First-morning Urine Samples Research Articles

Exploring diet as a source of plasticizers in pregnancy and implications for maternal second-trimester metabolic health

Background and objectivesDiet plays critical roles in modulating maternal metabolic health in pregnancy, but is also a source of metabolic-disrupting phthalates and their replacements. We aimed to evaluate whether the effects of better diet quality on favorable maternal metabolic outcomes could be partially explained by lower exposure to phthalates/replacements. MethodsAt 13 weeks gestation, 295 Illinois women (enrolled 2015–2018) completed a three-month food frequency questionnaire that we used to calculate the Alternative Healthy Eating Index (AHEI)-2010 to assess diet quality. We quantified 19 metabolites, reflecting exposure to 10 phthalates/replacements, in a pool of five first-morning urine samples collected monthly across pregnancy. We measured 15 metabolic biomarkers in fasting plasma samples collected at 17 weeks gestation, which we reduced to five uncorrelated principal components (PCs), representing adiposity, lipids, cholesterol, inflammation, and growth. We used linear regression to estimate associations of diet quality with [1] phthalates/replacements and [2] metabolic PCs, as well as [3] associations of phthalates/replacements with metabolic PCs. We estimated the proportion of associations between diet quality and metabolic outcomes explained by phthalates/replacements using a causal mediation framework. ResultsOverall, every 10-point improvement in AHEI-2010 score was associated with −0.15 (95% CI: −0.27, −0.04) lower adiposity scores, reflecting lower glucose, insulin, C-peptide, leptin, C-reactive protein, but higher adiponectin biomarker levels. Every 10-point increase in diet quality was also associated with 18% (95%CI: 7%, 28%) lower sum of di-2-ethylhexyl terephthalate urinary metabolites (∑DEHTP). Correspondingly, each 18% increase in ∑DEHTP was associated with 0.03 point (95% CI: 0.01, 0.05) higher adiposity PC scores. In mediation analyses, 21% of the inverse relationship between diet quality and adiposity PC scores was explained by lower ∑DEHTP. ConclusionsThe favorable impact of diet quality on maternal adiposity biomarkers may be partially attributed to lower metabolite concentrations of DEHTP, a plasticizer allowed to be used in food packaging materials.

Early menstrual cycle impacts of oestrogen and progesterone on the timing of the fertile window.

What is the effect of oestrogen and progesterone at the beginning of the menstrual cycle in delaying entry into the fertile window? Both oestrogen and progesterone contribute to a delay in the onset of the fertile window. Oestrogen enhances cervical mucus secretion while progesterone inhibits it. Observational study. Daily observation of 220 menstrual cycles contributed by 88 women with no known menstrual cycle disorder. Women recorded cervical mucus daily and collected first-morning urine samples for analysis of oestrone-3-glucuronide, pregnanediol-3-alpha-glucuronide (PDG), FHS, and LH. They underwent serial ovarian ultrasound examinations. The main outcome measure was the timing within the cycle of the onset of the fertile window, as identified by the appearance of mucus felt or seen at the vulva. Low oestrogen secretion and persistent progesterone secretion during the first week of the menstrual cycle both negatively affect mucus secretion. Doubling oestrogen approximately doubled the odds of entering the fertile window (OR: 1.82 95% CI=1.23; 2.69). Increasing PDG from below 1.5 to 4µg/mg creatinine was associated with a 2-fold decrease in the odds of entering the fertile window (OR: 0.51 95% CI=0.31; 0.82). Prolonged progesterone secretion during the first week of the menstrual cycle was also statistically significantly associated with higher LH secretion. Finally, the later onset of the fertile window was associated with statistically significant persistently elevated LH secretion during the luteal phase of the previous menstrual cycle. This post hoc study was conducted to assess the potential impact of residual progesterone secretion at the beginning of the menstrual cycle. It was conducted on an existing data set because of the scarcity of data available to answer the question. Analysis with other datasets with similar hormone results would be useful to confirm these findings. This study provides evidence for residual progesterone secretion in the early latency phase of some menstrual cycles, which may delay the onset of the fertile window. This progesterone secretion may be supported by subtly increased LH secretion during the few days before and after the onset of menses, which may relate to follicular waves in the luteal phase. Persistent progesterone secretion should be considered in predicting the onset of the fertile window and in assessing ovulatory dysfunction. The authors declare no conflicts of interest. No funding was provided for this secondary data analysis. N/A.

