Hippuric Acid Research Articles

Circulating gut microbiota-related metabolites influence endothelium plaque lesion formation in ApoE knockout rats.

Atherosclerosis is the main cause of cardiac and peripheral vessel infarction in developed countries. Recent studies have established that gut microbiota and their metabolites play important roles in the development and progression of cardiovascular disease; however, the underlying mechanisms remain unclear. The present study aimed to investigate endothelium plaque lesion formation in ApoE-deficient rats fed a normal chow diet under germ-free (GF) and specific-pathogen-free (SPF) conditions at various time points. There was no difference in serum cholesterol and triglyceride levels between SPF-rats and GF-rats. Histological studies revealed that the GF-rats developed endothelium plaques in the aorta from 26 to 52 weeks, but this was not observed in SPF-rats. GF-rat coronary arteries had moderate-to-severe endothelium lesions during this time period, but SPF-rat coronary arteries had only mild lesion formation. Immunohistochemical staining showed higher accumulation of CD68-positive and arginase-negative foamy-like macrophages on the arterial walls of GF-rats, and expression of TNF-α and IL-6 in foam cells was only observed in GF-rats. In addition, microbial metabolites, including equol derivatives, enterolactone derivatives, indole-3-propionate, indole-3-acrylic acid, cholic acid, hippuric acid, and isoquinolone, were significantly higher in the SPF group than in the GF group. In conclusion, our results indicate that gut microbiota may attenuate atherosclerosis development.

Occurrence of Polyphenols, Isoflavonoids, and Their Metabolites in Milk Samples from Different Cow Feeding Regimens

In this work, milk samples collected in a cohort of intensive dairy farms of the Po Valley (Italy) were screened for their (poly)-phenolic profile to check the occurrence of phenolic metabolites of biological interest. The selected dairy farms were previously classified on the basis of their cow feeding system, considering the utilization of corn silage as the main ingredient of the rations. Overall, ultra-high-pressure liquid chromatography coupled with mass spectrometry using an Orbitrap analyzer, followed by unsupervised and supervised statistics, allowed identifying clear different phenolic distributions in the milk samples. Accordingly, a great variability in the phenolic profiles of the different milk samples was observed, with two main phenolic clusters outlined by the unsupervised hierarchical clustering approach and not fully correlated to the nutritional strategy considered. The variables’ importance in the projection approach allowed selecting the most important metabolites, resulting in samples’ discrimination. Among the most discriminative compounds, we found phenolic metabolites (such as hippuric acid and 4-hydroxyhippuric acid), followed by lignans (such as enterolactone) and isoflavonoids (such as equol and O-desmethylangolensin). Taken together, our findings suggested that both the feeding systems and the ability of dairy cows to process parent phenolic compounds were the main factors providing the final (poly)-phenolic profile of the milk samples. Future targeted and ad hoc studies appear of great interest to evaluate the potential biological effects of these compounds on cow health.

Metabolomics analysis reveals molecular linkages for the impact of vitamin D on childhood allergic airway diseases.

Several studies have reported the relevance between serum vitamin D and allergic immunoglobulin E (IgE) responses and atopic diseases. However, a metabolomics-based approach to the impacts of vitamin D on allergic reactions remains unclear. A total of 111 children completed a 3-year follow-up were enrolled and classified based on longitudinal vitamin D status (≥ 30ng/ml, n=54; 20-29.9ng/ml, n=41; <20ng/ml, n=16). Urinary metabolomic profiling was performed using 1 H-Nuclear magnetic resonance (NMR) spectroscopy at age 3. Integrative analyses of their associations related to vitamin D levels, atopic indices, and allergies were performed, and their roles in functional metabolic pathways were also assessed. Six and five metabolites were identified to be significantly associated with vitamin D status and atopic diseases, respectively (FDR-adjusted p-value <.05). A further correlation analysis revealed that vitamin D-associated 3-hydroxyisobutyric acid and glutamine were positively correlated with atopic disease-associated succinic acid and alanine, respectively. Furthermore, hippuric acid was negatively correlated with atopic disease-associated formic acid, which was positively correlated with vitamin D level (p<.01). Absolute eosinophil count (AEC) was positively correlated with serum D.pteronyssinus- and D.farinae-specific IgE level (p<.01) but negatively correlated with vitamin D level (p<.05). Amino acid metabolisms were significantly associated with vitamin D related to childhood allergies. Integrative metabolomic analysis provides the link of vitamin D-associated metabolites with the gut microbiome and immunoallergic reactions related to childhood allergies.

