Polygenic scores for cardiovascular risk factors improve estimation of clinical outcomes in CCB treatment compared to pharmacogenetic variants alone.

Pharmacogenetic variants are associated with clinical outcomes during Calcium Channel Blocker (CCB) treatment, yet whether the effects are modified by genetically predicted clinical risk factors is unknown. We analyzed 32,000 UK Biobank participants treated with dihydropiridine CCBs (mean 5.9 years), including 23 pharmacogenetic variants, and calculated polygenic scores for systolic and diastolic blood pressures, body fat mass, and other patient characteristics. Outcomes included treatment discontinuation and heart failure. Pharmacogenetic variant rs10898815-A (NUMA1) increased discontinuation rates, highest in those with high polygenic scores for fat mass. The RYR3 variant rs877087 T-allele alone modestly increased heart failure risks versus non-carriers (HR:1.13, p = 0.02); in patients with high polygenic scores for fat mass, lean mass, and lipoprotein A, risks were substantially elevated (HR:1.55, p = 4 × 10-5). Incorporating polygenic scores for adiposity and lipoprotein A may improve risk estimates of key clinical outcomes in CCB treatment such as treatment discontinuation and heart failure, compared to pharmacogenetic variants alone.

The Triangulation WIthin a STudy (TWIST) framework for causal inference within pharmacogenetic research.

In this paper we review the methodological underpinnings of the general pharmacogenetic approach for uncovering genetically-driven treatment effect heterogeneity. This typically utilises only individuals who are treated and relies on fairly strong baseline assumptions to estimate what we term the 'genetically moderated treatment effect' (GMTE). When these assumptions are seriously violated, we show that a robust but less efficient estimate of the GMTE that incorporates information on the population of untreated individuals can instead be used. In cases of partial violation, we clarify when Mendelian randomization and a modified confounder adjustment method can also yield consistent estimates for the GMTE. A decision framework is then described to decide when a particular estimation strategy is most appropriate and how specific estimators can be combined to further improve efficiency. Triangulation of evidence from different data sources, each with their inherent biases and limitations, is becoming a well established principle for strengthening causal analysis. We call our framework 'Triangulation WIthin a STudy' (TWIST)' in order to emphasise that an analysis in this spirit is also possible within a single data set, using causal estimates that are approximately uncorrelated, but reliant on different sets of assumptions. We illustrate these approaches by re-analysing primary-care-linked UK Biobank data relating to CYP2C19 genetic variants, Clopidogrel use and stroke risk, and data relating to APOE genetic variants, statin use and Coronary Artery Disease.

SLCO1B1 Exome Sequencing and Statin Treatment Response in 64,000 UK Biobank Patients.

The solute carrier organic anion transporter family member 1B1 (SLCO1B1) encodes the organic anion-transporting polypeptide 1B1 (OATP1B1 protein) that transports statins to liver cells. Common genetic variants in SLCO1B1, such as *5, cause altered systemic exposure to statins and therefore affect statin outcomes, with potential pharmacogenetic applications; yet, evidence is inconclusive. We studied common and rare SLCO1B1 variants in up to 64,000 patients from UK Biobank prescribed simvastatin or atorvastatin, combining whole-exome sequencing data with up to 25-year routine clinical records. We studied 51 predicted gain/loss-of-function variants affecting OATP1B1. Both SLCO1B1*5 alone and the SLCO1B1*15 haplotype increased LDL during treatment (beta*5 = 0.08 mmol/L, p = 6 × 10-8; beta*15 = 0.03 mmol/L, p = 3 × 10-4), as did the likelihood of discontinuing statin prescriptions (hazard ratio*5 = 1.12, p = 0.04; HR*15 = 1.05, p = 0.04). SLCO1B1*15 and SLCO1B1*20 increased the risk of General Practice (GP)-diagnosed muscle symptoms (HR*15 = 1.22, p = 0.003; HR*20 = 1.25, p = 0.01). We estimated that genotype-guided prescribing could potentially prevent 18% and 10% of GP-diagnosed muscle symptoms experienced by statin patients, with *15 and *20, respectively. The remaining common variants were not individually significant. Rare variants in SLCO1B1 increased LDL in statin users by up to 1.05 mmol/L, but replication is needed. We conclude that genotype-guided treatment could reduce GP-diagnosed muscle symptoms in statin patients; incorporating further SLCO1B1 variants into clinical prediction scores could improve LDL control and decrease adverse events, including discontinuation.

