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Calcium orotate is one of the highly prescribed calcium salts. The orotic acid content is crucial for exhibiting its high bioavailability. There are no methods available to estimate orotic acid content in calcium orotate. The primary objective of this study was to develop and validate a robust method for the estimation of orotic acid from the bulk calcium orotate. A stability indicating RP-HPLC method was developed using an isocratic condition of mobile phase comprising 20 mM of monobasic potassium phosphate in water (pH 2.5) with phosphoric acid at a flow rate of 1 mL min-1 over C-18 (ODS, 250 × 4.6 mm) column at ambient temperature with UV detection at 214 nm. It was validated with respect to accuracy, specificity, precision, linearity, range, robustness and system suitability. Real sample analysis was also performed to test the applicability for regular analysis of calcium orotate bulk samples to assure its quality. [ABSTRACT FROM AUTHOR]

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The method of synthesis of unmodified and organo‐modified silica hydrogels and their composites with orotic acid as a model drug was developed. The hydrogels had a pH of 6.5–7.8. The particulate nature and highly porous structures of the hydrogel materials were revealed using scanning electron and optical microscopy methods. The content of aqueous phase in the hydrogels was 99% or more. In order to evaluate the possibility of their application as a basis for development of novel soft drug formulations and cosmetic compositions, rheological properties of the hydrogels and in vitro release kinetics of the drug were studied. The effects of synthesis conditions (increasing concentration of catalyst of silica sol formation, drug loading) and the silica matrix modification with various organic groups on the indicated properties were investigated. It was found that all synthesized hydrogels exhibited pseudoplasticity, thixotropy and controlled release of the drug, which are important for their potential application. However, in general, the indicated effects led to worsening the properties of the hydrogel materials in comparison with the unmodified silica hydrogels. [ABSTRACT FROM AUTHOR]

The drug–drug multicomponent crystals (cocrystal/salt) in pharmaceutical products may boost the major challenges of fixed-dose combinations (FDCs) in coming years. The challenges associated with FDCs such as drug incompatibility, solubility differences, and stability issues can be overcome by salt/cocrystallization. Herein, the authors report eight drug–drug/drug–nutraceutical salts of the anti-emetic drug domperidone (DOM) with various drug coformers such as nicotinic acid (NCA), DL -mandelic acid (MAA), salicylic acid (SLA), orotic acid (OTA), and aspirin (ASP). The single-crystal X-ray diffraction study revealed multiple solid forms in DOM–NCA and DOM–MAA salts which differ by the presence of water. The salts, namely, DOM–MAA, DOM–OTA and DOM–ASP crystallized as anhydrous salts. The salts DOM–NCA I, DOM–SLA, and DOM–MAA I were found to be similar in crystal packing based on CrystalCMP software. The prepared salts showed lower melting temperatures as compared to DOM and coformers except for DOM–OTA. The solubility of the prepared salts in three different buffer media revealed enhancement in the solubility of DOM in pH 6.8 and 1.2 buffers. The solubility of DOM in the salts was drastically reduced in the acetate buffer (pH 4.6) as compared to the parent DOM. The dissolution study in pH 6.8 buffer demonstrated the faster drug release in the salts. The prepared salts exhibited stability under accelerated humidity conditions (40 °C, 70–75% RH) for 30 days, demonstrating the possibility of further developing these drug–drug/drug–nutraceutical salts. [ABSTRACT FROM AUTHOR]

Polylactic acid (PLA) is a versatile and sustainable polymer used in various applications. This research explores the use of orotic acid (OA) and ethylene bis-stearamide (EBS) as nucleating agents to enhance the quiescent crystallization of PLA within the temperature range of 80 °C to 140 °C. Different blends were produced via melt processing before analyzing via DSC, XRD, and SEM. Our results show that both nucleating agents significantly accelerated the crystallization process and reduced the incubation time and the crystallization half-time. The most promising results were obtained with 1% EBS at 110 °C, achieving the fastest crystallization. The XRD analysis showed that at 80 °C, the disordered α'phase predominated, while more stable α phases formed at 110 °C and 140 °C. Combining the 1% nucleating agent and 110 °C promotes densely packed crystalline lamellae. The nucleated PLA exhibited a well-organized spherulitic morphology in agreement with the Avrami modeling of DSC data. Higher nucleating agent concentrations yielded smaller, more evenly distributed crystalline domains. Utilizing OA or EBS in PLA processing could offer enhanced properties, improved processability, and cost-efficiency, making PLA more competitive in various applications. [ABSTRACT FROM AUTHOR]

With orotic acid (H3OA) as building ligand, one one‐dimensional (1D) Zn(II) coordination polymer, [Zn2(HOA)(μ2‐OH)(μ3‐OH)(H2O)]n (1), and one tetra‐nuclear Cd(II) cluster, [Cd4(HOA)4(μ2‐OH2)2(H2O)10] (2), have been synthesized at 90 °C and room temperature, respectively. Single‐crystal X‐ray diffraction analyses reveal that complex 1 has an 1D stair‐stepping structure consisting of co‐edge bi‐rhomboid [Zn6] structural motif and μ3‐HOA2− linker, and 2 owns a quasi‐linear tetra‐nuclear [Cd4] cluster structure fastened by μ2‐OH2 and HOA2− fixators. The solid‐state photophysical properties were investigated and 1 and 2 display a distinct emission centered at 374 and 393 nm, respectively. [ABSTRACT FROM AUTHOR]

Orotic acid (OA) is an intermediate metabolite of pyrimidine nucleotide biosynthesis and represents a minor diet constituent. The measurement of urinary orotic acid is useful in confirming the diagnosis of hereditary metabolic diseases. Moreover, it could be of interest to know how the physiological concentration of this metabolite changes in relation to different conditions of clinical normality. The purpose of this study was to determine the orotic acid concentration in the urine of healthy patients, to observe normal oroticuria and to evaluate if the expression of pyrimidine intermediate biosynthesis differs between healthy males and females. The orotic acid concentration in urine was performed via the ICH M10-validated analytical method. Unexpectedly, females showed a greater oroticuria than males in pediatric age (0–10); conversely, we did not find significant differences until 70 years of age. The LC-MS/MS method was suitable for use in the differential diagnosis of hereditary metabolic disease and metabolic monitoring of anticancer drug-induced toxicity. The analytical protocol was found to be rapid and ideal, and was used in the routine analysis of a clinical chemistry laboratory. The biochemical aspects related to the expression of pyrimidine biosynthesis should be further investigated in light of the obtained results. [ABSTRACT FROM AUTHOR]

