Prediction of clinically significant prostate cancer through urine metabolomic signatures: A large-scale validated study.

PURPOSE: Currently, there are no accurate markers for predicting potentially lethal prostate cancer (PC) before biopsy. This study aimed to develop urine tests to predict clinically significant PC (sPC) in men at risk. METHODS: Urine samples from 928 men, namely, 660 PC patients and 268 benign subjects, were analyzed by gas chromatography/quadrupole time-of-flight mass spectrophotometry (GC/Q-TOF MS) metabolomic profiling to construct four predictive models. Model I discriminated between PC and benign cases. Models II, III, and GS, respectively, predicted sPC in those classified as having favorable intermediate risk or higher, unfavorable intermediate risk or higher (according to the National Comprehensive Cancer Network risk groupings), and a Gleason sum (GS) of ≥ 7. Multivariable logistic regression was used to evaluate the area under the receiver operating characteristic curves (AUC). RESULTS: In Models I, II, III, and GS, the best AUCs (0.94, 0.85, 0.82, and 0.80, respectively; training cohort, N = 603) involved 26, 24, 26, and 22 metabolites, respectively. The addition of five clinical risk factors (serum prostate-specific antigen, patient age, previous negative biopsy, digital rectal examination, and family history) significantly improved the AUCs of the models (0.95, 0.92, 0.92, and 0.87, respectively). At 90% sensitivity, 48%, 47%, 50%, and 36% of unnecessary biopsies could be avoided. These models were successfully validated against an independent validation cohort (N = 325). Decision curve analysis showed a significant clinical net benefit with each combined model at low threshold probabilities. Models II and III were more robust and clinically relevant than Model GS. CONCLUSION: This urine test, which combines urine metabolic markers and clinical factors, may be used to predict sPC and thereby inform the necessity of biopsy in men with an elevated PC risk.

Solar distillation of human urine to recover non-potable water and metal phosphate mineral.

Human urine is a readily available nutrient source that can complement commercial fertilizer production, which relies on finite mineral resources and global supply chains. This study evaluated the effectiveness of a simplified solar distillation process for urine to recover phosphorus (P) and nitrogen for agricultural use and water for non-potable purposes. Synthetic fresh, synthetic hydrolyzed, real fresh, and real hydrolyzed urine were exposed to direct sunlight for 6 h in a simple distillation apparatus, which produced distillation bottoms and distillate. Metal phosphate precipitation in the distillation bottoms was evaluated to recover P. The non-potable water was recovered as distillate. Hydrolyzed urine recovered more metal phosphate solid in the distillation bottoms and had a higher conductivity in the distillate than fresh urine. Hydrolyzed urine also achieved greater distillate volume recovery than fresh urine. Hydrolyzed urine had a greater presence of UV-absorbing organics in the distillate than fresh urine and therefore produced a lower-quality product water. There was no significant correlation between the daily high air temperature and the volume of distillate recovered. This study provides a comprehensive data set on simplified solar distillation of human urine considering the fate of nutrients and water for different types of urine.

Factors influencing the recovery of organic nitrogen from fresh human urine dosed with organic/inorganic acids and concentrated by evaporation in ambient conditions.

