Metabolites In Fluids Research Articles

Metabolomic profiles and microbiota of GDM offspring: The key for future perspective?

Gestational diabetes mellitus (GDM), or any degree of glucose intolerance recognized for the first time during pregnancy, is one of the diseases that most frequently aggravates the course of gestation. Missed or late diagnosis and inadequate treatment are associated with high maternal and fetal morbidity, with possible short- and long-term repercussions. Estimates on the prevalence of GDM are alarming and increasing by about 30% in the last 10–20 years. In addition, there is the negative influence of the SARS-CoV-2 emergency on the glycemic control of pregnant women, making the matter increasingly topical. To date, knowledge on the metabolic maturation of newborns is still incomplete. However, in light of the considerable progress of the theory of “developmental origins of health and disease,” the relevant role of the intrauterine environment cannot be overlooked. In fact, due to the high plasticity of the early stages of development, some detrimental metabolic alterations during fetal growth, including maternal hyperglycemia, are associated with a higher incidence of chronic diseases in adult life. In this context, metabolomic analysis which allows to obtain a detailed phenotypic portrait through the dynamic detection of all metabolites in cells, tissues and different biological fluids could be very useful for the early diagnosis and prevention of complications. Indeed, if the diagnostic timing is optimized through the identification of specific metabolites, the detailed understanding of the altered metabolic pathway could also allow better management and more careful monitoring, also from a nutritional profile, of the more fragile children. In this context, a further contribution derives from the analysis of the intestinal microbiota, the main responsible for the fecal metabolome, given its alteration in pregnancies complicated by GDM and the possibility of transmission to offspring. The purpose of this review is to analyze the available data regarding the alterations in the metabolomic profile and microbiota of the offspring of mothers with GDM in order to highlight future prospects for reducing GDM-related complications in children of mothers affected by this disorder.

446 Metabolites in early life bronchoalveolar fluid associate with future bronchiectasis risk in children with cystic fibrosis

446 Metabolites in early life bronchoalveolar fluid associate with future bronchiectasis risk in children with cystic fibrosis

Decreased cerebrospinal fluid kynurenic acid in epileptic spasms: A biomarker of response to corticosteroids.

SummaryBackgroundEpileptic (previously infantile) spasms is the most common epileptic encephalopathy occurring during infancy and is frequently associated with abnormal neurodevelopmental outcomes. Epileptic spasms have a diverse range of known (genetic, structural) and unknown aetiologies. High dose corticosteroid treatment for 4 weeks often induces remission of spasms, although the mechanism of action of corticosteroid is unclear. Animal models of epileptic spasms have shown decreased brain kynurenic acid, which is increased after treatment with the ketogenic diet. We quantified kynurenine pathway metabolites in the cerebrospinal fluid (CSF) of infants with epileptic spasms and explored clinical correlations.MethodsA panel of nine metabolites in the kynurenine pathway (tryptophan, kynurenine, kynurenic acid, 3-hydroxykynurenine, xanthurenic acid, anthranilic acid, 3-hydroxyanthranilic acid, quinolinic acid, and picolinic acid) were measured using liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS). CSF collected from paediatric patients less than 3 years of age with epileptic spasms (n=34, 19 males, mean age 0.85, median 0.6, range 0.3–3 yrs) were compared with other epilepsy syndromes (n=26, 9 males, mean age 1.44, median 1.45, range 0.3–3 yrs), other non-inflammatory neurological diseases (OND) (n=29, 18 males, mean age 1.47, median 1.6, range 0.1–2.9 yrs) and inflammatory neurological controls (n=12, 4 males, mean age 1.80, median 1.80, range 0.8–2.5 yrs).FindingsThere was a statistically significant decrease of CSF kynurenic acid in patients with epileptic spasms compared to OND (p<0.0001). In addition, the kynurenic acid/kynurenine (KYNA/KYN) ratio was lower in the epileptic spasms subgroup compared to OND (p<0.0001). Epileptic spasms patients who were steroid responders or partial steroid responders had lower KYNA/KYN ratio compared to patients who were refractory to steroids (p<0.005, p<0.05 respectively).InterpretationThis study demonstrates decreased CSF kynurenic acid and KYNA/KYN in epileptic spasms, which may also represent a biomarker for steroid responsiveness. Given the anti-inflammatory and neuroprotective properties of kynurenic acid, further therapeutics able to increase kynurenic acid should be explored.FundingFinancial support for the study was granted by Dale NHMRC Investigator grant APP1193648, Petre Foundation, Cerebral Palsy Alliance and Department of Biochemistry at the Children's Hospital at Westmead. Prof Guillemin is funded by NHMRC Investigator grant APP1176660 and Macquarie University.