Urea cycle deficiency is an inherited disorder of the ammonia detoxification pathway and arginine synthesis. They are caused by deficiencies in urea cycle enzymes. The urea cycle is the main metabolic pathway for purifying ammonia, a toxic waste product of the body’s protein catabolism. The clinical signs are therefore partly linked to ammonia intoxication. Diagnosis can be made at any age and requires the use of a range of investigative techniques, most of which are specific. The aim of our work is to review the pathophysiology and clinical features of urea cycle deficiencies and to present the various biological diagnostic methods currently available and used to investigate these deficiencies. [ABSTRACT FROM AUTHOR]

Inclusion body myositis (IBM) is an acquired inflammatory myopathy affecting proximal and distal muscles that leads to weakness in patients over 50. It is diagnosed based on clinical and histological findings in muscle related to inflammation, degeneration, and mitochondria. In relation to IBM, a shortage of validated disease models and a lack of biomarkers and effective treatments constitute an unmet medical need. To overcome these hurdles, we performed an omics analysis of multiple samples from IBM patients (saliva, fibroblasts, urine, plasma, and muscle) to gain insight into the pathophysiology of IBM. Degeneration was evident due to the presence of amyloid β peptide 1–42 (Aβ1–42) in the saliva of the analyzed IBM patients. The presence of metabolic disarrangements in IBM was indicated by an imbalanced organic acid profile in fibroblasts and urine. Specifically, abnormal levels of L-pyroglutamic and orotic acid were supported by the abnormal expression of related metabolites in plasma and urine (glutathione and pyrimidines) and the aberrant expression of upstream gene regulators (L2HGDH, IDH2, OPLAH, and ASL) in muscle. Combined levels of L-pyroglutamic and orotic acid displayed an outstanding biomarker signature in urine with 100% sensitivity and specificity. The confirmation of systemic metabolic disarrangements in IBM and the identification of novel biomarkers reported herein unveil novel insights that require validation in larger cohorts. [ABSTRACT FROM AUTHOR]

Pyrimidine itself or in different fused forms is an important chemical motif for every step of cell life. Alongside, it exists as a key pharmacophoric part in thiamine, alloxan, orotic acid, and other isoforms of nucleotide bases. It has emerged as a privileged scaffold in medicinal chemistry due to its various possible structural and pharmacological attributes. In a number of clinically used chemical agents, pyrimidine or its fused/clubbed form with other moieties act as a key pharmacophore. Structure activity relationship (SAR) studies of the explored derivatives demonstrated that slight modifications with various substitutions on the pyrimidine rings can deviate biological activity to a large extent. 2‐Pyrimidone and 2,4‐pyrimidione derivatives are largely explored by a number of research groups for various pharmacological targets and due to their similarity to nucleotide bases, it has a great impact on the biological profile of these molecules. The current review focuses on describing the recent advancements (2015 onward) in the development of pyrimidine‐based biologically active molecules along with drug candidates under different clinical trials. The aim of this review is to provide structural features associated with pyrimidine along with their SAR studies, which are believed to be valuable for the scientists working on the development of pyrimidine‐based small molecules in the aforementioned disease conditions. We believe that the results of this research will help to provide a structural idea that may be used in the design and development of new pyrimidine‐based small molecules that target a variety of receptors or enzymes. [ABSTRACT FROM AUTHOR]

Nilutamide (Nil), commercialized under the trade names Nilandron and Anandron, is a nonsteroidal antiandrogen drug used in prostate cancer treatment. The attempts to cocrystallize nilutamide with adenine (Adn) and orotic acid monohydrate (OMH) resulted in the formation of four novel solid forms of the drug, namely the [Nil + Adn] (1 : 1) cocrystal, two new polymorphs of Nil (forms IV and V), and the Nil hydrate ([Nil·H2O] (2 : 0.57)). The crystal structures of the solid forms were elucidated by single-crystal X-ray diffraction. New nilutamide polymorphic forms IV and V were found to contain multiple symmetry-independent molecules in the asymmetric unit whose Z′ = 3 (form IV) and Z′ = 2 (form V), while the three previously reported forms (form I, form II, form III) had Z′ = 1. Despite several attempts, the bulk phase-pure materials of form IV, form V, and [Nil·H2O] (2 : 0.57) could not be attained due to the concomitant crystallization of the solid forms and the poor reproducibility of the crystallization trials. Hence, only limited characterization studies were performed on these phases. In this work, structural elucidation, Hirshfeld, and conformational analysis were explored to understand the similarities and dissimilarities in the crystal packing environment for all the crystal structures. Thermal analysis reveals the phase transformation of [Nil·H2O] (2 : 0.57) to form II. Crystal structure analysis paved the way to understand the prominence of various intermolecular interactions along with the amide dimer and catemer formation. For the [Nil + Adn] (1 : 1) cocrystal, the thermodynamic solubility was evaluated using eutectic concentrations of the components and found to be higher than that of the parent Nil. The effect of hydroxypropyl methylcellulose on the dissolution behavior of the cocrystal was explored. [ABSTRACT FROM AUTHOR]

Chemically induced steatosis is characterized by lipid accumulation associated with mitochondrial dysfunction, oxidative stress and nucleus distortion. New approach methods integrating in vitro and in silico models are needed to identify chemicals that may induce these cellular events as potential risk factors for steatosis and associated hepatotoxicity. In this study we used high-content imaging for the simultaneous quantification of four cellular markers as sentinels for hepatotoxicity and steatosis in chemically exposed human liver cells in vitro. Furthermore, we evaluated the results with a computational model for the extrapolation of human oral equivalent doses (OED). First, we tested 16 reference chemicals with known capacities to induce cellular alterations in nuclear morphology, lipid accumulation, mitochondrial membrane potential and oxidative stress. Then, using physiologically based pharmacokinetic modeling and reverse dosimetry, OEDs were extrapolated from data of any stimulated individual sentinel response. The extrapolated OEDs were confirmed to be within biologically relevant exposure ranges for the reference chemicals. Next, we tested 14 chemicals found in food, selected from thousands of putative chemicals on the basis of structure-based prediction for nuclear receptor activation. Amongst these, orotic acid had an extrapolated OED overlapping with realistic exposure ranges. Thus, we were able to characterize known steatosis-inducing chemicals as well as data-scarce food-related chemicals, amongst which we confirmed orotic acid to induce hepatotoxicity. This strategy addresses needs of next generation risk assessment and can be used as a first chemical prioritization hazard screening step in a tiered approach to identify chemical risk factors for steatosis and hepatotoxicity-associated events. [ABSTRACT FROM AUTHOR]

