Bioactive Paper Sensor Based on the Acetylcholinesterase for the Rapid Detection of Organophosphate and Carbamate Pesticides

In many countries, people are becoming more concerned about pesticide residues which are present in or on food and feed products. For this reason, several methods have been developed to monitor the pesticide residue levels in food samples. In this study, a bioactive paper-based sensor was developed for detection of acetylcholinesterase (AChE) inhibitors including organophosphate and carbamate pesticides. Based on the Ellman colorimetric assay, the assay strip is composed of a paper support (1×10 cm), onto which a biopolymer chitosan gel immobilized in crosslinking by glutaraldehyde with AChE and 5,5′-dithiobis(2-nitrobenzoic) acid (DTNB) and uses acetylthiocholine iodide (ATChI) as an outside reagent. The assay protocol involves introducing the sample to sensing zone via dipping of a pesticide-containing solution. Following an incubation period, the paper is placed into ATChI solution to initiate enzyme catalyzed hydrolysis of the substrate, causing a yellow color change. The absence or decrease of the yellow color indicates the levels of the AChE inhibitors. The biosensor is able to detect organophosphate and carbamate pesticides with good detection limits (methomyl=6.16×10-4 mM and profenofos=0.27 mM) and rapid response times (~5 min). The results show that the paper-based biosensor is rapid, sensitive, inexpensive, portable, disposable, and easy-to-use

Penentuan Profil Ketebalan Sedimen Lintasan Kota Makassar Dengan Mikrotremor

Di kota metropolitan yang jauh dari fokus gempa besar misalnya Kota Makassar, getaran yang paling banyak mempengaruhi kontruksi bangunan adalah gempa mikro yang bersumber dari getaran mesin, angin, tumbuhan dan aktivitas manusia. Sumber getaran di permukaan bumi dapat menyebabkan tanah di sekitarnya beresonansi. Resonansi pada lapisan tanah yang terjadi secara periodik disebut gempa mikro atau mikrotremor. Hasil pengukuran mikrotremor dari arah Kabupaten Gowa ke Kota Makassar menjadi fokus analisis dalam penelitian ini. Studi mikrotremor ini bertujuan mengetahui berapa besar frekuensi resonansi, tingkat kerentanan seismik dan pendugaan ketebalan lapisan sedimen pada lintasan mikroteremor yang dibuat melewati daerah Gowa sampai kota Makassar. Frekuensi resonansi lintasan mikrotremor Gowa-Makassar sangat bervariasi mulai dari 0,647 – 11,698 Hz, dengan frekuensi resonansi rata-rata berada pada 6,29 Hz. Nilai indeks kerentanan seismik lintasan mikrotremor Gowa-Makassar berada pada interval nilai 0,15 – 30. Tingkat kerentanan seismik Gowa-Makassar cenderung semakin besar ke arah topografi yang lebih tinggi. Hasil perhitungan ketebalan sedimen lintasan mikrotremor Gowa-Makassar menggunakan teknik S/R adalah 6 – 66 m

Enhancement of Clay Compressibility and Strength Using Nano Magnesium Oxide

In order to satisfy the requirements of environmentally friendly construction projects and minimize greenhouse gas emissions, it was necessary to focus on utilizing nanomaterials instead of conventional materials to enhance clay soils; nano Magnesium Oxide was suggested for this study. The compression and unconfined compressive strength tests were achieved to assess the compressibility behavior and strength of silty clay treated using nano magnesium oxide. Additionally, an investigation into the alteration of the clay microstructure was conducted using field emission scanning electron microscopy (FE-SEM) tests. The silty clay utilized in this research is characterized by a compression index 0.23 and an undrained shear strength of 53 kPa. The soil was treated by adding different percentages of nano-MgO (0.4%, 0.5%, 0.6%, and 0.8%), and the curing period was seven days. The treated soil samples were prepared based on the optimal water content and maximal dry density of the natural soil. The results indicate that utilization of nano Magnesium Oxide leads to a reduction in both values of the index of compression (Cc) and recompression index (Cr), in addition to a decrease in the value of the coefficient of compressibility (av). Also, the test finding for unconfined compressive strength demonstrates a noticeable increase in strength as the percentage of nano-MgO increases, accompanied by a reduction in failure strain. Adding 0.6% (the optimum percentage) of nano Magnesium oxide enhances the strength by about 114% and decreases the value of the compression index by about 36% compared with natural soil

