Untreated Control Samples Research Articles

Rapid and Controllable Flame Reduction of TiO2 Nanowires for Enhanced Solar Water-Splitting

We report a new flame reduction method to generate controllable amount of oxygen vacancies in TiO2 nanowires that leads to nearly three times improvement in the photoelectrochemical (PEC) water-splitting performance. The flame reduction method has unique advantages of a high temperature (>1000 °C), ultrafast heating rate, tunable reduction environment, and open-atmosphere operation, so it enables rapid formation of oxygen vacancies (less than one minute) without damaging the nanowire morphology and crystallinity and is even applicable to various metal oxides. Significantly, we show that flame reduction greatly improves the saturation photocurrent densities of TiO2 nanowires (2.7 times higher), α-Fe2O3 nanowires (9.4 times higher), ZnO nanowires (2.0 times higher), and BiVO4 thin film (4.3 times higher) in comparison to untreated control samples for PEC water-splitting applications.

Mechanical properties of wood from Pinus sylvestris L. treated with Light Organic Solvent Preservative and with waterborne Copper Azole

Aim of study: To determine the effect on wood from Pinus sylvestris of treatment with preservatives on mechanical properties and to establish the relation between the penetration and compression strength.Area of study: SpainMaterial and Methods: 40 samples of defect-free wood from Pinus sylvestris L. were treated with Light Organic Solvent Preservative (Vacsol Azure WR 2601) and 50 with waterborne Copper Azole (Tanalith E 3492). 40 control samples were not treated (water or preservative). Mechanical resistance to static bending, modulus of elasticity and compression strength parallel to the grain were compared with untreated wood. Regression analysis between the penetration and compression strength parallel was done with the samples treated with waterborne preservative.Main results: The results indicate that the treated wood (with either product) presents a statistically significant increase in mechanical resistance in all three mechanical characteristics. The results obtained differ from earlier studies carried out by other authors.There was no correlation between parallel compression strength and the degree of impregnation of the wood with waterborne Copper Azole . The most probable explanation for these results concerns changes in pressure during treatment.The use of untreated control samples instead of samples treated only with water is more likely to produce significant results in the mechanical resistance studies.Research highlights: Treated wood presents a statistically significant increase in MOE, modulus of rupture to static bending and parallel compression strength.There was no correlation between parallel compression strength and the degree of impregnation with waterborne preservative.Keywords: Light Organic Solvent Preservative; MOE; parallel compression; static bending; waterborne Copper Azole; wood technology.

Inactivation of Penicillium digitatum and Penicillium italicum under in vitro and in vivo conditions by using UV-C light.

In this study, the effects of UV-C on two of the main wound pathogens of citrus fruits, Penicillium digitatum and Penicillium italicum, were investigated with different inoculation methods in vitro and on oranges. P. digitatum and P. italicum spores were inoculated onto the surface of potato dextrose agar or oranges using spread, spot, wound, and piercing inoculation methods. UV-C treatment for 1 min from a working distance of 8 cm reduced the numbers of P. italicum and P. digitatum by about 3.9 and 5.3 log units, respectively, following spread inoculation under in vitro conditions. Significant reductions were obtained after 1-min UV-C treatments of the tested fungi following inoculation using the spread and spot methods. With inoculation by the wound and piercing methods, the tested spores were not inactivated completely even after 10- and 20-min treatment times, respectively. The application of UV-C (7.92 kJ m(-2)) on oranges reduced the percentage of oranges infected at least threefold compared with the rate of infection in the untreated control samples. UV-C irradiation could effectively inactivate spores of P. italicum and P. digitatum inoculated by the spread plate and spot inoculation methods under in vitro and in vivo conditions. On the other hand, because of the low penetration ability of UV-C light, the tested fungi were not completely inactivated following inoculation with the wound and piercing methods. UV-C treatment has potential for use in surface decontamination of citrus fruits.

Effect of Xylopia aethiopica aqueous extract on antioxidant properties of refrigerated Roma tomato variety packaged in low density polyethylene bags.

Effects of Xylopia aethiopica (Dunal) A. Richard aqueous extract on the antioxidants of matured tomato fruits at red stage were investigated at 13 ± 2°C and 80 ± 5% relative humidity. A sample treated with sodium bicarbonate and untreated samples were included. Samples packaged in low density polyethylene (30μm thickness) bags were analysed at intervals of 5days. The treatments revealed statistically significant differences in ascorbic acid content of stored tomato fruits. Fruits treated with 5% X. aethiopica on day 5 of storage had 21.0mg/100g which was significantly (p < 0.05) higher than 18.2mg/100g in untreated control samples. At 15th day of storage, ascorbic acid was 10.0 and 14.2mg/100g in tomato fruits treated with sodium bicarbonate and 5% X. aethiopica respectively. The carotenoid and lycopene contents were lower in sodium bicarbonate-treated and the untreated control samples than in X. aethiopica-treated sample. The total phenolic contents were better retained in X. aethiopica-treated tomato than in control. Treatment of tomato fruits with X. aethiopica at 4 & 5% levels significantly retained the qualities evaluated.

Mineral nutrient imbalance, total antioxidants level and DNA damage in common bean (Phaseolus vulgaris L.) exposed to heavy metals.