Antioxidant Polyphenols of Antirhea borbonica Medicinal Plant and Caffeic Acid Reduce Cerebrovascular, Inflammatory and Metabolic Disorders Aggravated by High-Fat Diet-Induced Obesity in a Mouse Model of Stroke.

Metabolic disorders related to obesity and type 2 diabetes are associated with aggravated cerebrovascular damages during stroke. In particular, hyperglycemia alters redox and inflammatory status, leading to cerebral endothelial cell dysfunction, blood–brain barrier (BBB) disruption and brain homeostasis loss. Polyphenols constitute the most abundant dietary antioxidants and exert anti-inflammatory effects that may improve cerebrovascular complications in stroke. This study evaluated the effects of the characterized polyphenol-rich extract of Antirhea borbonica medicinal plant and its major constituent caffeic acid on a high-fat diet (HFD)-induced obesity mouse model during ischemic stroke, and murine bEnd3 cerebral endothelial cells in high glucose condition. In vivo, polyphenols administered by oral gavage for 12 weeks attenuated insulin resistance, hyperglycemia, hyperinsulinemia and dyslipidemia caused by HFD-induced obesity. Polyphenols limited brain infarct, hemorrhagic transformation and BBB disruption aggravated by obesity during stroke. Polyphenols exhibited anti-inflammatory and antioxidant properties by reducing IL-1β, IL-6, MCP-1, TNF-α and Nrf2 overproduction as well as total SOD activity elevation at the cerebral or peripheral levels in obese mice. In vitro, polyphenols decreased MMP-2 activity that correlated with MCP-1 secretion and ROS intracellular levels in hyperglycemic condition. Protective effects of polyphenols were linked to their bioavailability with evidence for circulating metabolites including caffeic acid, quercetin and hippuric acid. Altogether, these findings show that antioxidant polyphenols reduced cerebrovascular, inflammatory and metabolic disorders aggravated by obesity in a mouse model of stroke. It will be relevant to assess polyphenol-based strategies to improve the clinical consequences of stroke in the context of obesity and diabetes.

Nitrogen-rich palladium tetra amino-hippuric acid phthalocyanine complex and its hybrid composite with multi-walled carbon nanotubes for supercapacitor application

Supercapacitors are the most attractive alternative energy storage and conversion systems for the future generation to successfully overcome the global energy dilemma. An extensive number of studies have been conducted to enhance the performance of an electrode and to find an appropriate electrolyte for high power density and energy density. However, it is still challenging to develop a suitable electrode/electrolyte system to boost the performance of supercapacitors. Here, a novel nitrogen-rich palladium amide bridge hippuric acid phthalocyanine complex (PdTAHAPc) is synthesized in pure state. The purity and structural characteristics of the synthesized dark green-colored PdTAHAPc are analyzed by various analytical and spectroscopic techniques. To further enhance its performance, PdTAHAPc is combined with multi-walled carbon nanotubes to prepare the PdTAHAPc-MWCNTs hybrid composite. The supercapacitance behavior of the PdTAHAPc-MWCNTs is investigated with respect to that of PdTAHAPc. The hybrid composite electrode exhibits a superior specific capacitance of 245 F·g−1 and 221 F·g−1 at a current density of 0.2 and 1 A·g−1, respectively, in 1.0 M sulfuric acid electrolyte. Furthermore, the developed hybrid composite electrode exhibits an excellent stability towards 5000 electrochemical cycles and a higher capacitance value at 80 °C compared to room temperature. The combination of PdTAHAPc with MWCNTs provides an exceptionally high surface area, leading to a lesser charge transfer resistance and a superior charge storage capacity of the developed PdTAHAPc-MWCNTs hybrid composite.