To feed the world without transgressing regional and planetary boundaries for nitrogen and phosphorus, one promising strategy is to return nutrients present in domestic wastewater to farmland. This study tested a novel approach for producing bio-based solid fertilisers by concentrating source-separated human urine through acidification and dehydration. Thermodynamic simulations and laboratory experiments were conducted to evaluate changes in chemistry of real fresh urine dosed and dehydrated using two different organic and inorganic acids. The results showed that an acid dose of 1.36 g H2SO4 L-1, 2.86 g H3PO4 L-1, 2.53 g C2H2O4·2H2O L-1 and 5.9 g C6H8O7 L-1 was sufficient to maintain pH ≤3.0 and prevent enzymatic ureolysis in urine during dehydration. Unlike alkaline dehydration using Ca(OH)2 where calcite formation limits the nutrient content of fertiliser products (e.g. <15 % nitrogen), there is greater value proposition in acid dehydration of urine, as the products contain 17.9-21.2 % nitrogen, 1.1-3.6 % phosphorus, 4.2-5.6 % potassium and 15.4-19.4 % carbon. While the treatment recovered all phosphorus, recovery of nitrogen in the solid products was 74 % (±4 %). Follow-up experiments revealed that hydrolytic breakdown of urea to ammonia, chemically or enzymatically, was not the reason for the nitrogen losses. Instead, we posit that urea breaks down to ammonium cyanate, which then reacts with amino and sulfhydryl groups of amino acids excreted in urine. Overall, the organic acids evaluated in this study are promising for decentralised urine treatment, as they are naturally present in food and therefore already excreted in human urine.

An integrated strategy for nutrient harvesting from hydrolyzed human urine as high-purity products: Tracking of precipitation transformation and precise regulation.

The nutrient recovery from source-separated urine is of great significance for a sustainable and closed nutrient loop. However, common urine-processing techniques have several constraints, including inefficient recovery, low product purity and incapability of simultaneously harvesting multiple nutrients. In this study, an integrated strategy of P precipitation and N stripping was first proposed to harvest nutrients from hydrolyzed human urine as high-purity products via precisely regulating Ca/P dosing ratio. Ca(OH)2 was utilized to trigger Ca-P precipitation and elevate pH level. Different from the previously reported conventional struvite method, P recovery was oriented to calcium phosphate. P harvesting behavior was investigated as a function of key factors including initial P concentration and the dosing ratio. A thermodynamic model was constructed to unveil the precipitation transformation mechanism and visualize P recovery for an enhanced controllability. For N harvesting, Ca(OH)2 was dosed to increase the pH of the urine to converts ammonium to ammonia. The resulting ammonia was stripped and then adsorbed by H2SO4 as high-purity ammonium sulfate. Moreover, the sulfate derived from acidification treatment was recovered as calcium sulfate in the interests of material recycling and mitigating secondary contaminations. Results exhibited P recovery efficiency could reach 100 % and purity for calcium phosphate could be above 90 % within a Ca/P ratio range of 1.67-2.0. Further boosting pH to 12, over 85 % of S and 95 % of N was retrieved. The comprehensive scheme provides an efficient approach towards the precise P and N harvesting from hydrolyzed urine and advances the knowledge of precipitation transformation mechanism.

Optimization and kinetic modeling of phosphate recovery as struvite by electrocoagulation from source-separated urine.

Phosphorus recovery is indispensable due to the rapid depletion of its natural reserves and excessive utility in agriculture. Though human urine has high nutrient content including phosphate, nitrogen and potassium; direct use as a fertilizer is restricted due to hygienic, environmental, social and ethical issues. To overcome these limitations, the nutrients are precipitated by the external addition of magnesium (Mg) to form a slow-releasing fertilizer called struvite. The present study aims to maximize phosphate recovery through optimizing struvite production by an emerging electrocoagulation technique. A maximum of 95% phosphate recovery was achieved using inter-electrode distance of 0.5 cm, 2 A current from undiluted urine using Mg-Mg electrodes in a reaction time of 30 min. Further, kinetic modeling of phosphate recovery through electrocoagulation was conducted to comprehend the intended mechanism through the order of kinetics. The results revealed that the data best correlated with first-order kinetics with a correlation coefficient of 0.95. Electrocoagulation improved the supernatant quality by reducing the ion concentrations other than phosphate (30-50%), salinity (40-45%), and microbial population (99%). Qualitative assessment of the precipitate through sophisticated analysis further confirmed the presence of struvite crystals.