The present study aimed to analyze the biological effects induced by bioaccumulation of metals in common bean (Phaseolus vulgaris L.). Effects of mineral nutrient imbalance, total antioxidants level and DNA damage induced by accumulation of heavy metals, were investigated in bean seedlings treated with two selected metal concentrations for 7days. Metal content is analyzed by inductively coupled plasma - atomic emission spectrometer (ICP-AES), for total antioxidants level assessment the Ferric-Reducing Antioxidant Power (FRAP) assay is used and Random Amplified Polymorphic DNA (RAPD) method was applied for investigation of DNA damages. The increasing metal concentration in the treatment medium changed synchronously metal content in samples, and decreased total antioxidant activity in all samples with exception only for samples treated with Ni and Cd. The obtained "DNA fingerprints" demonstrated that the increasing metal concentrations induced changes in RAPD profiles (disappearance and/or appearance of bands in comparison with untreated control samples). The highest number of missing bands was observed in samples treated with zinc (total 4 bands) and nickel (total 4 bands) at both concentrations. These results suggested that mineral nutrient imbalance is involved in changes of antioxidant levels and DNA damages of the seedlings, which may help to understand the mechanism of metal toxicity in plants.

TREATABILITY OF PLANTATION TIMBER SPECIES WITH CCA (COPPER·CHROME·ARSENATE) PRESERVATIVES AND THEIR EFFECTIVENESS AGAINST INSECT AND FUNGAL ArrACK

This study was conducted to asscss the treatability of four plantation timber species (PiIlIlScariboca, 1/('\'('(/ brasiliensis, EII(,I1/I,/)IIIS gran.lis and EII('(/{y{)IIIS microcorvs) withCoppcr-Chromc-Arscn.uc (CC A) by using pressure impregnation full-cell process. Thetreatability of core wood and outer wood of the species were also investigated. Treatabilitywas evaluated by measuring the preservative retention (Net Dry Salt Rctcnt ion or NDSR)and dcpt h of pellet rat ion.According to the results. Pinus I'IIl'ih(/(,1I showed the highest treatability while treatabilityof //C'I'I'(/ hrasilicnsis, Em (/{YfiIIlS grandis and EII('(/II'f'IIIS microcorvs were respectivelylower. A significant negative corrckuion was observed between density of the timber andtill' treatability. Rased on depth of penetration of preservatives. Pinus (,(/I'i/)II(,(/ and 1/('\'('(/brasiliensis were classified as permeable to preservative treatment. E. grandis was resistantwhile E. microtorvs was extremely resistant to preservative treatment.Trc.unbilitv of outer wood of all the species were higher than that of core wood indicatingthe higher pcnncabiiity of outer S;IPWOOt! area compared with the inner core wood.Trc;lting schedule with initial vacuum of -O.X bar. pressure of 6.5 bar maintained for IXOminutes ami final vacuum of -(),X bar was effective treatment of l l evcu hrasilicnsis. Evenwith this pressure. it was unable to achieve depth of penetration levels as specified in theSri Lankan Standards for E. grandis and E, IIIicro 1'01'\'.1'.When wood samples were exposed to exterior ground contact in the grave yard test for fivemonths. none of the treated samples were infected, hut untreated control samples wereattacked hv fungi and termites. This indicates the effectiveness of the CCA treatment incontrolling fungal and insect attack.

The effect of transglutaminase on some physicochemical and sensory properties of the Turkish drinking yoghurt Ayran

This article presents research on the effect of microbial transglutaminase on the properties of Turkish drinking yoghurt Ayran. Transglutaminase (TGase) enzyme was added to Ayran milk at different production steps, after pasteurisation and together with starter culture addition, and two different incubation times (10 min and 1 h) were used. Three TGase‐treated Ayran samples and control sample were analysed on 1st, 10th and 20th days of storage. TGase addition did not cause significant changes on chemical properties of Ayran (P &gt; 0.05). However, physical properties of Ayran were improved by TGase throughout the storage period. Compared to the untreated control sample, pretreatment of milk with enzyme increased the viscosity dramatically (P &lt; 0.05) and prevented serum separation. Scanning electron microscopic studies revealed that transglutaminase treatment of milk led to more regular distribution of proteins in Ayran gel and decreased gel permeability. TGase treatment did not show any unfavourable effect on the sensory properties of the final product.

Comparative Studies on Effect of Pretreatment of Rice Husk for Enzymatic Digestibility and Bioethanol Production

Three common pretreatment processes based on dilute sulfuric acid, dilute sodium hydroxide and heat treatment (autoclaving) followed by enzymatic hydrolysis were evaluated to provide comparative performance data. Among them, the best result was obtained when the pretreatment of rice husk was carried out with 3% of NaOH solution. The pretreatment of rice husk with NaOH substantially increased the lignin removal, enzymatic digestibility of cellulose, the accessibility of cellulose and fermentable sugar production. The highest glucose concentration, glucose yield and ethanol concentration were 14.54 g/L, 59.6% and 6.22 g/L; which were 5.44, 3.77 and 6.15 times higher than the untreated control samples, respectively. Moreover, the SEM analysis of the pretreated sample illustrated significant physical changes of the rice husk after NaOH pretreatment.

Comparison of sugar content for ionic liquid pretreated Douglas-fir woodchips and forestry residues.

