Nutrient Exposure Research Articles

Macronutrient Intake in Pregnancy and Child Cognitive and Behavioural Outcomes

Prenatal nutrient exposures can impact on brain development and disease susceptibility across the lifespan. It is well established that maternal macronutrient intake during pregnancy influences foetal and infant development. Therefore, we hypothesise that macronutrient intakes during pregnancy are correlated with cognitive development during early childhood. The current study aimed to investigate the relationship between maternal macronutrient intake during pregnancy and child cognitive and behavioural outcomes at age 4 years. We analysed prospective data from a cohort of 64 Australian mother–child dyads. Maternal macronutrient intake was assessed using a validated 74-item food frequency questionnaire at 2 timepoints during pregnancy. Child cognition and behaviour were measured at age 4 years using the validated Wechsler Preschool and Primary Scale of Intelligence, 3rd version (WPPSI-III) and the Child Behaviour Checklist (CBC). Linear regression models were used to quantify statistical relationships and were adjusted for maternal age, education, pre-pregnancy BMI, breastfeeding duration and birthweight. Child Performance IQ was inversely associated with maternal starch intake (b = −11.02, p = 0.03). However, no other associations were found. Further research is needed to explore the association between different types of starch consumed during pregnancy and child cognitive development.

Carbohydrate Intake Prior to Oral Glucose Tolerance Testing.

With the emergence of glycated hemoglobin as a diagnostic test for diabetes, oral glucose tolerance tests (OGTTs) have become rare in endocrinology practice. As they have moved out of favor, the importance of patient instructions on preparation prior to OGTT has faded from memory. Decades-old literature, well-known to endocrinologists a generation ago, emphasized the importance of carbohydrate intake prior to OGTT. In this expert endocrine consult, we discuss an OGTT performed in a research setting without adequate carbohydrate intake at the evening meal prior to the OGTT. The resultant elevated plasma glucose levels at 1-hour and 2-hours mimicked the loss of first-phase insulin release seen in early type 1 and type 2 diabetes. With clinical concern that the research participant had evolving type 1 or type 2 diabetes, the volunteer was subjected to additional testing and experienced anxiety. Repeat OGTT was normal after adequate carbohydrate intake (>150 grams/day and >50 grams the evening prior to overnight fast for the study). The physiology of this phenomenon is explored and is likely mediated through beta cell adaptation and alteration in peripheral glucose uptake in response to nutrient exposure. The learnings of decades ago have clearly faded, and this literature should be revisited to ensure that OGTT results are not compromised when ordered for clinical or research purposes.

Exposure to elevated nutrient load results in structural and functional changes to microbial communities associated with riparian wetland plants Phalaris arundinaceae and Veronica anagallis-aquatica

The goal of this study was to better understand the resilience of rhizosphere microbial communities located in riparian wetlands subjected to high nutrient loads following effluent or runoff exposure. Specifically, this research monitored functional profiles of mesocosm communities during an artificial high nitrogen load exposure following adaptation to high or low water quality conditions. Based on prior research, two species of wetland plants were chosen, Phalaris arundinaceae and Veronica anagallis-aquatica, both located in local riparian zones, and planted in wetland mesocosms. Rhizosphere bacterial community response was monitored for approximately three weeks after exposure to elevated nutrient loads, following prior adaptation for several months to either high or low (poor) water quality conditions. The impact of prior acclimation conditions was then assessed. Changes to rhizosphere bacterial community structure and function were monitored using PCR-denaturing gel gradient electrophoresis (DGGE) and Biolog™ Ecoplates, respectively. Plants were also sampled to assess any impact on mycorrhizal associations over the exposure period. As an additional indicator of functionality and nitrogen utilization within the mesocosm system, inorganic intermediate N species were monitored at the outflow. Significant differences were observed over time following elevated nutrient exposure in bacterial structural diversity and richness, as well as functional diversity and richness (p < 0.001). The effects of interactions between time, plant species and water quality exposure were also significant for both structural (p = 0.002) and functional (p = 0.05) community diversity. Elevated nutrient exposure resulted in changes to microbial carbon source utilization profiles, specifically, bacterial usage of amino acids (p = 0.032), polymers (p = 0.029) and carboxylic and ketonic acids (p = 0.042). Carboxylic and ketonic acid utilization by bacterial communities was positively correlated with the removal of nitrate and nitrite from experimental mesocosms. Mycorrhizal associations as measured by percent colonization also decreased significantly in P. arundinaceae following elevated nutrient exposure (p < 0.001). Furthermore, abundance of nitrogenous compounds in outflow water differed between plant species treatments, with V. anagallis-aquatica removing more nitrate from the outflow (p = 0.005) while P. arundinaceae removed higher levels of ammonia (p = 0.03). This study provides insight into how microbial communities respond to transient nutrient loads such as that experienced by overland runoff following a high rain event and will assist in the design and rehabilitation of wetlands or riparian buffer strips to manage higher nutrient loads resulting from increasing human activity impacting our freshwater systems.