Metabolic study of trimetazidine using ultra-high performance liquid chromatography-tandem mass spectrometry

Abstract In the present study, the application of ultra-high performance liquid chromatography-tandem mass spectrometry allowed us to study of known-as well as hitherto unknown-trimetazidine (TMZ) metabolites in human urine and to propose their renal excretion profiles. Urine samples from a healthy volunteer were analyzed at baseline and at 0-4 h, 4-8 h, 8-12 h, and 12-24 h after a single dose of TMZ. A dilute-and-shoot procedure was used as sample treatment before separation. Full-scan spectra of possible metabolites were acquired. Additionally, product ion scan spectra of precursor ions of interest were also acquired at two collision energies. Intact TMZ was a major excretion product, with a maximum concentration at 4-8 h after administration. Moreover, five minor metabolites were observed, namely trimetazidine-N-oxide (M1), N-formyl trimetazidine (M2), desmethyl-trimetazidine O-sulfate (M3), desmethyl-trimetazidine O-glucuronide (M4), and desmethyl-trimetazidine-N-oxide-O-glucuronide (M5). Metabolite M5 has not previously been reported. Excretion curves were constructed based on the chromatographic peak areas of specific mass transitions (precursor ion > product ion) related to each of the detected metabolites

Donnan dialysis for phosphate recovery from diverted urine.

There is a critical need to shift from existing linear phosphorous management practices to a more sustainable circular P economy. Closing the nutrient loop can reduce our reliance on phosphate mining, which has well-documented environmental impacts, while simultaneously alleviating P pollution of aquatic environments from wastewater discharges that are not completely treated. The high orthophosphate, HxPO4(3-x)-, content in source-separated urine offers propitious opportunities for P recovery. This study examines the use of Donnan dialysis (DD), an ion-exchange membrane-based process, for the recovery of orthophosphates from fresh and hydrolyzed urine matrixes. H2PO4- transport against an orthophosphate concentration gradient was demonstrated and orthophosphate recovery yields up to 93% were achieved. By adopting higher feed to receiver volume ratios, DD enriched orthophosphate in the product stream as high as ≈2.5 × the initial urine feed concentration. However, flux, selectivity, and yield of orthophosphate recovery were detrimentally impacted by the presence of SO42- and Cl- in fresh urine, and the large amount of HCO3- rendered hydrolyzed urine practically unsuitable for P recovery using DD. The detrimental effects of sulfate ions can be mitigated by utilizing a monovalent ion permselective membrane, improving selectivity for H2PO4- transport over SO42- by 3.1 × relative to DD with a conventional membrane; but the enhancement was at the expense of reduced orthophosphate flux. Critically, widely available and low-cost/waste resources with sufficiently high Cl- content, such as seawater and waste water softening regenerant rinse, can be employed to improve the economic viability of orthophosphate recovery. This study shows the promising potential of DD for P recovery and enrichment from source-separated urine.

Studies on the mechanism of Panax Ginseng in the treatment of deficiency of vital energy dementia rats based on urine metabolomics.

Panax Ginseng (PG) has been used to strengthen memory and physique for thousands of years, because its main components ginsenosides (GS) and ginseng polysaccharides (GP) play a major role, but its mechanism is not clear. In this study, a rat model of dementia with vital energy deficiency (DED) was established through intraperitoneal injection with D-galactose and AlCl3 and combined with exhaustive swimming. Pharmacological studies and the urine metabolomics based on ultra-high-performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS) were employed for evaluation the efficacy of PG and exploring this treatment mechanism. Through urine metabolic profiling, it can be seen that DED rats after PG administration are close to normal group (NG) rats, and PG can regulate the in vivo status of DED rats which tend to NG. The results of behavioral, biochemical indicators and immunohistochemistry further verified the above results, and the mechanism of action of each component is refined. Ultimately, we believe that the mechanism of PG in the treatment of DED is that ginsenosides (GS) intervenes in phenylalanine tryptophan and tyrosine metabolism, stimulates dopamine production, inhibits Aß deposition and neuroinflammation; and that ginseng polysaccharides (GP) provides energy to strengthen the TCA cycle and improve immune capacity.