BackgroundThe development of affordable woody biomass feedstocks represents a significant opportunity in the development of cellulosic biofuels. Primary woodchips produced by forest mills are considered an ideal feedstock, but the prices they command on the market are currently too expensive for biorefineries. In comparison, forestry residues represent a potential low-cost input but are considered a more challenging feedstock for sugar production due to complexities in composition and potential contamination arising from soil that may be present. We compare the sugar yields, changes in composition in Douglas-fir woodchips and forestry residues after pretreatment using ionic liquids and enzymatic saccharification in order to determine if this approach can efficiently liberate fermentable sugars.ResultsThese samples were either mechanically milled through a 2 mm mesh or pretreated as received with the ionic liquid (IL) 1-ethyl-3-methylimidazolium acetate [C2mim][OAc] at 120°C and 160°C. IL pretreatment of Douglas-fir woodchips and forestry residues resulted in approximately 71-92% glucose yields after enzymatic saccharification. X-ray diffraction (XRD) showed that the pretreated cellulose was less crystalline after IL pretreatment as compared to untreated control samples. Two-dimensional nuclear magnetic resonance spectroscopy (2D-NMR) revealed changes in lignin and hemicellulose structure and composition as a function of pretreatment. Mass balances of sugar and lignin streams for both the Douglas-fir woodchips and forestry residues throughout the pretreatment and enzymatic saccharification processes are presented.ConclusionsWhile the highest sugar yields were observed with the Douglas-fir woodchips, reasonably high sugar yields were obtained from forestry residues after ionic liquid pretreatment. Structural changes to lignin, cellulose and hemicellulose in the woodchips and forestry residues of Douglas-fir after [C2mim][OAc] pretreatment are analyzed by XRD and 2D-NMR, and indicate that significant changes occurred. Irrespective of the particle sizes used in this study, ionic liquid pretreatment successfully allowed high glucose yields after enzymatic saccharification. These results indicate that forestry residues may be a more viable feedstock than previously thought for the production of biofuels.

Abstract 4097: Targeting of mTOR mediated signaling pathways in papillary thyroid cancer cell.

Abstract The mammalian target rapamycin (mTOR) signaling cascade is a key regulatory pathway controlling initiation of mRNA in mammalian cells. Torin2 is a potent, selective and orally available mTOR inhibitor for treatment of cancer. Dysregulation of mTOR signaling has been found in many cancers, however its role Papillary Thyroid Cancer (PTC) has not been explained. Therefore in this study, we investigated the role of mTOR and its associated molecules in PTC cell lines using MTT, flow cytometry and DNA fragmentation assays and western blotting. Our data showed that Torin2 caused a dose dependent growth inhibition in all cell lines studied. Cell cycle assay showed that the sub-G1 population in TPC-1 cell increased from 12.44% in untreated control sample to 31.50% after 200nM of Torin2 treatment and 44.08% after 400nM of Torin2 treatment for 48 hours. Similarly BCPAP cells increased from 8.47% in untreated control sample to 33.54% after 200nM of Torin2 treatment and 41.16% after 400nM of Torin2 treatment for 48 hours. Further confirmation using FITC-conjugated Annexin V and PI assay showed that apoptotic cell increased from 8.89% in untreated cells to 58.54% and 73.54% after 200nM and 400nM treatment in TPC-1 cells, and from 13.24% in control to 46.74% and 70.64% in BCPAP cells, indicating a dose dependent apoptosis induced in both cell lines. Treatment of PTC cell lines with Torin2 inactivated downstream targets of mTOR; p70S6 and 4E-BP1. Torin2 treatment of PTC cell lines also induced mitochondrial dependent apoptosis via activation and cleavage of caspase-9, and 3, and cleavage of PARP. Furthermore Torin2 positively regulated autophagy by disrupting the Beclin-1 complex, releasing Beclin-1 to induce autophagy. Our results suggest mTOR and its associated signaling pathway play a critical role in PTC tumorigenesis, therefore targeting of mTOR expression may be a viable therapeutic intervention for treatment of these cancers. Citation Format: Maqbool Ahmed, Azhar R. Hussain, Fouad Al Dayel, Shahab Uddin, Khawla S. Al-Kuraya. Targeting of mTOR mediated signaling pathways in papillary thyroid cancer cell. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 4097. doi:10.1158/1538-7445.AM2013-4097

AOSP21 GENETIC POLYMORPHISMS OF DRUG-METABOLISING ENZYMES AND CHEMOTHERAPY RESISTANCE IN PATIENTS WITH LYMPHOPROLIFERATIVE DISEASES