Nutrient Exposure Alters Microbial Composition, Structure, and Mercury Methylating Activity in Periphyton in a Contaminated Watershed.

The conversion of mercury (Hg) to monomethylmercury (MMHg) is a critical area of concern in global Hg cycling. Periphyton biofilms may harbor significant amounts of MMHg but little is known about the Hg-methylating potential of the periphyton microbiome. Therefore, we used high-throughput amplicon sequencing of the 16S rRNA gene, ITS2 region, and Hg methylation gene pair (hgcAB) to characterize the archaea/bacteria, fungi, and Hg-methylating microorganisms in periphyton communities grown in a contaminated watershed in East Tennessee (United States). Furthermore, we examined how nutrient amendments (nitrate and/or phosphate) altered periphyton community structure and function. We found that bacterial/archaeal richness in experimental conditions decreased in summer and increased in autumn relative to control treatments, while fungal diversity generally increased in summer and decreased in autumn relative to control treatments. Interestingly, the Hg-methylating communities were dominated by Proteobacteria followed by Candidatus Atribacteria across both seasons. Surprisingly, Hg methylation potential correlated with numerous bacterial families that do not contain hgcAB, suggesting that the overall microbiome structure of periphyton communities influences rates of Hg transformation within these microbial mats. To further explore these complex community interactions, we performed a microbial network analysis and found that the nitrate-amended treatment resulted in the highest number of hub taxa that also corresponded with enhanced Hg methylation potential. This work provides insight into community interactions within the periphyton microbiome that may contribute to Hg cycling and will inform future research that will focus on establishing mixed microbial consortia to uncover mechanisms driving shifts in Hg cycling within periphyton habitats.

Differential resistance and acclimation of two coral species to chronic nutrient enrichment reflect life‐history traits

Abstract The effects of nutrient pollution on coral reef ecosystems are multifaceted. Numerous experiments have sought to identify the physiological effects of nutrient enrichment on reef‐building corals, but the results have been variable and sensitive to choices of nutrient quantity, chemical composition and exposure duration. To test the effects of chronic, ecologically relevant nutrient enrichment on coral growth and photophysiology, we conducted a 5‐week continuous dosing experiment on two Hawaiian coral species, Porites compressa and Pocillopora acuta. We acclimated coral fragments to five nutrient concentrations (0.1–7 µM and 0.06–2.24 µM ) with constant stoichiometry 2.5:1 nitrate to phosphate) bracketing in situ observations from reefs throughout the Pacific. Nutrient enrichment linearly increased photophysiological performance of both species within 3 weeks. The effect of nutrients on P. acuta photochemical efficiency increased through time while a consistent response in P. compressa indicated acclimation to elevated nutrients within 5 weeks. Endosymbiont densities and total chlorophyll concentrations also increased proportionally with nutrient enrichment in P. acuta, but not in P. compressa, revealing contrasting patterns of host–symbiont acclimatization. The two species also exhibited contrasting effects of nutrient enrichment on skeletal growth. Calcification was enhanced at low nutrient enrichment (1 µM ) in P. acuta, but comparable to the control at higher concentrations, whereas calcification was reduced in P. compressa (30%–35%) above 3 µM . Stable isotope analysis revealed species‐specific nitrogen uptake dynamics in the coral–algal symbiosis. The endosymbionts of P. acuta exhibited increased nitrogen uptake (decreased δ15N) and incorporation (19%–31% decrease in C:N ratios) across treatments. In contrast, P. compressa endosymbionts maintained constant δ15N values and low levels of nitrogen incorporation (9%–11% decrease in C:N ratios). The inability of P. acuta to regulate endosymbiont nutrient uptake may indicate an emerging destabilization in the coral–algal symbiosis under nutrient enrichment that could compromise resistance to additional environmental stressors. Our results highlight species‐specific differences in the coral–algal symbiosis, which influence responses to chronic nutrient enrichment. These findings showcase how symbioses can vary among closely related taxa and underscore the importance of considering how life‐history traits modify species response to environmental change. A free Plain Language Summary can be found within the Supporting Information of this article.