Fluid intake, hydration status and body mass changes in U-15 judo athletes during a training day

Despite numerous studies related to dehydration there is still a lack of scientific literature presenting hydration status and fluid intake of judo athletes during different periods. Therefore, the aim of this study was to investigate, fluid intake, hydration status and body weight changes of young judo athletes during a typical day of training in preparation period. Twenty-two young judo athletes (age: 12 ± 0.7 y, experience: 3.5 ± 1.1) voluntarily participated in this study. Hydration status and weight were examined in the morning, before and immediately after the training. All athletes trained 90 min and they consumed fluids ad libitum during the exercise. According to morning urine specific gravity (USG) values, 81.2% of the athletes were dehydrated while only 18.8% of the athletes were euhydrated. Pre-training urine measurements showed that 63.64% of the athletes presented dehydration and 77.27% of the athletes completed the training in dehydrated condition despite fluid availability during the training. Mean body weight loss during training was -0.64 ± 0.66%. It can be concluded that young judo athletes presented high prevalence of dehydration as indicated by USG values. Most of the athletes were dehydrated during a typical training day and completed the training in more dehydrated conditions compared to pre training values despite ad libitum fluid intake. It is of great importance to evaluate hydration status of the athletes before training to refrain from common practice of fluid restriction for weight loss and adverse effects of a persistent state of fluid deficit on physical and health related state.

Governor Vessel Moxibustion Therapy Improves Microbiota Structure in Ankylosing Spondylitis Patients.

BACKGROUND: Clinical studies have shown that ankylosing spondylitis (AS) could be significantly improved by Governor Vessel moxibustion (GVM) therapy. OBJECTIVE: Study whether GVM therapy alleviates the clinical symptoms of AS by modulating intestinal microbiota. METHODS: A total of 9 AS patients and 9 paired healthy individuals were enrolled, and GVM therapy was provided to the AS patients. Stool, urine, and saliva samples from the healthy individuals and the AS patients before and after GVM therapy were collected, and 16S rRNA gene sequencing was performed for microbiota analysis. RESULTS: We found that GVM therapy can significantly alleviate the symptoms of AS, such as diarrhea, abdominal pain, and bloating. GVM therapy also decreased the abundances of Bacteroides and Prevotella while increasing the abundances of beneficial bacteria, such as Lactobacillus, in the gut microbiota of the AS patients. The analyses for AS clinical data and microbial abundances in AS patients revealed their multiple significant correlations (P < 0.01); for example, an unclassified crystal was positively correlated with AF12 and Delftia, monocyte had a negative correlation with Scardovia, and human leukocyte antigen-B27 was negatively correlated with Catenibacterium, Coprococcus, and Oscillospira. CONCLUSIONS: Overall, these findings demonstrate that GVM therapy can alleviate AS clinical symptoms, and at the same time, it improves the microbial structure of microbiota in AS patients. This trial is registered with Chinese Clinical Trial Registry ChiCTR2100051907.

Zootherapeutic uses of animals excreta: the case of elephant dung and urine use in Sayaboury province, Laos.