Expression profiles of urine exosomal tRNA-derived small RNAs and their potential roles in calcium oxalate stone disease

Background and objective: Exosomes have been confirmed to be implicated in the pathogenesis of calcium oxalate (CaOx) stones. tRNA-derived small RNAs (tsRNAs) are among the oldest small RNAs involved in exosome-mediated intercellular communication, yet their role in kidney stones remains unexplored. This pilot study aimed to identify differentially expressed tsRNAs (DEtsRNAs) in urine exosomes between CaOx stone patients and healthy controls and explore their potential roles in nephrolithiasis. Method: First-morning urine samples were collected from three CaOx stone patients and three healthy controls. Urinary exosomes were isolated and analyzed by high-throughput sequencing to generate the expression profiles of tsRNAs and detect DEtsRNAs. Predicted target genes of DEtsRNAs were subjected to functional enrichment analysis. The authors also combined the public dataset GSE73680 to investigate how DEtsRNAs were related to stone formation. Results: Four DEtsRNAs were significantly upregulated in CaOx stone patients compared to healthy controls. tRF-Lys-TTT-5005c was the most elevated, followed by tRF-Lys-CTT-5006c, tRF-Ala-AGC-5017b, and tRF-Gly-CCC-5004b. Bioinformatics analysis indicated that these four types of DEtsRNAs might serve distinct biological functions. Combined with data mining from the public dataset GSE73680, the authors assumed that exosomes carrying tRF-Lys-TTT-5005c and tRF-Lys-CTT-5006c could inhibit the expression of SMAD6, FBN1, and FZD1, thereby activating the BMP signaling pathway, which might induce an osteogenic-like transformation in target cells, resulting in the formation of Randall’s plaques and CaOx stones. Conclusion: The authors’ findings shed light on the potential roles of tsRNAs in the pathogenesis of CaOx stone disease, highlighting exosomal DEtsRNAs as promising diagnostic biomarkers and therapeutic targets in nephrolithiasis.

Secondhand Tobacco Exposure Assessed Using Urinary Cotinine Among 10-Year-Old Children in Japan: An 11-Year Repeated Cross-sectional Study

Abstract Introduction The emergence of heated tobacco products (HTPs) has made it important to monitor HTP-generated aerosols in addition to combustible cigarette (CC) smoke as a source of secondhand tobacco (SHT) exposure. We investigated the trend of SHT exposure in school-aged children and assessed whether SHT exposure depended on household tobacco use status. Aims and Methods This repeated cross-sectional study from 2011 to 2021 (15 927 participants) was based on data from an annual survey of fourth-grade students (aged 10 years) in Kumagaya City, Japan. In addition to a questionnaire which includes questions about household tobacco use status, we measured the urinary cotinine levels of each participant by their first-morning urine sample to objectively assess SHT exposure. We defined the participants with urinary cotinine levels ≥5.0 ng/mL as being exposed to SHT. Results The prevalence of SHT exposure decreased over the 11-year period from 18.6% in 2011 to 5.3% in 2021. It was significantly higher in households with tobacco users than without tobacco users (t-test p< .001). Prevalence of SHT exposure was 1.4% among the 68.1% of households not using tobacco, 22.9% among the 16.5% using only CC, 3.1% among the 12.3% using only HTP, and 27.6% among the 3.9% of households using CC and HTP. Conclusions While the prevalence of SHT exposure showed a decreasing trend from 2011 to 2021, the prevalence of SHT exposure was higher in children with household members using tobacco products, regardless of the type of tobacco product, than in those without tobacco users. Implications This study observed that the prevalence of SHT exposure was higher among children in households with tobacco users than among those without tobacco users, regardless of the type of tobacco product. Our findings highlight the importance of advocating that HTPs do not reduce the likelihood of SHT exposure to bystanders.

Improving hydration in underhydrated, free-living young adults results in better running performance

