Auxotrophy-Based Detection of Hyperornithinemia in Mouse Blood and Urine

Palanza, Kenneth M.; Nesta, Alex; Tumu, Renukanandan; Walton, Cherie M.; Davis, Michael A.; King, Thomas

doi:10.1177/2326409816649600

Cited by 2 publications

(3 citation statements)

References 35 publications

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“…This bioassay allows for rapid determination of amino acid concentrations within 60 min, a remarkably shorter time than necessary for any other conventional bioassays (Bertels et al 2012;Cardinal and Hedrick 1948;Chalova et al 2006;Erickson et al 2000;Kim et al 2010;Li and Ricke 2003;Palanza et al 2016;Steele et al 1949;Tamura et al 1952). The present results also demonstrate the high versatility of this bioassay in terms of both Fig.…”

Section: Discussionsupporting

confidence: 67%

“…Amino acid quantification is an important technology for a variety of fields and has applications in scientific research, healthcare (Hisamatsu et al 2012;Miyagi et al 2011;Palanza et al 2016), and the animal and human food industries (Chalova et al 2009;Mira de Orduna 2001). A variety of methods have been developed to meet these demands, such as instrumental methods (Kaspar et al 2009) and enzymatic assays Kameya et al 2013;Liu et al 2014;Matsui et al 2015).…”

Section: Introductionmentioning

confidence: 99%

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Translation-dependent bioassay for amino acid quantification using auxotrophic microbes as biocatalysts of protein synthesis

Kameya

Asano

2016

Appl Microbiol Biotechnol

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Bioassay for amino acid quantification is an important technology for a variety of fields, which allows for easy, inexpensive, and high-throughput analyses. Here, we describe a novel translation-dependent bioassay for the quantification of amino acids. For this, the gene encoding firefly luciferase was introduced into Lactococcus lactis auxotrophic to Glu, His, Ile, Leu, Pro, Val, and Arg. After a preculture where luciferase expression was repressed, the cells were mixed with analytes, synthetic medium, and an inducer for luciferase expression. Luminescence response to the target amino acid appeared just after mixing, and linear standard curves for these amino acids were obtained during 15-60-min incubation periods. The rapid quantification of amino acids has neither been reported in previous works on bioassays nor is it theoretically feasible with conventional methods, which require incubation times of more than 4 h to allow for the growth of the microbe used. In contrast, our assay was shown to depend on protein translation, rather than on cell growth. Furthermore, replacement of the luciferase gene with that of the green fluorescent protein (GFP) or β-galactosidase allowed for fluorescent and colorimetric detection of the amino acids, respectively. Significantly, when a Gln-auxotrophic Escherichia coli mutant was created and transformed by a luciferase expression plasmid, a linear standard curve for Gln was observed in 15 min. These results demonstrate that this methodology can provide versatile bioassays by adopting various combinations of marker genes and host strains according to the analytes and experimental circumstances.

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Section: Discussionsupporting

confidence: 67%

Section: Introductionmentioning

confidence: 99%

Translation-dependent bioassay for amino acid quantification using auxotrophic microbes as biocatalysts of protein synthesis

Kameya

Asano

2016

Appl Microbiol Biotechnol

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“…Additionally, proteinuria was determined using the Bradford assay on an aliquot of 24 h collected urine as previously described [25].…”

Section: Blood-urine Biochemical Analysismentioning

confidence: 99%

Reno-Protective Effect of GLP-1 Receptor Agonists in Type1 Diabetes: Dual Action on TRPC6 and NADPH Oxidases

et al. 2021

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Diabetic kidney disease (DKD), a serious diabetic complication, results in podocyte loss and proteinuria through NADPH oxidases (NOX)-mediated ROS production. DUOX1 and 2 are NOX enzymes that require calcium for their activation which enters renal cells through the pivotal TRPC channels. Hypoglycemic drugs such as liraglutide can interfere with this deleterious mechanism imparting reno-protection. Herein, we aim to investigate the reno-protective effect of GLP1 receptor agonist (GLP1-RA), via its effect on TRPC6 and NADPH oxidases. To achieve our aim, control or STZ-induced T1DM Sprague–Dawley rats were used. Rats were treated with liraglutide, metformin, or their combination. Functional, histological, and molecular parameters of the kidneys were assessed. Our results show that treatment with liraglutide, metformin or their combination ameliorates DKD by rectifying renal function tests and protecting against fibrosis paralleled by restored mRNA levels of nephrin, DUOX1 and 2, and reduced ROS production. Treatment with liraglutide reduces TRPC6 expression, while metformin treatment shows no effect. Furthermore, TRPC6 was found to be directly interacting with nephrin, and indirectly interacting with DUOX1, DUOX2 and GLP1-R. Our findings suggest that treatment with liraglutide may prevent the progression of diabetic nephropathy by modulating the crosstalk between TRPC6 and NADPH oxidases.

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Auxotrophy-Based Detection of Hyperornithinemia in Mouse Blood and Urine

Cited by 2 publications

References 35 publications

Translation-dependent bioassay for amino acid quantification using auxotrophic microbes as biocatalysts of protein synthesis

Translation-dependent bioassay for amino acid quantification using auxotrophic microbes as biocatalysts of protein synthesis

Reno-Protective Effect of GLP-1 Receptor Agonists in Type1 Diabetes: Dual Action on TRPC6 and NADPH Oxidases

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