BACKGROUND: Despite a widespread aversion towards faeces and urine, animal excreta are used in traditional medicine in many countries since centuries, but records are scattered and few therapeutic uses have been accurately documented while in the current context of emerging zoonoses such records may be of major interest. METHODOLOGY: In this study, we investigated the therapeutic uses that mahouts in Xayaboury province, Lao PDR make of elephant urine and faeces as well as of the brood chamber that beetles (Heliocopris dominus) fashion from elephant dung. Semi-structured interviews were conducted with mahouts on elephant diet, health problems and responses to disease, andwhether they use elephant products. Data were supplemented by interviews with traditional healers. RESULTS: Seven respondents reported the use of elephant urine in ethnoveterinary care for elephants and in human medicine in case of diabetes and otitis. 25 respondents reported therapeutic use of elephant faeces (EF) and elephant dung beetle brood chambers. The major indications are gastrointestinal and skin problems. Macerations or decoctions are drunk or used externally as a lotion. The mahouts attribute the therapeutic effectiveness of EFs to their content which includes the remains of many species from the elephant diet which they consider to be medicinal. DISCUSSION: The indications of these uses are consistent with pharmacological and clinical studies highlighting the properties of different animals' urine and faeces and their curative potential tested in vivo. The acknowledgement by the mahouts of medicinal properties of elephant faecal bolus contrasts with the rare justifications of animal material use recorded in zootherapeutic studies, which falls within the symbolic domain. However, numerous studies highlight the preponderant role of the microbiota in physiological processes, raising the hypothesis of a curative action of EF, by rebalancing the user's microbiota. CONCLUSION: The therapeutic uses of EF preparations despite their possible curative properties are a potential source of zoonotic transmission from elephants to humans. In the current context of globalisation of trade which favours the emergence of zoonoses and in relation with the issue of One Health, it becomes crucial to further document the zootherapeutic practices to prevent emerging diseases. As elephants and local related ethnoethological knowledge are threatened, documenting them is urgent to contribute to their preservation.

The influence of tart cherries (Prunus Cerasus) on vascular function and the urinary metabolome: a randomised placebo-controlled pilot study.

Montmorency tart cherries (MC) have been found to modulate indices of vascular function with interventions of varying duration. The objective of this preliminary study was to identify the chronic effects of MC supplementation on vascular function and the potential for urinary metabolomics to provide mechanistic evidence. We performed a placebo-controlled, double-blind, randomised study on 23 healthy individuals (18M, 7F) that consumed 30 ml MC or a placebo twice daily for 28 days. Whole body measures of vascular function and spot urine collections were taken at baseline and after supplementation. There were no significant changes to vascular function including blood pressure and arterial stiffness. Urinary metabolite profiling highlighted significant changes (P < 0⋅001) with putative discriminatory metabolites related to tryptophan and histidine metabolism. Overall, MC supplementation for 28 days does not improve indices of vascular function but changes to the urinary metabolome could be suggestive of potential mechanisms.

Effect of 7 vs 14 Days of Antibiotic Therapy on Resolution of Symptoms Among Afebrile Men With Urinary Tract Infection: A Randomized Clinical Trial.

Importance: Determination of optimal treatment durations for common infectious diseases is an important strategy to preserve antibiotic effectiveness. Objective: To determine whether 7 days of treatment is noninferior to 14 days when using ciprofloxacin or trimethoprim/sulfamethoxazole to treat urinary tract infection (UTI) in afebrile men. Design, Setting, and Participants: Randomized, double-blind, placebo-controlled noninferiority trial of afebrile men with presumed symptomatic UTI treated with ciprofloxacin or trimethoprim/sulfamethoxazole at 2 US Veterans Affairs medical centers (enrollment, April 2014 through December 2019; final follow-up, January 28, 2020). Of 1058 eligible men, 272 were randomized. Interventions: Participants continued the antibiotic prescribed by their treating clinician for 7 days of treatment and were randomized to receive continued antibiotic therapy (n = 136) or placebo (n = 136) for days 8 to 14 of treatment. Main Outcomes and Measures: The prespecified primary outcome was resolution of UTI symptoms by 14 days after completion of active antibiotic treatment. A noninferiority margin of 10% was selected. The as-treated population (participants who took ≥26 of 28 doses and missed no more than 2 consecutive doses) was used for the primary analysis, and a secondary analysis included all patients as randomized, regardless of treatment adherence. Secondary outcomes included recurrence of UTI symptoms and/or adverse events within 28 days of stopping study medication. Results: Among 272 patients (median [interquartile range] age, 69 [62-73] years) who were randomized, 100% completed the trial and 254 (93.4%) were included in the primary as-treated analysis. Symptom resolution occurred in 122/131 (93.1%) participants in the 7-day group vs 111/123 (90.2%) in the 14-day group (difference, 2.9% [1-sided 97.5% CI, -5.2% to ∞]), meeting the noninferiority criterion. In the secondary as-randomized analysis, symptom resolution occurred in 125/136 (91.9%) participants in the 7-day group vs 123/136 (90.4%) in the 14-day group (difference, 1.5% [1-sided 97.5% CI, -5.8% to ∞]) Recurrence of UTI symptoms occurred in 13/131 (9.9%) participants in the 7-day group vs 15/123 (12.9%) in the 14-day group (difference, -3.0% [95% CI, -10.8% to 6.2%]; P = .70). Adverse events occurred in 28/136 (20.6%) participants in the 7-day group vs 33/136 (24.3%) in the 14-day group. Conclusions and Relevance: Among afebrile men with suspected UTI, treatment with ciprofloxacin or trimethoprim/sulfamethoxazole for 7 days was noninferior to 14 days of treatment with regard to resolution of UTI symptoms by 14 days after antibiotic therapy. The findings support the use of a 7-day course of ciprofloxacin or trimethoprim/sulfamethoxazole as an alternative to a 14-day course for treatment of afebrile men with UTI. Trial Registration: ClinicalTrials.gov identifier: NCT01994538.

