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1.
Sci Rep ; 14(1): 14988, 2024 07 01.
Article in English | MEDLINE | ID: mdl-38951551

ABSTRACT

Breeding high yielding groundnut cultivars with 2-3 weeks of fresh seed dormancy, particularly in Spanish-type cultivars, enhances the sustainability of agriculture in groundnuts. In this context, we conducted a comprehensive phenotypic and genotypic evaluation of advanced breeding lines developed in the genetic background of Spanish types. By employing multi-phenotyping and marker data, we identified PBS 15044, 16004, 16013, 16015, 16016, 16017, 16020, 16021, 16026, 16031, 16035, 16037, 16038, 16039, 16041, and 16042 with 2-3 weeks dormancy (> 90%).The various parametric and non-parametric estimates identified the stable fresh dormant genotypes with one or more superior economic trait. PBS 16021, 15044, 16038, and 16039 identified with high hundred pod weight (HPW) were also reported having high intensity of dormancy (> 90% for up to 3 weeks); PBS 15044, 16016, PBS 16038 and PBS 16039 with high hundred kernel weight (HKW) also reported with up to 3 weeks fresh seed dormancy; and PBS 16013, 16031, and 16038 with up to 3 weeks fresh seed dormancy had high shelling percentage (SP). They can be used to develop lines with the desired level of dormancy, and high yields, by designing appropriate breeding strategies.


Subject(s)
Genotype , Phenotype , Plant Breeding , Plant Dormancy , Seeds , Plant Dormancy/genetics , Plant Breeding/methods , Seeds/genetics , Seeds/growth & development , Spain , Arachis/genetics , Crosses, Genetic
2.
Sci Rep ; 14(1): 2943, 2024 02 05.
Article in English | MEDLINE | ID: mdl-38316821

ABSTRACT

The global market has a high demand for premium edible grade groundnut, particularly for table use. India, in particular, exhibits significant potential for exporting confectionary grade large seeded groundnut. The environment plays a significant impact in influencing the expression of seed traits, which subsequently affects the confectionary quality of groundnut genotypes. The states of Gujarat and Rajasthan in India are prominent producers of high-quality groundnuts specifically used for confectionary purposes. The current study was conducted with 43 confectionery groundnut genotypes at Junagadh, Gujarat, and Bikaner, Rajasthan, with the goals of understanding genotype-by-environment interaction (GEI) effects and identifying stable, high yielding confectionery quality groundnut genotypes using AMMI and GGE biplot models. Pod yield per plant (PYP), number of pods per plant (NPP), hundred kernel weight (HKW), and shelling percent (SP) were estimated. The interplay between the environment and genotype has had a notable impact on the manifestation of confectionary grade characteristics in peanuts. The results from the Interaction Principal Component Analysis (IPCA) indicate that HKW contributed 76.68% and 18.95% towards the Global Environmental Index (GEI) through IPCA1 and IPCA2, respectively. Similarly, NPP contributed 87.52% and 8.65%, PYP contributed 95.87% and 2.1%, and SP contributed 77.4% and 16.22% towards GEI through IPCA1 and IPCA2, respectively. Based on the ranking of genotypes, the ideal genotypes were PBS 29079B for HKW, PBS 29230 for NPP. The genotypes PBS 29233 and PBS 29230 exhibited superior performance and stability in terms of pod yield, hundred kernel weight, number of pods per plant, and shelling percentage across various sites. These breeding lines have the potential to be developed for the purpose of producing confectionary grade groundnut with larger seeds, in order to fulfil the growing demand for export.