Dihydroorotate dehydrogenase (DHODH) is a central enzyme of the de novo pyrimidine biosynthesis pathway and is a promising drug target for the treatment of cancer and autoimmune diseases. This study presents the identification of a potent DHODH inhibitor by proteomic profiling. Cell-based screening revealed that NPD723, which is reduced to H-006 in cells, strongly induces myeloid differentiation and inhibits cell growth in HL-60 cells. H-006 also suppressed the growth of various cancer cells. Proteomic profiling of NPD723-treated cells in ChemProteoBase showed that NPD723 was clustered with DHODH inhibitors. H-006 potently inhibited human DHODH activity in vitro, whereas NPD723 was approximately 400 times less active than H-006. H-006-induced cell death was rescued by the addition of the DHODH product orotic acid. Moreover, metabolome analysis revealed that H-006 treatment promotes marked accumulation of the DHODH substrate dihydroorotic acid. These results suggest that NPD723 is reduced in cells to its active metabolite H-006, which then targets DHODH and suppresses cancer cell growth. Thus, H-006-related drugs represent a potentially powerful treatment for cancer and other diseases. [ABSTRACT FROM AUTHOR]

Functional cosmetics industries using skin microbiome screening and beneficial materials isolated from key microorganisms are receiving increasing attention. Since Epidermidibacterium keratini EPI-7T was first discovered in human skin, previous studies have confirmed that it can produce a new pyrimidine compound, 1,1′-biuracil, having anti-aging effects on human skin. Therefore, we conducted genomic analyses to judge the use value of E. keratini EPI-7T and provide up-to-date information. Whole-genome sequencing analysis of E. keratini EPI-7T was performed to generate new complete genome and annotation information. E. keratini EPI-7T genome was subjected to comparative genomic analysis with a group of closely-related strains and skin flora strains through bioinformatic analysis. Furthermore, based on annotation information, we explored metabolic pathways for valuable substances that can be used in functional cosmetics. In this study, the whole-genome sequencing (WGS) and annotation results of E. keratini EPI-7T were improved, and through comparative analysis, it was confirmed that the E. keratini EPI-7T has more metabolite-related genes than comparison strains. In addition, we annotated the vital genes for biosynthesis of 20 amino acids, orotic acid, riboflavin (B2) and chorismate. In particular, we were able to prospect that orotic acid could accumulate inside E. keratini EPI-7T under uracil-enriched conditions. Therefore, through a genomics approach, this study aims to provide genetic information for the hidden potential of E. keratini EPI-7T and the strain development and biotechnology utilization to be conducted in further studies. [ABSTRACT FROM AUTHOR]

Introduction: Hereditary orotic aciduria is an extremely rare, autosomal recessive disease caused by deficiency of uridinemonophosphate synthase. Untreated, affected individuals may develop refractory megaloblastic anemia, neurodevelopmental disabilities, and crystalluria. Newborn screening has the potential to identify and enable treatment of affected individuals before they become significantly ill. Methods: Measuring orotic acid as part of expanded newborn screening using flow injection analysis tandem mass spectrometry. Results: Since the addition of orotic acid measurement to the Israeli routine newborn screening program, 1,492,439 neonates have been screened. The screen has identified ten Muslim Arab newborns that remain asymptomatic so far, with DBS orotic acid elevated up to 10 times the upper reference limit. Urine organic acid testing confirmed the presence of orotic aciduria along with homozygous variations in the UMPS gene. Conclusion: Newborn screening measuring of orotic acid, now integrated into the routine tandem mass spectrometry panel, is capable of identifying neonates with hereditary orotic aciduria. [ABSTRACT FROM AUTHOR]

Orotate phosphoribosyltransferase (OPRT) exists as a bifunctional enzyme, uridine 5′-monophosphate synthase, in mammalian cells and plays an important role in pyrimidine biosynthesis. Measuring OPRT activity has been considered important for understanding biological events and development of molecular-targeting drugs. In this study, we demonstrate a novel fluorescence method for measuring OPRT activity in living cells. The technique utilizes 4-trifluoromethylbenzamidoxime (4-TFMBAO) as a fluorogenic reagent, which produces selective fluorescence for orotic acid. To perform the OPRT reaction, orotic acid was added to HeLa cell lysate, and a portion of the enzyme reaction mixture was heated at 80 °C for 4 min in the presence of 4-TFMBAO under basic conditions. The resulting fluorescence was measured using a spectrofluorometer, which reflects the consumption of orotic acid by the OPRT. After optimization of the reaction conditions, the OPRT activity was successfully determined in 15 min of enzyme reaction time without further procedures such as purification of OPRT or deproteination for the analysis. The activity obtained was compatible with the value measured by the radiometric method with [3H]-5-FU as the substrate. The present method provides a reliable and facile measurement of OPRT activity and could be useful for a variety of research fields targeting pyrimidine metabolism. [ABSTRACT FROM AUTHOR]

The present study investigated the effect of topical application of Epidermidibacterium Keratini (EPI-7) ferment filtrate, which is a postbiotic product of a novel actinobacteria, on skin aging, by performing a prospective randomized split-face clinical study on Asian woman participants. The investigators measured skin biophysical parameters, including skin barrier function, elasticity, and dermal density, and revealed that the application of the EPI-7 ferment filtrate-including test product resulted in significantly higher improvements in barrier function, skin elasticity, and dermal density compared to the placebo group. This study also investigated the influence of EPI-7 ferment filtrate on skin microbiome diversity to access its potential beneficial effects and safety. EPI-7 ferment filtrate increased the abundance of commensal microbes belonging to Cutibacterium, Staphylococcus, Corynebacterium, Streptococcus, Lawsonella, Clostridium, Rothia, Lactobacillus, and Prevotella. The abundance of Cutibacterium was significantly increased along with significant changes in Clostridium and Prevotella abundance. Therefore, EPI-7 postbiotics, which contain the metabolite called orotic acid, ameliorate the skin microbiota linked with the aging phenotype of the skin. This study provides preliminary evidence that postbiotic therapy may affect the signs of skin aging and microbial diversity. To confirm the positive effect of EPI-7 postbiotics and microbial interaction, additional clinical investigations and functional analyses are required. [ABSTRACT FROM AUTHOR]