s, 5th AOS and 9th Annual Conference of OOTR, 2013 / European Journal of Cancer 49S1 (2013) S1−S22 S13 AOSP19 ANTIMETASTATIC EFFECTS OF DIETHYLDITHIOCARBAMATE ON CHOLANGIOCARCINOMA CELLS P. Srikoona, R. Kariyaa, K. Vaeteewoottacharna,b, E. Kudoa, H. Gotoa, M. Tauraa, S. Wongkhamb, S. Okadaa *. aDivision of Hematopoiesis, Center for AIDS Research, Kumamoto University, Kumamoto, Japan, bDepartment of Biochemistry, Faculty of Medicine and Liver Fluke and Cholangiocarcinoma Research Center, Khon Kaen University, Khon Kaen, Thailand Introduction. Cholangiocarcinoma (CCA) is a malignant tumour of biliary ductswith poor prognosis andhighmortality rate. Highly constitutive activated transcription factor nuclear factor uB (NF-uB) is found generally in CCA cells. NF-uB is well known to play roles in different tumour genesis processes. Diethyldithiocarbamate (DDTC) has been reported to inhibit the NF-uB pathway, inducing cancer cell apoptosis. However, the effects of DDTC on cancer cell metastasis have not reported yet. Methods. Human CCA cell lines (KKU-M213, KKU-M214, and KKU-OCA17) were used to screen the inhibition of cell viability by DDTC treatment. Inhibitions of cell metastasis were performed using the KKU-M213 cell line. Cells treated with DDTC were compared with untreated (control) samples. Antimigration activities were investigated by wound healing assay and Transwell assay. Cell adhesion was observed by adhesion assay. NF-uB transcriptional activity was investigated by luciferase assay. Protein-DNA binding affinity was determined by electrophoresis mobility shift assay (EMSA). Western blot analysis was analysed to determine NF-uB protein levels. The levels of mRNA expression of NF-uB target genes related to cancer cellmetastasiswere determined by quantitative real-time PCR. Results. DDTC inhibited cell migration and adhesion in KKU-M213 cells in dose-dependent and time-dependent manners. Luciferase assay indicated that DDTC inhibit NF-uB transcriptional activity. The protein-DNA binding affinity was clearly suppressed, while protein levels showed the attenuated active p65 with DDTC treatment. The strong suppressionof intercellular adhesionmolecule-1 (ICAM1) gene expression suggested that CCA cell adhesion was inhibited by DDTC via the inhibition of NF-uB pathway. Discussion. These findings suggest that DDTC has an antimetastatic effect on CCA cells through the suppression of the NF-uB signalling pathway, which suggests therapeutic potential against human CCA metastasis. Funding. Grants-in-Aid for Science Research (No. 23107725) from the Ministry of Education, Science, Sports, and Culture of Japan. The authors declared no conflicts of interest. AOSP20 Abstract withdrawn AOSP21 GENETIC POLYMORPHISMS OF DRUG-METABOLISING ENZYMES AND CHEMOTHERAPY RESISTANCE IN PATIENTS WITH LYMPHOPROLIFERATIVE DISEASES E. Gorbachenkoa *, O. Gorevaa, A. Grishanovaa, E. Vigovskayab. aInstitute of Molecular Biology and Biophysics, Siberian Branch of the Russian Academy of Medical Sciences, Novosibirsk, Russian Federation, bMunicipal Clinical Hospital No. 2, Hematology Center, Novosibirsk, Russian Federation Introduction. One of the molecular mechanisms of multidrug resistance may be xenobiotic metabolism, including anticancer drugs. Cytochromes P450 (CYPs) and glutathioneS-transferases (GSTs) are important enzymes of phase I and phase II xenobiotic metabolism, respectively. CYPs and GSTP1 genes are polymorphic and can underlie the individual reactions of patients to chemotherapy. Our study analysed the association of functionally significant gene polymorphisms with chemotherapy resistance in patients with lymphoproliferative diseases. Methods. We determined CYP2C8*2, CYP2C8*3, CYP2C9*2, CYP2C9*3, CYP2C19*2, CYP2C19*3, CYP3A5*3, GSTP1 313A→G and GSTP1 341C→T genetic polymorphisms in blood samples of 79 patients (mean age 57 years, SD 13) with lymphoproliferative diseases (non-Hodgkin lymphoma, chronic lymphocytic leukaemia) by PCR-RFLP assays. All patients received chemotherapy (cyclophosphamide, doxorubicin, vincristine, and prednisolone). The odds ratio (OR) was used to estimate association of CYP genotypes with chemotherapy resistance risk and was calculated for the combined group (mutant homozygous and heterozygous) in comparison with the wild genotype group. Results.Wehave found the followingmutant allele frequencies in patients with chronic lymphoproliferative diseases: CYP2C8*2, 0.00; CYP2C8*3, 0.14; CYP2C9*2, 0.11; CYP2C9*3, 0.05; CYP2C19*2, 0.13; CYP2C19*3, 0.00; CYP3A5*3, 0.93; GSTP1 313A→G, 0.21; and GSTP1 341C→T, 0.12. Patients were divided in two groups according to the response to chemotherapy: effect was estimated as positive (n=57) in cases of complete or partial remission or stabilisation and as negative (n=22) in cases of progression of disease. The risk of resistance to chemotherapy for patients had one or two CYP2C8*3 or CYP2C9*2 allele was 7.05-fold significantly (p=0.002) and 4.1fold significantly (p=0.02) higher than for wild genotype carriers, respectively. There were no significant associations of CYP2C9*3 (OR 0.85, p=0.99), CYP2C19*2 (OR 0.88, p=0.99), CYP3A5*3 (OR 1.58, p=0.72), GSTP1 313A→G (OR 1.38, p=0.74), and GSTP1 341C→T (OR 1.21, p=0.97) genetic polymorphisms with chemotherapy response. Discussion. Our results suggest that CYP2C8*3 and CYP2C9*2 polymorphisms are associated with chemotherapy resistance in patients with lymphoproliferative diseases. Funding. None declared. The authors declared no conflicts of interest.