The circulating plasma metabolome of Neoparamoeba perurans-infected Atlantic salmon (Salmo salar)

Metabolomics can provide insights into the dynamic small-molecule fluctuations occurring in response to infection and has become a valuable tool in studying the pathophysiology of diseases in recent years. However, its application in fish disease research is limited. Here, we report the circulating plasma metabolome of Atlantic salmon (Salmo salar) experimentally infected with Neoparamoeba perurans—the causative agent of amoebic gill disease (AGD). Plasma samples were collected from fish with varying degrees of infection inferred from an external gross morphological score of gill pathology (i.e., gill score [GS] 1 -- GS3), where a higher GS indicates advanced infection stage. Uninfected fish (GS0) served as the control. Typical pathologies associated with AGD infection, such as hyperplastic lesions and lamellar fusion, were evident in infected gill samples. Plasma metabolites were identified by ultra-performance liquid chromatography coupled with a high-resolution quadrupole-orbitrap mass spectrometer. Identification of compounds were performed at four levels of certainty, where level 1 provided the most accurate compound identity. A total of 900 compounds were detected in the samples of which 143 were annotated at level 3, 68 on level 2b, 74 on level 2a, and 66 on level 1. Versus GS0, GS1 showed the highest number of significantly affected metabolites (104), which decreased with a higher GS. Adrenaline and adenosine were the two Level 1 compounds significantly affected by AGD regardless of GS, with the former increasing and the latter decreasing in infected fish. Hippuric acid significantly increased in GS1 and GS2, while the tryptophan metabolite indole-3-lactic acid decreased in response to the initial stage of infection but returned to basal levels at a higher GS. There were ten significantly affected metabolic pathways: Eight of which were significantly downregulated while two were downregulated in GS1 relative to GS0. The super-pathway of purine nucleotide salvage was enriched both within the upregulated metabolites in GS1vsGS0 and the down-regulated metabolites in GS3vsGS1. This is the first report on the circulating plasma metabolome of AGD infected salmon, and the results show that low infection levels resulted in a more dramatic metabolomic dysregulation than advanced infection stages. The metabolites identified are potential biological markers for the systemic physiological impact of AGD.

Blood serum levels of selected biomarkers of oxidative stress among printing workers occupationally exposed to low-levels of toluene and xylene.

Printing workers (PWs) are exposed to a mixture of solvents, yet the health risks associated with such exposuer are unknown. This study aimed to compare the serum levels of selected biomarkers of oxidative stress among occupationally exposed PWs to low-level of toluene and xylenes and unexposed controls. Associations between levels of such biomarkers and occupational exposures to toluene and xylene were also investigated. Urinary levels of hippuric acid (HA) and methyl hippuric acids (MHAs) as exposure biomarkers of toluene and xylenes, respectively, and serum levels of oxidative stress biomarkers, including total antioxidant capacity (TAC), malondialdehyde (MDA), and the antioxidant enzymes superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx), were measured among the 84 subjects, comprising 44 PWs and 40 unexposed subjects. Mean concentrations of urinary HA and MHAs of PWs showed a significant increase compared with the unexposed controls. Although levels of urinary biomarkers of exposure to toluene (HA) and xylenes (MHAs) were well below the biological exposure indices (BEIs; ACGHI), PWs presented significantly increased serum levels of MDA, and significantly decreased serum activities of SOD and GPx compared to the unexposed controls. However, for serum TAC and CAT activity, no significant difference was observed between the two groups. Correlation analyses indicated that urinary levels of HA and MHAs were positively correlated with MDA levels and negatively correlated with GPx and SOD. Our study suggested that the alterations evidenced in serum levels of MDA, SOD, and GPx could be involved in the oxidative stress caused by co-exposure to low levels of toluene and xylene. Further investigation is needed to clarify the effect of low-level occupational exposure to solvents among PWs.

A new method for electrochemical determination of Hippuric acid based on molecularly imprinted copolymer

This study introduces easy detection of Hippuric Acid (HA) through electrochemical polymerization technique. The molecularly imprinted polymer (MIP) was immobilized on a glassy carbon electrode (GCE) through electropolymerization of monomers, including m-dihydroxy benzene (m-DB) and o-aminophenol (o-AP), in presence of HA as the template. Moreover, it was subsequently delineated using electrochemical probe of hexacyanoferrate redox, atomic force microscopy (AFM), fourier-transform infrared spectroscopy (FTIR), scanning electron microscopy (SEM) and electrochemical techniques such as cyclic voltammetry (CV), differential pulse voltammetry (DPV) and electrochemical impedance spectroscopy (EIS). This sensor exhibited limit of detection (0.012 nM) with a good linear range of 0.05–40 nM and 40–500 nM. The developed sensor was properly employed to determine HA in real samples with acceptable results.

