Rapid electrochemical lateral flow device for the detection of Δ9-tetrahydrocannabinol.

Cannabis is a plant that is harmful and beneficial because it contains more than 400 bioactive compounds, and the main compounds are Δ9 tetrahydrocannabinol (THC) and cannabidiol (CBD). Currently, cannabis extracts are used in medicine, but the amount of THC as a main psychoactive component is strictly regulated. Therefore, the ability to rapidly and accurately detect THC is important. Herein, we developed a sensitive electrochemical method combining a rapid lateral flow assay (LFA) to detect THC rapidly. An electrochemical LFA device was constructed by attaching a screen-printed electrode inside a lateral-flow device to exploit the remarkable binding of THC to the cannabinoid type 2 (CB2) receptor in the test zone. The ferrocene carboxylic acid attached to the monoclonal THC antibody acts as an electroactive species when it binds to the THC in the sample before it flows continuously to the CB2 receptor region on the electrode. Under optimal conditions, the detection time is within 6 min and the devise shows excellent performance with a detection limit of 1.30 ng/mL. Additionally, the device could be applied to detect THC in hemp extract samples. The results obtained from this sensor are similar to the standard method (HPLC) for detecting THC. Therefore, this proposed device is useful as an alternative device for the on-site determination of THC because it is inexpensive, portable, and exhibits high sensitivity.

Paper-based electrochemical immunosensor for the determination of symmetric dimethylarginine.

In this work, we present the development of an immunosensor for the direct, selective, and sensitive determination of symmetric dimethylarginine (SDMA) in urine, in view of the emerging role of this molecule as a biomarker for renal disease. SDMA is almost completely excreted by the kidneys, hence in renal dysfunction, the excretion is decreased, resulting in accumulation in plasma. Reference values for plasma or serum have already been established in small animal practice. Values < 15 µg/dL are considered normal, 15-19 µg/dL are values of concern, and at values > 20 µg/dL kidney disease is likely. The proposed electrochemical paper-based sensing platform uses anti-SDMA antibodies for targeted detection of SDMA. Quantification is related to a decrease in the signal of a redox indicator due to the formation of an immunocomplex that interferes with electron transfer. Square wave voltammetry measurements showed a linear correlation of the peak decline for 50 nM - 1 µM SDMA with a detection limit of 15 nM. The influence of common physiological interferences caused no significant peak reduction, indicating excellent selectivity. The proposed immunosensor was successfully applied for the quantification of SDMA in human urine of healthy individuals. Surveillance of SDMA concentration in urine could prove to be very valuable in the diagnosis or monitoring of renal disease.

Electrochemical flow injection approach for routine screening of lipase activity in pancreatic preparations.

A state-of-the-art strategy for the determination of lipase activity in pancreatic preparations using flow injection analysis (FIA) with electrochemical detection (FIA-ED) is described. The procedure is based on the enzymatic reaction of a specific substrate (1,3-dilinoleoyl-glycerol) with lipase from porcine pancreas and determination of enzymatically formed linoleic acid (LA) at +0.4 V by applying a cobalt (II) phthalocyanine-multiwalled carbon-nanotubes modified carbon paste electrode (Co(II)PC/MWCNT/CPE). In order to get a high-performance analytical method, sample preparation, flow system, and electrochemical conditions were optimized. Under optimized conditions the lipase activity of porcine pancreatic lipase was calculated to be 0.47 units per mg lipase protein based on the definition that 1 unit hydrolyses 1 microequivalent linoleic acid from 1,3-dilinoleoyl-glycerol per 1 min at pH 9 and 20 °C (kinetic measurement: 0-25 min). Moreover, the developed procedure was shown to be easily adaptable for the fixed-time assay (incubation time 25 min) as well. In this case, linear correlation between flow signal and lipase activity was found in the range from 0.8 to 18 U L-1. LOD and LOQ were determined to be 0.3 U L-1 and 1 U L-1, respectively. The kinetic assay was further preferred for the determination of lipase activity in commercially available pancreatic preparations. The lipase activities of all preparations obtained by the present method were found to be in good correlation with those obtained by the titrimetric method and declared by manufacturers.