Subject(s)
Ammi , Gene-Environment Interaction , Plant Breeding/methods , India , Genotype
3.
Sci Rep ; 13(1): 12705, 2023 08 05.
Article in English | MEDLINE | ID: mdl-37543669

ABSTRACT

Water/drought stress experiments are frequently conducted under imposed stress or rainout shelters, while natural drought hot-spot investigations are rare. The "drought hot spot" in Anantapur, Andhra Pradesh, India, is appropriate for drought stress evaluation due to its hot, arid environment, limited rainfall, with over 50% rainfall variability. According to reports, 30 out of 200 groundnut cultivars in India are supposed to possess drought-tolerant characteristics. However, these cultivars are yet to be evaluated in areas that are prone to drought. This study tested these drought-tolerant genotypes in naturally drought-prone areas of Anantapur under rainfed conditions from Kharif 2017 to 2019. Pod yield and rainfall-use-efficiency (RUE) were measured for these genotypes. Genotype and genotype*environment interactions affected pod yield and RUE (GEI). The AMMI model exhibits significant season-to-season variability within the same area with environmental vectors > 90° angles. GGE biplot suggested the 2018 wet season for drought-resistant cultivar identification. Kadiri5 and GPBD5 were the most drought-tolerant cultivars for cultivation in Anantapur and adjacent regions. These types could also be used to generate drought-tolerant groundnut variants for drought-prone regions.


Subject(s)
Droughts , Genotype , Seasons , Base Sequence , India
4.
Mol Biol Rep ; 50(7): 6201-6216, 2023 Jul.
Article in English | MEDLINE | ID: mdl-37294468

ABSTRACT

BACKGROUND: Climate change has had a tremendous impact on the environment in general as well as agricultural crops grown in these situations as time passed. Agricultural production of crops is less suited and of lower quality due to disturbances in plant metabolism brought on by sensitivity to environmental stresses, which are brought on by climate change. Abiotic stressors that are specific to climate change, including as drought, extremes in temperature, increasing CO2, waterlogging from heavy rain, metal toxicity, and pH changes, are known to negatively affect an array of species. Plants adapt to these challenges by undergoing genome-wide epigenetic changes, which are frequently accompanied by differences in transcriptional gene expression. The sum of a cell's biochemical modifications to its nuclear DNA, post-translational modifications to histones, and variations in the synthesis of non-coding RNAs is called an epigenome. These modifications frequently lead to variations in gene expression that occur without any alteration in the underlying base sequence. EPIGENETIC MECHANISMS AND MARKS: The methylation of homologous loci by three different modifications-genomic (DNA methylation), chromatin (histone modifications), and RNA-directed DNA methylation (RdDM)-could be regarded as epigenetic mechanisms that control the regulation of differential gene expression. Stresses from the environment cause chromatin remodelling, which enables plant cells to adjust their expression patterns temporarily or permanently. EPIGENOMICS' CONSEQUENCES FOR GENOME STABILITY AND GENE EXPRESSION: DNA methylation affects gene expression in response to abiotic stressors by blocking or suppressing transcription. Environmental stimuli cause changes in DNA methylation levels, either upward in the case of hypermethylation or downward in the case of hypomethylation. The type of stress response that occurs as a result also affects the degree of DNA methylation alterations. Stress is also influenced by DRM2 and CMT3 methylating CNN, CNG, and CG. Both plant development and stress reactions depend on histone changes. Gene up-regulation is associated with histone tail phosphorylation, ubiquitination, and acetylation, while gene down-regulation is associated with de-acetylation and biotinylation. Plants undergo a variety of dynamic changes to histone tails in response to abiotic stressors. The relevance of these transcripts against stress is highlighted by the accumulation of numerous additional antisense transcripts, a source of siRNAs, caused by abiotic stresses. The study highlights the finding that plants can be protected from a range of abiotic stresses by epigenetic mechanisms such DNA methylation, histone modification, and RNA-directed DNA methylation. TRANSGENERATIONAL INHERITANCE AND SOURCES OF EPIGENETIC VARIATION: Stress results in the formation of epialleles, which are either transient or enduring epigenetic stress memory in plants. After the stress is gone, the stable memory is kept for the duration of the plant's remaining developmental cycles or passed on to the next generations, leading to plant evolution and adaptability. The bulk of epigenetic changes brought on by stress are temporary and return to normal after the stress has passed. Some of the modifications, however, might be long-lasting and transmitted across mitotic or even meiotic cell divisions. Epialleles often have genetic or non-genetic causes. Epialleles can arise spontaneously due to improper methylation state maintenance, short RNA off-target effects, or other non-genetic causes. Developmental or environmental variables that influence the stability of epigenetic states or direct chromatin modifications may also be non-genetic drivers of epigenetic variation. Transposon insertions that change local chromatin and structural rearrangements, such copy number changes that are genetically related or unrelated, are two genetic sources of epialleles. EPIGENOMICS IN CROP IMPROVEMENT: To include epigenetics into crop breeding, it is necessary to create epigenetic variation as well as to identify and evaluate epialleles. Epigenome editing or epi-genomic selection may be required for epiallele creation and identification. In order to combat the challenges given by changing environments, these epigenetic mechanisms have generated novel epialleles that can be exploited to develop new crop types that are more climate-resilient. Numerous techniques can be used to alter the epigenome generally or at specific target loci in order to induce the epigenetic alterations necessary for crop development. Technologies like CRISPR/Cas9 and dCas, which have recently advanced, have opened up new avenues for the study of epigenetics. Epialleles could be employed in epigenomics-assisted breeding in addition to sequence-based markers for crop breeding. CONCLUSIONS AND FUTURE PROSPECTUS: A few of the exciting questions that still need to be resolved in the area of heritable epigenetic variation include a better understanding of the epigenetic foundation of characteristics, the stability and heritability of epialleles, and the sources of epigenetic variation in crops. Investigating long intergenic non-coding RNAs (lincRNAs) as an epigenetic process might open up a new path to understanding crop plant's ability to withstand abiotic stress. For many of these technologies and approaches to be more applicable and deployable at a lower cost, technological breakthroughs will also be necessary. Breeders will probably need to pay closer attention to crop epialleles and how they can affect future responses to climate changes. The development of epialleles suitable for particular environmental circumstances may be made possible by creating targeted epigenetic changes in pertinent genes and by comprehending the molecular underpinnings of trans generational epigenetic inheritance. More research on a wider variety of plant species is required in order to fully comprehend the mechanisms that produce and stabilise epigenetic variation in crops. In addition to a collaborative and multidisciplinary effort by researchers in many fields of plant science, this will require a greater integration of the epigenomic data gathered in many crops. Before it may be applied generally, more study is required.