AOSP19 ANTIMETASTATIC EFFECTS OF DIETHYLDITHIOCARBAMATE ON CHOLANGIOCARCINOMA CELLS

s, 5th AOS and 9th Annual Conference of OOTR, 2013 / European Journal of Cancer 49S1 (2013) S1−S22 S13 AOSP19 ANTIMETASTATIC EFFECTS OF DIETHYLDITHIOCARBAMATE ON CHOLANGIOCARCINOMA CELLS P. Srikoona, R. Kariyaa, K. Vaeteewoottacharna,b, E. Kudoa, H. Gotoa, M. Tauraa, S. Wongkhamb, S. Okadaa *. aDivision of Hematopoiesis, Center for AIDS Research, Kumamoto University, Kumamoto, Japan, bDepartment of Biochemistry, Faculty of Medicine and Liver Fluke and Cholangiocarcinoma Research Center, Khon Kaen University, Khon Kaen, Thailand Introduction. Cholangiocarcinoma (CCA) is a malignant tumour of biliary ductswith poor prognosis andhighmortality rate. Highly constitutive activated transcription factor nuclear factor uB (NF-uB) is found generally in CCA cells. NF-uB is well known to play roles in different tumour genesis processes. Diethyldithiocarbamate (DDTC) has been reported to inhibit the NF-uB pathway, inducing cancer cell apoptosis. However, the effects of DDTC on cancer cell metastasis have not reported yet. Methods. Human CCA cell lines (KKU-M213, KKU-M214, and KKU-OCA17) were used to screen the inhibition of cell viability by DDTC treatment. Inhibitions of cell metastasis were performed using the KKU-M213 cell line. Cells treated with DDTC were compared with untreated (control) samples. Antimigration activities were investigated by wound healing assay and Transwell assay. Cell adhesion was observed by adhesion assay. NF-uB transcriptional activity was investigated by luciferase assay. Protein-DNA binding affinity was determined by electrophoresis mobility shift assay (EMSA). Western blot analysis was analysed to determine NF-uB protein levels. The levels of mRNA expression of NF-uB target genes related to cancer cellmetastasiswere determined by quantitative real-time PCR. Results. DDTC inhibited cell migration and adhesion in KKU-M213 cells in dose-dependent and time-dependent manners. Luciferase assay indicated that DDTC inhibit NF-uB transcriptional activity. The protein-DNA binding affinity was clearly suppressed, while protein levels showed the attenuated active p65 with DDTC treatment. The strong suppressionof intercellular adhesionmolecule-1 (ICAM1) gene expression suggested that CCA cell adhesion was inhibited by DDTC via the inhibition of NF-uB pathway. Discussion. These findings suggest that DDTC has an antimetastatic effect on CCA cells through the suppression of the NF-uB signalling pathway, which suggests therapeutic potential against human CCA metastasis. Funding. Grants-in-Aid for Science Research (No. 23107725) from the Ministry of Education, Science, Sports, and Culture of Japan. The authors declared no conflicts of interest. AOSP20 Abstract withdrawn AOSP21 GENETIC POLYMORPHISMS OF DRUG-METABOLISING ENZYMES AND CHEMOTHERAPY RESISTANCE IN PATIENTS WITH LYMPHOPROLIFERATIVE DISEASES E. Gorbachenkoa *, O. Gorevaa, A. Grishanovaa, E. Vigovskayab. aInstitute of Molecular Biology and Biophysics, Siberian Branch of the Russian Academy of Medical Sciences, Novosibirsk, Russian Federation, bMunicipal Clinical Hospital No. 2, Hematology Center, Novosibirsk, Russian Federation Introduction. One of the molecular mechanisms of multidrug resistance may be xenobiotic metabolism, including anticancer drugs. Cytochromes P450 (CYPs) and glutathioneS-transferases (GSTs) are important enzymes of phase I and phase II xenobiotic metabolism, respectively. CYPs and GSTP1 genes are polymorphic and can underlie the individual reactions of patients to chemotherapy. Our study analysed the association of functionally significant gene polymorphisms with chemotherapy resistance in patients with lymphoproliferative diseases. Methods. We determined CYP2C8*2, CYP2C8*3, CYP2C9*2, CYP2C9*3, CYP2C19*2, CYP2C19*3, CYP3A5*3, GSTP1 313A→G and GSTP1 341C→T genetic polymorphisms in blood samples of 79 patients (mean age 57 years, SD 13) with lymphoproliferative diseases (non-Hodgkin lymphoma, chronic lymphocytic leukaemia) by PCR-RFLP assays. All patients received chemotherapy (cyclophosphamide, doxorubicin, vincristine, and prednisolone). The odds ratio (OR) was used to estimate association of CYP genotypes with chemotherapy resistance risk and was calculated for the combined group (mutant homozygous and heterozygous) in comparison with the wild genotype group. Results.Wehave found the followingmutant allele frequencies in patients with chronic lymphoproliferative diseases: CYP2C8*2, 0.00; CYP2C8*3, 0.14; CYP2C9*2, 0.11; CYP2C9*3, 0.05; CYP2C19*2, 0.13; CYP2C19*3, 0.00; CYP3A5*3, 0.93; GSTP1 313A→G, 0.21; and GSTP1 341C→T, 0.12. Patients were divided in two groups according to the response to chemotherapy: effect was estimated as positive (n=57) in cases of complete or partial remission or stabilisation and as negative (n=22) in cases of progression of disease. The risk of resistance to chemotherapy for patients had one or two CYP2C8*3 or CYP2C9*2 allele was 7.05-fold significantly (p=0.002) and 4.1fold significantly (p=0.02) higher than for wild genotype carriers, respectively. There were no significant associations of CYP2C9*3 (OR 0.85, p=0.99), CYP2C19*2 (OR 0.88, p=0.99), CYP3A5*3 (OR 1.58, p=0.72), GSTP1 313A→G (OR 1.38, p=0.74), and GSTP1 341C→T (OR 1.21, p=0.97) genetic polymorphisms with chemotherapy response. Discussion. Our results suggest that CYP2C8*3 and CYP2C9*2 polymorphisms are associated with chemotherapy resistance in patients with lymphoproliferative diseases. Funding. None declared. The authors declared no conflicts of interest.