An Updated Review of Recent Advances in Neurosyphilis.

Neurosyphilis is caused by Treponema pallidum invading the central nervous system, of which the incidence is increasing worldwide. Due to its variable clinical manifestations, diagnosis of neurosyphilis remains challenging, especially the asymptomatic form. This review focuses on recent advances in neurosyphilis, including epidemiology, clinical manifestations, laboratory findings, comorbidities, diagnosis, treatment, prognosis, and basic research. The expansion of men who have sex with men and the infection of human immunodeficiency virus mainly accounted for the increasing incidence of neurosyphilis. The rate of some historically described forms of neurosyphilis in the pre-antibiotic era declined significantly; atypical features are more prevalent. Neurosyphilis, regarded as a great mimicker for neuro-ophthalmic, audio-vestibular, and psychiatric disorders, often presents concomitantly with other diseases, including metabolic disorders. Studies on long non-coding RNAs, miRNAs, chemokines, and metabolites in peripheral blood and cerebrospinal fluid may facilitate exploring the pathogenesis and identifying novel biomarkers of neurosyphilis. The drug resistance of Treponema pallidum to penicillin has not been reported; ceftriaxone was proposed to be more effective than penicillin, whereas few randomized controlled trials supported this view. This study may pave the way for further research, especially the diagnosis and treatment of neurosyphilis.

A Comprehensive Analysis of Microflora and Metabolites in the Development of Ulcerative Colitis into Colorectal Cancer Based on the Lung-Gut Correlation Theory.

The lungs and large intestine can co-regulate inflammation and immunity through the lung–gut axis, in which the transportation of the gut microbiota and metabolites is the most important communication channel. In our previous study, not only did the composition of the gut microbiota and metabolites related to inflammation change significantly during the transition from ulcerative colitis (UC) to colorectal cancer (CRC), but the lung tissues also showed corresponding inflammatory changes, which indicated that gastrointestinal diseases can lead to pulmonary diseases. In order to elucidate the mechanisms of this lung–gut axis, metabolites in bronchoalveolar lavage fluid (BALF) and lung tissues were detected using UHPLC–Q-TOF-MS/MS technology, while microbiome characterization was performed in BALF using 16S rDNA sequencing. The levels of pulmonary metabolites changed greatly during the development of UC to CRC. Among these changes, the concentrations of linoleic acid and 7-hydroxy-3-oxocholic acid gradually increased during the development of UC to CRC. In addition, the composition of the pulmonary microbiota also changed significantly, with an increase in the Proteobacteria and an obvious decrease in the Firmicutes. These changes were consistent with our previous studies of the gut. Collectively, the microbiota and metabolites identified above might be the key markers related to lung and gut diseases, which can be used as an indication of the transition of diseases from the gut to the lung and provide a scientific basis for clinical treatment.

A study of connective tissue metabolites in biological fluids of patients with various types of glaucoma

PURPOSE. To quantify the content of connective tissue metabolites reflecting the exchange of its main components in biological fluids in low tension glaucoma (LTG) and primary open-angle glaucoma (POAG), and to conduct a comparative analysis of the obtained data. METHODS. The studies were conducted in 33 patients with low tension glaucoma aged 51 to 80 years old (mean age 69 ± 10.4 years), 20 patients with POAG aged 53 to 80 years old (mean age 67 ± 10.1 years) and 15 clinically healthy volunteers aged 52 to 78 years old (mean age 68 ± 9.2 years). The protein components of the fibrous part of the connective tissue were analyzed for daily excretion of oxyproline in urine by the method involving para-Dimethylamin-obenzaldehyde modified by P.N. Sharaev. Serum glycosaminoglycans (GAG) were studied by analyzing carbohydrate components of connective tissue. The concentration of GAG in the blood serum was determined by the colorimetric method, using Dische's carbazole reaction. RESULTS. The amount of GAG in the blood serum of LTG patients was 3.26 ± 1.57 mmol/L, while in the control group consisting of clinically healthy volunteers — 2.92 ± 1.25 mmol/L. The differences were not statistically significant (p &gt; 0.1). The amount of daily excretion of oxyproline in the urine of LTG patients was 17.17 ± 8.64 mmol/day, and in the group of clinically healthy individuals — 13.96 ± 5.61 mmol/day. The differences were statistically significant (p &lt; 0.05). The amount of GAG in the blood serum of POAG patients was 3.97 ± 1.17 mmol/L. The differences in relation to LTG were statistically significant (p &lt; 0.05). The amount of daily excretion of oxyproline in the urine of POAG patients was 32.71 ± 19.79 mmol/day. The differences were statistically significant in relation to LTG (p &lt; 0.05). The amount of GAG and oxyproline in biological fluids tends to increase with the advancement of LTG and POAG. CONCLUSION. Destruction processes in the connective tissue do occur in LTG, but are less pronounced in comparison to POAG and more pronounced in comparison to healthy individuals. The indicator that most clearly reflects the activity of destructive processes in the connective tissue is the daily excretion of oxyproline in urine.