Subject(s)
Epigenomics , Histones , Histones/genetics , Histones/metabolism , Climate Change , Plant Breeding , DNA Methylation/genetics , Epigenesis, Genetic , Chromatin , Crops, Agricultural/genetics , RNA/metabolism
5.
Physiol Mol Biol Plants ; 29(5): 725-737, 2023 May.
Article in English | MEDLINE | ID: mdl-37363420

ABSTRACT

Peanut is mostly grown in calcareous soils with high pH which are deficient in available iron (Fe2+) for plant uptake causing iron deficiency chlorosis (IDC). The most pertinent solution is to identify efficient genotypes showing tolerance to limited Fe availability in the soil. A field screening of 40 advanced breeding lines of peanut using NRCG 7472 and ICGV 86031 as IDC susceptible and tolerant checks, respectively, was envisaged for four years. PBS 22040 and 29,192 exhibited maximum tolerance while PBS 12215 and 12,185 were most susceptible. PBS 22040 accumulated maximum seed resveratrol (5.8 ± 0.08 ppm), ferulic acid (378.6 ± 0.31 ppm) and Fe (45.59 ± 0.41 ppm) content. Enhanced chlorophyll retention (8.72-9.50 µg ml-1), carotenoid accumulation (1.96-2.08 µg ml-1), and antioxidant enzyme activity (APX: 35.9-103.9%; POX: 51- 145%) reduced the MDA accumulation (5.61-9.11 µM cm-1) in tolerant lines. The overexpression of Fe transporters IRT1, ZIP5, YSL3 was recorded to the tune of 2.3-9.54; 1.45-3.7; 2.20-2.32- folds respectively in PBS 22040 and 29,192, over NRCG 7472. PBS 22040 recorded the maximum pod yield (282 ± 4.6 g/row), hundred kernel weight (55 ± 0.7 g) and number of pods per three plants (54 ± 1.7). The study thus reports new insights into the roles of resveratrol, ferulic acid and differential antioxidant enzyme activities in imparting IDC tolerance. PBS 22040, being the best performing line, can be the potent source of IDC tolerance for introgression in high yielding but susceptible genotypes under similar edaphic conditions. Supplementary Information: The online version contains supplementary material available at 10.1007/s12298-023-01321-9.