Calcium-channel blockers: Clinical outcome associations with reported pharmacogenetics variants in 32 000 patients.

AIMS: Pharmacogenetic variants impact dihydropyridine calcium-channel blockers (dCCBs; e.g., amlodipine) treatment efficacy, yet evidence on clinical outcomes in routine primary care is limited. Reported associations in pharmacogenomics knowledge base PharmGKB have weak supporting evidence. We aimed to estimate associations between reported pharmacogenetic variants and incident adverse events in a community-based cohort prescribed dCCB. METHODS: We analysed up to 32 360 UK Biobank participants prescribed dCCB in primary care (from UK general practices, 1990-2017). We investigated 23 genetic variants. Outcomes were incident diagnosis of coronary heart disease, heart failure (HF), chronic kidney disease, oedema and switching antihypertensive medication. RESULTS: Participants were aged 40-79 years at first dCCB prescription. Carriers of rs877087 T allele in RYR3 had increased risk of hazard ratio (HF 1.13: 95% confidence interval 1.02 to 1.25, P = .02). Although nonsignificant after multiple testing correction, the association is consistent with prior evidence. We estimated that if rs877087 T allele could experience the same treatment effect as noncarriers, the incidence of HF in patients prescribed dCCB would reduce by 9.2% (95% confidence interval 3.1 to 15.4). In patients with a history of heart disease prior to dCCB (n = 2296), rs877087 homozygotes had increased risk of new coronary heart disease or HF compared to CC variant. rs10898815 in NUMA1 and rs776746 in CYP3A5 increased likelihood of switching to an alternative antihypertensive. The remaining variants were not strongly or consistently associated with studied outcomes. CONCLUSION: Patients with common genetic variants in NUMA1, CYP3A5 and RYR3 had increased adverse clinical outcomes. Work is needed to establish whether outcomes of dCCB prescribing could be improved by prior knowledge of pharmacogenetics variants supported by clinical evidence of association with adverse events.

Statin treatment effectiveness and the SLCO1B1*5 reduced function genotype: Long-term outcomes in women and men.

OBJECTIVE: To estimate the effect of rs4149056 (SLCO1B1*5) genotype (decreases statin transport) on cholesterol control and treatment duration in male and female primary care patients prescribed common statin medications. METHODS AND ANALYSIS: This study comprised 69 185 European-ancestry UK Biobank cohort participants prescribed simvastatin or atorvastatin (aged 40-79 years at first prescription, treatment duration 1 month to 29 years, mean 5.7 years). Principal outcomes were clinically high total cholesterol (>5 mmol/L) at baseline, plus treatment discontinuation. RESULTS: A total of 48.4% of 591 females homozygous for SLCO1B1*5 decreased function genotype had raised cholesterol vs 41.7% of those with functioning SLCO1B1 (odds ratio 1.31, 95% confidence interval [CI] 1.1-1.55, P = .001). Fewer males had high cholesterol and the genotype effect was attenuated. In primary care prescribing, females homozygous for SLCO1B1*5 were more likely to stop receiving these statins (29.5%) than women with normal SLCO1B1 (25.7%) (hazard ratio [HR] 1.19, 95% CI 1.03-1.37, P = .01), amounting to five discontinuations per 100 statin-years in the SLCO1B1*5 group vs four in the normal SLCO1B1 function group. This remained significant after the first year of treatment (HR for discontinuing >1 year after first prescription 1.3, 95% CI 1.08-1.56, P = .006). In men SLCO1B1*5 was only associated with treatment discontinuation in the first year. CONCLUSIONS: In this large community sample of patients on commonly prescribed statins, the SLCO1B1*5 decreased function variant had much larger effects on cholesterol control and treatment duration in women than in men. Efforts to improve the effectiveness of statin therapy in women may need to include SLCO1B1*5 genotype-guided statin selection.