Comparison of metabolites in rumen fluid, urine, and feces of dairy cow from subacute ruminal acidosis model measured by proton nuclear magnetic resonance spectroscopy.

In this study, metabolites that changed in the rumen fluid, urine and feces of dairy cows fed different feed ratios were investigated. Eight Holstein cows were used in this study. Rumen fluid, urine, and feces were collected from the normal concentrate diet (NCD) (Italian ryegrass 80%: concentrate 20% in the total feed) and high concentrate diet (HCD) groups (20%: 80%) of dairy cows. Metabolite analysis was performed using proton nuclear magnetic resonance (NMR) identification, and statistical analysis was performed using Chenomx NMR software 8.4 and Metaboanalyst 4.0. The two groups of rumen fluid and urine samples were separated, and samples from the same group were aggregated together. On the other hand, the feces samples were not separated and showed similar tendencies between the two groups. In total, 160, 177, and 188 metabolites were identified in the rumen fluid, urine, and feces, respectively. The differential metabolites with low and high concentrations were 15 and 49, 14 and 16, and 2 and 2 in the rumen fluid, urine, and feces samples, in the NCD group. As HCD is related to rumen microbial changes, research on different metabolites such as glucuronate, acetylsalicylate, histidine, and O-Acetylcarnitine, which are related to bacterial degradation and metabolism, will need to be carried out in future studies along with microbial analysis. In urine, the identified metabolites, such as gallate, syringate, and vanillate can provide insight into microbial, metabolic, and feed parameters that cause changes depending on the feed rate. Additionally, it is thought that they can be used as potential biomarkers for further research on subacute ruminal acidosis.

The role of analytical pharmacotoxicology in adressing the main functions of the National Early Warning System on NPS

In compliance with the European Council Decision 2005/387/JHA, the Italian AntiDrug Policies Department designated the National Center for Addiction and Doping of the Istituto Superiore di Sanità (ISS) to organize and manage the National Early Warning System (NEWS) on New Psychoactive Substances (NPS) to promote a rapid exchange of information on NPS within Italy and between Italy and the EU. The analytical pharmacotoxicology Unit of the Center supports NEWS with screening and confirmation of NPS presence in conventional and non-conventional biological matrices from acute and fatal intoxication cases. Furthermore, yearly, pure standards of new NPS are tested and provided to Laboratories from collaborative centres and proficiency testing on NPS in oral fluid and hair are organized. Different analytical technologies, including high-sensitivity gas chromatography-mass spectrometry (GC-MS) and tandem mass spectrometry (GC-MS-MS), liquid chromatography-tandem mass spctrometry (LC-MS-MS) and ultra-high-performance liquid chromatography-high-resolution mass spectrometry (UHPLC-HRMS) are used for a screening and confirmation analysis of classic drugs, prescription medicines and new psychoactive substances in and their metabolites in blood, plasma, urine, hair and oral fluid from intoxication cases and clinical trials. Sample preparation involves both liquid-liquid or solid phase extraction. Dilute and shoot methodology is also applied. During 2021 and first months of 2022, around 80 different psychoactive substances, including new synthetic opioids and/or metabolites were identified in urine from 296 spanish patients with a history of opioid use disorder; 12 classic drugs and NPS were detected in hair from 300 mexican pregnant women; cocaine, THC, heroin, methadone, ketamine and mephedrone and metabolites were determined in urine from 76 italian prisoners; JWH-122 and JWH-210, UR-144 were quantified in oral fluid from consumers and finally some cases of intoxications with fentanyl, norfentanyl and analogs were also disclosed. Furthermore, two exercises of Proficiency testing for NPS in oral fluid and hair were organized. Samples from both matrices were prepared in cooperation with Comedical (Trento, Italy). For the 2021 round classic drugs together with 5-fluoro CUMYL-PeGACLONE, 5-fluoro MDMB-PICA, AB-FUBINACA,5F-ADB, Ethylone, Euthylone, Furanylfentanyl, Ocfentanyl, Isobutyryl fentanyl, Fentanyl and Isotonitazene were included. For the 2022 round, classic drugs plus 5-MMPA, Methoxpropamine, Brorphine, Butonitazene, Etodesnitazene, Flunitazene, 4-fluoro MDMB-BUTICA, 5-fluoro CUMYL—PICA, 5-fluoro EDMB-PICA, 3-Methylmethcathinone, ADB-4en-PINACA, MDMB-4en-PICA were comprises. About 20 different pharmacotoxicological national laboratories participated in the exercised with very variable quali-quantitative results. Although the NPS phenomenon is not uniform, it is well known as a global problem, concerning all the region of the world. Due to the mutating nature of the phenomenon, we believe that investments in toxicological and forensic analytical data sources are strongly needed. Differently from the past when psychotropic drugs occupied the fingers of two hands, in the 21th century, more than 1000 NPS have been identified. Not all of them found the same diffusion in the illicit market, but many of them are temporary adulterant of substitute of controlled compounds or finally fake drugs. The values of accurate and immediately available toxicological and forensic data are crucial to the implementation of national and international early warning system on NPS. Indeed, the drug related severe intoxications and fatalities are seldom associated with the consumption of one substance alone. Drug consumption patterns are often characterized by polyconsumption of both classical drugs and NPS and laboratories supporting emergency rooms, forensic toxicologies and police forces must be enabled to identify the substance/s causing the acute or chronic intoxication or a fatality.