6.
PLoS One ; 18(4): e0282438, 2023.
Article in English | MEDLINE | ID: mdl-37098075

ABSTRACT

India imports the most edible oils because domestic demand exceeds production. Horizontally expanding groundnut production in non-traditional areas especially in the potato-paddy rice-fallow system is possible for increasing production and it requires trait-specific cultivars. Only 1% of oilseeds are grown in non-traditional regions. Nine interspecific groundnut derivatives were tested in potato-fallow system at Deesa, Gujarat, and Mohanpura, West Bengal, and non-potato fallow areas in Junagadh during Kharif 2020 to examine their performance and adaptability. Genotype-by-environment (G×E) interaction significantly affected pod yield and its components in the combined ANOVA. "Mean vs. stability" showed that the interspecific derivative NRCGCS 446 and variety TAG 24 were the most stable and valuable genotypes. GG 7 yielded more pods in Junagadh, whereas NRCGCS 254 yielded more in Mohanpur. Low heritability estimates and strong G×E interaction for flowering days showed complicated inheritance and environmental effects. The shelling percentage was significantly correlated with days to 50% blooming, days to maturity, SCMR, HPW, and KLWR, demonstrating negative connections between maturity, component characteristics, and seed size realisation.


Subject(s)
Fabaceae , Solanum tuberosum , Arachis/genetics , Solanum tuberosum/genetics , Genotype , Phenotype
7.
Sensors (Basel) ; 22(11)2022 Jun 01.
Article in English | MEDLINE | ID: mdl-35684847

ABSTRACT

Several illnesses that are chronic and acute are becoming more relevant as the world's aging population expands, and the medical sector is transforming rapidly, as a consequence of which the need for "point-of-care" (POC), identification/detection, and real time management of health issues that have been required for a long time are increasing. Biomarkers are biological markers that help to detect status of health or disease. Biosensors' applications are for screening for early detection, chronic disease treatment, health management, and well-being surveillance. Smart devices that allow continual monitoring of vital biomarkers for physiological health monitoring, medical diagnosis, and assessment are becoming increasingly widespread in a variety of applications, ranging from biomedical to healthcare systems of surveillance and monitoring. The term "smart" is used due to the ability of these devices to extract data with intelligence and in real time. Wearable, implantable, ingestible, and portable devices can all be considered smart devices; this is due to their ability of smart interpretation of data, through their smart sensors or biosensors and indicators. Wearable and portable devices have progressed more and more in the shape of various accessories, integrated clothes, and body attachments and inserts. Moreover, implantable and ingestible devices allow for the medical diagnosis and treatment of patients using tiny sensors and biomedical gadgets or devices have become available, thus increasing the quality and efficacy of medical treatments by a significant margin. This article summarizes the state of the art in portable, wearable, ingestible, and implantable devices for health status monitoring and disease management and their possible applications. It also identifies some new technologies that have the potential to contribute to the development of personalized care. Further, these devices are non-invasive in nature, providing information with accuracy and in given time, thus making these devices important for the future use of humanity.