Opening of Astrocytic Mitochondrial ATP-Sensitive Potassium Channels Upregulates Electrical Coupling between Hippocampal Astrocytes in Rat Brain Slices

Astrocytes form extensive intercellular networks through gap junctions to support both biochemical and electrical coupling between adjacent cells. ATP-sensitive K+ (KATP) channels couple cell metabolic state to membrane excitability and are enriched in glial cells. Activation of astrocytic mitochondrial KATP (mitoKATP) channel regulates certain astrocytic functions. However, less is known about its impact on electrical coupling between directly coupled astrocytes ex vivo. By using dual patch clamp recording, we found that activation of mitoKATP channel increased the electrical coupling ratio in brain slices. The electrical coupling ratio started to increase 3 min after exposure to Diazoxide, a mitoKATP channel activator, peaked at 5 min, and maintained its level with little adaptation until the end of the 10-min treatment. Blocking the mitoKATP channel with 5-hydroxydecanoate, inhibited electrical coupling immediately, and by 10-min, the ratio dropped by 71% of the initial level. Activation of mitoKATP channel also decreased the latency time of the transjunctional currents by 50%. The increase in the coupling ratio resulting from the activation of the mitoKATP channel in a single astrocyte was further potentiated by the concurrent inhibiting of the channel on the recipient astrocyte. Furthermore, Meclofenamic acid, a gap-junction inhibitor which completely blocked the tracer coupling, hardly reversed the impact of mitoKATP channel's activation on electrical coupling (by 7%). The level of mitochondrial Connexin43, a gap junctional subunit, significantly increased by 70% in astrocytes after 10-min Diazoxide treatment. Phospho-ERK signals were detected in Connexin43 immunoprecipitates in the Diazoxide-treated astrocytes, but not untreated control samples. Finally, inhibiting ERK could attenuate the effects of Diazoxide on electrical coupling by 61%. These findings demonstrate that activation of astrocytic mitoKATP channel upregulates electrical coupling between hippocampal astrocytes ex vivo. In addition, this effect is mainly via up-regulation of the Connexin43-constituted gap junction coupling by an ERK-dependent mechanism in the mitochondria.

Effect of Hydrocolloid Pre-Treatment on Instrumental and Sensory Texture Attributes of Frozen Carrot (Daucus carota)

The effect of different hydrocolloids (pectin, carboxy methylcellulose, xanthan gum, and sodium alginate) pre-treatment used at different concentrations (0.2–0.4%) on instrumental texture parameters as well as on the sensory attributes of frozen-thawed pre-cut carrots was evaluated. Instrumental texture profile analysis of frozen-thawed carrot showed a significant (P < 0.05) increase in cohesiveness with an increase in the concentration of all the hydrocolloids. Nevertheless, the increase in springiness was not significant (P > 0.05) over the untreated control samples, whereas chewiness did not show consistent results with an increase of any of the hydrocolloids. As the concentration of hydrocolloids increased, the firmness values obtained by cutting the frozen-thawed carrot samples with a Warner Bratzler Blade were also found to increase. Higher texture retention after freezing and thawing was observed in xanthan gum (0.4%) pre-treated samples than other hydrocolloids used as well as than the control samples. It increased the firmness up to 196.2% than the control samples. However, 0.3% xanthan pre-treatment gave the best sensory results. Sensory acceptability increased on increasing the pectin and carboxy methylcellulose concentrations, while lower concentration levels of alginate pre-treatment showed higher sensory preference than the higher concentration levels. Based on the results, untreated frozen samples suffered from a detrimental texture and sensory deterioration during 15 months of frozen storage. The carrot tissue integrity was well retained during frozen storage in the hydrocolloid pre-treated samples.

Tratamento da Superfície de Cateteres de Poliamida 11 por Plasma de Oxigênio

Polyamide 11, as well as the great majority of polymers used in the construction of minimally invasive devices for medical use, shows no significant interaction with polar liquids, which limits its use in some applications, since most of internal biological tissues have hydrophilic character (high affinity for water). In this study, an investigation was made of the influence of oxygen plasma treatment of polyamide 11 surface on the composition, roughness and hydrophilicity. Through the plasma treatment, the polymer characteristics were changed to promote adhesion of a hydrophilic polymer layer on its surface, increasing the hydrophilicity and, consequently, the biocompatility of the polymeric device. Polyamide 11 catheters were submitted to different times of exposure to plasma, generated in an oxygen atmosphere with pressure, temperature and constant voltage. The treated and untreated (control) samples were then characterized using infrared spectroscopy, scanning electron microscope and contact angle measurements with water. Both the roughness and contact angle increased with the plasma treatment, indicating a possible improvement of the adherence of the hydrophilic layer coating the polymer.