Use of solid phase microextraction in tips (SPME-Tips) for analysis of cocaine and biotransformation products in oral fluid by liquid chromatography tandem mass spectrometry (LC-MS/MS)

To develop and validate a method for analyzing cocaine and its metabolites using SPME-Tips and LC-MS/MS. Among the biological matrices used for analytical toxicology, oral fluid has advantages such as simplicity and non-invasive collection, the difficulty of adulteration, the low level of contamination risk and the possibility of recent abuse evaluation. Recently, nearly two million users have used cocaine in the US/Europe. When abused, cocaine is rapidly metabolized, mainly into methylecgonine and benzoylecgonine. Cocaethylene is formed in the concomitant use of cocaine with ethanol, and anhydroecgonine methyl ester (AEME) (in the crack abuse). Currently, in (green) analytical toxicology there is a growing interest in the use of new sample preparation techniques that are fast, efficient, automatable, and ecologically correct. Thus, methods based on microextraction processes have been developed, such as solid phase microextraction (SPME), a miniaturization of solid phase extraction. A variation of SPME technique was recently developed, consisting of a microfiber coated with extractor phase installed in a micropipette tip. In SPME-Tips, the extraction of analytes occurs in a extractor phase, allowing the direct immersion of the tip in biological fluids, causing the analytes to be adsorbed on the fiber and allowing the desorption to be carried out using small volume of more polar and less toxic solvents. The procedure was optimized, studying parameters such as salting-out effect, extraction time, sample agitation speed, elution solvent volume, and constitution. For extraction, SPME-Tips were immersed in 150 μL of oral fluid (collected with Quantisal) and kept under agitation at 500 rpm for 80 min. For the elution, 125 μL of methanolic solution with 0.1% formic acid was used, kept under agitation at 500 rpm for 30 min. The elution content was taken to LC-MS/MS, in which 5 μL were analyzed. The analyses were performed in a Nexera-UHPLC chromatographic system, coupled with a LCMS8060 mass spectrometer. The chromatographic separation was performed with Atlantis-T3 column (150 × 3.0 mm, 3.0 μm), maintained at 40 °C. The mobile phase consisted of ultra-pure water (A) and methanol (B), both containing formic acid (0.1%) and 2 mmol/L ammonium formate. The flow rate was set to 0.4 mL/min, and the elution gradient used started with 10%B for 1 min, followed by a linear increase to 100%B for 2 min and then maintained at 100%B for 2 min and returning to the proportion start in 0.1 min. This condition was maintained for another 3 min, completing a total of 8 min of chromatographic run. With this optimization, the method validation began, as recommended by the AAFS Standard Practices for Method Validation in Forensic Toxicology. For cocaine, benzoylecgonine, and cocaethylene, the correlation coefficient r 2 was greater than 0.98 (50 to 500 ng/mL). Good accuracy and precision results were obtained for low, medium and high QC. In this way, low extraction efficiency was observed to AEME and methylecgonine in C18 SPME-Tips. For the other analytes, the precision and accuracy values were in accordance to the criteria of the method validation guideline. Also, to prove the applicability of the method in real situations, 10 samples of oral fluid collected from individuals who participated in university parties will be analyzed. The samples were already collected and are stored in the Laboratory. A green analytical toxicology method was developed using SPME-Tips for the analysis of cocaine and its metabolites in oral fluid.