Metabolomics has expanded from cellular to subcellular level to elucidate subcellular compartmentalization. By applying isolated mitochondria to metabolome analysis, the hallmark of mitochondrial metabolites has been unraveled, showing compartment-specific distribution and regulation of metabolites. This method was employed in this work to study a mitochondrial inner membrane protein Sym1, whose human ortholog MPV17 is related to mitochondria DNA depletion syndrome. Gas chromatography–mass spectrometry-based metabolic profiling was combined with targeted liquid chromatography–mass spectrometry analysis to cover more metabolites. Furthermore, we applied a workflow employing ultra-high performance liquid chromatography–quadrupole time of flight mass spectrometry with a powerful chemometrics platform, focusing on only significantly changed metabolites. This workflow highly reduced the complexity of acquired data without losing metabolites of interest. Consequently, forty-one novel metabolites were identified in addition to the combined method, of which two metabolites, 4-guanidinobutanal and 4-guanidinobutanoate, were identified for the first time in Saccharomyces cerevisiae. With compartment-specific metabolomics, we identified sym1Δ cells as lysine auxotroph. The highly reduced carbamoyl-aspartate and orotic acid indicate a potential role of the mitochondrial inner membrane protein Sym1 in pyrimidine metabolism. [ABSTRACT FROM AUTHOR]

Introduction: Hereditary orotic aciduria is an extremely rare, autosomal recessive disease caused by deficiency of uridine monophosphate synthase. Untreated, affected individuals may develop refractory megaloblastic anemia, neurodevelopmental disabilities, and crystalluria. Newborn screening has the potential to identify and enable treatment of affected individuals before they become significantly ill.Methods: Measuring orotic acid as part of expanded newborn screening using flow injection analysis tandem mass spectrometry.Results: Since the addition of orotic acid measurement to the Israeli routine newborn screening program, 1,492,439 neonates have been screened. The screen has identified ten Muslim Arab newborns that remain asymptomatic so far, with DBS orotic acid elevated up to 10 times the upper reference limit. Urine organic acid testing confirmed the presence of orotic aciduria along with homozygous variations in the UMPS gene.Conclusion: Newborn screening measuring of orotic acid, now integrated into the routine tandem mass spectrometry panel, is capable of identifying neonates with hereditary orotic aciduria.

In this study, a supramolecular hydrogel was fabricated with orotic acid (OA) modified chitosan (OACS) and 2,6-diaminopurine (DAP). The obtained OACS-DAP supramolecular hydrogels have dual responsiveness to temperature and pH. Phase transition experiments indicate this is a temperature-dependent thermoreversible supramolecular hydrogel. Rheological experiments proved the formation of the supramolecular hydrogel and its thixotropic properties. FTIR spectra confirmed that hydrogen bonds and π-π interactions are the main driving forces for OACS and DAP to form hydrogels through intermolecular self-assembly. XRD pattern confirmed the amorphous morphology of OACS-DAP hydrogels. The hydrogel has excellent electrical conductivity with a conductivity of 9.48 μ S·cm−1. And can achieve the precise release of gastrointestinal drugs. OACS-DAP hydrogel is expected to have better applications in the field of gastrointestinal drug release. [ABSTRACT FROM AUTHOR]

Cystinosis is a rare, devastating hereditary disease secondary to recessive CTNS gene mutations. The most commonly used diagnostic method is confirmation of an elevated leukocyte cystine level; however, this method is expensive and difficult to perform. This study aimed to identify candidate biomarkers for the diagnosis and follow-up of cystinosis based on multiomics studies. The study included three groups: newly-diagnosed cystinosis patients (patient group, n = 14); cystinosis patients under treatment (treatment group, n = 19); and healthy controls (control group, n = 30). Plasma metabolomics analysis identified 10 metabolites as candidate biomarkers that differed between the patient and control groups [L-serine, taurine, lyxose, 4-trimethylammoniobutanoic acid, orotic acid, glutathione, PE(O-18:1(9Z)/0:0), 2-hydroxyphenyl acetic acid, acetyl-N-formil-5-metoxikinuramine, 3-indoxyl sulphate]. As compared to the healthy control group, in the treatment group, hypotaurine, phosphatidylethanolamine, N-acetyl-d-mannosamine, 3-indolacetic acid, p-cresol, phenylethylamine, 5-aminovaleric acid, glycine, creatinine, and saccharic acid levels were significantly higher, and the metabolites quinic acid, capric acid, lenticin, xanthotoxin, glucose-6-phosphate, taurine, uric acid, glyceric acid, alpha-D-glucosamine phosphate, and serine levels were significantly lower. Urinary metabolomic analysis clearly differentiated the patient group from the control group by means of higher allo-inositol, talose, glucose, 2-hydroxybutiric acid, cystine, pyruvic acid, valine, and phenylalanine levels, and lower metabolite (N-acetyl-L-glutamic acid, 3-aminopropionitrile, ribitol, hydroquinone, glucuronic acid, 3-phosphoglycerate, xanthine, creatinine, and 5-aminovaleric acid) levels in the patient group. Urine metabolites were also found to be significantly different in the treatment group than in the control group. Thus, this study identified candidate biomarkers that could be used for the diagnosis and follow-up of cystinosis. [ABSTRACT FROM AUTHOR]

In an era of antibiotic resistance where natural antibiotic substitutes are considered essential, the antimicrobial and antibiofilm activities of Citrus limon extract on strains of pathogenic Escherichia coli isolated from pork were evaluated. The strains which form biofilms were more resistant (MIC50 = 2.5 mgml−1) compared to non-biofilm forming strains (MIC50 = 1.25 mgml−1). Use of C. limon extract at 20 mgml−1 concentration has resulted in inhibition of biofilm formation by 53.96%. Cyclobarbital, 5, 8-dimethoxycumarin, orotic acid and 3-methylsalicylhydrazide were the major phytochemicals in C. limon extract with highest docking affinities against the biofilm associated proteins in E. coli. The results of simulation studies have clearly illustrated the energy stability of the protein-ligand complexes. Absorption, distribution, metabolism, excretion and toxicity (ADMET) profiles revealed that the phytochemicals in C. limon could be used in the drug design studies to preferentially target the specific receptors to combat biofilms associated with E. coli. [ABSTRACT FROM AUTHOR]