133 Effects of Benzoic Acid and Sodium Benzoate Supplementation on Growth Performance of Nursery Pigs

Abstract This study was to investigate the effects of dietary supplementation of benzoic acid and sodium benzoate on growth performance of nursery pigs. A total of 432 pigs (6.9 ± 0.9 kg BW) were assigned to 8 treatments (6 pigs/pen, n = 9) in a randomized complete block design with initial BW as a block and fed for 41 d in 3 phases (7/17/17 d, respectively). Treatments were 1) basal diet (BD), 2) BD + antibiotic, 3) BD + 0.30% sodium benzoate, 4) BD + 0.40% sodium benzoate, 5) BD + 0.60% sodium benzoate, 6) BD + 0.25% benzoic acid, 7) BD + 0.35% benzoic acid, and 8) BD + 0.50% benzoic acid. Diets contained nutrients meeting the requirement. Growth performance and fecal score were measured for each phase. Pigs representing each pen were euthanized to collect stomach and intestinal digesta to measure pH, and urine to measure pH and hippuric acid content. Data were analyzed using contrasts in Proc MIXED and slope ratio in Proc GLM of SAS. Increasing supplemental sodium benzoate showed a quadratic response (P &amp;lt; 0.05, max at 339 g/d) on ADG and linear increase (P &amp;lt; 0.05, 514 to 578 g/d) in ADFI. Increasing supplemental benzoic acid tended to have a quadratic response on ADG (P = 0.094, max at 323 g/d), but no difference in ADFI. Urine pH linearly decreased (P &amp;lt; 0.05, 7.5 to 5.7) with increasing supplemental benzoic acid, but was not affected by supplemental sodium benzoate. Increasing both supplemental sodium benzoate and benzoic acid linearly increased (P &amp;lt; 0.05) hippuric acid content in urine (3.0 to 10.4 and 3.0 to 9.2 g/L, respectively). With the slope-ratio assay using ADG and urine hippuric acid as dependent variables and benzoic acid intake as an independent variable, the estimates of bioavailability of benzoic acids were not different between sources. In conclusion, supplementation of sodium benzoate and benzoic acid increased growth performance without differences in efficacy between the sources.

Evaluation of cytokines in exhaled breath condensate in an occupationally exposed population to pneumotoxic pollutants.

The quarrying is considered a precarious occupation with high toxicity, is an informal economic activity that employs low technology, limited protection, and poses a risk to workers and their families. In quarrying, silica dust is generated and there is also occupational exposure to significant mixtures of pneumotoxic pollutants, including mineral dust (crystalline silica, carbon or cement, polycyclic aromatic hydrocarbons (PAHs), solvents, and others, which are aggravated by the lack of use of protective equipment, causing irreversible damage to the worker's respiratory health. Thus, the objective of this work focused on the evaluation of the respiratory health of artisan stonemasons in San Luis Potosí, Mexico through the study of exhaled breath condensate (EBC) (pH, pro-inflammatory cytokines) as well as the study of the exposure to pollutants present in the work area (PAHs, toluene, and 2.5µm particulate matter) through biomarkers of exposure (hippuric acid and hydroxylated metabolites of PAHs). The results show the presence of crystalline SiO2 in 100% of the samples analyzed; the PM2.5 concentrations were 5 to 10 times the permitted levels. Regarding exposure to PAHs, all the stonemasons presented urine concentrations of at least 5 of the OH-PAHs evaluated; 9-OH-FLU occurred at higher concentrations of 171.2 (122.7-279.4) µg L-1; hippuric acid, which was present in 100% of the workers evaluated in concentrations of 283.4 (27.72-1119) mg L-1, 100% of which were above the values established for occupational scenarios. The pH values obtained for the EBC samples were presented at an average of 7.07 (6.33-7.66). Pro-inflammatory cytokines were present in 86.1% of the study population. The cytokine that was found in higher concentrations was IL-2, with a mean of 178.01pgmL-1 and 3124.01pgmL-1 for the pH < 7 and pH > 7 groups, respectively. Some correlations between the cytokines and the exposure biomarkers were presented. Stonemasons are highly exposed to pneumotoxic pollutants and markers of inflammation at the pulmonary level; in addition, a high risk of developing silicosis. Quarrying should be addressed as a carcinogenic activity, which would imply the design of monitoring and control strategies for these pollutants that our country currently lacks, particularly in precarious occupations. It is necessary to develop strategies to protect the health of precarious workers.