Quality response of table grapes (Vitis vinifera L.) during cold storage to postharvest cap stem excision and hot water treatments

SummaryBerry quality, total phenol, sensory characteristics and decay incidence of grapes (V. vinifera cvs. Müşküle and Red Globe) as affected by prestorage hot water immersion with or without cap stem excision treatments were investigated with 7‐day interval during storage at 1 °C for 21 days. Weight loss occurred mainly up to the second week and was more pronounced for the untreated control samples subjected to the stem excision. In Müşküle, the least weight loss value was obtained from hot water–treated grapes (0.62%), while the highest loss occurred in control grapes of stem‐excised berries (0.96%). Immersion of stem‐retained grapes in hot water was the best applications for maintenance of overall storage quality for both cultivars. According to panellist scores, it is evident that hot water has positive effect on maintaining the minimally processed table grapes without altering the flavour and taste of the commodity. Stem‐retained berries of hot water treatment had the highest score for both Müşküle and Red Globe, with their values 3.8 and 4.2, respectively, while the least values were obtained from stem‐excised berries of untreated control group of both the cultivars (1.9 and 3.0, respectively). At the end of three‐week storage, the least decay rates were determined in stem‐retained berries that received hot water treatment (5.3% for Müşküle and 1.1% for Red Globe). Attenuate influence of brief high temperature exposure on ripening parameters (SSC, TA, MI, pH) was also observed. Postharvest hot water treatment may be used as a nonchemical means to extent storage life of minimally processed grapes with its alleviating effect on physiological disorders. Cap stem retaining in minimal processing can be recommended to maintain the quality of grapes.

Effects of chemical preservative treatments on durability of wood flour/HDPE composites

The main objective of this study was to determine the durability of chemical preservatives treated wood flour/high density polyethylene (HDPE) composites to white-rot fungus (Coriolus versicolor). Specimens, containing treated and untreated poplar wood flour (PF, 60%), were mixed with HDPE (38%) as polymer matrix and maleic anhydride grafted polyethylene (MAPE, 2wt.%) as coupling agent. Two fungicide materials, namely 3-iodo-2-propynyl butylcarbamate (IPBC, 0.3, 0.6 and 0.9wt.%) and 2 thiazol-4-yl-1H-benzoimidazole (TBZ, 0.3, 0.6 and 0.9wt.%) were used in preparation of wood plastic composites (WPCs). Then, treated and untreated composites were exposed to the fungal decay for 12weeks according to the European Union (EN) 113 standard. Mechanical and physical properties of the composites were evaluated before and after fungal incubation. The experimental results indicated that treated composites were more resistant to decay, with strength losses significantly lower than the untreated (control) sample. Physical properties in terms of water absorption and thickness swelling were improved by the incorporation of fungicide agents, but no significant differences were observed between the treaded samples. Weight losses for the various treated composites ranged from 1.1% to 4.5%. In addition, IPBC treated samples showed slightly lower weight loss compared with the treated composites with TBZ. The highest weight loss corresponds to the control. Accordingly, IPBC and TBZ can be effectively used as preservatives for WPC. However, IPBC showed superior results compared to the TBZ and it is recommended for the WPCs preservation.

Microrna-34a As a Marker for Fludarabine Resistance and Impairment of p53-Pathway in Chronic Lymphocytic Leukemia