Development of Molecularly Imprinted Magnetic Amino Acid-Based Nanoparticles for Voltammetric Analysis of Lead Ions in Honey.

Lead (Pb) is a hazardous metal that poses a significant threat to both the environment and human health. The presence of Pb in food products such as honey can pose a significant risk to human health and is therefore important to detect and monitor. In this study, we propose a voltammetric detection method using molecularly imprinted polymer (MIP) electrodes to detect Pb (II) ions in honey. Pb (II) ion-imprinted amino acid-based nanoparticles with magnetic properties on a carbon paste electrode (MIP-CPE) were designed to have high sensitivity and selectivity towards Pb (II) ions in the honey sample. Zetasizer measurements, electron spin resonance, and scanning electron microscopy were used to characterize magnetic polymeric nanoparticles. The results showed that the voltammetric detection method using MIP-CPE was able to accurately detect Pb (II) ions in honey samples with a low detection limit. The proposed method offers a simple, rapid, cost-effective solution for detecting Pb (II) ions in honey. It could potentially be applied to other food products to ensure their safety for human consumption. The MIP-CPE sensor was designed to have high sensitivity and selectivity towards Pb (II) ions in the honey sample. The results showed that the technique was able to deliver highly sensitive results since seven different concentrations were prepared and detected to obtain an R2 of 0.9954, in addition to a low detection limit (LOD) of 0.0912 µM and a low quantification limit (LOQ) of 0.276 µM. Importantly, the analysis revealed no trace of Pb (II) ions in the honey samples obtained from Cyprus.

Development of a Plasmonic Sensor for a Chemotherapeutic Agent Cabazitaxel.

Drug dosage is a crucial subject in both human and animal treatment. Administering less drug dosage may prevent treatment or make it less effective, and high drug dosage may cause a heightened risk of adverse effects, or in some cases, cost a patient's life. Also, even when the dosage is administered carefully, metabolic differences may cause different effects on different patients. Because of these considerations, monitoring drug dosage in the body is a critical and significant requirement in the health industry. Within the scope of this study, a reusable surface plasmon resonance (SPR) chip with fast response, high selectivity, and no pretreatment is produced for the chemotherapeutic agent cabazitaxel. A cabazitaxel-imprinted nanofilm was synthesized on the sensor chip surface and characterized by atomic force microscopy, ellipsometry, and contact angle measurements. Standard cabazitaxel solution and an artificial plasma sample were used for the kinetic analysis. Docetaxel, methylprednisolone, and dexamethasone were analyzed for their selectivity experiment. In addition, the repeatability and storage durability of the sensor were also evaluated. As a result of the adsorption studies, the limit of detection and limit of quantitation values were found to be 0.012 and 0.036 µg/mL, respectively. High-performance liquid chromatography analysis was used to validate the response of the cabazitaxel-imprinted sensor.

Preparation of Immobilised 17ß-Estradiol-Imprinted Nanoparticles onto Bacterial Cellulose Nanofibres to Use for the Removal of 17ß-Estradiol from Wastewater.