The article provides data on current stress loads in the population of Ukraine, which have significantly increased as a result of emotional overstrain (stress) and the reduction of periods of positive emotional states. The presence of active military operations in the country and, as a result, the appearance of the population with the status of “displaced persons”; the part of the population that is/was in the zone of temporary occupation; the appearance of the wounded and disabled persons among the civilian population led the completely new tasks for the organism adaptive systems, which it is not always ready to overcome them. Over the last two year, the life of a person in Ukraine has been affected by a combination of two “dark forces”, namely, the war and the COVID-19 pandemic, and this influence is still continuing. The war for almost a year led to the layering of acute stress on already existing problems, deterioration of the social situation; forced change of place/country of residence (often repeated changes); interruptions in the supply of quality food products, medicines, dietary supplements, etc.; violation of “doctor-patient” contacts and, as a result, the impossibility of receiving timely and high-quality consultations and treatment. On the background of these problems there are significant risks of wartime for pregnant women, which can hinder the favorable course and termination of pregnancy. Among the most significant difficulties is a severe psychological trauma. The problem of the impact of the war on the pregnant women and the results of childbirth in our country began to be considered since 2014, when as a result of Russia’s aggression in the east of the country and in the Crimea a completely new part of the population appeared – internally displaced persons (IDPs) from these regions. To date, the significantly increased contingent of IDPs has also been joined by those persons who displaced to other countries because of the war from February 24, 2022. The stay of a pregnant woman in conditions of acute or long-term chronic stress negatively affects the course of pregnancy, increasing the number of complications and negatively affecting the condition of the fetus and newborn. Pregnancy can also be considered a significant adaptive load, which requires the woman’s body to mobilize mechanisms for synchronization between indicators of the basic systems and a favorable course of the gestation and childbirth process. Even during its physiological course it has a certain influence on the psychological condition of a woman, and some authors consider it even as a crisis situation, or as a “period of psychological uncertainty”. Individual features of response to stress cannot be dismissed either. Yes, the degree of reaction of a pregnant woman can be completely different depending on the specifics of the situation. One of the effective and available ways to prevent the effects of long-term stress during pregnancy is magnesium (Mg) supplementation in its oral form, taking into account bioavailability and the daily dose for pregnant women. Magnesium is indispensable in maintaining the conduction of nerve impulses, serves as a natural muscle relaxant that relaxes smooth and skeletal muscles, has a positive effect on the functioning of the placenta and endothelium, ensures the full development of the fetus in synergy with other useful substances, in particular B vitamins. The preparations based on magnesium salts with organic acids (orotic acid, vitamin B6), in which the anion of the acid serves as a “magnesium carrier” (ligand) into the cell, are characterized by a high bioavailability. It is important in the context of the impact on the central nervous system during the war that magnesium, vitamin B6 and orotic acid potentiate each other’s anti-stress effect. Our experience allows us to recommend a magnesium preparation for long-term use, starting with pre-gravid preparation, throughout pregnancy and lactation, which includes the chelated form of marine magnesium «Simag 55» (purified natural marine mineral extract) with two conductors (vitamin B6 and orotic acid) to ensure the highest bioavailability. [ABSTRACT FROM AUTHOR]

Background: Citrullinemia type I (CTLN1) is a rare urea cycle disorder (UCD) with few adult cases described so far. Diagnosis of late-onset CTLN1 is difficult, and delayed treatment may increase the risk of severe hyperammonemia. Pregnancy is an important risk factor for women with CTLN1. However, the clinical manifestations of CTLN1 in a pregnant woman may be mistaken for pregnancy side effects and ultimately delay a timely diagnosis. Case presentation: A 34-year-old woman developed vomiting and disturbance of consciousness after 12 weeks of gestation. A blood test showed hyperammonemia (454 μg/dL) with normal liver function tests. She fell into a deep coma, and her serum ammonia level increased to 800 μg/dL. Continuous renal replacement therapy (CRRT) was administered as a diagnostic treatment for UCD and serum ammonia. This patient's case was complicated by co-infection; her dependents decided to withdraw life support and the patient died. She was diagnosed with CTLN1 by analyses of plasma amino acids, urinary orotic acid, and second-generation gene sequencing. Discussion and conclusion: When a patient displays symptoms of emesis and disturbance of consciousness in early pregnancy, blood ammonia should be monitored, and UCD should be considered, particularly for patients with hyperammonemia in the absence of severe liver function abnormalities. [ABSTRACT FROM AUTHOR]

Polylactic acid (PLA) is a bio-based biodegradable polymer and is considered to be an environmentally friendly alternative to petroleum-based polymers for various applications. Neat PLA requires an extended period at elevated temperatures to attain its maximum crystallinity, which can be mitigated by the addition of nucleating agents. Orotic acid is a natural heterocyclic nucleating agent in PLA. The effect of orotic acid on the crystallization behavior of a commercial, high-purity PLA was studied in detail. A differential scanning calorimetry (DSC) technique was utilized for this purpose. A new protocol for the quantitative characterization of crystallization kinetics from DSC data was developed. It was found that the total crystallinity increased from 26% to 63% at 80 °C with 1% content of orotic acid. Meanwhile, the crystallization rate of PLA-OA blends increased by ~10 times as compared to neat PLA. The addition of orotic acid also reduced the incubation time by >17% under quiescent conditions. Injection molding experiments showed that highly crystallized (>50%) PLA samples could be fabricated with a 1% addition of orotic acid. The required mold temperature was reduced from the 120 °C recommended by the supplier to 80 °C. [ABSTRACT FROM AUTHOR]

[Display omitted] • Escherichia coli was engineered to produce industrial-yields of orotic acid. • A plasmid-free, uninduced and nonauxotrophic orotic acid strain was constructed. • Metabolic flux for orotic acid biosynthesis was optimized by systematic engineering. • A novel "Chaos to Order Cycles" fermentation method was developed. • The highest production titer, productivity and yield of orotic acid was achieved. Orotic acid is widely used in healthcare and cosmetic industries. However, orotic acid-producing microorganisms are auxotrophic, which results in inefficient microbial production. Herein, a plasmid-free, uninduced, non-auxotrophic orotic acid hyperproducer was constructed from Escherichia coli W3110. Initially, the orotic acid degradation pathway was blocked and the carbamoyl phosphate supply was enriched. Subsequently, pyr operon from Bacillus subtilis F126 was heterologously expressed and precursors' supply was optimized. Thereafter, pyrE was dynamically regulated to reconstruct the non-auxotrophic pathway. Employing fed-batch cultivation, orotic acid titer, yield, and productivity of strain Ora21 reached 182.5 g/L, 0.58 g/g, and 3.80 g/L/h, respectively, the highest levels reported so far. Finally, a novel "Chaos to Order Cycles (COC)" fermentation was developed, which effectively increased the yield to 0.63 g/g. This research is a remarkable achievement in orotic acid production by microbial fermentation and has vast potential for industrial applications. [ABSTRACT FROM AUTHOR]