A novel immunosensor based on cobalt oxide nanocomposite modified single walled carbon nanohorns for the selective detection of aflatoxin B1.

We developed a novel, sensitive, and selective platform for the specific determination of aflatoxin B1 (AFB1). Single-walled carbon nanohorns decorated by a cobalt oxide composite and gold nanoparticles were created to provide facile electron transfer and improve the sensor's sensitivity. In addition, we attributed the selectivity of the proposed sensor to the specific binding property of the anti-aflatoxin B1 antibody. We clarified the specific interaction of the proposed immunosensor to AFB1 using homology modeling combined with molecular docking. In the presence of AFB1, the current signal of the modified electrode reduced; this involved specific antibody-antigen binding, including hydrophobic hydrogen bonding and pi-pi stack interactions. The new AFB1 sensor platform showed two linearity ranges of 0.01-1 ng mL-1 and 1-100 ng mL-1, with the limit of detection at 0.0019 ng mL-1. We investigated the proposed immunosensor in real samples, including peanuts, certified reference material of a peanut sample (labeled 206 µg kg-1 AFB1), corn, and chicken feed. The sensor's accuracy was 86.1-104.4% recovery, which agrees with the reference HPLC technique using paired t-test analysis. The present work shows excellent performance for AFB1 detection and could be applied for food quality control or modified to detect other mycotoxins.

An electrochemical sensor for the voltammetric determination of artemisinin based on carbon materials and cobalt phthalocyanine.

An electrochemical sensor for the determination of artemisinin has been developed based on a glassy carbon electrode modified with hybrid nanocomposites of cobalt phthalocyanine, graphene nanoplatelets, multi-walled carbon nanotubes and ionic liquids (IL). To improve the sensitivity and selectivity of the sensor, cobalt phthalocyanine (CoPc) was used as an effective redox mediator to promote and catalyze the artemisinin reduction. Furthermore, the graphene nanoplatelets and multi-walled carbon nanotubes were used as excellent conducting supporting materials to improve the sensitivity of the electrochemical sensor. Moreover, IL with a surface charge was also employed to prevent aggregation of the graphene nanoplatelets and multi-walled carbon nanotubes. The analytical signal was generated from the reduction of Co(III)Pc generated by artemisinin. The proposed electrochemical sensor was applied to the detection of artemisinin using differential pulse voltammetry and provided a signal with wide linearity ranging from 1.5-60 µM and 60-600 µM and a detection limit of 0.70 µM (3SD/m). Furthermore, the proposed sensor displayed good repeatability and reproducibility of 2.9-3.0 and 3.1-4.4% RSD, respectively. Applications of the sensor to drug and plant samples demonstrated accuracy in a range of 105-116% recoveries. In addition, the results were in good agreement with those obtained from the HPLC method as a reference technique. Thus, the proposed electrochemical sensor provides a new alternative platform for sensitive and selective determination of artemisinin in the analysis of pharmaceuticals with good precision and accuracy.

Voltammetric lipase activity assay based on dilinolein and a modified carbon paste electrode.

In this work, a novel electrochemical assay for characterizing both lipases and lipase inhibitors as well as for the determination of lipase activity is described. It is based on a carbon paste electrode, modified with cobalt(II)phthalocyanine, and multi-walled carbon nanotubes (MWCNTs). As reaction media, a sodium borate buffer was used (0.1 M, pH 9). The measurements were carried out in a batch system using differential pulse voltammetry (DPV) and 1,3-dilinolein as standard substrate. The activity assay showed a linearity for porcine pancreas lipase activity in a range between 20 and 300 U L-1 (per min) with a limit of detection (LOD) of 7 U L-1 and a limit of quantification (LOQ) of 20 U L-1. The kinetic behavior of the lipase reaction was investigated, resulting in a KM value of 0.29 mM. The applicability of the activity assay could be shown by investigating the activity of lipases from Aspergillus oryzae and Candida rugosa, and the results were confirmed by a reference method. The inhibitory effects were characterized with Orlistat.