Structural, Metabolic and Evolutionary Comparison of Bacterial Endospore and Exospore Formation.

Sporulation is a specialized developmental program employed by a diverse set of bacteria which culminates in the formation of dormant cells displaying increased resilience to stressors. This represents a major survival strategy for bacteria facing harsh environmental conditions, including nutrient limitation, heat, desiccation, and exposure to antimicrobial compounds. Through dispersal to new environments via biotic or abiotic factors, sporulation provides a means for disseminating genetic material and promotes encounters with preferable environments thus promoting environmental selection. Several types of bacterial sporulation have been characterized, each involving numerous morphological changes regulated and performed by non-homologous pathways. Despite their likely independent evolutionary origins, all known modes of sporulation are typically triggered by limited nutrients and require extensive membrane and peptidoglycan remodeling. While distinct modes of sporulation have been observed in diverse species, two major types are at the forefront of understanding the role of sporulation in human health, and microbial population dynamics and survival. Here, we outline endospore and exospore formation by members of the phyla Firmicutes and Actinobacteria, respectively. Using recent advances in molecular and structural biology, we point to the regulatory, genetic, and morphological differences unique to endo- and exospore formation, discuss shared characteristics that contribute to the enhanced environmental survival of spores and, finally, cover the evolutionary aspects of sporulation that contribute to bacterial species diversification.

The Cancer Prevention Study-3 FFQ Is a Reliable and Valid Measure of Nutrient Intakes among Racial/Ethnic Subgroups, Compared with 24-Hour Recalls and Biomarkers

BackgroundValid assessment of dietary intake in diverse populations is important for studies of chronic disease risk in the United States. ObjectivesWe evaluated the reproducibility and validity of a food frequency questionnaire (FFQ) modified for the American Cancer Society’s Cancer Prevention Study-3 (CPS-3) prospective cohort, among a racially/ethnically diverse subgroup. MethodsThe Diet Assessment Substudy included 677 CPS-3 participants (64% women; 61% non-Hispanic white, 24% non-Hispanic black, 15% Hispanic), aged 31–70 y, who completed 2 FFQs 1 y apart (FFQ1, FFQ2), 4–6 telephone-administered 24-h dietary recalls (24HRs), and 2 fasting blood samples and 24-h urine collections ∼6 mo apart in the interim. Spearman rank correlation coefficients (ρ) were used to evaluate FFQ reproducibility and validity compared with 24HRs for 67 nutrient exposures. For 18 of these nutrients, we used the method of triads to calculate validity coefficients (VCs, ρ) from pairwise correlations of FFQ2, 24HRs, and biomarkers. Analyses were stratified by sex, race/ethnicity, education, and BMI. ResultsMean (range) FFQ reproducibility correlations were ρ = 0.65 (0.50–0.91) for men and ρ = 0.63 (0.37–0.89) for women; mean (range) energy-adjusted, deattenuated correlations of FFQ2 with 24HRs were ρ = 0.60 (0.33–0.84) for men and ρ = 0.55 (0.21–0.79) for women. FFQ2 VCs (ρ) among men ranged from 0.42 for β-cryptoxanthin to 0.91 for omega-3 (n–3) fatty acids and, among women, from 0.41 for sodium to 0.79 for total vitamin D. Mean FFQ reproducibility and validity were highest among whites (ρ = 0.68, ρ = 0.58, respectively) and slightly lower among blacks (ρ = 0.57, ρ = 0.49, respectively) and Hispanics (ρ = 0.59, 0.55, respectively). FFQ reproducibility and validity were slightly lower among those with less than a 4-y college degree, and those with a BMI ≥30 kg/m2. ConclusionsReproducibility and validity of the CPS-3 FFQ were comparable with similar studies for most nutrients, among all subgroups. These findings support future dietary analyses in the contemporary CPS-3 cohort and other similar cohorts.