Dehydration impairs endurance exercise performance, especially in the heat. However, it is unclear how exercise performance is affected by improved hydration status in underhydrated individuals. Therefore, this study examined the impact of a one-week hydration intervention on endurance running performance. We hypothesized better hydration would lead to a faster 2-mile running time. Eighty-nine healthy and moderately fit individuals completed a 2-mile run as fast as possible before (PRE) and after (POST) a 1-week hydration intervention. Performance was incentivized by providing prize money based on ranked average times. First-morning urine samples were collected on the day of each run. If PRE urine specific gravity was >1.020, the subjects were instructed to increase their water intake for one week before the second run. The remaining subjects were instructed to maintain their water intake. All subjects were provided with a filter water bottle. Of the 89 subjects that completed both runs, 44 (18% female, age: 21±2 y, weight: 73.3±12.5 kg, height: 1.76±0.10 m, body mass index: 23.6±3.2 kg/m2) were included in the analysis. Subjects were classified as improved hydration (IMP) if UOsm decreased between PRE and POST by ≥100 mmol/kg and POST was <800 mmol/kg. Subjects were classified as diminished hydration (DIM) if UOsm increased between PRE and POST by ≥100 mmol/kg and POST was >500 mmol/kg. The subjects were removed from the analysis if they met any of the following three conditions: a) had a small UOsm change (<100 mmol/kg) between PRE and POST, b) lowered UOSM but it was still >800 mmol/kg, and c) increased UOsm but it was still <500 mmol/kg. The 2-mile runs took place in Phoenix, Arizona, during the morning (0700 hours) with a maximum ambient temperature of 22±2℃, relative humidity of 37±10%, and WBGT of 16±2℃. UOsm significantly decreased for IMP from PRE: 791±197 to POST: 419±136 mmol/kg (p<0.05) and increased for DIM from PRE: 532±283 to POST: 799±235 mmol/kg (p<0.05). The 2-mile run time significantly improved in IMP from PRE: 13.5±1.8 to POST: 13.3±1.8 min (-11.4±25.8 sec, P=0.03), but did not change in DIM from PRE: 14.2±2.7 to POST: 14.3±2.6 min (+4.8±19.9 sec, P>0.05). The results suggest that improving hydration in underhydrated subjects via one week of increased water intake may improve 2-mile running performance in moderately fit individuals. This study was funded by a grant from the Arizona State University’s Global Sport Institute. Brita donated the filter water bottles. This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

Investigation of Urinary Metabolites of Organophosphate Esters in Hanoi, Vietnam: Assessment Exposure and Estimated Daily Intake.

In recent years, organophosphate esters (OPEs) have become one of the most common additives in various consumer products worldwide, therefore the exposure and impact of OPEs on human health are drawing a lot of attention. In this study, three metabolites of OPEs including bis(1,3-dichloro-2-propyl) phosphate (BDCIPP), diphenyl phosphate (DPhP) and diethyl phosphate (DEP) were investigated in first-morning void urine samples taken from a population (age range: 3-76years old) in Hanoi, Vietnam. The most dominant urinary OPE metabolite was DEP with the geometric mean of specific gravity adjust (SG-adjusted) concentration were 1960ngmL-1 and detected frequency (DF) of 98%. Followed by DPhP (8.01ngmL-1, DF: 100%) and BDCIPP (2.18ngmL-1, DF: 51%). The results indicated that gender and age might have associations with the OPE metabolites variation in urine samples. The levels of OPE metabolites in urine samples from females were slightly higher than in males. An increase in age seems to have an association with a decrease in DPhP levels in urine. Exposure doses of parent OPEs were evaluated from the unadjusted urinary concentration of corresponding OPE metabolite. The estimated exposure doses of triethyl phosphate (TEP) (mean: 534,000ngkg-1d-1) were significantly higher than its corresponding reference dose, suggesting the high potential risk from the current exposure doses of TEP to human health. The results of this work provided the initial information on the occurrence of three OPE metabolites in urine from Hanoi, Vietnam and estimated exposure dose of corresponding parent OPEs.

Simultaneous detection of mycotoxins and pesticides in human urine samples: A 24-h diet intervention study comparing conventional and organic diets in Spain

Pesticides and mycotoxins, prominent chemical hazards in the food chain, are commonly found in plant-based foods, contributing to their pervasive presence in the human body, as evidenced by biomonitoring programs. Despite this, there is limited knowledge about their co-occurrence patterns. While intervention studies have demonstrated that organic diets can significantly reduce pesticide levels, their impact on mycotoxin exposure has been overlooked.To address this gap, this study pursued two objectives: first, to characterize the simultaneous presence of mycotoxins and pesticides in human urine samples by means of the control of the biomarkers of exposure, and second, to investigate the influence of consuming organic foods on these co-exposure patterns. A pilot study involving 20 healthy volunteers was conducted, with participants consuming either exclusively organic or conventional foods during a 24-h diet intervention in autumn 2021 and spring 2022 to account for seasonal variability. Participants provided detailed 24-h dietary records, and their first-morning urine samples were collected, minimally treated and analysed using LC-Q-ToF-MS by means of a multitargeted method in order to detect the presence of these residues.Results indicated that among the 52 screened compounds, four mycotoxins and seven pesticides were detected in over 25% of the samples. Deoxynivalenol (DON) and the non-specific pesticide metabolite diethylphosphate (DEP) exhibited the highest frequency rates (100%) and concentration levels. Correlations were observed between urine levels of mycotoxins (DON, ochratoxin alpha [OTα], and enniatin B [ENNB]) and organophosphate pesticide metabolites DEP and 2-diethylamino-6-methyl-4-pyrimidinol (DEAMPY). The pilot intervention study suggested a reduction in ENNB and OTα levels and an increase in β-zearalenol levels in urine after a short-term replacement with organic food. However, caution is advised due to the study's small sample size and short duration, emphasizing the need for further research to enhance understanding of the human chemical exposome and refine chemical risk assessment.