Comparison of urinary biomarkers concentrations in exposed and non-exposed petrol station workers in the Eastern Economic Corridor (EEC), Thailand.

The Thai government has been developing its Eastern Economic Corridor (EEC), which spans three provinces, with the aim of improving connections with other Asian nations. Since this strategic development, the number of trucks, private car, and passenger car registrations have continued to grow, with a corresponding increase in related to benzene, toluene, ethylbenzene, and xylene (BTEX). This study aims to compare the levels of trans, trans-muconic acid (t, t MA); toluene (TU); mandelic acid (MA); and methyl hippuric acid (MHA) in the urine of gas station employees, considering demographic and occupational factors. These employees worked either near or away from the fuel dispenser, and there 100 people in each group. Data were collected using interviews and testing environmental air and urine samples for benzene, toluene, ethyl benzene, and xylene (BTEX). The results showed that BTEX concentrations were just detectable in all 200 cases (100%). The mean (±SD) urine level of t, t MA was 449.28 (±213.32) μg/g creatinine, while the median (min-max) was 428.23 (95.58-1202.56) μg/g creatinine. The mean TU was 0.011 (0.001) mg/L, while the median (min-max) was 0.011 (0.010-0.013) mg/L. MA levels were higher inside the pollution control zone than they were outside the zone (p=.009). Employees who practiced poor personal hygiene had relatively high urinary toluene and MHA levels (p=.009) and those who did not wear personal protective equipment (PPE) had relatively high MA levels (p=.040). The results of this study revealed statistically significant biomarkers influencing the levels of t, t MA; TU; MA; and MHA in urine. The recommendations of this study are that employers should provide their employees with suitable PPE, check regularly to ensure that it is worn, and strongly encourage employees to take care of their sanitation. Employees should take appropriate breaks and days off to minimize their exposure to BTEX.

Investigation of kidney function changes in sea salt workers during harvest season in Thailand.

Occupational factors have previously been mentioned as contributing to decreased kidney function and the development of chronic kidney disease of unknown cause. Sea salt workers are one of the occupations facing high outdoor temperatures and a highly, intensive workload. The purpose of the study was to examine whether the kidney function of sea salt workers at the beginning of the harvest season differs from kidney function at the end of the harvest season and to identify factors that can predict the change of kidney function. Data were collected from salt workers (n=50) who were between 18–60 years of age without hypertension, diabetes, and kidney disease in Samut Sakhon province, Thailand. Urine specific gravity (USG) was used for hydration status and the estimated glomerular filter rate (eGFR) was used to measure kidney function. The mixed model was used to find differences over the harvest season and prediction of factors. On average, the eGFR was estimated to decrease by 15.2 ml/min/1.73 m2 over the harvest season. The decline in eGFR of sea salt workers with moderate and heavy workloads were significantly faster than their light workload counterparts after controlling for other covariates. Similarly, dehydration (USG ≥ 1.030) significantly accelerated the rate of kidney function loss. Our study confirmed exposure to heat over the harvest season leads to decreased eGFR in sea salt workers. The rate of change of eGFR could be predicted by workload and hydration status. Workers with dehydration who performed medium to heavy workloads in farms showed faster kidney function decline than those who performed light workload.

Weaning age influences indicators of rumen function and development in female Holstein calves

BackgroundPrenatal and postnatal conditions are crucial for the development of calves. Primiparous cows are still maturing during pregnancy, thus competing with the nutritional needs of their offspring. Therefore, mature cows might provide a superior intrauterine condition. Furthermore, weaning calves at an older age might affect them positively as well by reducing stress and offering time for various organs and their functions to develop. We aimed to evaluate effects of mothers’ parity and calves’ weaning age on gastrointestinal development and corresponding acid–base balance. Fifty-nine female German Holstein calves (about 8 days old) were investigated in a 2 × 2 factorial experiment with factors weaning age (7 vs. 17 weeks) and parity of mother (primiparous vs. multiparous). Calves were randomly assigned to one of these four groups. Animal behavior that was observed included resting, chewing and active behavior.ResultsBehavioral patterns were interactively affected by time and weaning age. Rumen sounds per 2 min increased in early-weaned calves during their weaning period. In late-weaned calves a consistently increase in rumen sounds was already recorded before their weaning period. Urinary N-containing compounds (creatinine, hippuric acid, uric acid, urea, allantoin) were interactively affected by time and weaning age. Concentrations of all measured compounds except urea increased during early weaning. All except hippuric acid concentration decreased in early-weaned calves after weaning. In late-weaned calves allantoin and uric acid increased before weaning and did not change during weaning.ConclusionThese results suggest that late-weaned calves developed adequate rumen functions and acid–base balance, whereas early-weaned calves might have suffered from ruminal acidosis and catabolism. Weaning calves at 7 weeks of age might be too early for an adequate rumen development.

