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A one-step analysis method was developed for four types of amino acids using a microfluidic paper-based analytical device fabricated from chromatography filtration paper and laminate films. Aminoacyl-tRNA synthetase was used to detect each amino acid. The obtained laminated paper-based analytical device (LPAD) contained four enzymatic reaction areas. Colorimetric detection was performed based on the molybdenum blue reaction. A model method for the simple, easy, and simultaneous detection of several amino acid concentrations was suggested, in contrast to the conventional methods such as HPLC or LC–MS. The method provided a selective quantification at the ranges of 3.6–100 μM for tryptophan, 10.1–100 μM for glycine, 5.9–100 μM for histidine and 5.6–100 μM for lysine with a detection limit of 1.1 μM, 3.3 μM, 1.9 μM and 1.8 μM, respectively. LPAD fabrication was considerably simple, and the subsequent detection process was easy and required a short period of time (within 15 min). [ABSTRACT FROM AUTHOR]

A laminated paper-based analytical device (LPAD) for histidine detection was fabricated from a chromatography filtration paper and laminate films. Histidine recognition was effected by histidyl-tRNA synthetase (HisRS), and its detection was signaled colorimetrically based on the molybdenum blue reaction. The analytical conditions and detectable concentration range of histidine were examined. The method provided selective quantification from 1 to 100 μM histidine. LPAD fabrication is considerably simple, involving only the craft-cutting of the chromatography filtration paper and laminate film, and is cost-effective. [ABSTRACT FROM AUTHOR]

PG16 is a broadly neutralizing antibody to the human immunodeficiency virus (HIV). A crystal structure of PG16 revealed that the unusually long 28-residue complementarity determining region (CDR) H3 forms a unique subdomain, referred to as a "hammerhead", that directly contacts the antigen. The hammerhead apparently governs the function of PG16 while a previous experimental assay showed that the mutation of TyrH100Q to Ala, which does not directly contact the antigen, decreased the neutralization ability of PG16. However, the molecular mechanism by which a remote mutation from the hammerhead or contact paratope affects the neutralization potency has remained unclear. Here, we performed molecular dynamics simulations of the wild-type and variants (TyrH100Q to Ala, and TyrH100Q to Phe) of PG16, to clarify the effects of these mutations on the dynamics of CDR-H3. Our simulations revealed that the structural rigidity of the CDR-H3 in PG16 is attributable to the hydrogen bond interaction between TyrH100Q and ProH99, as well as the steric support by TyrH100Q. The loss of both interactions increases the intrinsic fluctuations of the CDR-H3 in PG16, leading to a conformational transition of CDR-H3 toward an inactive state. [ABSTRACT FROM AUTHOR]

Wastewater is discharged during washing processes in the production of biodiesel fuel (BDF) using alkaline catalysts. It can be recycled as a liquid fertilizer for hydroponics by adding essential components for plant growth. The effects of the liquid fertilizer on plant growth were investigated. Liquid fertilizer containing a smaller amount of the BDF wastewater had a similar effect on plant growth as the standard nutrient solutions. This result reveals that BDF wastewater can be recycled for use as a liquid fertilizer for hydroponics. However, fertilizer with a larger amount of the BDF wastewater showed poor and varied plant growth due to the growth of microorganisms in the contaminated wastewater. Hence, when BDF wastewater becomes contaminated during storage, sterilization is necessary to recycle it as a liquid fertilizer. Moreover, contamination during storage should be avoided for successful recycling. [ABSTRACT FROM AUTHOR]

Abnormal concentrations of amino acids in blood and urine can be indicative of several diseases, including cancer and diabetes. Therefore, analyses that examine amino acid concentrations are useful for the diagnosis of such diseases. In this study, we developed an enzyme-immobilized, small reactor column for flow analysis of amino acid concentrations. For the recognition of asparagine and lysine, asparaginyl-tRNA synthetase and lysyl-tRNA synthase were immobilized onto microparticles, respectively, and coupled with coloration reagents for spectrophotometric detection. This assay has some advantages in the analytical field, such as the ability to detect small amounts of analyte, allowing for the use of a small reaction volume, and ensuring a rapid and efficient reaction rate. This approach provided selective quantitation of up to 480 μM of asparagine and lysine in 200 mM Tris-HCl buffer (pH 8.0). [ABSTRACT FROM AUTHOR]

Analysis of the concentration of free amino acids in biological samples is useful in clinical diagnostics. However, currently available methods are time consuming, potentially delaying diagnosis. Therefore, the development of more rapid analytical tools is needed. In this study, a chemiluminescence detection method for amino acids was developed, and the conditions for the enzyme reaction and assay were examined. For the recognition of each amino acid (here, serine, proline, glycine, asparagine, leucine, and histidine), the corresponding aminoacyl-tRNA synthetase (aaRS) was employed, and multiple enzymatic reactions were combined with a luminol chemiluminescence reaction. This method provided selective quantification from 1 to 20 μM for serine, proline, glycine, and leucine; 1 to 60 μM for asparagine; and 1 to 150 μM for histidine. This assay, which utilized aaRSs for the detection of amino acids, could be useful for simple and rapid analysis of amino acids in clinical diagnostics. [ABSTRACT FROM AUTHOR]

The measurement of amino acid content is useful for the diagnosis of several types of diseases, including cancer and diabetes. In this study, a microfluidic method for the analysis of serine using enzymatic reactions coupled with spectrophotometric detection was developed. The assay system has some advantages in the analytical field, such as the ability to detect small amounts of analyte and reaction solution and a rapid and efficient reaction. For the specific detection of serine, seryl-tRNA synthetase was coupled with the generation of hydrogen peroxide, which was then detected by the Trinder reagent spectrophotometric method. Seryl- and other aminoacyl-tRNA synthetases are involved in the biosynthesis of peptides and proteins in the human body and should allow precise recognition of the corresponding amino acids. This approach provided selective quantitation of up to 250 μM serine in 100 mM Tris-HCl buffer (pH 8.0) in a semiautomatic system. [ABSTRACT FROM AUTHOR]

A phosphate-selective molecularly imprinted polymer was prepared using 1-allyl-2-thiourea as a functional monomer, and a method of sensing for phosphate was suggested using an ion-sensitive field-effect transistor (ISFET) or pH meter as a transducer. The phosphate-selective polymer bound to phosphate selectively, and the ion concentration change or the pH shift in the analyte solutions after incubation with the imprinted polymer were measured in these systems. In using ISFET electrode, concentration of phosphate from 10 to 40 μM could be measured selectively. [ABSTRACT FROM AUTHOR]

Molecularly imprinted polymers for indoleacetic acid were prepared by co-polymerizing N, N-dimethylaminoethyl methacrylate, 2-hydroxyethyl methacrylate (HEMA), and ethylene glycol dimethacrylate. The dependence of the affinity and selectivity of the imprinted polymers on HEMA content was evaluated chromatographically. The affinity was improved by increasing the HEMA content; the selectivity of the imprinted polymer was best when the HEMA content was approximately 30%, irrespective of monomer content. [ABSTRACT FROM AUTHOR]