Association of urinary chlorpyrifos, paraquat, and cyproconazole levels with the severity of fatty liver based on MRI

BackgroundThe objective of this study was to detect the urinary levels of chlorpyrifos, paraquat, and cyproconazole in residents living in Fuyang City and to analyze the correlation between these urinary pesticides levels and the severity of fatty liver disease (FLD).MethodsAll participants’ fat fraction (FF) values were recorded by MRI (Magnetic resonance imaging). First-morning urine samples were collected from 53 participants from Fuyang Peoples’Hospital. The levels of three urinary pesticides were measured using β-glucuronidase hydrolysis followed by a. The results were analyzed by using Pearson correlation analysis and binary logistic regression analysis to reveal the correlation between three urinary pesticides and the severity of fatty liver.Results53 individuals were divided into 3 groups based on the results from MRI, with 20 cases in the normal control group, 16 cases in the mild fatty liver group, and 17 cases in the moderate and severe fatty liver group. Urinary chlorpyrifos level was increased along with the increase of the severity of fatty liver. Urinary paraquat level was significantly higher both in the low-grade fatty liver group and moderate & serve grade fatty liver group compared with the control group. No significant differences in urinary cyproconazole levels were observed among the three groups. Furthermore, urinary chlorpyrifos and paraquat levels were positively correlated with FF value. And chlorpyrifos was the risk factor that may be involved in the development of FLD and Receiver Operating Characteristic curve (ROC curve) analysis showed that chlorpyrifos and paraquat may serve as potential predictors of FLD.ConclusionThe present findings indicate urinary chlorpyrifos and paraquat were positively correlated with the severity of fatty liver. Moreover, urinary chlorpyrifos and paraquat have the potential to be considered as the predictors for development of FLD. Thus, this study may provide a new perspective from the environmental factors for the diagnosis, prevention, and treatment of FLD.

First-morning urine samples to study the urobiome.

First-morning urine samples to study the urobiome.

Difference in urinary protein-to-creatinine ratio using the benzethonium chloride and pyrogallol red methods in children.

The appropriate reference value of the urinary protein-to-creatinine ratio (PCR) for proteinuria may change when the urinary pyrogallol red (PR) protein assay method is changed to the benzethonium chloride method (BC). This study aimed to evaluate the difference between BC-based PCR (BC-PCR) and PR-based PCR (PR-PCR) values in children. We compared the BC-PCR and PR-PCR values in the same first-morning urine samples without significant proteinuria in school urine screening settings. The upper limit of the reference values was set at the 97.5th percentile. Notably, 133 samples from 124 individuals (female: 62%, age: median 12.3 years, range 6.3-16.9 years) were collected between August 2020 and October 2022. The diagnoses included 34 normal individuals and 99 with asymptomatic hematuria. The urinary protein (UP) concentrations measured using the BC (BC-UP) and PR (PR-UP) methods were in a linear relationship; however, the BC-UP concentrations were higher than the PR-UP concentrations (mean of differences: 11.2, 95% confidence interval (CI): 11.0-13.4 mg/dL). Also, the BC-PCR values were higher than the PR-PCR values (mean of differences: 0.090, 95% CI: 0.082-0.098 g/gCr). The BC-PCR showed a body-size-related decrease, reflecting a body-size-related urinary creatinine increase. The suggested BC-PCR reference values for proteinuria were 0.25 and 0.17 g/gCr for elementary (6-12.4 years) and junior high school students (12.5-16 years), respectively. These values were higher than those of the PR-PCR and need further studies. When evaluating PCR results, the urinary protein assay should be stated.

Associations of urinary non-persistent endocrine disrupting chemical biomarkers with early-to-mid pregnancy plasma sex-steroid and thyroid hormones