Characterization of Metabolites in Plasma, Urine and Feces of Healthy Participants after Taking Brahmi Essence for Twelve Weeks Using LC-ESI-QTOF-MS Metabolomic Approach.

Brahmi essence, developed from Bacopa monnieri (L.) Wettst. standardized extract and mulberry juice, was proven to improve the memory speed of healthy participants aged 55-80 years old, following a 12-week dietary program. However, the metabolites have not yet been reported. Our objective was to characterize the altered metabolites in the plasma, urine, and feces of healthy volunteers after consumption of Brahmi essence for 12 weeks, using the LC-MS metabolomics approach. The altered metabolites were selected from OPLS-DA S-plots; 15 metabolites in the plasma, 7 in the urine, and 17 in the feces samples were tentatively identified by comparison with an online database and literature. The metabolites in the plasma samples were in the classes of amino acids, acylcarnitine, and phospholipids. Benzeneactamide-4-O-sulphate and 3-hydroxyhippuric acid were found in urine samples. The metabolites in the class of amino acids, together with jujubogenin and pseudojujubogenin, were identified in the fecal samples. The aminoacyl-tRNA, aromatic amino acids, and branched-chain amino acid biosynthetic pathways were mainly related to the identified metabolites in all three samples. It could be implied that those metabolites and their pathways might be linked with the effect of Brahmi essence on memory speed.

Effects of Lactobacillus plantarum CJLP55 on Clinical Improvement, Skin Condition and Urine Bacterial Extracellular Vesicles in Patients with Acne Vulgaris: A Randomized, Double-Blind, Placebo-Controlled Study.

Lactobacillus plantarum CJLP55 has anti-pathogenic bacterial and anti-inflammatory activities in vitro. We investigated the dietary effect of CJLP55 supplement in patients with acne vulgaris, a prevalent inflammatory skin condition. Subjects ingested CJLP55 or placebo (n = 14 per group) supplements for 12 weeks in this double-blind, placebo-controlled randomized study. Acne lesion count and grade, skin sebum, hydration, pH and surface lipids were assessed. Metagenomic DNA analysis was performed on urine extracellular vesicles (EV), which indirectly reflect systemic bacterial flora. Compared to the placebo supplement, CJLP55 supplement improved acne lesion count and grade, decreased sebum triglycerides (TG), and increased hydration and ceramide 2, the major ceramide species that maintains the epidermal lipid barrier for hydration. In addition, CJLP55 supplement decreased the prevalence of Proteobacteria and increased Firmicutes, which were correlated with decreased TG, the major skin surface lipid of sebum origin. CJLP55 supplement further decreased the Bacteroidetes:Firmicutes ratio, a relevant marker of bacterial dysbiosis. No differences in skin pH, other skin surface lipids or urine bacterial EV phylum were noted between CJLP55 and placebo supplements. Dietary Lactobacillus plantarum CJLP55 was beneficial to clinical state, skin sebum, and hydration and urine bacterial EV phylum flora in patients with acne vulgaris.