Simultaneous determination and pharmacokinetics study of valsartan, sacubitril and its major metabolite in human plasma, urine and peritoneal dialysis fluid in patients with end-stage renal disease by UPLC–MS/MS

Sacubitril/valsartan was indicated for the treatment of heart failure and hypertension in patients with end-stage renal disease on peritoneal dialysis therapy. Herein, a rapid, sensitive and robust method based on ultra-liquid chromatography–tandem mass spectrometry (UPLC–MS/MS) was developed and validated for the determination of the concentrations of valsartan, sacubitril and its major metabolite LBQ657 in plasma, urine and peritoneal dialysis fluid. Samples were extracted from various biological fluids using protein precipitation. Extracts were subjected to UPLC–MS/MS with electrospray ionization in positive-ion mode. The developed method was fully validated over a concentration range of 8.00–8000 ng/mL for valsartan, 2.00–2000 ng/mL for sacubitril and 30.0–30,000 ng/mL for LBQ657 in plasma, 2.00–1000 ng/mL for valsartan, 1.00–500 ng/mL for sacubitril and 20.0–10000 ng/mL for LBQ657 in urine, and 0.200–100 ng/mL for valsartan, 0.0400–20.0 ng/mL for sacubitril and 2.00–1000 ng/mL for LBQ657 in peritoneal dialysis fluid. The intra- and inter-day precisions for all analytes in various matrices were less than 12.3%, and the intra- and inter-day accuracies results were all between 88.0% and 109.2%. All analytes were stable for at least 8 h at room temperature (25°C), five freeze–thaw cycles, and 37 days at −40°C and −80°C in plasma, urine and peritoneal dialysis fluid. In conclusion, this developed method is reliable, sensitive and specific for determining the concentrations of valsartan, sacubitril and LBQ657 in plasma, urine and peritoneal dialysis fluid. The assay was available to pilot the pharmacokinetics study of sacubitril/valsartan in patients with end-stage renal disease on peritoneal dialysis, and it could provide evidence that peritoneal dialysis had an effect on the clearance of sacubitril/valsartan.

Effect of transformed sunflower husk on enzymatic processes in the rumen &lt;I&gt;in vitro&lt;/I&gt;

Fermentation processes in the rumen of ruminants when using sunflower husk subjected to ultrasound treatment in combination with phytobiotics were studied. During the experiment, the formation of terminal metabolites in the ruminal fluid as well as the formation of methane were determined. The object of the study was the rumen fluid, which was selected from bulls of Kazakh whiteheaded breed at the age of 12 months with the average weight of 230-235 kg. Chronic rumen fistula (n = 3) was identified according to the method of A.A. Aliev. The following samples were incubated by in vitro method using the ANKOM Daisy II device (modifications D200 and D200I) according to a specialized technique: control (sample N 1): Mechanically ground husk + hydromodulus (water) + ultrasound treatment at 20 °C, 15 min, 27 kHz; sample N 2: sample N 1 + gamma-octalactone (0.25 ml); sample N 3: sample N 1 + quercetin (10.0 mg); sample N 4: sample N 1 + 7-hydroxycoumarin (2.0 mg). Each experiment was conducted in five replications. The digestibility of dry matter, concentration of volatile fatty acids, formation of nitrogenous metabolites and methane concentration were determined. The data obtained were statistically analyzed using software. New data were obtained on the effect of sunflower husk together with phytobiotics on the formation of final metabolites in the rumen fluid. The addition of biologically active substances of gamma-octalactone, quercetin, 7-hydroxycoumarin was found to increase the digestibility of dry matter relative to control by 2.0 (p ≤ 0.05), 3.1 (p ≤ 0.01) and 4.3% (p ≤ 0.05), respectively. An increase in the concentration of volatile fatty acids and nitrogenous fractions was noted when using these substances. The level of methane concentration was 10% lower than that of the control with 7-hydroxycoumarin in sample N 4 (p ≤ 0.01).

Modulation of expression of estrus, steroidogenesis and embryo development following peri-artificial insemination nutrient restriction in beef heifers