Dose- and Sex-Dependent Changes in Hemoglobin Oxygen Affinity by the Micronutrient 5-Hydroxymethylfurfural and α-Ketoglutaric Acid.

5-Hydroxymethylfurfural (5-HMF) is known to increase hemoglobin oxygen affinity (Hb-O2 affinity) and to induce a left shift of the oxygen dissociation curve (ODC). It is under investigation as a therapeutic agent in sickle cell anemia and in conditions where pulmonary oxygen uptake is deteriorated or limited (e.g., various clinical conditions or altitude exposure). The combination of 5-HMF and α-ketoglutaric acid (αKG) is commercially available as a nutritional supplement. To further elucidate dose effects, ODCs were measured in vitro in venous whole blood samples of 20 healthy volunteers (10 female and 10 male) after the addition of three different doses of 5-HMF, αKG and the combination of both. Linear regression analysis revealed a strong dose-dependent increase in Hb-O2 affinity for 5-HMF (R2 = 0.887; p < 0.001) and the commercially available combination with αKG (R2 = 0.882; p < 0.001). αKG alone increased Hb-O2 affinity as well but to a lower extent. Both the combination (5-HMF + αKG) and 5-HMF alone exerted different P50 and Hill coefficient responses overall and between sexes, with more pronounced effects in females. With increasing Hb-O2 affinity, the sigmoidal shape of the ODC was better preserved by the combination of 5-HMF and αKG than by 5-HMF alone. Concerning the therapeutic effects of 5-HMF, this study emphasizes the importance of adequate dosing in various physiological and clinical conditions, where a left-shifted ODC might be beneficial. By preserving the sigmoidal shape of the ODC, the combination of 5-HMF and αKG at low (both sexes) and medium (males only) doses might be able to better maintain efficient oxygen transport, particularly by mitigating potentially deteriorated oxygen unloading in the tissue. However, expanding knowledge on the interaction between 5-HMF and Hb-O2 affinity in vitro necessitates further investigations in vivo to additionally assess pharmacokinetic mechanisms.

Antioxidant Solution in Combination with Angiotensin-(1-7) Provides Myocardial Protection in Langendorff-Perfused Rat Hearts.

As progressive organ shortage in cardiac transplantation demands extension of donor criteria, effort is needed to optimize graft survival. Reactive oxygen and nitrogen species, generated during organ procurement, transplantation, and reperfusion, contribute to acute and late graft dysfunction. The combined application of diverse substances acting via different molecular pathways appears to be a reasonable approach to face the complex mechanism of ischemia reperfusion injury. Thus, an antioxidant solution containing α-ketoglutaric acid, 5-hydroxymethylfurfural, N-acetyl-L-methionine, and N-acetyl-selenium-L-methionine was combined with endogenous angiotensin-(1-7). Its capacity of myocardial protection was investigated in isolated Langendorff-perfused rat hearts subjected to warm and cold ischemia. The physiological cardiac parameters were assessed throughout the experiments. Effects were evaluated via determination of the oxidative stress parameters malondialdehyde and carbonyl proteins as well as immunohistochemical and ultrastructural tissue analyses. It was shown that a combination of 20% (v/v) antioxidant solution and 220 pM angiotensin-(1-7) led to the best results with a preservation of heart tissue against oxidative stress and morphological alteration. Additionally, immediate cardiac recovery (after warm ischemia) and normal physiological performance (after cold ischemia) were recorded. Overall, the results of this study indicate substantial cardioprotection of the novel combination with promising prospective for future clinical use.

Drug interactions in the treatment of rheumatoid arthritis and psoriatic arthritis.