Adipose Triglyceride Lipase Loss Promotes a Metabolic Switch in A549 Non–Small Cell Lung Cancer Cell Spheroids

Cancer cells undergo complex metabolic adaptations to survive and thrive in challenging environments. This is particularly prominent for solid tumors, where cells in the core of the tumor are under severe hypoxia and nutrient deprivation. However, such conditions are often not recapitulated in the typical 2D in vitro cancer models, where oxygen as well as nutrient exposure is quite uniform. The aim of this study was to investigate the role of a key neutral lipid hydrolase, namely adipose triglyceride lipase (ATGL), in cancer cells that are exposed to more tumor-like conditions. To that end, we cultured lung cancer cells lacking ATGL as multicellular spheroids in 3D and subjected them to comprehensive proteomics analysis and metabolic phenotyping. Proteomics data are available via ProteomeXchange with identifier PXD021105. As a result, we report that loss of ATGL enhanced growth of spheroids and facilitated their adaptation to hypoxia, by increasing the influx of glucose and endorsing a pro-Warburg effect. This was followed by changes in lipid metabolism and an increase in protein production. Interestingly, the observed phenotype was also recapitulated in an even more “in vivo like” setup, when cancer spheroids were grown on chick chorioallantoic membrane, but not when cells were cultured as a 2D monolayer. In addition, we demonstrate that according to the publicly available cancer databases, an inverse relation between ATGL expression and higher glucose dependence can be observed. In conclusion, we provide indications that ATGL is involved in regulation of glucose metabolism of cancer cells when grown in 3D (mimicking solid tumors) and as such could be an important factor of the treatment outcome for some cancer types. Finally, we also ratify the need for alternative cell culture models, as the majority of phenotypes observed in 3D and spheroids grown on chick chorioallantoic membrane were not observed in 2D cell culture.

Process-based modeling to assess the nutrient removal efficiency of two endangered hydrophytes: Linking nutrient-cycle with a multiple-quotas approach

Hydrophytes have been widely used to reduce nutrient levels in aquatic ecosystems, but only limited species with high nutrient removal efficiencies have been implemented. Thus, it is necessary to continually explore new candidate species with high nutrient removal efficiencies. To effectively explore the nutrient removal ability of hydrophytes, a new process-based model combining the multiple-quotas approach and nutrient-cycle model was developed. The multiple-quotas approach provides a theoretical framework to conceptually explain the uptake and response of autotrophs to multiple nutrients. The developed process-based model was validated using observational data from microcosm experiments with two emergent hydrophytes, Menyanthes trifoliata and Cicuta virosa. The results showed that both M. trifoliata and C. virosa effectively reduced nitrogen (N) and phosphorus (P) in both water and sediment layers, but M. trifoliata showed a higher removal efficiency for both nutrients than C. virosa, particularly for total ammonia + ammonium-nitrogen (NHx-N) and nitrate-nitrogen (NO3-N) in the sediment layer (M. trifoliata: 0.579–0.976 for NHx-N, 0.567–0.702 for NO3-N; C. virosa: 0.212–0.501 for NHx-N, 0.466–0.560 for NO3-N). In addition, M. trifoliata achieved the maximum removal efficiency for N and P at higher nutrient exposure levels than C. virosa (M. trifoliata: exposure level of 0.725–0.775; C. virosa: exposure level of 0.550–0.575). The developed model well simulated the species-specific growth patterns of hydrophytes depending on the nutrient exposure level as well as the N and P dynamics in the water and sediment layers. The approach adopted in this study provides a useful tool for discovering candidate species to improve hydrophyte diversity and effectively remove nutrients from aquatic ecosystems.