Estradiol, a phenolic steroid oestrogen, is one of the endocrine-disrupting chemicals (EDCs) found in natural and tap waters. The detection and removal of EDCs is attracting attention daily as they negatively affect animals' and humans' endocrine functions and physiological conditions. Therefore, developing a fast and practical method for the selective removal of EDCs from waters is essential. In this study, we prepared 17ß-estradiol (E2)-imprinted HEMA-based nanoparticles onto bacterial cellulose nanofibres (E2-NP/BC-NFs) to use for the removal of E2 from wastewater. FT-IR and NMR confirmed the structure of the functional monomer. The composite system was characterised by BET, SEM, µCT, contact angle, and swelling tests. Additionally, the non-imprinted bacterial cellulose nanofibres (NIP/BC-NFs) were prepared to compare the results of E2-NP/BC-NFs. Adsorption of E2 from aqueous solutions was performed in batch mode and investigated via several parameters for optimisation conditions. The effect of pH studies was examined in the 4.0-8.0 range using acetate and phosphate buffers and a concentration of E2 of 0.5 mg/mL. The maximum E2 adsorption amount was 254 µg/g phosphate buffer at 45 °C. The experimental data show that the Langmuir is a relevant isotherm model for E2 adsorption. Additionally, the relevant kinetic model was the pseudo-second-order kinetic model. It was observed that the adsorption process reached equilibrium in less than 20 min. The E2 adsorption decreased with the increase in salt at varying salt concentrations. The selectivity studies were performed using cholesterol and stigmasterol as competing steroids. The results show that E2 is 46.0 times more selective than cholesterol and 21.0 times more selective than stigmasterol. According to the results, the relative selectivity coefficients for E2/cholesterol and E2/stigmasterol were 8.38 and 86.6 times greater for E2-NP/BC-NFs than for E2-NP/BC-NFs, respectively. The synthesised composite systems were repeated ten times to assess the reusability of E2-NP/BC-NFs.

Removal of amoxicillin via chromatographic monolithic columns: comparison between batch and continuous fixed bed.

This study presented a hydrophobic interaction-based poly(HEMA-MATrp) monolithic chromatographic column (MCC) to remove amoxicillin from aqueous solutions. In addition to their porous structure, monolithic-filled columns offer superior properties without loss of performance, which is one of the points that make them unique. The specific surface area of the monolithic column synthesized by the bulk polymerization of 2-hydroxyethyl methacrylate and N-Methacryloyl-L-tryptophan. Also, poly(HEMA-MATrp) MCC has been characterized via FTIR, SEM, and elemental analysis. According to BET analysis, the specific surface area of the poly(HEMA-MATrp) monolithic chromatographic column (MCC) is 14.2 mg/g. The adsorption and desorption of amoxicillin in an aqueous solution were investigated comparatively in both continuous fixed bed and batch adsorption. The highest adsorption value of amoxicillin was determined at pH 7 in the presence of PBS as 62.11 mg/g. The appropriate adsorption isotherm for the adsorption of amoxicillin was Langmuir, and the reaction kinetics was pseudo-second-order. No significant loss was observed for the adsorption capacity of poly(HEMA-MATrp) MCC after the 5 cycles of adsorption-desorption studies. Also, the loss for the adsorption capacity of the monolithic column is just %5.2 after 6-month storage, proving the reusability and storability of the monolithic column.

Simple and Fast Pesticide Nanosensors: Example of Surface Plasmon Resonance Coumaphos Nanosensor.

Here, a molecular imprinting technique was employed to create an SPR-based nanosensor for the selective and sensitive detection of organophosphate-based coumaphos, a toxic insecticide/veterinary drug often used. To achieve this, UV polymerization was used to create polymeric nanofilms using N-methacryloyl-l-cysteine methyl ester, ethylene glycol dimethacrylate, and 2-hydroxyethyl methacrylate, which are functional monomers, cross-linkers, and hydrophilicity enabling agents, respectively. Several methods, including scanning electron microscopy (SEM), atomic force microscopy (AFM), and contact angle (CA) analyses, were used to characterize the nanofilms. Using coumaphos-imprinted SPR (CIP-SPR) and non-imprinted SPR (NIP-SPR) nanosensor chips, the kinetic evaluations of coumaphos sensing were investigated. The created CIP-SPR nanosensor demonstrated high selectivity to the coumaphos molecule compared to similar competitor molecules, including diazinon, pirimiphos-methyl, pyridaphenthion, phosalone, N-2,4(dimethylphenyl) formamide, 2,4-dimethylaniline, dimethoate, and phosmet. Additionally, there is a magnificent linear relationship for the concentration range of 0.1-250 ppb, with a low limit of detection (LOD) and limit of quantification (LOQ) of 0.001 and 0.003 ppb, respectively, and a high imprinting factor (I.F.4.4) for coumaphos. The Langmuir adsorption model is the best appropriate thermodynamic approach for the nanosensor. Intraday trials were performed three times with five repetitions to statistically evaluate the CIP-SPR nanosensor's reusability. Reusability investigations for the two weeks of interday analyses also indicated the three-dimensional stability of the CIP-SPR nanosensor. The remarkable reusability and reproducibility of the procedure are indicated by an RSD% result of less than 1.5. Therefore, it has been determined that the generated CIP-SPR nanosensors are highly selective, rapidly responsive, simple to use, reusable, and sensitive for coumaphos detection in an aqueous solution. An amino acid, which was used to detect coumaphos, included a CIP-SPR nanosensor manufactured without complicated coupling methods and labelling processes. Liquid chromatography with tandem mass spectrometry (LC/MS-MS) studies was performed for the validation studies of the SPR.