BACKGROUND: Treating patients with inflammatory joint diseases (rheumatoid arthritis, psoriatic arthritis) according to established treatment algorithms often requires the simultaneous use of three or more medications to relieve symptoms and prevent long-term joint damage as well as disability. OBJECTIVE: To assess and give an overview on drug-drug interactions in the pharmacotherapy of inflammatory joint diseases with regards to their clinical relevance. METHODS: All possible drug combinations were evaluated using three commercially available drug interaction programs. In those cases where only limited/no data were found, a comprehensive hand search of Pubmed was carried out. Finally, the drug-drug interactions of all possible combinations were classified according to evidence-based medicine and a specifically generated relevance-based system. RESULTS: All three interaction software programs showed consistent results. All detected interactions were combined in clearly structured tables. CONCLUSION: A concise overview on drug-drug interactions is given. Especially in more sophisticated cases extensive knowledge of drug interactions supports optimisation of therapy and results in improved patient safety.

Electrochemical detection of NOx gas based on disposable paper-based analytical device using a copper nanoparticles-modified screen-printed graphene electrode.

A disposable gas-sensing paper-based device (gPAD) was fabricated in origami design which integrates the gas adsorbent and the electrochemical detection zone in a single device. The gPAD for the determination of NOx gas uses a screen-printed graphene electrode modified with copper nanoparticles (CuNP/SPGE) to achieve high sensitivity and selectivity. The gPAD detects both, NO and NO2 (as NOx) with same current responses. The measurement could be performed directly through differential pulse voltammetry (DPV) with a detection limit as low as 0.23 vppm and 0.03 vppm with exposure times of 25 min and 1 h, respectively. The reproducibility in terms of relative standard deviation was less than 5.1% (n = 7 devices) at 25, 75 and 125 vppm NO2 and the life-time of this device was more than 30 days. The gPAD was applied to detect NOx in air and exhaust gases from cars. In comparison with spectrophotometry, there are no significant differences between both methods using a paired t-test of the results on a 95% confidence level. The designed gPAD can provide a new template model for other gas sensors with features of disposability and portability for fieldwork analysis at low cost.

Investigation of antioxidative effects of a cardioprotective solution in heart tissue.

A multi-component solution, containing α-ketoglutaric acid (α-KG), 5-hydroxymethylfurfural (5-HMF), N-acetyl-seleno-L-methionine (NASeLM), and N-acetyl-L-methionine (NALM) as active ingredients, has been tested considering its supposed antioxidative effect with respect to heart transplantations. Oxidative stress was induced on isolated rat hearts through occlusion of a coronary artery and in chicken heart tissue through hydrogen peroxide. Both heart types were analyzed and the oxidative stress markers malondialdehyde (MDA) and carbonyl proteins (CPs) were determined via HPLC/UV-Vis. In both approaches, it was found that treatment with the multi-component solution led to a lower amount of MDA and CPs compared to a negative control treated with Krebs-Ringer solution (KRS). Further investigation on chicken heart tissue identified α-KG as antioxidative component in these experiments. However, numerous factors like arrhythmia, vessel dilatation, and minimization of oxidative stress effects play an important role for successful transplantation. Therefore, the investigated multi-component solution might be a novel approach against oxidative stress situations, for example at ischemia reperfusion injury during heart transplantations.

Development of a cobalt(II) phthalocyanine- MWCNT modified carbon paste electrode for the detection of polyunsaturated fatty acids.

