Development of a biosensor for urea assay based on amidase inhibition, using an ion-selective electrode

A biosensor for urea has been developed based on the observation that urea is a powerful active-site inhibitor of amidase, which catalyzes the hydrolysis of amides such as acetamide to produce ammonia and the corresponding organic acid. Cell-free extract from Pseudomonas aeruginosa was the source of amidase (acylamide hydrolase, EC 3.5.1.4) which was immobilized on a polyethersulfone membrane in the presence of glutaraldehyde; anion-selective electrode for ammonium ions was used for biosensor development. Analysis of variance was used for optimization of the biosensorresponse and showed that 30 mu L of cell-free extract containing 7.47 mg protein mL(-1), 2 mu L of glutaraldehyde (5%, v/v) and 10 mu L of gelatin (15%, w/v) exhibited the highest response. Optimization of other parameters showed that pH 7.2 and 30 min incubation time were optimum for incubation ofmembranes in urea. The biosensor exhibited a linear response in the range of 4.0-10.0 mu M urea, a detection limit of 2.0 mu M for urea, a response timeof 20 s, a sensitivity of 58.245 % per mu M urea and a storage stability of over 4 months. It was successfully used for quantification of urea in samples such as wine and milk; recovery experiments were carried out which revealed an average substrate recovery of 94.9%. The urea analogs hydroxyurea, methylurea and thiourea inhibited amidase activity by about 90%, 10% and 0%, respectively, compared with urea inhibition

Taking stock of 10 years of published research on the ASHA programme: Examining India’s national community health worker programme from a health systems perspective

Background: As India’s accredited social health activist (ASHA) community health worker (CHW) programme enters its second decade, we take stock of the research undertaken and whether it examines the health systems interfaces required to sustain the programme at scale. Methods: We systematically searched three databases for articles on ASHAs published between 2005 and 2016. Articles that met the inclusion criteria underwent analysis using an inductive CHW–health systems interface framework. Results: A total of 122 academic articles were identified (56 quantitative, 29 mixed methods, 28 qualitative, and 9 commentary or synthesis); 44 articles reported on special interventions and 78 on the routine ASHA program. Findings on special interventions were overwhelmingly positive, with few negative or mixed results. In contrast, 55% of articles on the routine ASHA programme showed mixed findings and 23% negative, with few indicating overall positive findings, reflecting broader system constraints. Over half the articles had a health system perspective, including almost all those on general ASHA work, but only a third of those with a health condition focus. The most extensively researched health systems topics were ASHA performance, training and capacity-building, with very little research done on programme financing and reporting, ASHA grievance redressal or peer communication. Research tended to be descriptive, with fewer influence, explanatory or exploratory articles, and no predictive or emancipatory studies. Indian institutions and authors led and partnered on most of the research, wrote all the critical commentaries, and published more studies with negative results. Conclusion: Published work on ASHAs highlights a range of small-scale innovations, but also showcases the challenges faced by a programme at massive scale, situated in the broader health system. As the programme continues to evolve, critical comparative research that constructively feeds back into programme reforms is needed, particularly related to governance, intersectoral linkages, ASHA solidarity, and community capacity to provide support and oversight

Construction of an acetylcholinesterase-choline oxidase biosensor for aldicarb determination

Biosensors have increasingly attracted attention in several areas due to their various advantages over conventional analytical techniques. They are currently used in the detection of various substances in medicine, in the control of industrial processes, in agriculture, environmental monitoring, and defence. Detection of the pesticides in environmental samples is an important concern due to the high toxicity of these compounds. Biosensors; are good alternatives for chromatographic methods in pesticide detection since they do not require tedious sample pretreatment, can be used in on-site detection and are economical

Construction of an acetylcholinesterase-choline oxidase biosensor for aldicarb determination

In this study, acetylcholinesterase and choline oxidase were co-immobilized on poly(2-hydroxyethyl methacrylate) membranes and the change in oxygen consumption upon aldicarb introduction was measured. Immobilization of the enzymes was achieved either by entrapment or by surface attachment via a hybrid immobilization method including epichlorohydrin and Cibacron Blue F36A activation. Immobilized enzymes had a long-storage stability (only 15% activity decrease in 2 months in wet storage and no activity loss in dry storage). Aldicarb detection studies showed that a linear working range of 10-500 and 10-250 ppb aldicarb could be achieved by entrapped and surface immobilized enzymes, respectively. Enzymes immobilized on membrane surfaces responded to aldicarb presence more quickly than entrapped enzymes. Aldicarb concentrations as low as 23 and 12 ppb could be detected by entrapped and surface immobilized enzymes, respectively, in 25 min

Immobilization of acetylcholinesterase and choline oxidase in/on pHEMA membrane for biosensor construction

In this study, acetylcholinesterase (AChE) and choline oxidase (ChO) were co-immobilized on poly(2-hydroxyethyl methacrylate) (pHEMA) membranes with the aim of using them in biosensor construction. pHEMA membranes were prepared with the addition of different salts in different HEMA : aqueous solution ratios and characterized in terms of porosity, thickness, permeability, and mechanical properties. Membranes prepared in the presence of SnCl4 were found to be superior in terms of porosity and permeability and were chosen as the immobilization matrix. Immobilization of the enzymes was achieved both by entrapment and surface attachment via epichlorohydrin (Epi) and Cibacron Blue F36A (CB) activation. The effect of immobilization on enzyme activity was evaluated by the comparison of K-m and V-max values for the free and immobilized bi-enzyme systems. The increase in K-m was negligible (1.08-fold) for the bi-enzyme system upon immobilization on surface but was 2.12-fold upon entrapment. Specific activity of the free enzyme system was found to be 0,306 mV s(-1) mug(-1) ChO while it was 0.069 (4.43-fold decrease) for entrapped and 0.198 (1.54 fold decrease) for CB-Epi immobilized enzymes. The performance of immobilized enzymes in different buffer types, pH, and temperature conditions were evaluated. The best enzyme activity was obtained at pH 9.0. Activity of the enzymes was found to increase with increasing temperature (in the range 25-40degreesC)