Application of Fe3O4@TbBd nanobeads in microextraction by packed sorbent (MEPS) for determination of BTEXs biomarkers by HPLC–UV in urine samples

A relatively new adsorbent based on covalent organic frameworks (COFs) was employed for the first time to extract and determine Trans, trans-muconic acid (tt-MA), Mandelic acid (MA), Hippuric acid (HA), and 3-Methylhippuric acid (m-MHA) in urine. For this purpose, microextraction was performed using the packed sorbent (MEPS) method. Following the extraction process, the prepared samples were specified via the high-performance liquid chromatography-ultraviolet detector system. The precipitation polymerization was applied to synthesize the Fe3O4@TbBd nanobeads, and the morphological and dimensional structures of the products were specified with FE-SEM images. Some key variables affecting the extraction efficiency (i.e., sample volume, elution volume, condition and washing solvents, type and volume of elution solvent, extraction cycles, temperature, and pH of the sample solution) were investigated. In ideal conditions, the limit of detection (LOD) was obtained from 0.02 µg/ml for tt-MA to 0.5 µg/ml for MA. Calibration curves (at five-point) were plotted in the range 0.05–5 µg/ml for tt-MA to 1–300 µg/ml for MA (R2 > 0.98). Moreover, intra- and inter-day precision values were 3.1–5.5 and 4.6–9.8%, respectively. The developed method was successfully employed to determine four analytes in three concentrations (low, medium, and high QCs). The results showed a satisfactory recovery (70–87%). COF-MEPS technique is a rapid, easy, user-friendly, and environment-friendly method for separating the minimum values of all BTEXs chief biomarkers from urine samples without using complicated processes and only with one adsorbent. Also, it can be a good alternative for biomonitoring the workers exposed to BTEX compounds in occupational and environmental access.

Exploring the Contribution of (Poly)phenols to the Dietary Exposome Using High Resolution Mass Spectrometry Untargeted Metabolomics.

This study presents a workflow to construct a Dietary Exposome Library (DEL) comprised of phytochemicals and their metabolites derived from host and gut microbiome metabolism for use in peak identification/annotation of untargeted metabolomics datasets. An evidence mapping initiative established target analytes related to the consumption of phytochemical-rich foods. Analytes were confirmed by ultra-performance liquid chromatography-mass spectrometry (UPLC-MS(n)) analysis of human biospecimens from dietary intervention studies of (poly)phenol-rich diets. One hundred and sixty six verified compounds were subsequently analyzed on an untargeted metabolomics platform to acquire chromatographic and high-resolution mass spectral data for construction of a DEL. The DEL facilitated identification/annotation of 123 metabolites associate with exposure to (poly)phenol enriched diets, which included aromatic ketones, benzoic acids, ellagic acids, caffeoylquinic acids, catecholamines, coumarins, hippuric acid, hydroxytoluenes, phenylamines, stilbenes, urolithins, valerolactones, and xanthonoids, in untargeted metabolomics datasets acquire from human plasma and urine reference materials. The DEL focusing on (poly)phenols and their metabolites of dietary exposure facilitated identification/annotation of ingested food components and their associated pathways in untargeted metabolomics datasets acquired from human biospecimens. The DEL continues to expand with the aim to provide evidence-based data for dietary metabolites in exposome research and inform the development of dietary intervention strategies.

Pre- and Post-Resection Urine Metabolic Profiles of Bladder Cancer Patients: Results of Preliminary Studies on Time Series Metabolomics Analysis.