In this work the development of an electrochemical sensor for the determination of polyunsaturated fatty acids (PUFAs), in particular linoleic acid, in commercially available safflower oil as complex matrix is described. The sensor consists of a carbon paste electrode with cobalt(II) phthalocyanine, Co(II)Pc, as mediator and multiwalled carbon-nanotubes (MWCNT) as nanomaterial. As carrier medium a sodium borate buffer (0.1 M, pH 9) was used. PUFAs were detected at a working voltage of 0.35-0.45 V (vs. Ag/AgCl). The sensor development was carried out in a batch system with differential pulse voltammetry (DPV) and cyclic voltammetry (CV). Sensor specification was tested by using various fatty acids (stearic acid, oleic acid, linoleic acid, linolenic acid and arachidonic acid) as well as esterified fatty acids, resulting in a specific applicability towards PUFAs, especially linoleic acid (LAH). The optimized sensor was applied in a flow injection analysis system (FIA) for the analysis of PUFAs in complex matrix. Linoleic acid was used as standard substrate to determine the analytical parameters. The linearity ranges between 7.5 and 200 µg mL-1 LAH, while the limit of detection was determined to be 2.5 µg mL-1 and the limit of quantification is approximately 7.5 µg mL-1 LAH. The LAH content was successfully detected in commercially available safflower oil via standard addition method and the results could be confirmed by a reference method. The PUFA content was calculated as LAH-equivalent.

A review of factors associated with maintenance of sinus rhythm after elective electrical cardioversion for atrial fibrillation.

Atrial fibrillation is the most common heart-rhythm disorder, affecting about 1.5% to 2% of the population with an increased risk of mortality and morbidity due to stroke, thromboembolism, and heart failure. If the conversion back to sinus rhythm does not happen spontaneously, pharmacological or electrical cardioversion (ECV) is the next available treatment options for some patients. However, the long-term success following ECV is variable. This review describes the factors that are associated with maintenance of sinus rhythm following ECV and proposes a clinical strategy based on the available evidence.

Biosensor for the characterisation of hMAO B inhibitors and the quantification of selegiline.

A novel human monoamine oxidase B (hMAO B) based biosensor for inhibitory measurements was developed. It allows both the characterisation of the type of enzyme inhibition and the sensitive and simple determination of inhibitors like selegiline hydrochloride. The sensor consists of a screen printed carbon working electrode modified with 20% manganese dioxide (MnO2) and the enzyme hMAO B, which was immobilised on the electrode via a dialysis membrane (regenerated cellulose, molecular weight cut-off 14000). Inhibition of hMAO B is evaluated by adding different concentrations of the inhibitor selegiline hydrochloride to the enzyme and applying a defined amount of the hMAO B substrate phenylethylamine (PEA). The enzymatically formed H2O2 is amperometrically detected at 0.4V vs. Ag/AgCl in a flow injection analysis (FIA) system. With 100µM PEA the sensor showed a linear correlation between peak height and inhibitor concentration in a range of 0.51-3.25µg/mL selegiline hydrochloride. LOD and LOQ were determined to be 0.15 and 0.51µg/mL, respectively. The sensor showed a repeatability of 3.7% and an intermediate precision of 8.1%. The inhibition-based biosensor was successfully employed to quantify selegiline hydrochloride in pharmaceutical samples. Kinetic studies via Lineweaver-Burk plot and enzyme quantity vs. current plot revealed that the inhibition is irreversible.

Unexpected deaths in cardiology outpatients - what can we learn from case review?

OBJECTIVES: A proportion of cardiac patients managed at a cardiology outpatient clinic will die between clinic visits. This study aimed to identify the cause of death, to determine if case review occurred and if a formal review of such cases might be useful. DESIGN: Single-centre retrospective cohort study. SETTING: A remote regional centre in the North of Scotland. PARTICIPANTS: All patients who had been removed from the cardiology outpatient clinic due to death in the community. MAIN OUTCOME MEASURES: Cause of death, comorbidities and treatments were collected from hospital records and the national register of deaths. Chi-squared test and Student's t-test were used with significance taken at the 5% level. RESULTS: Of 10,606 patients who attended the cardiology outpatient clinic, 75 (0.7%) patients died in the community. The majority (57.0%) died from a non-cardiac cause. Eleven patients (14.9%) died due to an unexpected cardiac death. A detailed case note review was undertaken. In only two (18.2%) cases was any note made as to the cause of death in the hospital records and in only one was there details of post mortem discussion between primary and secondary care. CONCLUSIONS: A small proportion of patients attending a cardiology outpatient clinic died in the community. Documentation of the death in the hospital notes was very poor and evidence of post mortem communication between primary and secondary care was absent in all but one case. Better documentation and communication between primary and secondary care would seem desirable.