PCR in Comparison with Culture Methods for The Diagnosis of Candida albicans Responsible for Candidemia in Leukemic Patients

Background: Currently, candidemia infections represent an increasing cause of morbidity and mortality in seriously ill hospitalized patients. Because the accurate diagnosis of candidiasis remains difficult, fast and reliable assay for characterization of fungal pathogens is critical for the early initiation of adequate antifungal therapy and/or for introduction of preventive measures. Objective: To detect candidemia in leukemic patients by molecular methods in comparing with golden standard method (culture method). Materials and methods: A total of 60 leukemic patients were included in this study. Clinical type and other demographic data were recorded. Blood samples were taken from each patient, culture; germ tube formation and carbohydrate fermentation were done for each sample. DNA extraction and polymerase chain reaction (PCR) were used for detection of Candida albicansin cultured bottles. This study was conducted on leukemic patients admitted to four different hospitals in Baghdad city from September 2010 to March 2011.  Sixty patients suffering from acute lymphoid (ALL) and myeloid (AML) leukemia were included in this study. The age of patients were ranging between 3-46 years old. Twenty five apparently healthy individuals were enrolled in this study as control group. Three milliliters of blood were collected from each patient; 1.5 ml was inoculated in 20 ml Brain heart infusion broth (Cruikshank. 1975). The rest of blood (1.5ml) was stored in -20ᴼC for further analysis. Blood cultures were incubated at 37°C for 10 days, and examined daily for growth. DNA purification kit was purchased from QIAGENE®Company. This method was used for the purification of genomic DNA from fresh or frozen samples of 1 ml overnight yeast cultures by using the GentraPuregene Yeast/Bact. Kit. PCR was performed to detect Candida albicans species through the amplification of specific gene (αINT1) Results: Only one positive culture result out of 60 samples was obtained for Candida sp., (1.7%).  PCR results showed that there were only three out of sixty were positive for C. albicans (5%). In this study we obtained only one positive sample according to culture, while three samples only gave positive results according to PCR method. These results suggest that molecular analysis of candidemia is more sensitive and less time consuming than culture and other conventional methods. Conclusions: we concluded the following: The rate of candidemia was 1.7% among leukemic patients, according to culture results, only 5% of blood cultures was positive according to PCR. Results showed 100% sensitivity and 96.6% specificity and it is rapid, easy, reliable and also applicable in clinical laboratory for identification of medically important Candida sp

Toxicity Assessment of Buprofezin, Lufenuron, and Triflumuron to the Earthworm Aporrectodea caliginosa

Earthworms are particularly important soil macroinvertebrates and are often used in assessing the general impact of pesticide pollution in soil. The present study was conducted in order to investigate the toxicity of three insect growth regulators (IGRs) buprofezin, lufenuron, and triflumuron, at different application rates and exposure times toward mature earthworms Aporrectodea caliginosa. The effects of these pesticides on the growth rate in relation to the activities of acetylcholinesterase (AChE) and glutathione S-transferase (GST) as biochemical indicators were evaluated to elucidate the mechanisms of action. Toxicity studies indicated that lufenuron was the most harmful pesticide to mature earthworms, followed in descending order by buprofezin and triflumuron. A reduction in growth rate in all pesticide-treated worms was dose-dependent over the 28-day exposure period, which was accompanied by a decrease in AChE and GST activities. Relationships between growth rate, AChE, and GST provided strong evidence for the involvement of pesticidal contamination in the biochemical changes in earthworms, which can be used as a bioindicator of soil contamination by pesticides