Controlled release of Aldicarb from carboxymethylcellulose microcapsules

Kok, Fatma Nese/0000-0002-5093-6013WOS: A1997XF27900005Sodium carboxymethyl cellulose was converted into microspheres by crosslinking with aluminum chloride. Various microsheres with different amounts of crosslinker, biopolymer of various concentrations and molecular weights, and with different pesticide (Aldicarb) contents and pesticide to polymer ratios were prepared. The pesticide encapsulation efficiencies and aldicarb release kinetics of the resultant microcapsules were investigated. It was possible to modify the release behaviour by varying the above parameters and to have drug release with half-lives longer than 100 hours in aqueous media. The release kinetics were described by first order and zero order kinetics

Controlled release of aldicarb from carboxymethyl cellulose microspheres: in vitro and field applications

Kok, Fatma Nese/0000-0002-5093-6013WOS: 000084918700010Aldicarb is a carbamate pesticide that is widely used throughout the world in the protection Of crops (eg cotton, nuts, potatoes, onion, tobacco, sugar beet and sugar cane). In Turkey, especially in the Cukurova region, it is used for the control of the cotton white fly (Bemisia tabaci) which attacks cotton plants cultivated in this region. Aldicarb contamination in surface and ground water is a serious problem in several countries, partly due to its high water solubility. It is also highly toxic to mammals. In order to overcome these problems, microspheres of aldicarb were prepared using carboxymethyl cellulose (CMC) as the biodegradable support material cross-linked with aluminium chloride. A strong hysteresis behaviour was observed upon drying and reswelling. Encapsulation efficiency was in the range 12-23% and aldicarb contents of 5.7-10.3 mg per 100mg of microspheres was achieved. In vitro release was distinctly Fickian, and Higuchi constants were very close to 0.5. Release in pots revealed that only one sample had a release capability for more than four weeks. In the cotton plot much longer durations of release (more than seven weeks) were observed while a commercial granular formulation released its content immediately. It was thus possible to construct a controlled pesticide release system that prolonged the bioavailability to about eight weeks. (C) 1999 Society of Chemical Industry

Biodegradation of aldicarb in a packed-bed reactor by immobilized Methylosinus

Kok, Fatma Nese/0000-0002-5093-6013WOS: 000078929100008Carboxymethylcellulose microspheres cross-linked via aluminum ions were used as a support material for immobilization of Methylosinus isolated from soil contaminated with aldicarb. The degradation capacity of immobilized bacteria, different parameters such as substrate concentration (50-800 ppm), flow rate (10-60 ml h(-1)), and continuous contact with reaction medium (flow rate, 20 mi h(-1) and concentration, 100 ppm) that affect aldicarb degradation were investigated in a packed-bed reactor. Increases in the flow rate decreased the conversion of aldicarb into its metabolites. On the other hand, increasing the substrate concentration up to 400 ppm led to an increase in the amount of aldicarb converted (max 16%). Beyond this, the proportion of aldicarb that converted was decreased, reaching approximately 7% at 800 ppm. The apparent kinetic parameters, K-m' and V-max, were determined to be as 310.11 ppm and 2.29 X 10(-2) ppm s(-1), respectively. Operation of the bioreactor in the recycled mode was much more efficient, degrading 50% of the aldicarb in 24 h and 100% in four days. (C) 1999 Elsevier Science Inc

Controlled release of aldicarb from lignin loaded ionotropic hydrogel microspheres

Wilkins, Richard M/0000-0001-8984-4960; Kok, Fatma Nese/0000-0002-5093-6013WOS: 000082217000006PubMed: 10499841Aldicarb loaded microspheres of carboxymethyl cellulose, CMC, of various compositions were prepared with the crosslinking action of AI(III). Into these microspheres, lignin was introduced as a filler to further regulate the rate of release. The encapsulation efficiency was low (4.15-13.00%) due to the high water solubility of aldicarb. Release into aqueous media was quite rapid and almost complete within 24h. Initial stages of the release (60% of AS released) was investigated to determine the release mechanism and it was found that release from the microspheres was governed by Fickian diffusion. Among the various parameters, the ratio of lignin to CMC and aldicarb to CMC were found to be influential on the rate of release from the crosslinked CMC-lignin microspheres. The soil behaviour confirmed the delayed release effect of the controlled release formulation compared to Temik (a commercial preparation of aldicarb), especially when lignin was incorporated

Controlled release of aldicarb from lignin loaded ionotropic hydrogel microspheres

Aldicarb loaded microspheres of carboxymethyl cellulose, CMC, of various compositions were prepared with the crosslinking action of AI(III). Into these microspheres, lignin was introduced as a filler to further regulate the rate of release. The encapsulation efficiency was low (4.15-13.00%) due to the high water solubility of aldicarb. Release into aqueous media was quite rapid and almost complete within 24h. Initial stages of the release (60% of AS released) was investigated to determine the release mechanism and it was found that release from the microspheres was governed by Fickian diffusion. Among the various parameters, the ratio of lignin to CMC and aldicarb to CMC were found to be influential on the rate of release from the crosslinked CMC-lignin microspheres. The soil behaviour confirmed the delayed release effect of the controlled release formulation compared to Temik (a commercial preparation of aldicarb), especially when lignin was incorporated