Simple SummaryBladder cancer is one of the most frequently diagnosed cancers worldwide and due to non-specific symptoms, it is often detected at a late stage. For this reason, possible diagnostic alternatives that could be used for non-invasive screening are still being sought. In recent years, metabolomics approach has been frequently used for this type of research, using urine or blood collected from two groups: patients with a given disease and healthy volunteers. Usually, to minimize the impact of between-subject differences, participants of the study are matched in terms of age, gender, or BMI. Another way to rule out the impact of this variability is to analyze samples taken at intervals from the same patient. Therefore, the aim of our study was to validate results obtained using the traditional approach on a small group of patients, from whom samples were taken before and after resection of the bladder tumor, in a given time frame.The incidence of bladder cancer (BCa) has remained high for many years. Nevertheless, its pathomechanism has not yet been fully understood and is still being studied. Therefore, multiplatform untargeted urinary metabolomics analysis has been performed in order to study differences in the metabolic profiles of urine samples collected at three time points: before transurethral resection of bladder tumor (TURBT), the day after the procedure and two weeks after TURBT. Collected samples were analyzed with the use of high-performance liquid chromatography hyphenated with time-of-flight mass spectrometry detection (HPLC-TOF/MS) and gas chromatography coupled with triple quadrupole mass spectrometry detection (GC-QqQ/MS, in a scan mode). Levels of metabolites selected in our previous study were assessed in order to confirm their potential to differentiate the healthy and diseased samples, regardless of the risk factors and individual characteristics. Hippuric acid, pentanedioic acid and uridine confirmed their potential for sample differentiation. Based on the results of statistical analysis for the paired samples (comparison of metabolic profiles of samples collected before TURBT and two weeks after), a set of metabolites belonging to nucleotide metabolism and methylation processes was also selected. Longitudinal studies proved to be useful for the evaluation of metabolic changes in bladder cancer.

Untargeted Metabolomics Identify a Panel of Urinary Biomarkers for the Diagnosis of Urothelial Carcinoma of the Bladder, as Compared to Urolithiasis with or without Urinary Tract Infection in Dogs.

Urothelial carcinoma (UC), the most common urologic cancer in dogs, is often diagnosed late because the clinical signs are shared by other non-malignant lower urinary tract disorders (LUTD). The urine-based BRAFV595E test for UC is highly effective only in certain breeds; hence additional non-invasive biomarkers of UC are needed. Here, urine from dogs with UC (n = 27), urolithiasis (n = 8), or urolithiasis with urinary tract infection (UTI) (n = 8) were subjected to untargeted metabolomics analyses, using GC-TOF-MS for primary metabolites, QTOF-MS for complex lipids, and HILIC-QTOF MS for secondary and charged metabolites. After adjusting for age and sex, we identified 1123 known metabolites that were differentially expressed between UC and LUTD. Twenty-seven metabolites were significant (1.5 ≤ log2FC ≤ −1.5, adjusted p-value < 0.05); however, 10 of these could be attributed to treatment-related changes. Of the remaining 17, 6 (hippuric acid, N-Acetylphenylalanine, sarcosine, octanoylcarnitine, N-alpha-methylhistamine, glycerol-3-galactoside) discriminated between UC and LUTD (area under the ROC curve > 0.85). Of the 6 metabolites, only hippuric acid and N-alpha-methylhistamine were discriminatory in both male (n = 20) and female (n = 23) dogs, while sarcosine was an effective discriminator in several breeds, but only in females. Further investigation of these metabolites is warranted for potential use as non-invasive diagnostic biomarkers of dogs with UC that present with LUTD-related clinical signs.

Maternal microbiota-derived metabolic profile in fetal murine intestine, brain and placenta

BackgroundThe maternal microbiota affects the development of the offspring by microbial metabolites translocating to the fetus. To reveal the spectrum of these molecular mediators of the earliest host-microbe interactions, we compared placenta, fetal intestine and brain from germ-free (GF) and specific pathogen free (SPF) mouse dams by non-targeted metabolic profiling.ResultsOne hundred one annotated metabolites and altogether 3680 molecular features were present in significantly different amounts in the placenta and/or fetal organs of GF and SPF mice. More than half of these were more abundant in the SPF organs, suggesting their microbial origin or a metabolic response of the host to the presence of microbes. The clearest separation was observed in the placenta, but most of the molecular features showed significantly different levels also in the fetal intestine and/or brain. Metabolites that were detected in lower amounts in the GF fetal organs included 5-aminovaleric acid betaine, trimethylamine N-oxide, catechol-O-sulphate, hippuric and pipecolic acid. Derivatives of the amino acid tryptophan, such as kynurenine, 3-indolepropionic acid and hydroxyindoleacetic acid, were also less abundant in the absence of microbiota. Ninety-nine molecular features were detected only in the SPF mice. We also observed several molecular features which were more abundant in the GF mice, possibly representing precursors of microbial metabolites or indicators of a metabolic response to the absence of microbiota.ConclusionsThe maternal microbiota has a profound impact on the fetal metabolome. Our observations suggest the existence of a multitude of yet unidentified microbially modified metabolites which pass through the placenta into the fetus and potentially influence fetal development.