The purpose of this study was to evaluate the effect of 6 different feeding systems (based on corn silage as the main ingredient) on the chemical composition of milk and to highlight the potential of untargeted metabolomics to find discriminant marker compounds of different nutritional strategies. Interestingly, the multivariate statistical analysis discriminated milk samples mainly according to the high-moisture ear corn (HMC) included in the diet formulation. Overall, the most discriminant compounds, identified as a function of the HMC, belonged to AA (10 compounds), peptides (71 compounds), pyrimidines (38 compounds), purines (15 compounds), and pyridines (14 compounds). The discriminant milk metabolites were found to significantly explain the metabolic pathways of pyrimidines and vitamin B 6. Interestingly, pathway analyses revealed that the inclusion of HMC in the diet formulation strongly affected the pyrimidine metabolism in milk, determining a significant up-accumulation of pyrimidine degradation products, such as 3-ureidopropionic acid, 3-ureidoisobutyric acid, and 3-aminoisobutyric acid. Also, some pyrimidine intermediates (such as l -aspartic acid, N-carbamoyl- l -aspartic acid, and orotic acid) were found to possess a high discrimination degree. Additionally, our findings suggested that the inclusion of alfalfa silage in the diet formulation was potentially correlated with the vitamin B 6 metabolism in milk, being 4-pyridoxic acid (a pyridoxal phosphate degradation product) the most significant and up-accumulated compound. Taken together, the accumulation trends of different marker compounds revealed that both pyrimidine intermediates and degradation products are potential marker compounds of HMC-based diets, likely involving a complex metabolism of microbial nitrogen based on total splanchnic fluxes from the rumen to mammary gland in dairy cows. Also, our findings highlight the potential of untargeted metabolomics in both foodomics and foodomics-based studies involving dairy products. [ABSTRACT FROM AUTHOR]

Background and Objectives: Platelet concentrates (PCs) contaminated with Staphylococcus aureus can escape detection during PC screening, causing septic transfusion reactions. This study aimed to determine the impact of S. aureus contamination on platelet metabolism and functionality during PC storage. Materials and Methods: Targeted metabolomics (N = 3) was performed on non‐spiked PCs and PCs inoculated with 10–20 colony‐forming units (CFU)/bag of S. aureus. Metabolites were quantified at 0, 48 and 144 h using high‐performance mass spectrometry (MS). Additionally, PCs spiked with approximately 20 CFU/bag of S. aureus were sampled every 24 h for up to 144 h to evaluate platelet functionality using flow cytometry (N = 2). Results: Eight metabolites had significantly different levels in spiked PCs (log2 fold‐change ≤ or ≥±1) versus non‐spiked units at 48 and 144 h. Xanthine, uridine, serine, glutamine and threonine were increased, whereas orotic acid, dihydroorotic acid and aspartic acid were decreased. Flow cytometry showed a significant decrease in expression of GPIIb while P‐selectin expression was significantly increased in spiked PCs after 72 h of storage when S. aureus concentration was ≥10E+08 CFU/ml. Additionally, phosphatidylserine exposure was significantly increased after 48 h of PC storage, when S. aureus had reached a concentration of 2E+06. Conclusion: Contamination with S. aureus exacerbates platelet storage lesions in contaminated PCs but only when the bacterium has reached clinically significant levels. [ABSTRACT FROM AUTHOR]

Targeting dendritic cells (DCs) metabolism-related pathways and in-situ activation of DCs have become a new trend in DC-based immunotherapy. Studies have shown that Lycium barbarum polysaccharide can promote DCs function. This study is aimed at exploring the mechanism of LBP affecting DCs function from the perspective of metabolomics. MTT method was used to detect the activity of DC2.4 cells. ELISA kit method was used to detect the contents of IL-6, IL-12, and TNF-α in the supernatant of cells. Ultra-performance liquid chromatography-quadrupole-time-of-flight mass spectrometry (UPLC-Q-TOF/MS) was used to detect general changes in DC2.4 cell metabolism. And then multidistance covariates and bioinformatics, partial least squares-discriminant analysis (PLS-DA) were used to analyze differential metabolites. Finally, metabolic pathway analysis was performed by MetaboAnalyst v5.0. The results showed that LBP had no significant inhibitory effect on the activity of DC2.4 cells at the experimental dose of 50-200 μg/ml. LBP (100 μg/ml) could significantly stimulate DC2.4 cells to secrete IL-6, TNF-α, and IL-12. Moreover, 20 differential metabolites could be identified, including betaine, hypoxanthine, L-carnitine, 5'-methylthioadenosine, orotic acid, sphingomyelin, and L-glutamine. These metabolites were involved 28 metabolic pathways and the top 5 metabolic pathways were aspartate metabolism, pyrimidine metabolism, phenylacetate metabolism, methionine metabolism, and fatty acid metabolism. These results suggest that the effect of LBP on DCs function is related to the regulation of cell metabolism. [ABSTRACT FROM AUTHOR]

Many molecular drugs exist as liquid materials, however this state is not necessarily the most viable or stable means to store pharmaceuticals. Crystal engineering can be utilized to isolate these liquid materials in the solid state. Herein, we describe the liquid-state to solid-state transformation of one of the most widely consumed liquid alkaloid drugs; nicotine. Nicotine is known to be highly unstable with a multitude of degradation pathways. To isolate nicotine in the solid-state, crystallization was utilized with the generally recognized as safe (GRAS) salt former orotic acid. The melting point of the isolated crystalline salt was determined along with the photo-stabilizing effects the solid state offers for liquid pharmaceuticals such as nicotine. [ABSTRACT FROM AUTHOR]