Maternal selenium deficiency in mice promotes sex-specific changes to urine flow and renal expression of mitochondrial proteins in adult offspring.

Selenium deficiency during pregnancy can impair fetal development and predispose offspring to thyroid dysfunction. Given that key selenoproteins are highly expressed in the kidney and that poor thyroid health can lead to kidney disease, it is likely that kidney function may be impaired in offspring of selenium-deficient mothers. This study utilized a mouse model of maternal selenium deficiency to investigate kidney protein glycation, mitochondrial adaptations, and urinary excretion in offspring. Female C57BL/6 mice were fed control (>190 µg selenium/kg) or low selenium (<50 µg selenium/kg) diets four weeks prior to mating, throughout gestation, and lactation. At postnatal day (PN) 170, offspring were placed in metabolic cages for 24 hr prior to tissue collection at PN180. Maternal selenium deficiency did not impact selenoprotein antioxidant activity, but increased advanced glycation end products in female kidneys. Male offspring had reduced renal Complex II and Complex IV protein levels and lower 24 hr urine flow. Although renal aquaporin 2 (Aqp2) and arginine vasopressin receptor 2 (Avpr2) mRNA were not altered by maternal selenium deficiency, a correlation between urine flow and plasma free T4 concentrations in male but not female offspring suggests that programed thyroid dysfunction may be mediating impaired urine flow. This study demonstrates that maternal selenium deficiency can lead to long-term deficits in kidney parameters that may be secondary to impaired thyroid dysfunction. Considering the significant burden of renal dysfunction as a comorbidity to metabolic diseases, improving maternal selenium intake in pregnancy may be one simple measure to prevent lifelong disease.

Acceptability and Feasibility of a 13-Week Pilot Randomised Controlled Trial Testing the Effects of Incremental Doses of Beetroot Juice in Overweight and Obese Older Adults.

Nitrate-rich food can increase nitric oxide production and improve vascular and brain functions. This study examines the feasibility of a randomised controlled trial (RCT) testing the effects of prolonged consumption of different doses of dietary nitrate (NO3-) in the form of beetroot juice (BJ) in overweight and obese older participants. A single-blind, four-arm parallel pilot RCT was conducted in 62 overweight and obese (30.4 ± 4 kg/m2) older participants (mean ± standard deviation (SD), 66 ± 4 years). Participants were randomized to: (1) high-NO3- (HN: 2 × 70 mL BJ/day) (2) medium-NO3- (MN: 70 mL BJ/day), (3) low-NO3- (LN: 70 mL BJ on alternate days) or (4) Placebo (PL: 70 mL of NO3--depleted BJ on alternate days), for 13 weeks. Compliance was checked by a daily log of consumed BJ, NO3- intake, and by measuring NO3- and NO2- concentrations in plasma, saliva, and urine samples. Fifty participants completed the study. Self-reported compliance to the interventions was >90%. There were significant positive linear relationships between NO3- dose and the increase in plasma and urinary NO3- concentration (R2 = 0.71, P < 0.001 and R2 = 0.46 P < 0.001, respectively), but relationships between NO3- dose and changes in salivary NO3- and NO2- were non-linear (R2 = 0.35, P = 0.002 and R2 = 0.23, P = 0.007, respectively). The results confirm the feasibility of prolonged BJ supplementation in older overweight and obese adults.

The chemical profiling of aqueous soluble fraction from Lagopsis supina and its diuretic effects via suppression of AQP and RAAS pathways in saline-loaded rats.