Background/ObjectivesPregnant women are exposed to numerous endocrine disrupting chemicals (EDCs) that can affect hormonal pathways regulating pregnancy outcomes and fetal development. Thus, we evaluated overall and fetal sex-specific associations of phthalate/replacement, paraben, and phenol biomarkers with sex-steroid and thyroid hormones. MethodsIllinois women (n = 302) provided plasma for progesterone, estradiol, testosterone, free T4 (FT4), total T4 (TT4), and thyroid stimulating hormone (TSH) at median 17 weeks gestation. Women also provided up-to-five first-morning urine samples monthly across pregnancy (8–40 weeks), which we pooled to measure 19 phthalate/replacement metabolites (reflecting ten parent compounds), three parabens, and six phenols. We used linear regression to evaluate overall and fetal sex-specific associations of biomarkers with hormones, as well as weighted quantile sum and Bayesian kernel machine regression (BKMR) to assess cumulative associations, non-linearities, and chemical interactions. ResultsIn women of relatively high socioeconomic status, several EDC biomarkers were associated with select hormones, without cumulative or non-linear associations with progesterone, FT4, or TT4. The biomarker mixture was negatively associated with estradiol (only at higher biomarker concentrations using BKMR), testosterone, and TSH, where each 10% mixture increase was associated with −5.65% (95% CI: −9.79, −1.28) lower testosterone and −0.09 μIU/mL (95% CI: −0.20, 0.00) lower TSH. Associations with progesterone, testosterone, and FT4 did not differ by fetal sex. However, in women carrying females, we identified an inverted u-shaped relationship of the mixture with estradiol. Additionally, in women carrying females, each 10% increase in the mixture was associated with 1.50% (95% CI: −0.15, 3.18) higher TT4, whereas in women carrying males, the mixture was associated with −1.77% (95% CI: −4.08, 0.58) lower TT4 and −0.18 μIU/mL (95% CI: −0.33, −0.03) lower TSH. We also identified select chemical interactions. ConclusionSome biomarkers were associated with early-to-mid pregnancy hormones. There were some sex-specific and non-linear associations. Future studies could consider how these findings relate to pregnancy/birth outcomes.

Detection and quantification of Dioctophyme renale eggs in dog urine after nephrectomy.

Dioctophyme renale is a zoonotic nematode that parasitizes mainly right kidney of domestic and wild canines, and can affect humans, and its eggs are eliminated in urine. The duration of egg dissemination after surgical treatment is unknown. Therefore, the objective of this study was to identify, quantify, and verify the duration of the elimination of D. renale eggs in dog urine after the surgical removal of nematodes. The study involved 15 dogs in which female specimens of D. renale were detected in kidneys. Urine samples, preferably first-morning urine samples, were collected before and for the first ten days after nephrectomy. For egg quantification, 40 µL samples of urinary sediment were analyzed in triplicate. In laboratory analyses, between 900 and > 6,000 eggs/urine sample were detected in 86.7% of the dogs prior to surgery, and in 40% of the dogs on postoperative day 1. Of the 15 dogs evaluated, 14 (93.3%) eliminated D. renale eggs on each of the first ten postoperative days. Egg elimination peaked on postoperative day 1. Our results indicate that dogs can continue to be sources of D. renale infection even after the helminths have been removed from a parasitized kidney, underscoring the novelty of these findings and their importance for the One Health approach.

Microbiome Profiling in Bladder Cancer Patients Using the First-morning Urine Sample

BackgroundSeveral studies support the interplay between the urinary microbiome (ie, urobiome) and bladder cancer (BCa). Specific urinary bacteria may be responsible for chronic inflammation, which in turn promotes carcinogenesis. Different signatures of urobiome in BCa patients were identified depending on tumor type, geographical area, age, and sex. ObjectiveWe explored the urobiome in BCa patients undergoing transurethral resection of bladder tumor (TURBT), to identify possible predictive biomarkers of cancer. Design, setting, and participantsThe urobiome analysis was conducted in 48 patients (13 females) undergoing TURBT, of whom 30 with BCa (five females) and 18 with benign bladder tumor, analyzing bacterial 16S rRNA by next-generation sequencing in first-morning (FM) urine samples. Forty-three cancer-free individuals and 17 prostate cancer patients were used as controls. Outcome measurements and statistical analysisFirst, we identified the better urine collection procedure to perform the urobiome analysis, comparing bacterial composition between catheterized (CAT) and FM urine samples in TURBT patients. Successively, we observed a specific urobiome in BCa patients rather than controls. A combined pipeline including the DESeq2 and linear discriminant analysis effect size tests was used to identify differential urinary taxa, strictly associated with BCa patients. Results and limitationsThe bacterial composition of CAT and FM urine samples was comparable, so the latter was used for the following analyses. An increased abundance of Porphyromonas and Porphyromonas somerae was found in BCa patients compared with controls. This signature seems to be more related (p <0.05) to male BCa patients over 50 yr old. Owing to the low biomass of urinary microbiota, several samples were excluded from the study, reducing the number of BCa patients considered. ConclusionsFM urine samples represent a manageable specimen for a urobiome analysis; P. somerae is a specific biomarker of BCa risk. Patient summaryOur study showed an increased abundance of Porphyromonas and Porphyromonas somerae in male bladder cancer (BCa) patients, supporting the use of a first-morning urine sample, a less invasive and low-cost collection method, for the urobiome analysis of patients at risk of BCa.

Sex Differences in Branched-chain Amino Acid and Tryptophan Metabolism and Pathogenesis of Youth-onset Type 2 Diabetes.