Seasonal periphyton response to low-level nutrient exposure in a least disturbed mountain stream, the Buffalo River, Arkansas

Like most streams located in the Ozark Plateaus, the Buffalo River in Arkansas generally has excellent water quality. Water-quality conditions in Big Creek, however, a major tributary of the middle Buffalo River, have been less favorable than that of other Buffalo River tributaries. Concerns regarding the influence of water quality in Big Creek on the Buffalo River magnified in 2013 when a large confined animal feeding operation (CAFO) began operating in the watershed. In response to these concerns, the U.S. Geological Survey compared monthly nutrient concentrations and seasonal periphyton assemblage metrics of a site on Big Creek downstream of the CAFO, two Buffalo River control sites upstream of the confluence with Big Creek, and three Buffalo River test sites downstream of the confluence with Big Creek. In addition to identifying potential nutrient patterns and periphyton responses along a low-level nutrient exposure gradient, the study determined how nutrient contributions from Big Creek (and the CAFO) are affecting ecological conditions and consequent ecosystem services in the Buffalo River. Nutrient and periphyton data exhibited more temporal than spatial variability. Nutrient concentrations were generally highest of all sites at the Big Creek site. Concentrations at the five sites on the Buffalo River were typically low (near laboratory reporting limits), and concentrations at the three test sites rarely exceeded those of the two control sites. An index developed with three ecologically relevant periphyton metrics (oligotrophic taxa and Homoeothrix percent relative abundance and mesotrophic diatoms percent taxa richness) suggested that nutrient uptake at sites downstream of the Big Creek-Buffalo River confluence resulted in subtle shifts in downstream periphyton assemblages. The periphyton index of biological integrity at control sites was slightly and generally more favorable compared to test sites. Even so, when periphyton data were considered in conjunction with both hydrology and water-quality data, the negative consequences of antecedent high flows and associated scouring exceeded the potential positive effects that low-level nutrients had on algal productivity. These findings emphasize the importance of comparing biological and chemical data across extended temporal scales, particularly when working with low-level nutrient gradients.

Risk Factors Associated with Carbapenemase-Producing Enterobacterales (CPE) Positivity in the Hospital Wastewater Environment.

Hospital wastewater is an increasingly recognized reservoir for resistant Gram-negative organisms. Factors involved in establishment and persistence of Klebsiella pneumoniae carbapenemase-producing organisms (KPCOs) in hospital wastewater plumbing are unclear. This study was conducted at a hospital with endemic KPCOs linked to wastewater reservoirs and robust patient perirectal screening for silent KPCO carriage. Over 5 months, both rooms occupied and rooms not occupied by KPCO-positive patients were sampled at three wastewater sites within each room (sink drain, sink P-trap, and toilet or hopper). Risk factors for KPCO positivity were assessed using logistic regression. Whole-genome sequencing (WGS) identified environmental seeding by KPCO-positive patients. A total of 219/475 (46%) room sampling events were KPCO positive in at least one wastewater site. KPCO-positive patient exposure was associated with increased risk of environmental positivity for the room and toilet/hopper. Previous positivity and intensive care unit room type were consistently associated with increased risk. Tube feeds were associated with increased risk for the drain, while exposure to patients with Clostridioides difficile was associated with decreased risk. Urinary catheter exposure was associated with increased risk of P-trap positivity. P-trap heaters reduced risk of P-trap and sink drain positivity. WGS identified genomically linked environmental seeding in 6 of 99 room occupations by 40 KPCO-positive patients. In conclusion, KPCO-positive patients seed the environment in at least 6% of opportunities; once positive for KPCOs, wastewater sites are at greater risk of being positive subsequently. Increased nutrient exposure, e.g., due to tube food disposal down sinks, may increase risk; frequent flushing may be protective.IMPORTANCEKlebsiella pneumoniae carbapenemase-producing organisms (KPCOs) are bacteria that are resistant to most antibiotics and thus are challenging to treat when they cause infections in patients. These organisms can be acquired by patients who are hospitalized for other reasons, complicating their hospital stay and even leading to death. Hospital wastewater sites, such as sink drains and toilets, have played a role in many reported outbreaks over the past decade. The significance of our research is in identifying risk factors for environmental positivity for KPCOs, which will facilitate further work to prevent transmission of these organisms to patients from the hospital environment.