Impact of organic amendments addition to sandy clay loam soil and sandy loam soil on leaching process of chlorantraniliprole insecticide and bispyribac-sodium herbicide

The leaching of two pesticides, cholantraniliprole (CAP) and bispyribac-sodium (BPS) in sandy clay loam soil (soil A) and sandy loam soil (soil B) were examined, by soil columns under laboratory conditions. In addition, the effect of adding 5% biochar and wheat straw to the soils on the leaching of CAP and BPS was studied. It is clear from the results that BPS is more leachable than CAP in all soil columns, and more leached to soil B. It was found that the addition of biochar and wheat straw has a significant effect on the movements of these pesticides and can be used to reduce the environmental impact. For CAP, 71 to 97% and 84 to 97% were recovered from the soil A columns and soil B columns, respectively, while for BPS, 94 to 99% were recovered from columns of soil A and soil B

Effect of soil organic amendments on sorption behavior of two insecticides and two herbicides

The effect of biochar, compost, peat and wheat straw at 1 and 5% on adsorption isotherm of chlorantraniliprole, dinotefuran, bispyribac-sodium, and metribuzin was studied in clay loam soil and sandy loam soil. Biochar, compost, peat and wheat straw (at a rate of 1 % in soil) improved the adsorption capacity of chlorantraniliprole and metribuzin in sandy loam soil. The sorption coefficients are higher for chlorantraniliprole and metribuzin whereas lower for dinotefuran and bispyribac-sodium in amended soil compared to unamended sandy loam soil. There is not a clear direct correlation between Freundlich parameters as well as Kd or Koc and type of organic amendment. The sorption of all tested pesticides on biochar was increased, whereas on compost was decreased. The order of pesticides sorption in soils and different organic amendments is generally inversely proportional to their aqueous solubilites. Adsorption of chlorantraniliprole increases on the sandy loam soil amendment at the rate of 1% in the following order: peat > compost > biochar > original soil. Also, the magnitude of adsorption on soil A amendment at the rate of 5% can be arranged for dinotefuran in the order; peat > biochar > compost > original soil and for bispyribac-sodium and metribuzin peat = wheat straw > biochar > original soil

Experimental modeling design to study the effect of different soil treatments on the dissipation of metribuzin herbicide with effect on dehydrogenase activity

The dissipation and side-effect of metribuzin (MBZ) were studied with various factors; two soil types (clay loam and sandy loam), soil amendment (wheat straw and without amendment), two temperature levels (25 and 50°C), sterilization (sterilized and unsterilized soil) and time of incubation (15 and 30 days) and designed by Windows version of MINITAB software package to reduce the time and the cost as well as increased the precision. Determination of MBZ by HPLC with recoveries ranged from 50.85 to 108.09%. The MBZ residues were detected in all samples up to 60 days of storage, respectively with decline in their concentrations with the time of incubation. The clay loam soil showed higher dissipation than the sandy loam soil. The different factors in the present study confirmed that the wheat straw amendment, non-sterilization and incubation at 50°C caused higher dissipation of MBZ than without wheat straw, sterilization and incubation at 25°C. The dissipation was described mathematically by a first order equation with t0.5 was ranged from 9.62 to 16.82 days in clay loam soil and from 10.01 to 16.04 days in sandy loam soil. The side-effect of MBZ was tested on soil dehydrogenase activity that can be considered as an indicator of the biological activity and microbial degradation. The result proved that the enzyme activity was significantly decreased in all treatments compared with the controls at 1 and 3 days of incubation then it was gradually increased at 7, 10, 15 and 30 days of incubation. Treatments of wheat straw, non-sterilized and incubated at 25°C or 50°C showed the lowest enzyme inhibition among all treatments

Modeling study of adsorption isotherms of chlorantraniliprole and dinotefuran on soil