Insulin resistance is associated with elevations in plasma branched-chain amino acids (BCAAs). BCAAs compete with aromatic amino acids including tryptophan for uptake into β cells. To explore relationships between BCAAs and tryptophan metabolism, adiposity, and glucose tolerance, we compared urine metabolites in overweight/obese youth with type 2 diabetes (T2D) with those in nondiabetic overweight/obese and lean youth. Metabolites were measured in 24-hour and first-morning urine samples of 56 nondiabetic adolescents with overweight/obesity, 42 adolescents with T2D, and 43 lean controls, aged 12 to 21 years. Group differences were assessed by Kruskal Wallis or ANOVA. Groups were comparable for age, pubertal status, and ethnicity. Youth with T2D were predominantly female and had highest percent body fat. BCAAs, branched-chain ketoacids (BCKAs), tryptophan, and kynurenine were higher in urine of subjects with T2D. There were no differences between lean controls and nondiabetic youth with overweight/obesity. T2D was associated with diversion of tryptophan from the serotonin to the kynurenine pathway, with higher urinary kynurenine/serotonin ratio and lower serotonin/tryptophan and 5-HIAA/kynurenine ratios. Urinary BCAAs, BCKAs, tryptophan, and ratios reflecting diversion to the kynurenine pathway correlated positively with metrics of body fat and hemoglobin A1c. Increases in these metabolites in the obese T2D group were more pronounced and statistically significant only in adolescent girls. Increases in urinary BCAAs and BCKAs in adolescent females with T2D are accompanied by diversion of tryptophan metabolism from the serotonin to the kynurenine pathway. These adaptations associate with higher risks of T2D in obese adolescent females than adolescent males.

Renal Function Impairment in Children With Congenital Cytomegalovirus Infection: A Cross-sectional Study.

We aimed to determine the prevalence and severity of glomerular and tubular renal dysfunction by means of urinalysis in infants and toddlers with congenital cytomegalovirus infection (cCMV) and their association with cCMV disease, viruria and antiviral treatment. This cross-sectional study was done using the Spanish Registry of Congenital Cytomegalovirus Infection. First-morning urine samples were collected from January 2016 to December 2018 from patients <5 years old enrolled in Spanish Registry of Congenital Cytomegalovirus Infection. Samples were excluded in case of fever or other signs or symptoms consistent with acute infection, bacteriuria or bacterial growth in urine culture. Urinary protein/creatinine and albumin/creatinine ratios, urinary beta-2-microglobulin levels, hematuria and CMV viruria were determined. A 0.4 cutoff in the urinary albumin/protein ratio was used to define tubular (<0.4) or glomerular (>0.4) proteinuria. Signs and symptoms of cCMV at birth, the use of antivirals and cCMV-associated sequelae at last available follow-up were obtained from Spanish Registry of Congenital Cytomegalovirus Infection. Seventy-seven patients (37 females, 48.1%; median [interquartile range] age: 14.0 [4.4-36.2] months) were included. Symptom-free elevated urinary protein/creatinine and albumin/creatinine ratios were observed in 37.5% and 41.9% of patients, respectively, with tubular proteinuria prevailing (88.3%) over glomerular proteinuria (11.6%). Proteinuria in the nephrotic range was not observed in any patients. In multivariate analysis, female gender was the only risk factor for tubular proteinuria (adjusted odds ratio = 3.339, 95% confidence interval: 1.086-10.268; P = 0.035). cCMV disease at birth, long-term sequelae, viruria or the use of antivirals were not associated with urinalysis findings. Mild nonsymptomatic tubular proteinuria affects approximately 40% of infants and toddlers with mostly symptomatic cCMV in the first 5 years of life.

Association of urine glyphosate levels with renal injury biomarkers in children living close to major vegetable-producing regions in China

Glyphosate (GLY)-based herbicides exposure contributes to renal dysfunction in experimental conditions, but the effects on humans are rarely reported. Biomonitoring is practically relevant for evaluating the association of urine GLY levels and renal damage in children living close to vegetable-cultivating regions. In this study, we collected the first-morning void urine samples of 239 healthy children (aged 3–12, 48.12 % boys) living near major vegetable-producing regions in March–May and August 2023 in Shandong Province, China. Urine levels of GLY and kidney injury-associated biomarkers were determined using ELISA kits to assess their correlation. GLY was detected in 92.05 % of urine samples (220 out of 239 participants) and the geometric concentration (GM) was 7.429 μg/L (range: 0.625 to 38.267 μg/L). Binary logistic regression and multivariate regression analysis revealed GLY detectability and levels positively correlated with home ventilation and self-producing vegetable intake of the subjects, as well as sampling periods. Moreover, a statistically significant concentration association with urine GLY was found for kidney injury-associated biomarkers (NGAL and KIM-1) (R2 = 0.923 and 0.855, respectively). Additionally, risk assessment revealed that the maximum value of probable daily intake was 0.150 mg/kg bw/day, accounting for 30.1 % of the established Acceptable Daily Intake of GLY. This study unveils a positive correlation between continuous GLY-based herbicide exposure and renal injury biomarkers of children. A large-scale epidemiological study is warranted for comprehensively assessing the effects of GLY-based herbicides on kidney function of the entire public.