Nutritional changes immediately after insemination cause increased embryonic mortality, but the mechanisms controlling this are not well known. Our objective was to evaluate the impact of nutritional change on estrus expression, steroid concentrations, peripheral and uterine luminal fluid metabolites, and embryo quality in beef heifers. Heifers (n = 139) were assigned to one of two pre-artificial insemination (AI) dietary treatments: LOW (≤ 90% NEm) or HIGH (≥ 139% NEm). Heifers were on treatment for 33–36 days before AI (d0) when half of the heifers in each treatment were randomly reassigned to generate four treatments; HIGH-HIGH, HIGH-LOW, LOW-HIGH, and LOW-LOW. Heifers remained on treatments until embryo collection (d 6–8). Negative energy balance was achieved among LOW heifers as demonstrated by body weight loss and increased NEFA concentrations (P < 0.05). Pre-AI treatment influenced expression of estrus (P = 0.05; HIGH 80.4 ± 4.0% vs. LOW 69.4 ± 4.2%). Estradiol concentrations and interval to estrus were not affected by treatment (P > 0.55); however, progesterone concentrations were reduced among LOW compared to HIGH (3.57 ± 0.27, 4.64 ± 0.26 ng/mL, respectively; P = 0.004), and heifers maintained on the HIGH pre-AI diet had consistently greater concentrations of progesterone from d 0 to d 8 (P = 0.014). Pre-AI treatment influenced embryo stage (P = 0.05; HIGH 3.61 ± 0.32 vs. LOW 2.72 ± 0.30). Post-AI treatment affected embryo grade (P = 0.02; HIGH 1.78 ± 0.23 vs. LOW 2.64 ± 0.27). In summary, pre-AI nutrient restriction caused decreased expression of estrus, reduced progesterone concentrations after AI, and negatively impacted embryo development, while post-AI restriction hindered embryo quality.

Progressive microbial adaptation of the bovine rumen and hindgut in response to a step-wise increase in dietary starch and the influence of phytogenic supplementation.

Microbial composition and activity in the gastrointestinal tract (GIT) of cattle has important implications for animal health and welfare, driving the focus of research toward ways to modify their function and abundance. However, our understanding of microbial adaption to nutritional changes remains limited. The aim of this study was to examine the progressive mechanisms of adaptation in the rumen and hindgut of cattle receiving increasing amounts of starch with or without dietary supplementation of a blended phytogenic feed additive (PFA; containing menthol, thymol and eugenol). We used 16S rRNA gene amplicon sequencing to assess the microbial composition and predicted metabolic pathways in ruminal solid and liquid digesta, and feces. Furthermore, we employed targeted liquid chromatography-mass spectrometry methods to evaluate rumen fluid metabolites. Results indicated a rapid microbial adaptation to diet change, starting on the second day of starch feeding for the particle associated rumen liquid (PARL) microbes. Solid rumen digesta- and feces-associated microbes started changing from the following day. The PARL niche was the most responsive to dietary changes, with the highest number of taxa and predicted pathways affected by the increase in starch intake, as well as by the phytogenic supplementation. Despite the differences in the microbial composition and metabolic potential of the different GIT niches, all showed similar changes toward carbohydrate metabolism. Metabolite measurement confirmed the high prevalence of glucose and volatile fatty acids (VFAs) in the rumen due to the increased substrate availability and metabolic activity of the microbiota. Families Prevotellaceae, Ruminococcaceae and Lachnospiraceae were found to be positively correlated with carbohydrate metabolism, with the latter two showing wide-ranging predicted metabolic capabilities. Phytogenic supplementation affected low abundant taxa and demonstrated the potential to prevent unwanted implications of feeding high-concentrate diet, such as reduction of microbial diversity. The inclusion of 50% concentrate in the diet caused a major shift in microbial composition and activity in the GIT of cattle. This study demonstrated the ability of microorganisms in various GIT niches to adjust differentially, yet rapidly, to changing dietary conditions, and revealed the potential beneficial effects of supplementation with a PFA during dietary adaptation.

Recent Advances in Garcinia cambogia Nutraceuticals in Relation to Its Hydroxy Citric Acid Level. A Comprehensive Review of Its Bioactive Production, Formulation, and Analysis with Future Perspectives.

Garcinia cambogia (Gaertn.) Desr. (known as Malabar tamarind) is a popular traditional herbal medicine and is one of the well-known folk medicines reported for the treatment of obesity and incorporated in several nutraceuticals worldwide. These effects are mediated by a myriad of bioactive compounds with most effects attributed to its hydroxy citric acid (HCA) content. This review aims to present a holistic overview on novel trends in the production of G. cambogia bioactive components and how extraction optimization is important to ensure best product quality with its reported nanoformulations with particular emphasis on HCA content. Further, an overview of the different analytical approaches used for quality control assessment of G. cambogia plant and its nutraceuticals is presented highlighting both advantages and limitations. Moreover, analytical approaches for detecting G. cambogia metabolites in biological fluids with emphasis on HCA level to determine its pharmacokinetics and proof of efficacy are presented for the first time.