ETHNOPHARMACOLOGICAL RELEVANCE: Lagopsis supina (Steph.) Ik. -Gal. ex Knorr. has been widely used as a remedy treatment for diuresis and edema in China over 2500 years. Our previous results showed that the aqueous soluble fraction from L. supina (LSB) possessed acute diuretic effect. AIM OF THE STUDY: The aim of this study was to appraise the acute (6 h) and prolonged (7 d) diuretic effects, underlying mechanisms, and chemical profiling of LSB. MATERIALS AND METHODS: The chemical profiling of LSB was performed by ultra-high-performance liquid chromatography-quadrupole time-of-flight tandem mass spectrometry (UHPLC-qTOF-MS/MS). Then, oral administration of LSB (40, 80, 160 and 320 mg/kg) and furosemide (10 mg/kg) once daily for 7 consecutive days to evaluate the diuretic effects in saline-loaded rats. The body weight, food consumption, and water intake were recorded once daily. The urinary volume, pH and electrolyte concentrations (Na+, K+, Cl-, and Ca2+) were measured after administration drugs for acute and prolonged diuretic effects. In addition, the serum levels of Na+-K+-ATPase, angiotensin II (Ang II), anti-diuretic hormone (ADH), aldosterone (ALD), atriopeptin (ANP), aquaporins (AQPs)-1, 2 and 3 were determined by ELISA kits. The mRNA expressions and protein levels of AQPs-1, 2 and 3 were analyzed by real-time quantitative PCR and Western blot assays, respectively. RESULTS: 30 compounds were identified in LSB based on accurate mass and MS/MS fragmentation compared to literature, among which phenylpropanoids and flavonoids could be partly responsible for the major diuretic effect. Daily administration of LSB (160 or 320 mg/kg) prominently increased urinary excretion volume after the 2 h at the first day of treatment, remaining until the 7th day. LSB did not cause Na+ and K+ electrolyte abnormalities, and has minor effect on Cl- and Ca2+ concentrations at 320 mg/kg. Furthermore, LSB observably suppressed renin-angiotensin-aldosterone system (RAAS) activation, including decreased serum levels of Ang II, ADH, and ALD, and prominently increased serum level of ANP in rats. LSB treatment significantly down-regulated the serum levels, mRNA expressions and protein levels of AQP1, AQP2, and AQP3. CONCLUSION: LSB has a prominent acute and prolonged diuretic effects via suppression of AQP and RAAS pathways in saline-loaded rats, and support the traditional folk use of this plant. Taken together, LSB might be a potential diuretic agent.

Gut Microbial and Metabolic Profiling Reveal the Lingering Effects of Infantile Iron Deficiency Unless Treated with Iron.

SCOPE: Iron deficiency (ID) compromises the health of infants worldwide. Although readily treated with iron, concerns remain about the persistence of some effects. Metabolic and gut microbial consequences of infantile ID were investigated in juvenile monkeys after natural recovery (pID) from iron deficiency or post-treatment with iron dextran and B vitamins (pID+Fe). METHODS AND RESULTS: Metabolomic profiling of urine and plasma is conducted with 1 H nuclear magnetic resonance (NMR) spectroscopy. Gut microbiota are characterized from rectal swabs by amplicon sequencing of the 16S rRNA gene. Urinary metabolic profiles of pID monkeys significantly differed from pID+Fe and continuously iron-sufficient controls (IS) with higher maltose and lower amounts of microbial-derived metabolites. Persistent differences in energy metabolism are apparent from the plasma metabolic phenotypes with greater reliance on anaerobic glycolysis in pID monkeys. Microbial profiling indicated higher abundances of Methanobrevibacter, Lachnobacterium, and Ruminococcus in pID monkeys and any history of ID resulted in a lower Prevotella abundance compared to the IS controls. CONCLUSIONS: Lingering metabolic and microbial effects are found after natural recovery from ID. These long-term biochemical derangements are not present in the pID+Fe animals emphasizing the importance of the early detection and treatment of early-life ID to ameliorate its chronic metabolic effects.