Role of luteinizing hormone urinary levels in the diagnostic and therapeutic management of female central precocious puberty

BackgroundDiagnosing central precocious puberty (CPP) requires an integrated approach based on clinical, biochemical and instrumental data. The diagnostic gold standard is represented by GnRH (gonadotropin-releasing hormone) stimulation test. Some undoubted limitations of this procedure led the international scientific community to look for cheaper and less invasive alternative diagnostic methods, such as luteinizing hormone urinary levels (uLH) measurement. This study aims to define the reliability of urinary LH levels as a biomarker of pubertal development, both concerning the initial diagnostic management and the monitoring of patients with central precocious puberty undergoing therapy with GnRH analogues. Furthermore, the study plans to detect the potential association between LH peak serum (pLH) and urinary LH in patients undergoing diagnostic tests with GnRH and to identify a possible cut-off of uLH that may be suggestive of ensued successful hormonal stimulation.MethodsThe study includes 130 female patients with suspected precocious puberty or in follow-up during suppressive therapy. After the collection of the informed consent, the patients underwent clinical evaluation, auxological assessment, and hormone assays (basal levels of LH, FSH, and oestradiol; GnRH stimulating test in patients with suspected precocious puberty; urinary LH assay on the first-morning urine sample, collected after waking up).ResultsTwo uLH cut-off values have been identified: the first of 0.25 UI/L [C.I. 95% 0.23–0.27], able to distinguish between pubertal and pre-pubertal patients, the second of 0.45 UI/L [C.I. 95% 0,20 − 0,70] suggestive of occurred hormonal stimulation in patients with diagnosis of CPP at GnRH test. All 30 patients with CPP in follow-up during suppressive therapy presented uLH values ≤ 0.45 IU/L (pU < 0.05), and uLH collected in prepubertal group control.ConclusionsuLH assays on the first morning urine specimen could be considered a low-cost and minimally invasive tool for precocious puberty diagnosing and monitoring, making possible to be easily performed even by a general pediatrician. Thus, this could help referring only selected patients to pediatric endocrinologists. After an appropriate validation, this approach could reasonably reduce hospital attendance and costs of performing more invasive procedures, with a more significant emotional impact on the pediatric patient.

Assessment of acrylamide exposure in Spain by human biomonitoring: Risk and predictors of exposure

Acrylamide (AA), a chemical compound currently classified as “reasonably anticipated to be a human carcinogen”, is formed through the Maillard reaction in processed carbohydrate-rich foods and is also present in tobacco smoke. The primary sources of AA exposure in the general population are dietary intake and inhalation. Within a 24-h period, humans eliminate approximately 50% of AA in the urine, predominantly in the form of mercapturic acid conjugates such as N-acetyl-S-(2-carbamoylethyl)-L-cysteine (AAMA), N-acetyl-S-(2-carbamoyl-2- hydroxyethyl)-L-cysteine (GAMA3), and N-acetyl-3-[(3-amino-3-oxopropyl)sulfinyl]-L-alanine (AAMA-Sul). These metabolites serve as short-term biomarkers for AA exposure in human biomonitoring studies. In this study, we analysed first-morning urine samples from the adult population (aged 18–65 years) residing in the Valencian Region, Spain, (n = 505). AAMA, GAMA-3 and AAMA-Sul were quantified in 100% of the analysed samples, with geometric means (GM) of 84, 11 and 26 μg L−1, respectively, while the estimated daily intake of AA in the studied population ranged from 1.33 to 2.13 μg·kg-bw−1·day−1 (GM). Statistical analysis of the data indicated that the most significant predictors of AA exposure were smoking and the amount of potato fried products and, biscuits and pastries consumed last 24 h. Based on risk assessment approaches conducted, the findings suggest that exposure to AA could pose a potential health risk. Therefore, it is crucial to closely monitor and continuously evaluate AA exposure to ensure the well-being of the population.

