities were obtained with F. trogii as 2219 Ul⁻¹  ± 176.14^. and 513 ± 4.12 Ul⁻¹, respectively.     

Dephenolisation of olive mill wastewater using adapted Trametes versicolor

Olive mill wastewater (OMW) is an effluent of the olive oil extraction process. The large volumes involved, along with the high phenolic content and chemical oxygen demand, cause major environmental problems. The presence of phenolics limits the effectiveness of aerobic or anaerobic treatment of this wastewater. In most of the studies performed on OMW, the concentration of phenolics is reduced by diluting the OMW prior to biological treatment, which leads to an increase in waste volume. Therefore, the aim of this work was to investigate the possibility of reducing the phenolic content without dilution and without any addition of nutrients or pretreatment by using the white-rot fungi Trametes versicolor FPRL 28A INI. Through an adaptation process, the fungus was able to remove 78% of total phenolics in shake flask experiments and 39% in static culture using undiluted OMW medium. In continuously stirred tank reactor (CSTR) conditions, 70% of total phenolics removal was achieved. Analysis with GC–MS showed that all simple phenolics disappeared from the medium after the 8th day of cultivation at an 0.25 vvm aeration rate. The maximum activities of phenol degrading enzymes laccase and manganese peroxidase (MnP) obtained under these conditions were 762.14 ± 42.11 and 97.80 ± 8.11 U l^?1 respectively.