Determination of Angiotensin-(1-7) with HPLC/Fluorescence-Detection.

Angiotensin-(1-7) is an important active component in the renin-angiotensin-system. Due to its cardio protective effects it is now under investigation in combination with antioxidants as a reperfusion solution. The combination showed impressive effects on isolated hearts of male Wistar rats after induced ischemia. In this work a high performance liquid chromatography method with fluorescence detection was developed for the first time for in-process measurements as well as for stability tests of the peptide in the novel antioxidant-containing Karal® solution. For fluorescence detection of angiotensin-(1-7) fluorescamine as derivatization dye was applied. Under optimized conditions the method showed linearity over the range of 50 to 5000 ng/mL with R(2) of 0.9988 and an overall precision better than 5.0 %. LOD and LOQ were determined to be in the femtomol range on column. It was found that stability of angiotensin-(1-7) could be significantly improved in the antioxidant containing preparation compared to aqueous solutions.

Simultaneous quantitation of alpha-ketoglutaric acid and 5-hydroxymethylfurfural in plasma by HPLC with UV and fluorescence detection.

Alpha-ketoglutaric acid (KG) and hydroxymethylfurfural (HMF) are currently being investigated in clinical trials as an approach in targeted cancer therapy. Hence, a method for the simultaneous determination of KG and HMF in plasma has been developed. Due to the strongly discriminative chemical properties of KG and HMF, SPE purification is performed using an ion-exchange cartridge to separate KG, and a hydrophobic polymeric cartridge to separate HMF. The cartridges are connected together for several steps, thus resulting in a quicker approach for the purification of plasma samples. The derivatization step is based on the reaction of the carbonyl groups of KG and HMF with dansylhydrazine (DNSH) catalyzed by trifluoroacetic acid. The formed derivatives could be separated by reversed-phase LC on a C8-column, and analyzed by UV and fluorescence detection in a single run using a gradient program. The obtained results show good reproducibility, specificity, and detection limits down to the low picomole range.

Bis-pentafluorobenzyl derivatives of N-acetyl-L-methionine and N-acetyl-L-selenomethionine for the quantitative determination in human plasma by gas chromatography-negative ion chemical ionisation mass spectrometry.

Targeted anti-cancer combination therapy with infusion of N-acetyl-L-methionine (NALM) and N-acetyl-L-selenomethionine (NASeLM) shows promising results in cancer treatment. Selenium has been recognised as a valuable additive in cancer therapeutics due to its ability to minimise side effects of chemotherapy and its role in cancer prevention and therapy. Due to the promising results of this new therapeutic approach evaluation of pharmacokinetic data for NALM and NASeLM is of ultimate importance. We have therefore elaborated a method for the quantitative measurement of these compounds in human plasma based on GC-negative ion chemical ionisation-MS. The derivatisation sequence elaborated can be regarded as a novel strategy for the chemical modification of delicate sulphur- and selenium-containing compounds, and underlines the enhanced reactivity of selenium-analogues of sulphur-containing amino acids. The target compounds were extracted from plasma with ethyl acetate and converted to the S/Se-pentafluorobenzyl-homocysteine pentafluorobenzyl ester derivative. Reaction conditions were optimised for derivative yield. Calibration graphs were established in the range of 2.938-481.105 ng/0.5 mL plasma (NALM) and 0.233-59.543 ng/0.5 mL plasma (NASeLM). Accuracy, precision and stability data were elaborated. The method was applied to pharmacokinetic profiling of the compounds after infusion into human volunteers.