Electrochemical Detection of Cortisol by Silver Nanoparticle-Modified Molecularly Imprinted Polymer-Coated Pencil Graphite Electrodes.

The sensitive cortisol detection by an electrochemical sensor based on silver nanoparticle-doped molecularly imprinted polymer was successfully improved. This study describes the method development for cortisol detection in both aqueous solution and biological samples using molecularly imprinted poly(hydroxyethyl methacrylate-N-methacryloyl-(l)-histidine methyl ester)-coated pencil graphite electrodes modified with silver nanoparticles (AgNPs) by differential pulse voltammetry (DPV). The cortisol-imprinted pencil graphite electrode (PGE) has a large surface area because of doped AgNPs with enhanced electroactivity. The prepared molecularly imprinted polymer was characterized by scanning electron microscopy. The DPV response of the synthesized electrode with outstanding electrical conductivity was clarified. Cortisol-imprinted polymer-coated PGEs (MIP), cortisol-imprinted polymer-coated PGEs with AgNPs (MIP@AgNPs), and nonimprinted polymer-coated PGEs with AgNPs (NIP@AgNPs) were evaluated for sensitive and selective detection of cortisol in aqueous solution. Five different cortisol concentrations (0.395, 0.791, 1.32, 2.64, and 3.96 nM) were applied to the MIP@AgNPs, and signal responses were detected by the DPV with a regression coefficient (R2) value of 0.9951. The modified electrode showed good electrocatalytic activity toward cortisol for the linear concentration range from 0.395 to 3.96 nM, and a low limit of detection was recorded as 0.214 nM. The results indicate that the MIP@AgNPs sensor has great potential for sensitive and selective cortisol determination in biological samples.

Inspirations of Biomimetic Affinity Ligands: A Review.

Affinity chromatography is a well-known method dependent on molecular recognition and is used to purify biomolecules by mimicking the specific interactions between the biomolecules and their substrates. Enzyme substrates, cofactors, antigens, and inhibitors are generally utilized as bioligands in affinity chromatography. However, their cost, instability, and leakage problems are the main drawbacks of these bioligands. Biomimetic affinity ligands can recognize their target molecules with high selectivity. Their cost-effectiveness and chemical and biological stabilities make these antibody analogs favorable candidates for affinity chromatography applications. Biomimetics applies to nature and aims to develop nanodevices, processes, and nanomaterials. Today, biomimetics provides a design approach to the biomimetic affinity ligands with the aid of computational methods, rational design, and other approaches to meet the requirements of the bioligands and improve the downstream process. This review highlighted the recent trends in designing biomimetic affinity ligands and summarized their binding interactions with the target molecules with computational approaches.

Computational Investigation of the Monomer Ratio and Solvent Environment for the Complex Formed between Sulfamethoxazole and Functional Monomer Methacrylic Acid.