Constraints on the utilization of cereal straw in lactating dairy cows: A review from the perspective of systems biology.

Cereal straw, a human inedible crop byproduct, can be used as a roughage source in ruminants. However, the nutrition density and palatability are very low, limiting its efficient utilization in animal production. This review aims to systematically provide an overview of the limitations of cereal straws, which is crucial for developing new strategies to enhance the efficient use of cereal straws by lactating dairy cows. Evolutionary molecular biology makes it possible to comprehensively understand the limitations of using cereal straw as a roughage source in dairy cows by different techniques, e.g., multi-omics. Main constraints for utilization of cereal straw and stover in lactating dairy cows include low contents of easily fermented carbohydrates (pectin) and essential amino acids (Met, Phe, and branched-chain amino acids), high content of lignin and silica, and low nutrient digestibility. These cause insufficient supply of the precursors for milk synthesis and result in increased loss of nutrients in feces and urine. Several molecular mechanisms are revealed by multi-omics techniques, including changed amino acid and glucose metabolism, altered rumen microbial composition and function, and differential expression of miRNAs, mRNA, and protein in multi-organs that are associated with milk synthesis. These can be targets of approaches to improve the utilization of cereal straw by dairy cows. In addition, much attention should be given to the efficient countermeasures, including pretreatments by fibrolytic enzymes or steam explosion, dietary formulations such as supplement of pectin, methionine, and branched-chain amino acids, and feeding with other functional feedstuffs, which may improve the feeding and economic value of cereal straw for lactating dairy cows. The newly revealed functional genes (such as BAG3 in the rumen, PC in the liver, CSN1S2 in the mammary gland) and biomarkers (hippuric acid) as well as the integrative signaling and metabolic pathways (phenylalanine metabolism) related to the shortages of cereal straws could be used as nutritional or genetic regulatory targets to improve dairy cow production.

Assessment of Fruit and Vegetables Intake with Biomarkers in Children and Adolescents and Their Level of Validation: A Systematic Review.

Fruit and vegetables (FV) are part of a healthy diet and should be frequently consumed already at a young age. However, intake of FV is difficult to assess in children and adolescents due to various misreporting aspects. Thus, measurement of dietary biomarkers may be a promising alternative to assess FV intake more objectively at young age. To date, dietary biomarkers have been primarily studied in adults, and research focused on their usefulness in children is scarce. However, clinical studies have revealed important differences between children and adults, most importantly in their gut microbiome composition, resulting in differences in postprandial metabolism, as well as in food choices and meal compositions that may influence individual biomarker levels. Therefore, the present review aimed to identify biomarkers of FV intake (BFVI) currently available in children and adolescents and to explore whether there are any differences in the BFVI profile above between children and adolescents and adults. In addition, the current level of validation of BFVI in children and adolescents was examined. In total, 28 studies were eligible for this review, and 18 compounds were identified as potential biomarkers for FV intake in children and adolescents. Carotenoid concentration in skin was a valuable biomarker for total FV intake for both children and adult populations. Common BFVI in blood in adults (e.g., carotenoids and vitamin C) showed inconsistent results in children and adolescents. Biomarkers particularly useful in children included urinary hippuric acid as a biomarker of polyphenolic compound intake originating from FV and the combination of N-methylnicotinic acid and acetylornithine as a biomarker of bean intake. Further studies are needed to assess their kinetics, dose–response, and other validation aspects. There is limited evidence so far regarding valid BFVI in children and adolescents. Thus, to put BFVI into practice in children and adolescents, further studies, particularly based on metabolomics, are needed to identify and validate BFVI that can be used in future epidemiological studies.