Abstract 4222: Glyphosate and AMPA exposure: Associations with dietary intake and other factors

Abstract Background: Animal and epidemiologic studies suggest that exposure to the weed killer, glyphosate, and its primary metabolite aminomethylphosphonic acid (AMPA), is associated with increased risk for cancer, including breast cancer and non-Hodgkin's lymphoma. The International Agency for Research on Cancer (IARC) has categorized glyphosate as a “probable carcinogen,” yet glyphosate is the most highly used pesticide in the world. Biomonitoring studies assessing levels of human glyphosate and AMPA exposure in the United States have been limited. Thus, we examined urinary levels of these chemicals in a cohort of postmenopausal women residing in Southern California and evaluated associations with demographics, dietary intake, and other behavioral factors. Methods: 338 postmenopausal women provided two first-morning urine samples and at least one paired 24-hour dietary recall. Participants also reported on demographic and other factors. Urinary glyphosate and AMPA were measured using LC-MS/MS. Results: Glyphosate was detected in nearly 90% of urine samples and AMPA in 67.2%. Urinary glyphosate and AMPA levels were lower in women who reported high levels of organic eating (p=0.04 and p=0.01, respectively). Urinary glyphosate was lower in women who reported higher levels of physical activity (p=0.03). Multivariate analysis showed that grain consumption, particularly refined grains, was significantly associated with higher urinary glyphosate levels (adjusted p=0.004). Alcohol consumption (adjusted p=0.001) and frequency of eating fast-food (adjusted p=0.006) were associated with higher urinary AMPA levels, while fruit (adjusted p=0.03) and corn (adjusted p=0.008) consumption were associated with lower AMPA. Conclusions: In the largest study to date examining paired dietary recall data and measurements of urinary glyphosate and AMPA, the vast majority of women sampled had detectable levels. Consumption of certain foods, including grains and alcoholic beverages, were associated with higher urinary levels of glyphosate and AMPA. Citation Format: Rachel McFarland Lucia, Xiyue Liao, Wei-Lin Huang, Danielle Forman, Alexis Kim, Argyrios Ziogas, Trina M. Norden-Krichmar, Deborah Goodman, Andrea Alvarez, Irene Masunaka, Khyatiben Pathak, Marissa McGilvrey, Victoria David-Dirgo, Apurva Hegde, Patrick Pirrotte, Hannah Lui Park. Glyphosate and AMPA exposure: Associations with dietary intake and other factors. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 4222.

Kisspeptin as potential biomarker of environmental chemical mixture effect on reproductive hormone profile: A pilot study in adolescent males

BackgroundKisspeptin has been proposed as an effect biomarker to understand the mechanisms by which some environmental chemicals adversely affect the human reproductive system. ObjectiveTo ascertain whether kisspeptin serum protein and DNA methylation levels are associated with exposure to several environmental chemicals (individually and as a mixture) and serum reproductive hormone levels in adolescent males. MethodsThree phenols (bisphenol A [BPA], methyl-paraben [MPB], and benzophenone-3 [BP3]); two toxic metals (arsenic and cadmium); and four metabolites of non-persistent pesticides, including insecticides (2-isopropyl-6-methyl-4-pyrimidinol [IMPy], malathion diacid [MDA], and dimethylcyclopropane carboxylic acid [DCCA]) and fungicides (ethylene thiourea [ETU]) were measured in first-morning urine samples of 133 adolescent males aged 15–17 years from the INMA-Granada cohort. In blood samples collected on the same day, KISS1 gene DNA methylation was measured at four CpGs from the Exon IV, as well as serum levels of kiss54 protein, total testosterone (T), estradiol (E2), sex hormone binding-globulin, dehydroepiandrosterone sulfate, luteinizing hormone (LH), and follicle-stimulating hormone (FSH). Multiple linear regression and mixture (quantile g-computation) models were fit. ResultsUrinary MDA and DCCA concentrations were associated with higher kiss54 levels [% change (95%CI) for each log-unit increase in concentration = 2.90 (0.32;5.56), and 1.93 (0.45,3.43), respectively]; IMPy with lower DNA methylation percentage at CpG1 and total CpGs [% change (95%CI) = −1.15 (−1.96;-0.33): −0.89 (−1.73;-0.01), respectively]; and BP3 and DCCA with lower total CpGs methylation [−0.53 (−1.04;-0.01) and − 0.69 (−1.37;-0.01), respectively]. The pesticide mixture and the whole chemical mixture were associated with higher kiss54 [% change (95%CI) = 9.09 (3.29;15.21) and 11.61 (3.96;19.82), respectively] and lower methylation levels at several CpGs. Additionally, serum kiss54 in the third tertile was associated with higher LH levels [% change (95%CI) = 28.69 (3.75–59.63)], and third-tertile CpG1, CpG2, and total CpG methylation percentages were associated with lower FSH and E2. ConclusionThe findings of the present study and the negative correlation between serum kiss54 levels and KISS1 DNA methylation percentages suggested that kisspeptin may be a promising effect biomarker.