Photoperiod is an important factor that stimulates the reproductive performance of broiler breeder roosters. However, the mechanism by which photoperiod affects the reproductive performance of broiler breeder roosters has not been fully studied. To study the effects of different photoperiods on the reproductive performance of broiler breeder roosters, 120 Arbor Acres broiler breeder roosters aged 20 weeks were randomly assigned to three groups (n = 40), and the three groups were treated with different photoperiod regimes: control (CTR; 12.5 h of light and 11.5 h of dark, 12.5 L: 11.5 D), short day (SD; 16 L: 8 D) and long day (LD; 8 L: 16 D). Serum and testes were collected after 4 weeks of feeding, and testosterone‐related indices were detected. We found that testosterone synthesis in the testes of broiler roosters was boosted with prolonged of photoperiod. Subsequently, metabonomics was used to identify the differential endogenous metabolites that may affect the function of the testes in breeder roosters. We found compared with other groups, the concentrations of creatine, uridine monophosphate, phosphoribosyl pyrophosphate, dCMP, α‐D‐glucose and citric acid in the SD group decreased significantly (p < 0.05), and glyoxylic acid, D‐ribose 5‐phosphate, deoxyuridine and orotic acid in the SD group increased significantly (p < 0.05), while the CTR group and LD group showed no significant difference (p > 0.05). The concentrations of linoleic acid and α‐linolenic acid in the LD group were increased significantly (p < 0.05) than those in the CTR and SD groups. Compared with the CTR group, the concentrations of histamine in the SD and LD groups were significant increased (p < 0.05). The 13 of the different metabolites could be used as candidate biomarkers for different photoperiods affecting testosterone synthesis, may be used to molecular breeding of high reproductive performance broiler roosters. [ABSTRACT FROM AUTHOR]

Abstract Alpha‐tricalcium phosphate (α‐TCP), which can be converted to calcium‐deficient hydroxyapatite (HA) by hydrolysis reaction, has been researched as a bone filling. Although the composites of α‐TCP with various organic molecules can exhibit a novel functionality, the reaction based on the addition of the interactive organic molecules has not been understood. Investigation of the hydrolysis conversion process under various conditions can lead to the elucidation of the biomineralization process. In this study, we investigated the hydrolysis conversion of α‐TCP to HA in the presence of orotic acid (Ort). As a result, the conversion rate from α‐TCP to HA at the reaction time of 3 h was improved to be more than five times as compared to that in the absence of Ort. Moreover, the addition of Ort promoted the crystal growth and suppressed the crystal growth in the m‐plane direction. Furthermore, the mechanism for promoting HA formation is assumed to be related to the molecular structure of Ort, which is capable of forming the interactions between carboxylate and Ca2+ ions.

The terahertz (THz) absorption spectrum of orotic acid monohydrate in the crystalline phase was experimentally obtained by using THz time-domain spectroscopy and computationally simulated by using density functional theory. Four distinct peaks were observed within the range of 12–128 cm–1, and were computationally reproduced by simulations using the Perdew–Burke–Ernzerhof functional. A comparison of the experimental and calculated data indicated that the measured peaks mostly originated from intermolecular forces in which the interactions between orotic acid molecules dominated. In addition, the feature located at 110.2 cm–1 was attributed to the interactions between orotic acid and water molecules. These findings demonstrate that THz spectroscopy can be used to monitor molecular dehydration during industrial production. [ABSTRACT FROM AUTHOR]

Orotic acid (OA) is an intermediate of the pyrimidine biosynthesis with high industrial relevance due to its use as precursor for production of biochemical pyrimidines or its use as carrier molecule in drug formulations. It can be produced by fermentation of microorganisms with engineered pyrimidine metabolism. In this study, we surprisingly discovered the yeast Yarrowia lipolytica as a powerful producer of OA. The overproduction of OA in the Y. lipolytica strain PO1f was found to be caused by the deletion of the URA3 gene which prevents the irreversible decarboxylation of OA to uridine monophosphate. It was shown that the lack of orotidine‐5′‐phosphate decarboxylase was the reason for the accumulation of OA inside the cell since a rescue mutant of the URA3 deletion in Y. lipolytica PO1f completely prevented the OA secretion into the medium. In addition, pyrimidine limitation in the cell massively enhanced the OA accumulation followed by secretion due to intense overflow metabolism during bioreactor cultivations. Accordingly, supplementation of the medium with 200 mg/L uracil drastically decreased the OA overproduction by 91%. OA productivity was further enhanced in fed‐batch cultivation with glucose and ammonium sulfate feed to a maximal yield of 9.62 ± 0.21 g/L. Y. lipolytica is one of three OA overproducing yeasts described in the literature so far, and in this study, the highest productivity was shown. This work demonstrates the potential of Y. lipolytica as a possible production organism for OA and provides a basis for further metabolic pathway engineering to optimize OA productivity. [ABSTRACT FROM AUTHOR]

Thermoduric bacteria are known to affect the quality of Cheddar cheese, with manifested defects including slits, weak body, and blowing. Thermoduric bacteria are likely to increase in numbers during cheese-making, as in-process conditions are conducive to proliferation. The present study was conducted to track thermoduric bacterial progression during an 18- to 20-h Cheddar cheese production run and during ripening when the pasteurizer was washed at midway through the production day. This study also correlated a broad range of chemical changes to the growth of thermoduric bacteria during ripening. Three independent cheese trials were performed at 3.5- ± 0.5-mo intervals. Samples were drawn in duplicates at 4 different times of the day: at the start of the run (vat 1), prior to a midday wash of the pasteurizer (vat 20), after the midday wash of the pasteurizer (vat 21), and at the end of the run (vat 42) for raw milk, pasteurized milk, and cheese. Cheeses were also tested during ripening for 6 mo. Results showed that raw milk total bacterial counts comprised 0.24% thermoduric mesophiles (TM) and 0.12% thermoduric thermophiles (TT). The thermoduric thermophilic bacterial counts increased by log 10 1.23 during the pasteurizer run of 9 to 10 h, indicating a buildup of thermoduric thermophilic bacteria during the pasteurization process itself. Midday washing reduced thermophilic counts by log 10 1.36, as evident by pre- and post-midday wash counts. However, a thermophilic buildup during post-midday wash was again noticed near the end of the 20-h run. We found that TT bacteria decreased in the first 60 d of ripening, whereas TM bacteria increased during the same period. However, TT bacteria increased later during 60 to 180 d of ripening. Bacillus licheniformis was the most frequently isolated bacteria in this study and was recovered at all production stages sampled during the cheese-making and ripening. We observed a significant increase in the level of orotic and uric acids in the vat made at the end of the day. No significant difference in the overall chemical composition, proteolysis, sugar, or other organic acids was observed in cheese made at the start versus the end of the production run. [ABSTRACT FROM AUTHOR]