Knowledge of pesticide adsorption characteristics is essential to predict their behavior in soil. The adsorption equilibrium isotherms of two insecticides chlorantraniliprole (CAP) and dinotefuran (DNF) on two common Egyptian soil types, clay loam and sandy loam were studied and modeled. To predict the adsorption isotherms and to determine the adsorption parameters, ten isotherm models: Langmuir (five linear forms), Freundlich, Temkin, Dubinin-Radushkevich, Elovick, Fowler-Guggenheim, Kiselev, Jovanoic, Harkins-Jura, and Halsey were applied on experimental data. The results revealed that the adsorption isotherm models fitted the data in the order of Halsey > Freundlich > Jovanoic > Langmuir isotherme. The models of Harkins-Jura, Elovich, Temkin, and Fowler-Guggenheim are not applicable to predict the adsorption isotherms of the tested insecticides. In order to determine the best-fit isotherm, the correlation coefficient (R2), comparing the experimental (exp) and calculated (cal) adsorption data, and a normalized standard deviation (Δg%) were used to evaluate the data. Therefore, the isotherm models Halsey and Freundlich could be used to predict the adsorption characteristics of CAP and DNF in the common Egyptian soil types, clay loam and sandy loam. Consequently, the mathematical models Halsey, Freundlich, and Jovanoic can describe the fate of CAP and DNF and can be used to control Egyptian soil contamination

Protein Isoaspartate Methyltransferase Prevents Apoptosis Induced by Oxidative Stress in Endothelial Cells: Role of Bcl-Xl Deamidation and Methylation

BACKGROUND:Natural proteins undergo in vivo spontaneous post-biosynthetic deamidation of specific asparagine residues with isoaspartyl formation. Deamidated-isomerized molecules are both structurally and functionally altered. The enzyme isoaspartyl protein carboxyl-O-methyltransferase (PCMT; EC 2.1.1.77) has peculiar substrate specificity towards these deamidated proteins. It catalyzes methyl esterification of the free alpha-carboxyl group at the isoaspartyl site, thus initiating the repair of these abnormal proteins through the conversion of the isopeptide bond into a normal alpha-peptide bond. Deamidation occurs slowly during cellular and molecular aging, being accelerated by physical-chemical stresses brought to the living cells. Previous evidence supports a role of protein deamidation in the acquisition of susceptibility to apoptosis. Aim of this work was to shed a light on the role of PCMT in apoptosis clarifying the relevant mechanism(s). METHODOLOGY/PRINCIPAL FINDINGS:Endothelial cells transiently transfected with various constructs of PCMT, i.e. overexpressing wild type PCMT or negative dominants, were used to investigate the role of protein methylation during apoptosis induced by oxidative stress (H(2)O(2); 0.1-0.5 mM range). Results show that A) Cells overexpressing "wild type" human PCMT were resistant to apoptosis, whereas overexpression of antisense PCMT induces high sensitivity to apoptosis even at low H(2)O(2) concentrations. B) PCMT protective effect is specifically due to its methyltransferase activity rather than to any other non-enzymatic interactions. In fact negative dominants, overexpressing PCMT mutants devoid of catalytic activity do not prevent apoptosis. C) Cells transfected with antisense PCMT, or overexpressing a PCMT mutant, accumulate isoaspartyl-containing damaged proteins upon H(2)O(2) treatment. Proteomics allowed the identification of proteins, which are both PCMT substrates and apoptosis effectors, whose deamidation occurs under oxidative stress conditions leading to programmed cell death. These proteins, including Hsp70, Hsp90, actin, and Bcl-xL, are recognized and methylated by PCMT, according to the general repair mechanism of this methyltransferase. CONCLUSION/SIGNIFICANCE:Apoptosis can be modulated by "on/off" switch partitioning the amount of specific protein effectors, which are either in their active (native) or inactive (deamidated) molecular forms. Deamidated proteins can also be functionally restored through methylation. Bcl-xL provides a case for the role of PCMT in the maintenance of functional stability of this antiapoptotic protein