In this study, the molecularly imprinted polymers (MIPs) that will be formed by the sulfamethoxazole (SMX) molecule and methacrylic acid (MAA) molecule were examined theoretically. The most stable interaction region between the two molecules was determined in solvent environments (ethanol, acetonitrile, and dimethylsulfoxide), and monomer ratios (SMX/MAA; 1:1, 1:2, and 1:3) were examined to form the most stable geometry. The number and length of the hydrogen bonds formed between the template molecule and the functional monomer and the interaction between the atoms were determined. Geometry optimizations of the molecules were calculated by the DFT method at the M06-2X/ccpVTZ level, and single-point energy calculations were carried out at the B2PLYP-D3/ccpVDZ level. In addition to the theoretical studies, the experimental Fourier-transform infrared spectroscopy (FTIR) spectrum of the complex formed between SMX and MAA was compared with the theoretical FTIR spectrum. As a result of the studies, the monomer ratio and solvent environment in which the stable complex was formed were determined in the MIP studies carried out with the SMX template molecule and MAA monomer. The most stable template molecule-monomer ratio of the complex between SMX and MAA was determined to be 1:3, and the solvent medium in which the most stable geometry was formed was acetonitrile.

Gold-Modified Molecularly Imprinted N-Methacryloyl-(l)-phenylalanine-containing Electrodes for Electrochemical Detection of Dopamine.

A molecularly imprinted polymer-based pencil graphite electrode (MIP PGE) sensor, modified with gold nanoparticles, was utilized for the detection of dopamine in the presence of other biochemical compounds using cyclic voltammetry (CV) and differential pulse voltammetry (DPV), depending on its strong electroactivity function. The pulse voltammetry methods recorded the highest response. In addition to the high oxidation rate of DA and the other biomolecule interferences available in the sample matrix used, which cause overlapping voltammograms, we aimed to differentiate them in a highly sensitive limit of detection range. The calibration curves for DA were obtained using the CV and DPV over the concentration range of 0.395-3.96 nM in 0.1 M phosphate buffer solution (PBS) at pH 7.4 with a correlation coefficient of 0.996 and a detection limit of 0.193 nM. The electrochemical technique was employed to detect DA molecules quantitatively in human blood plasma selected as real samples without applying any pre-treatment processes. MIP electrodes proved their ability to detect DA with high selectivity, even with epinephrine and norepinephrine competitor molecules and interferences, such as ascorbic acid (AA). The high level of recognition achieved by molecularly imprinted polymers (MIPs) is essential for many biological and pharmaceutical studies.

Analysis of CYP2C19 genetic variants with ischaemic events in UK patients prescribed clopidogrel in primary care: a retrospective cohort study.

OBJECTIVE: To determine whether CYP2C19 loss-of-function (LoF) alleles increase risk of ischaemic stroke and myocardial infarction (MI) in UK primary care patients prescribed clopidogrel. DESIGN: Retrospective cohort analysis. SETTING: Primary care practices in the UK from January 1999 to September 2017. PARTICIPANTS: 7483 European-ancestry adults from the UK Biobank study with genetic and linked primary care data, aged 36-79 years at time of first clopidogrel prescription. INTERVENTIONS: Clopidogrel prescription in primary care, mean duration 2.6 years (range 2 months to 18 years). MAIN OUTCOME MEASURE: Hospital inpatient-diagnosed ischaemic stroke, MI or angina while treated with clopidogrel. RESULTS: 28.7% of participants carried at least one CYP2C19 LoF variant. LoF carriers had higher rates of incident ischaemic stroke while treated with clopidogrel compared with those without the variants (8 per 1000 person-years vs 5.2 per 1000 person-years; HR 1.53, 95% CIs 1.04 to 2.26, p=0.031). LoF carriers also had increased risk of MI (HR 1.14, 95% CI 1.04 to 1.26, p=0.008). In combined analysis LoF carriers had increased risk of any ischaemic event (stroke or MI) (HR 1.17, 95% CI 1.06 to 1.29, p=0.002). Adjustment for aspirin coprescription produced similar estimates. In lifetables using observed incidence rates, 22.5% (95% CI 14.4% to 34.0%) of CYP2C19 LoF carriers on clopidogrel were projected to develop an ischaemic stroke by age 79 (oldest age in the study), compared with 15.4% (95% CI 11.4% to 20.5%) in non-carriers, that is, 7.1% excess stroke incidence in LoF carriers by age 79. CONCLUSIONS: A substantial proportion of the UK population carry genetic variants that reduce metabolism of clopidogrel to its active form. In family practice patients on clopidogrel, CYP2C19 LoF variants are associated with substantially higher incidence of ischaemic events. Genotype-guided selection of antiplatelet medications may improve outcomes in patients carrying CYP2C19 genetic variants.