AbstractAbstract 3883 Background.MicroRNAs (miRNAs) are known to be involved in the pathogenesis of chronic lymphocytic leukemia (CLL) and affect its clinical course (Calin et al. NEJM, 2005; Mraz et al. Blood, 2012). Moreover, we and others have shown that several miRNAs are down-regulated in the aggressive CLL subtype harboring p53 aberration (Mraz et al. Leukemia, 2009). The role of miRNAs in primary or acquired resistance to therapy in CLL, however, is poorly understood. Aim.In this study, we screened for miRNAs that are induced by fludarabine-mediated apoptosis in vitro, and we suggested that differences in the expression of one of the identified miRNAs (miR-34a) can be used to distinguish patients with impairment of the p53-apoptotic pathway. Results.Ten primary CLL B-cell samples (purity>95% of CD5+19+ cells) were treated in vitro with fludarabine dose (IC50 dose of 3.5 ug/mL, 48hrs), and the expression of 750 miRNAs was subsequently profiled (TaqMan miRNA Cards, ABI). In comparison with untreated control samples, 15 miRNAs were induced by fludarabine (fold change>1.5, SAM FDR<0.05). The most prominently up-regulated miRNA was miR-34a (fold change 3.7, P=0.003), which is a known p53 down-stream target (He et al. Nature, 2007). We then compared miR-34a up-regulation post fludarabine treatment to the decrease in cell viability (wt-p53 samples, N=20). This revealed that miR-34a induction was significantly higher in CLL samples more sensitive to fludarabine and suggested its role in the apoptotic effects of fludarabine in B-cells. Moreover, the up-regulation of miR-34a was also observed in vivo in samples obtained from fifty FCR-treated CLL patients (fold change 2.2, P<0.0001, analyzed at day 0 and 3 of FCR). These data encouraged us to develop an assay for absolute quantification of miR-34a which would allow determining the copy numbers of miR-34a, defining precise cut-offs, and comparing miR-34a levels during the course of the disease in one patient. We designed synthetic RNA oligos that were used to construct standard curves for both miR-34a and a normalization gene (RNU48). Using this assay, we profiled the expression of miR-34a in a cohort of CLL patients (N=200) to define a cut-off value that would discriminate therapy resistant cases. The distribution of miR-34a expression in the cohort ranged from 1 to 81820 molecules (per 10e6 copies of RNU48). Significantly, miR-34a levels below 2500 copies (N=47) were correlated with shorter overall survival (9.6 years vs. not-reached, HR 2.2 [CI 1.1–4.5], P=0.03). Subsequently, the expression of miR-34a was compared in samples stratified by known prognostic markers: chromosomal aberrations (del17p13, del11q23, tris.12, and del13q14), IgHV status, expression of CD38, CD49d, age, gender, and Rai stage. The lower miR-34a levels were only associated with the deletion of 17p13 locus that includes the p53 gene (N=18, fold change −3.4, P=0.003). Remarkably, CLL samples with sole p53 mutation not accompanied by p53 deletion (N=13) also expressed low levels of miR-34a compared to wt-p53 (P=0.005, fold change −2.7). Most notably, 77% (10/13) of these samples had miR-34a levels below the cut-off of 2500 copies. We further validated our observations and assay by analyzing miR-34a expression in paired samples from 12 CLL cases that acquired p53 aberration during the course of the disease. This emphasized that miR-34a expression decreased in all cases after occurrence of p53 mutation (P=0.0008, fold change −6.1). Additionally, the effect of miR-34a up-regulation on therapy response is currently being investigated in a cohort of FCR-treated patients (N=50). Conclusions.Our data provide complex evidence for the use of miR-34a as a marker of fludarabine-resistant disease. MicroRNA-34a quantification can identify p53 mutated cases that would not be recognized by FISH (mutation not accompanied by del17p13), and miR-34a down-regulation can be used as a sensor for acquisition of p53 abnormality during the course of the disease. This can be accomplished without treatment of cells with gamma-irradiation, which was previously used to identify functional impairment of the p53-pathway (Pettitt et al. Blood, 2001; Mous et al. Leukemia, 2009; Lin et al. CCR, 2012). The described assay for absolute quantification of miR-34a encourages further inter-laboratory validation.IGA MZCR NT11218–6/2010, CZ.1.07/2.3.00/20.0045, CZ.1.05/1.1.00/02.0068 Disclosures:No relevant conflicts of interest to declare.

English

The antifungal effect of pumpkin (Cucurbita moschata) fruit aqueous extract againstAspergillus flavus was investigated in vitro. The result showed that incubation of different pumpkin aqueous extract (0.5 to 2%) with living mass of the fungus has suppressive effect on the fungal growth after 6 days compared with untreated control sample. However 2% of pumpkin aqueous extract was the most effective in inhibiting the fungal growth. The protective efficacy of pumpkin fruit aqueous extract against either A. flavus fungus infection or aflatoxin B1 (AFB1) toxicity induced renal damage in rats was also investigated. A. flavus and AFB1 were administered intraperitoneally (0.1 ml/100 g of body weight) for 15 consecutive days. The result revealed that infection of rats with A. flavus or intoxication with AFB1 significantly induced renal damage as indicated by marked increased levels of serum urea, uric acid and creatinine as well as histopathological pictures compared with normal healthy rats. Oral co-administration of aqueous extract of pumpkin fruits (1.0 mg / kg of body weight) to either rat groups infected with A. flavus or intoxicated with AFB1 for 20 consecutive days effectively normalized the serum kidney function biomarkers and confirmed by histomorphologic pictures which showed normal histological structure. Key words: Aspergillus flavus, aflatoxin B1, antifungal, renal damage.

Analysis of microRNA expression profile induced by AICAR in mouse hepatocytes

AMP-activated protein kinase (AMPK) has been proposed to act as a key energy sensor mediating the metabolism of glucose and lipids, and pharmacological activation of AMPK may provide a new strategy for the management of type 2 diabetes. MicroRNAs (miRNAs) are a group of endogenous noncoding RNA that play important roles in many biological processes including energy metabolism. Whether miRNAs mediate AMPK action in regulating metabolic process is not clear. In this study, 0.5mM 5-aminoimidazole-4-carboxamide-1-β-d-ribofuranoside (AICAR) was added to increase activation of AMPK in 8week old C57BL/6 mice primary hepatocytes. MiRNA microarray was performed to compare the miRNA expression profiles of hepatocytes treated with or without AICAR. We discovered that 41 miRNAs were significantly altered in AICAR-treated sample (fold change: >2) compared with untreated control sample. Among them, 19 miRNAs were upregulated. MiRNA targets were predicted by TargetScan. Further bioinformatic analysis indicated that these predicted targets might be mainly involved in pathways of cellular metabolism and tumor pathogenesis. FUNDO analysis suggested that these predicted targets were enriched in cancer, diabetes mellitus, hypertension, obesity and heart failure (P<0.01). A series of miRNAs could be regulated by the activation of AMPK and might mediate the action of AMPK during metabolic processes and tumor pathogenesis. Predicted target genes discovered in this study and pathway analysis provide new insights into hepatic metabolism and tumor pathogenesis regulated by AMPK signaling and clues to the possible molecular mechanism underlying the effect of AMPK.