Subject(s)
Wearable Electronic Devices , Aged , Biomarkers , Disease Management , Health Status , Humans , Monitoring, Physiologic
8.
Mol Biol Rep ; 49(6): 5483-5494, 2022 Jun.
Article in English | MEDLINE | ID: mdl-35478296

ABSTRACT

The fundamental concepts of the genetics, race classification and epidemiology of the Wheat spike blast causing fungus Magnaporthe oryzae pathotype Triticum (MoT) are still evolving despite of its discovery in 1985 in Brazil for the first time. The fungus seems to defy the research progress that is being made globally by continuously evolving into pathotypes which have already overcome the much celebrated 2NS resistance in wheat lines as well as few of the initially effective fungicides. The compartmentalized i.e. two speed genome of the MoT, conferring the fungus an evolutionary advantage, has emerged as a challenge for the wheat spike blast researchers complicating its already difficult management. The airborne fungus with a range of alternative hosts is finding new geographical niches situated on different continents and is a matter of great apprehension among the nations whose food security is primarily dependent on wheat. The wheat blast outbreak in Bangladesh during 2016 was attributed to an isolate from Latin America escaping through a seed import consignment while the latest Zambian outbreak is still to be studied in detail regarding its origin and entry. The challenges in dealing wheat spike blast are not only on the level of genetics and epidemiology alone but also on the levels of policy making regarding international seed movement and research collaborations. The present review deals with these issues mainly concerning the effective management and controlling the international spread of this deadly disease of wheat, with a particular reference to India. We describe the origin, taxonomy, epidemiology and symptomology of MoT and briefly highlight its impact and management practices from different countries. We also discuss the advances in genomics and genome editing technologies that can be used to develop elite wheat genotypes resistant against different stains of wheat spike blast.


Subject(s)
Magnaporthe , Triticum , Ascomycota , Genetic Engineering , Magnaporthe/genetics , Plant Diseases/genetics , Plant Diseases/microbiology , Triticum/genetics , Triticum/microbiology
9.
Int J Biol Macromol ; 164: 3589-3602, 2020 Dec 01.
Article in English | MEDLINE | ID: mdl-32882275

ABSTRACT

Salt stress is a major constrain to the productivity of nutritionally rich pigeonpea, an important legume of SE Asia and other parts of the world. The present study provides a comprehensive insight on integrated proteomic and transcriptomic analysis of root and shoot tissues of contrasting pigeonpea varieties (ICP1071- salt-sensitive; ICP7- salt-tolerant) to unravel salt stress induced pathways. Proteome analysis revealed 82 differentially expressed proteins (DEPs) with ≥±1.5 fold expression on 2-Dimensional (2D) gel. Of these, 25 DEPs identified through MALDI-TOF/TOF were classified using Uniprot software into functional categories. Pathways analyses using KAAS server showed the highest abundance of functional genes regulating metabolisms of carbohydrate followed by protein folding/degradation, amino acids and lipids. Expression studies on six genes (triosephosphate isomerase, oxygen evolving enhancer protein 1, phosphoribulokinase, cysteine synthase, oxygen evolving enhancer protein 2 and early nodulin like protein 2) with ≥±3 fold change were performed, and five of these showed consistency in transcript and protein expressions. Transcript analysis of root and shoot led to positive identification of 25 differentially expressed salt-responsive genes, with seven genes having ≥±5 fold change have diverse biological functions. Our combinatorial analysis suggests important role of these genes/proteins in providing salt tolerance in pigeonpea.