Metabolomics: A New Approach in the Evaluation of Effects in Human Beings and Wildlife Associated with Environmental Exposition to POPs

Human beings and wild organisms are exposed daily to a broad range of environmental stressors. Among them are the persistent organic pollutants that can trigger adverse effects on these organisms due to their toxicity properties. There is evidence that metabolomics can be used to identify biomarkers of effect by altering the profiles of endogenous metabolites in biological fluids or tissues. This approach is relatively new and has been used in vitro studies mainly. Therefore, this review addresses those that have used metabolomics as a key tool to identify metabolites associated with environmental exposure to POPs in wildlife and human populations and that can be used as biomarkers of effect. The published results suggest that the metabolic pathways that produce energy, fatty acids, and amino acids are commonly affected by POPs. Furthermore, these pathways can be promoters of additional effects. In the future, metabolomics combined with other omics will improve understanding of the origin, development, and progression of the effects caused by environmental exposure.

Metabolic discrimination of synovial fluid between rheumatoid arthritis and osteoarthritis using gas chromatography/time-of-flight mass spectrometry

Rheumatoid arthritis (RA) and osteoarthritis (OA) are clinicopathologically different. We aimed to assess the feasibility of metabolomics in differentiating the metabolite profiles of synovial fluid between RA and OA using gas chromatography/time-of-flight mass spectrometry. We first compared the global metabolomic changes in the synovial fluid of 19 patients with RA and OA. Partial least squares-discriminant, hierarchical clustering, and univariate analyses were performed to distinguish metabolites of RA and OA. These findings were then validated using synovial fluidsamples from another set of 15 patients with RA and OA. We identified 121 metabolites in the synovial fluid of the first 19 samples. The score plot of PLS-DA showed a clear separation between RA and OA. Twenty-eight crucial metabolites, including hypoxanthine, xanthine, adenosine, citrulline, histidine, and tryptophan, were identified to be capable of distinguishing RA metabolism from that of OA; these were found to be associated with purine and amino acid metabolism. Our results demonstrated that metabolite profiling of synovial fluid could clearly discriminate between RA and OA, suggesting that metabolomics may be a feasible tool to assist in the diagnosis and advance the comprehension of pathological processes for diseases.

Preovulatory follicular fluid and serum metabolome profiles in lactating beef cows with thin, moderate, and obese body condition.

Extremes in body condition reduce fertility and overall productivity in beef cattle herds, due in part to altered systemic metabolic conditions that influence the intrafollicular and uterine environment. Follicular fluid and serum metabolome profiles are influenced by body composition in women and dairy cattle; however, such information is lacking in beef cattle. We hypothesized that body condition score (BCS)-related alterations in the metabolome of preovulatory follicular fluid and serum may influence oocyte maturation while impacting the oviductal or uterine environment. Therefore, we performed a study with the objective to determine the relationship between BCS and the metabolome of follicular fluid and serum in lactating beef cattle. We synchronized the development of a preovulatory follicle in 130 cows of varying BCS. We collected blood and performed transvaginal follicle aspirations to collect follicular fluid from the preovulatory follicle ~18 h after gonadotropin-releasing hormone administration to stimulate the preovulatory gonadotropin surge. We then selected follicular fluid and serum samples from cows with BCS 4 (Thin; n = 14), BCS 6 (Moderate; n = 18), or BCS >8 (Obese; n = 14) for ultra-high performance liquid chromatography-high resolution mass spectrometry. We identified differences in the follicular fluid or serum of thin, moderate, and obese animals based on multiple linear regression. MetaboAnalyst 5.0 was used for enrichment analysis of significant metabolites. We identified 38 metabolites in follicular fluid and 49 metabolites in serum. There were no significant differences in follicular fluid metabolite content among BCS classifications. There were 5, 22, and 1 serum metabolites differentially abundant between thin-obese, moderate-thin, and moderate-obese classifications, respectively (false discovery rate [FDR] < 0.10). These metabolites were enriched in multiple processes including "arginine biosynthesis," "arginine/proline metabolism," and "D-glutamine/D-glutamate metabolism" (FDR < 0.04). Pathways enriched with serum metabolites associated with BCS indicate potentially increased reactive oxygen species (ROS) in serum of thin cows. ROS crossing the blood follicular barrier may negatively impact the oocyte during oocyte maturation and contribute to the reduced pregnancy rates observed in thin beef cows.

Preovulatory serum estradiol concentration is positively associated with oocyte ATP and follicular fluid metabolite abundance in lactating beef cattle.