Molecularly imprinted polymer integrated plasmonic nanosensor for cocaine detection.

A molecularly imprinted polymeric nanofilm was prepared for cocaine detection and applied to plasmonic nanosensor for real-time kinetic, selectivity and reusability analyses. The sensing polymeric surface was fabricated by synthesizing a selective and specific nanofilm on the gold plasmonic nanosensor surface. After characterization experiments with atomic force microscopy, ellipsometer, and contact angle measurements, the kinetic studies of cocaine detection in aqueous solutions in a wide concentration range between 0.2-100 µg/mL were applied to plasmonic nanosensor system at 24 °C with a low limit of detection (0.1 µg/L) and quantification values (0.3 µg/L) and the results showed that this molecularly imprinted polymeric nanofilm integrated plasmonic nanosensor is providing a model for the fastest, most accurate and most precise identification of the cocaine molecule which constitutes a large part of the workload of forensic laboratories.

A dye-affinity cryogel membrane for malate dehydrogenase purification from Saccharomyces cerevisiae.

Cibacron blue F3GA functionalized poly(hydroxyethyl methacrylate) cryogel membranes were prepared and applied for a simple purification of malate dehydrogenase (MDH) from crude extract of Saccharomyces cerevisiae. Swelling tests, scanning electron microscopy, surface area measurements and Fourier transform infrared (FTIR) spectroscopy techniques were used for the characterization of dye-affinity cryogel membranes. Following cell homogenization and extraction, MDH was purified using the dye-affinity cryogel membranes at a high yield of 80.5% with 54-fold purification. Maximum MDH adsorption amount was determined to be 267.7 mg/g of membranes at pH 7.4, 25 °C and a flow rate of 1.0 mL/min. Interestingly, the cryogel membranes were used for several purification runs without any significant decrease in MDH adsorption capacity. Sodium dodecyl sulfate polyacrylamide gel electrophoresis was carried out to assess the purity of the eluted MDH. The obtained results highlight the dye-affinity cryogel membranes as powerful dye affinity adsorbents for MDH purification from S. cerevisiae.

Synthesis of cholesterol imprinted polymeric particles.

The aim of this study is to prepare cholesterol-imprinted polymeric particles. N-Methacryloyl-(L)-tyrosinemethylester (MAT) was chosen as the complexing monomer. In the first step, functional monomer MAT was synthesized by the reaction of L-tyrosine methylester and methacryloyl chloride and characterized by FTIR and NMR. Then, cholesterol was complexed with MAT in different mol ratios and the cholesterol-imprinted poly(2-hydroxyethyl methacrylate-N-methacryloyl-(L)-tyrosine methylester) [MIP] particles were synthesized by bulk polymerization. After that, the template molecules (i.e., cholesterol) were removed using chloroform. MIP particles were characterized by elemental analysis, FTIR, SEM, swelling tests and surface area measurements. Cholesterol adsorption experiments were performed in a batch experimental set-up. Adsorption medium was methanol or intestinal mimicking solution. Stigmasterol and estradiol were used as competing molecules in selectivity tests. Obtained results were as follows: swelling ratio of MIP and non-imprinted (NIP) particles were 60.8% and 44.1% in water. With the increase in the amount of MAT in the polymerization medium, incorporation of MAT was increased (16.6-78.0 micromol/g). SEM photographs showed the surface roughness and porosity. Specific surface area of NIP and MIP particles were found as 19.2 and 31.5 m(2)/g, respectively. Template molecules (i.e., cholesterol) were removed from the polymer structure in the ratio of 76-84% of the initial concentration. Cholesterol adsorption increased with the increase in cholesterol concentration up to 1.5 mg/mL. MIP particles prepared using higher amounts of cholesterol exhibit significantly higher capacity to the NIP particles (i.e., control polymer). MIP particles were 3.09 and 3.60 times selective with respect to the stigmasterol and estradiol, respectively. Reusability of MIP particles was also investigated. MIP particles showed negligible loss in the cholesterol adsorption capacity after five adsorption-desorption cycles with the same adsorbent.