Subject(s)
Cajanus/genetics , Plant Proteins/genetics , Salt Stress/genetics , Transcriptome/genetics , Gene Expression Regulation, Plant/genetics , Plant Proteins/classification , Proteome/genetics , Proteomics/methods , Salt Stress/physiology , Salt Tolerance
10.
Indian J Ophthalmol ; 68(8): 1569-1572, 2020 08.
Article in English | MEDLINE | ID: mdl-32709778

ABSTRACT

Purpose: To evaluate the astigmatism correcting effect of penetrating arcuate keratotomy (AK) done during femtosecond laser-assisted cataract surgery (FLACS). Methods: In this nonrandomized prospective study, 80 eyes of 70 patients were studied. The study included patients who underwent combined FLACS and AK, with corneal astigmatism ranging from 0.4 to 1.5 diopters (D). Femtosecond laser-assisted penetrating arcuate keratotomies were created at 8 mm optical zone at 80% depth and were centered at the limbus. Keratometric astigmatism was measured prior to and 3 months post-surgery. Vector analysis was performed using Power vector analysis method. Results: The mean preoperative keratometric astigmatism without accounting for axis was 0.85 ± 0.27 D, which reduced significantly to 0.47 ± 0.27 D at 3-month follow-up. The mean astigmatism correction attained without accounting for axis was 0.38 ± 0.32 D. The vector corrected mean preoperative astigmatism was 0.85 ± 0.27 D which reduced significantly to 0.50 ± 0.31 D postoperatively (P < 0.001, 95% CI). Vector corrected mean astigmatism correction attained was 0.35 ± 0.38 D. There were no significant intraoperative or postoperative complications. Conclusion: Preexisting astigmatism can be tackled effectively with penetrating AK during FLACS although under correction is observed with present nomograms. Further refinements may achieve better correction.


Subject(s)
Astigmatism , Cataract , Astigmatism/surgery , Cataract/complications , Cataract/diagnosis , Cornea/surgery , Corneal Topography , Humans , Keratoplasty, Penetrating , Lasers , Prospective Studies , Refraction, Ocular , Retrospective Studies , Visual Acuity
11.
PLoS One ; 14(12): e0226252, 2019.
Article in English | MEDLINE | ID: mdl-31830093

ABSTRACT

Peanut (Arachis hypogaea L.) is an important nutrient-rich food legume and valued for its good quality cooking oil. The fatty acid content is the major determinant of the quality of the edible oil. The oils containing higher monounsaturated fatty acid are preferred for improved shelf life and potential health benefits. Therefore, a high oleic/linoleic fatty acid ratio is the target trait in an advanced breeding program. The two mutant alleles, ahFAD2A (on linkage group a09) and ahFAD2B (on linkage group b09) control fatty acid composition for higher oleic/linoleic ratio in peanut. In the present study, marker-assisted backcrossing was employed for the introgression of two FAD2 mutant alleles from SunOleic95R into the chromosome of ICGV06100, a high oil content peanut breeding line. In the marker-assisted backcrossing-introgression lines, a 97% increase in oleic acid, and a 92% reduction in linoleic acid content was observed in comparison to the recurrent parent. Besides, the oleic/linoleic ratio was increased to 25 with respect to the recurrent parent, which was only 1.2. The most significant outcome was the stable expression of oil-content, oleic acid, linoleic acid, and palmitic acid in the marker-assisted backcrossing-introgression lines over the locations. No significant difference was observed between high oleic and normal oleic in peanuts for seedling traits except germination percentage. In addition, marker-assisted backcrossing-introgression lines exhibited higher yield and resistance to foliar fungal diseases, i.e., late leaf spot and rust.