Cattle induced to ovulate a small, physiologically immature preovulatory follicle had reduced oocyte developmental competence that resulted in decreased embryo cleavage and day 7 embryo quality compared with animals induced to ovulate a more advanced follicle. RNA-sequencing was performed on oocytes and their corresponding cumulus cells approximately 23 h after gonadotropin-releasing hormone (GnRH) administration to induce the preovulatory gonadotropin surge suggested reduced capacity for glucose metabolism and oxidative phosphorylation in the cumulus cells and oocytes from follicles ≤11.7 mm, respectively. We hypothesized that induced ovulation of a small, physiologically immature preovulatory follicle results in a suboptimal follicular microenvironment and reduced oocyte metabolic capacity. We performed a study with the objective to determine the impact of preovulatory follicle diameter and serum estradiol concentration at GnRH administration on oocyte metabolic competence and follicular fluid metabolome profiles. We synchronized the development of a preovulatory follicle and collected the follicle contents via transvaginal aspiration approximately 19 h after GnRH administration in lactating beef cows (n = 319). We determined ATP levels and mitochondrial DNA (mtDNA) copy number in 110 oocytes and performed ultra-high-performance liquid chromatography–high resolution mass spectrometry metabolomic studies on 45 follicular fluid samples. Intraoocyte ATP and the amount of ATP produced per mtDNA copy number were associated with serum estradiol concentration at GnRH and time from GnRH administration to follicle aspiration (P < 0.05). mtDNA copy number was not related to follicle diameter at GnRH, serum estradiol concentration at GnRH, or any potential covariates (P > 0.10). We detected 90 metabolites in the aspirated follicular fluid. We identified 22 metabolites associated with serum estradiol concentration at GnRH and 63 metabolites associated with follicular fluid progesterone concentration at the time of follicle aspiration (FDR < 0.10). Pathway enrichment analysis of significant metabolites suggested altered proteinogenesis, citric acid cycle, and pyrimidine metabolism in follicles of reduced estrogenic capacity pre-gonadotropin surge or reduced progesterone production by the time of follicle aspiration.

Ruminal Bacterial Communities and Metabolome Variation in Beef Heifers Divergent in Feed Efficiency

This study determined potential microbial and metabolic biomarkers of feed efficiency in Angus heifers. Seventeen ruminal cannulated Angus heifers underwent a 70-day feed efficiency trial. Residual feed intake was used to determine high and low feed efficient heifers. On day 70 of the trial, rumen content and blood were collected and used for microbial and metabolomic analyses, respectively. Bacterial populations were examined by targeting the V4 region of the 16S rRNA gene and analyzed using QIIME and SAS. Rumen fluid and serum metabolites were analyzed using MetaboAnalyst. No microbial taxa differed after false discovery rate correction, but seven did differ (p ≤ 0.05) prior to correction, including Lachnospiraceae (Other), Desulfobulbaceae, Neisseriaceae, Shuttleworthia, Corynebacterium, p-75-a5, and L7A-E11. No differences were observed in alpha diversity metrics. Beta diversity utilizing unweighted UniFrac distances analyzed via PERMANOVA was significant (p = 0.03). Several metabolites in rumen fluid metabolites were correlated with bacteria that differed by feed efficiency phenotype. The metabolites correlated with bacteria were primarily involved in nutrient signaling and microbial crude protein availability. These data suggest variation in the availability of nutrients, primarily amino acids, as well as a relationship among microbiota, metabolome, and host feed efficiency phenotypes in heifers.

An Overview on the Application of Solid Phase Microextraction in the Analysis of Volatile Organic Compounds as Potential Biomarkers of Cancer

The search for biomarkers of diseases, especially for the early detection of cancer, is one of the most popular research fields in biomedicine. The development of non-invasive screening techniques for the early detection of cancer is one of the greatest scientific challenges of the 21st century. Because various diseases cause to generation of biomarkers in the body, early diagnosis of diseases can be performed by analyzing these biomarkers in all body feces. Detection of volatile organic compounds (VOCs) as biomarkers of the body's metabolic processes is a new frontier in the fast, sensitive, selective and non-invasive analysis and medical diagnosis of human diseases. VOCs as biomarkers in the human body can be distinct and specific to metabolic conditions or diseases. VOCs produced as end products of cellular metabolism and are released through a variety of biological matrices such as breath, blood, saliva, urine and feces. The study of volatile metabolites in the body fluids of the patients is performed to establish "potential biomarkers" for early diagnosis and prognosis of the disease. Various genetic, epigenetic, proteomic, and metabolic markers are evaluated for early detection of cancer. One of the promising emerging methods for early detection of cancer is the analysis of VOCs. The purpose of this review is to provide insights for recent research work on the analysis of VOC-biomarker of cancers with extraction by solid phase microextraction (SPME), and how to detect VOCs in different body matrices as a potential biomarker of disease.