Plastic antibody based surface plasmon resonance nanosensors for selective atrazine detection.

This study reports a surface plasmon resonance (SPR) based affinity sensor system with the use of molecular imprinted nanoparticles (plastic antibodies) to enhance the pesticide detection. Molecular imprinting based affinity sensor is prepared by the attachment of atrazine (chosen as model pesticide) imprinted nanoparticles onto the gold surface of SPR chip. Recognition element of the affinity sensor is polymerizable form of aspartic acid. The imprinted nanoparticles were characterized via FTIR and zeta-sizer measurements. SPR sensors are characterized with atomic force microscopy (AFM), scanning electron microscopy (SEM), Fourier transform infrared spectrophotometry (FTIR) and contact angle measurements. The imprinted nanoparticles showed more sensitivity to atrazine than the non-imprinted ones. Different concentrations of atrazine solutions are applied to SPR system to determine the adsorption kinetics. Langmuir adsorption model is found as the most suitable model for this affinity nanosensor system. In order to show the selectivity of the atrazine-imprinted nanoparticles, competitive adsorption of atrazine, simazine and amitrole is investigated. The results showed that the imprinted nanosensor has high selectivity and sensitivity for atrazine.

Synthesis of tentacle-type magnetic beads as immobilized metal-chelate affinity support for cytochrome c adsorption.

Magnetic poly(2-hydroxyethylmethacrylate) (mPHEMA) beads with an average diameter of 100-140 microm were produced by suspension polymerization in the presence of magnetite particles (i.e. Fe3O4). Specific surface area and average pore size of the magnetic beads was found to be 50 m2/g and 819 nm, respectively. Ester groups in the mPHEMA structure were converted to imine groups by reacting with poly(ethyleneimine) (PEI) in the presence of NaH. Amino (-NH2) content of PEI-attached mPHEMA beads was determined as 102 mg PEI/g. Then, Cu2+ ions were chelated on the magnetic beads in the range of 20-793 micromol Cu2+/g. Cytochrome c (cyt c) adsorption was performed on the metal chelating beads from aqueous solutions containing different amounts of cyt c at different pHs, Cu2+ loadings and temperatures. Cyt c adsorption on the mPHEMA/PEI beads was 4.6 mg/g. Cu2+ chelation increased the cyt c adsorption significantly (40.1 mg/g). Adsorption capacity increased with Cu2+ loading and then reached a saturation value. Cyt c adsorption decreased with increasing temperature. Cyt c molecules could be reversibly adsorbed and eluted ten times with the magnetic adsorbents without noticeable loss in their cyt c adsorption capacity. The applicability of two kinetic models including pseudo-first order and pseudo-second order model was estimated on the basis of comparative analysis of the corresponding rate parameters, equilibrium capacity and correlation coefficients. Results suggest that chemisorption processes could be the rate-limiting step in the adsorption process. In the last part of this article, cyt c adsorption experiments were performed in a magnetically stabilized fluidized bed (MSFB) system at optimum conditions determined from the batch experiments. The adsorption capacity decreased significantly from 46.8 to 15.4 mg/g polymer with the increase of the flow-rate from 0.5 to 4.0 ml/min. The resulting magnetic chelator beads possessed excellent long-term storage stability.