Subject(s)
Arachis/metabolism , Fatty Acid Desaturases/metabolism , Germination , Mutation , Oleic Acid/metabolism , Seedlings/metabolism , Seeds/metabolism , Alleles , Arachis/genetics , Arachis/growth & development , Biomarkers/analysis , Fatty Acid Desaturases/genetics , Peanut Oil/analysis , Plant Breeding , Plant Proteins/genetics , Plant Proteins/metabolism , Seedlings/genetics , Seedlings/growth & development , Seeds/genetics , Seeds/growth & development
12.
Indian J Biochem Biophys ; 43(2): 98-104, 2006 Apr.
Article in English | MEDLINE | ID: mdl-16955758

ABSTRACT

3alpha-Hydroxysteroid dehydrogenase (3alpha-HSD) from Pseudomonas testosteronei and diaphorase (lipoyl dehydrogenase) from Clostridium spp were immobilized individually onto alkylamine glass beads through glutaraldehyde coupling. A cost-effective enzymic colorimetric method for determination of bile acid in the serum and bile was developed employing mixture of the immobilized enzymes. The method was based upon measurement of NADH generated from NAD+ during oxidation of bile acid by immobilized 3alpha-HSD with a color reagent consisting of nitrobluetetrazolium (NBT) chloride salt and immobilized diaphorase in 0.065 M sodium phosphate buffer (pH 7.0). The minimum detection limit of the method was 4.8 pmol/L in the serum and 19.5 micromol/L in bile. The per cent recovery of added bile acid in the serum and bile was 89.1 and 95.0, respectively. Within and between batch coefficients of variation (CV) for bile acid determination were <1.0% and <0.2% in the serum and <0.2% and <0.6% in bile, respectively. A good correlation for bile acid in the serum (r1= 0.95) and in bile (r2 = 0.93) was obtained by a standard chemical method (a commonly used method in India) and the present method. The mixture of immobilized 3alpha-HSD and diaphorase lost 30% of its initial activity after 4 months of regular use. The cost of bile acid determination for 100 the serum and bile samples by the present method was found to be lower than by a commercially available method (Sigma kit 450-A).


Subject(s)
3-alpha-Hydroxysteroid Dehydrogenase (B-Specific) , Bile Acids and Salts/analysis , Bile/metabolism , Dihydrolipoamide Dehydrogenase , Enzymes, Immobilized , Glass , Adolescent , Adult , Bile Acids and Salts/blood , Child , Humans , Middle Aged
13.
Anal Biochem ; 332(1): 32-7, 2004 Sep 01.
Article in English | MEDLINE | ID: mdl-15301946

ABSTRACT

3Alpha-hydroxysteroid dehydrogenase (3-HSD) from Pseudomonas testosteroni and diaphorase (lipoyl dehydrogenase) from Clostridium spp. have been immobilized individually onto arylamine glass beads through diazotization. A cost-effective enzymic colorimetric method for determination of bile acid in serum and bile employing a mixture of these immobilized enzymes was developed. The method is based on measurement of reduced nicotinamide adenine dinucleotide generated from bile acid in serum/bile by immobilized 3alpha-HSD with a color reagent consisting of nitro blue tetrazolium chloride salt, oxidized nicotinamide adenine dinucleotide, and immobilized lipoyl dehydrogenase in 0.065 M sodium phosphate buffer, pH 7.0. Analytical recovery of added bile acid (50 and 200 micromol/L) was 95.57 and 85.46% in serum and 97.6 and 91.6% in bile, respectively. Within- and between-batch coefficients of variation (CV) for bile acid determination were <1.2 and <0.2% in serum and >0.1 and <0.1% in bile, respectively. Good correlations for bile acid in serum (r1=0.92) and in bile (r2=0.97) were obtained by use of a standard chemical method and the present method. The mixture of immobilized 3alpha-HSD dehydrogenase and lipoyl dehydrogenase lost 50% of its initial activity after 6 months of regular use. The cost of bile acid determination in 100 serum and bile samples by the present method has been compared with that of the Sigma kit method.


Subject(s)
3-alpha-Hydroxysteroid Dehydrogenase (B-Specific)/metabolism , Bile Acids and Salts/analysis , Bile/metabolism , Dihydrolipoamide Dehydrogenase/metabolism , Glass , Bile Acids and Salts/blood , Bile Acids and Salts/metabolism , Humans , Hydrogen-Ion Concentration , Taurodeoxycholic Acid/analysis
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