Impairment of pancreatic β cells is a principal driver of the development of diabetes. Restoring normal insulin release from the β cells depends on the ATP produced by the intracellular mitochondria. In maintaining mitochondrial function, the tumor suppressor p53 has emerged as a novel regulator of metabolic homeostasis and participates in adaptations to nutritional changes. In this study, we used orotic acid, an intermediate in the pathway for de novo synthesis of the pyrimidine nucleotide, to reduce genotoxicity. Administration of orotic acid reduced p53 activation of MIN6 β cells and subsequently reduced β cell death in the db/db mouse. Orotic acid intake helped to maintain the islet size, number of β cells, and protected insulin secretion in the db/db mouse. In conclusion, orotic acid treatment maintained β cell function and reduced cell death, and may therefore, be a future therapeutic strategy for the prevention and treatment of diabetes. • Orotic acid, an intermediate of the pyrimidine nucleotide pathway, reduced DNA damage. • Orotic acid ameliorated the p53 activation of MIN6 β cells. • Orotic acid intake maintained pancreatic β cell function of the db/db mouse. [ABSTRACT FROM AUTHOR]

Since ancient times, sheep's milk cheeses have been a part of a human diet. Currently, their consumption is of great interest due to its nutritional and health values. The aim of the article was to review the chemical composition of sheep's milk cheeses and its main bioactive ingredients in the context of nutritional and health values. Sheep's milk cheeses are rich in functionally and physiologically active compounds such as: vitamins, minerals, fatty acids, terpenes, sialic acid, orotic acid and L-carnitine, which are largely originate from milk. Fermentation and maturation process additionally enrich them in other bioactive substances as: bioactive peptides, γ-aminobutyric acid (GABA) or biogenic amines. Studies show that sheep's milk cheese consumption may be helpful in the prevention of civilization diseases, i.e. hypertension, obesity or cancer. However, due to the presence of biogenic amines, people with metabolic disorders should be careful of their intake. [ABSTRACT FROM AUTHOR]

Orotate phosphoribosyltransferase (OPRT) exists as a bifunctional enzyme, uridine 5′-monophosphate synthase, in mammalian cells and plays an important role in pyrimidine biosynthesis. Measuring OPRT activity has been considered important for understanding biological events and development of molecular-targeting drugs. In this study, we demonstrate a novel fluorescence method for measuring OPRT activity in living cells. The technique utilizes 4-trifluoromethylbenzamidoxime (4-TFMBAO) as a fluorogenic reagent, which produces selective fluorescence for orotic acid. To perform the OPRT reaction, orotic acid was added to HeLa cell lysate, and a portion of the enzyme reaction mixture was heated at 80 °C for 4 min in the presence of 4-TFMBAO under basic conditions. The resulting fluorescence was measured using a spectrofluorometer, which reflects the consumption of orotic acid by the OPRT. After optimization of the reaction conditions, the OPRT activity was successfully determined in 15 min of enzyme reaction time without further procedures such as purification of OPRT or deproteination for the analysis. The activity obtained was compatible with the value measured by the radiometric method with [3H]-5-FU as the substrate. The present method provides a reliable and facile measurement of OPRT activity and could be useful for a variety of research fields targeting pyrimidine metabolism.

Alpha‐tricalcium phosphate (α‐TCP), which can be converted to calcium‐deficient hydroxyapatite (HA) by hydrolysis reaction, has been researched as a bone filling. Although the composites of α‐TCP with various organic molecules can exhibit a novel functionality, the reaction based on the addition of the interactive organic molecules has not been understood. Investigation of the hydrolysis conversion process under various conditions can lead to the elucidation of the biomineralization process. In this study, we investigated the hydrolysis conversion of α‐TCP to HA in the presence of orotic acid (Ort). As a result, the conversion rate from α‐TCP to HA at the reaction time of 3 h was improved to be more than five times as compared to that in the absence of Ort. Moreover, the addition of Ort promoted the crystal growth and suppressed the crystal growth in the m‐plane direction. Furthermore, the mechanism for promoting HA formation is assumed to be related to the molecular structure of Ort, which is capable of forming the interactions between carboxylate and Ca2+ ions. [ABSTRACT FROM AUTHOR]

A new hydrated molecular salt of GABA was crystallized, and its crystal structure was solved. This is the first example of a hydrated molecular salt of an achiral amino acid with L -tartaric acid and also the first communication on a hydrated molecular salt of GABA with a chiral molecule. The structure contains molecular bi-layers of L -tartaric acid anions connected together by R 44 (12) motifs based on GABA cations and water molecules. The role of water in the 3D hydrogen bonded network was analyzed. Structural similarity between the monohydrates of GABA with L -tartaric and orotic acid was found. We can suppose that the general R 44 (12) motif including 2 GABA cations and 2 water molecules would be common for other hydrated molecular salts of GABA with other co-formers, which will be obtained in the future. The stability on storage, thermal behavior and dehydration process for the novel molecular compound were investigated. [ABSTRACT FROM AUTHOR]

Pathophysiology of visceral leishmaniasis (VL) is not fully understood and it has been widely accepted that the parasitic components and host immune response both contribute to the perpetuation of the disease. Host alterations during leishmaniasis is a feebly touched area that needs to be explored more to better understand the VL prognosis and diagnosis, which are vital to reduce mortality and post-infection sequelae. To address this, we performed untargeted metabolomics of Leishmania donovani (Ld) infected, uninfected and treated BALB/c mice's tissues and biofluids to elucidate the host metabolome changes using gas chromatography–mass spectrometry. Univariate and multivariate data treatments provided numerous significant differential hits in several tissues like the brain, liver, spleen and bone marrow. Differential modulations were also observed in serum, urine and fecal samples of Ld-infected mice, which could be further targeted for biomarker and diagnostic validations. Several metabolic pathways were found to be upregulated/downregulated in infected (TCA, glycolysis, fatty acids, purine and pyrimidine, etcetera) and treated (arginine, fumaric acid, orotic acid, choline succinate, etcetera) samples. Results also illustrated several metabolites with different pattern of modulations in control, infected and treated samples as well as in different tissues/biofluids; for e.g. glutamic acid identified in the serum samples of infected mice. Identified metabolites include a range of amino acids, saccharides, energy-related molecules, etcetera. Furthermore, potential biomarkers have been identified in various tissues—arginine and fumaric acid in brain, choline in liver, 9-(10) EpOME in spleen and bone marrow, N-acetyl putrescine in bone marrow, etcetera. Among biofluids, glutamic acid in serum, hydrazine and deoxyribose in urine and 3-Methyl-2-oxo pentanoic acid in feces are some of the potential biomarkers identified. These metabolites could be further looked into for their role in disease complexity or as a prognostic marker. The presented profiling approach allowed us to attain a metabolic portrait of the individual tissue/biofluid modulations during VL in the host and represent a valuable system readout for further studies. Our outcomes provide an improved understanding of perturbations of the host metabolome interface during VL, including identification of many possible potential diagnostic and therapeutic targets. [ABSTRACT FROM AUTHOR]