Identifying environmental risk factors for inflammatory bowel diseases: a Mendelian randomization study

Several studies have examined environmental factors and inflammatory bowel diseases (IBD) using traditional approaches; however, provided results are still conflicting. Our aim was to determine whether lifestyle and nutrient exposures, related to IBD in observational meta-analyses, influence IBD risk using a Mendelian randomization (MR) approach. A two-sample MR approach was applied on summary-level genome-wide association results. Genetic variants strongly associated with measures of tobacco smoking, obesity and fat distribution, physical activity, and blood levels of vitamins and fatty acids were evaluated on genetic data from international IBD consortia including a total of 25,042 IBD cases (12,194 cases of Crohn’s disease (CD) and 12,366 cases of ulcerative colitis (UC)) and 34,915 controls. Our results indicated that, among lifestyle exposures, being a smoker was positively associated with CD (OR 1.13, P = 0.02), but it was not associated with UC risk (OR 0.99, P = 0.88). Body-mass index (BMI) and body fat percentage were positively associated with CD (OR 1.11, P = 0.02, per standard deviation (SD) of 4.6 kg/m2; and OR 1.50, P = 3 × 10–10, per SD of 6.6%; respectively); while for UC, BMI was inversely associated (OR 0.85, P = 5 × 10–5; per SD) and body fat percentage showed a OR of 1.11 (P = 0.11; per SD). Additionally, among nutrient exposures, omega-3 fatty acids levels were inversely associated with CD (OR 0.67, P = 2 × 10–6). Our MR results did not support a protective effect for being a smoker on UC risk; however, they are compatible with a risk effect for higher body fat proportion and a protective role for higher levels of omega-3 fatty acids on CD etiology.

Dietary patterns among pregnant women in the SELMA study

Background: Nutrition is important during pregnancy both for the health of the mother and fetal development. There are nutritional guidelines for pregnant women in order to get the nutrients that are needed for healthy pregnancy outcomes. Our objective was to identify dietary patterns among pregnant women. Methods: We used data from 1,770 pregnant women participating in the Swedish Environmental Longitudinal Mother and child Asthma and allergy (SELMA) study. Food intake was collected through a 30-day recall food-frequency questionnaire during 2nd trimester of pregnancy. Dietary intake was calculated to grams per day. We performed principal component (PC) analysis to identify different food patterns. Results: Overall, women were 31 years of age, of normal weight (BMI=25), a college degree or higher (65%), and were mainly non-smokers (95%). We found 11 principal components (PC) with eigenvalues above 1 explaining 61% of the dietary variation. PC1, representing 13% of the variance, showed a varied diet with higher positive loadings for meat, fish, vegetables, fruits, cakes, and sweets. PC2 (8% variance) had higher loadings on cakes and sweets and lower on vegetables, whereas PC3 (7% variance) had negative loadings on fish. The group in PC4 (6% variance) had a less varied diet with positive loadings on sweets and vegetables but negative loadings on dairy, cereals, meat and fish. Women in PC8 (4% variance) ate more dairy, meat and fish but no sweets, bread or cereals. These dietary patterns are currently being analyzed with a nutrition index based on nutritional guidelines and with prenatal exposure to endocrine disruptors (EDCs) in a mixture analysis. Conclusion: Results provide a depiction of women’s dietary patterns during mid-pregnancy. They will be further analyzed with prenatal nutrient intake and exposure to EDCs in order to get a better understanding of the relationship between food patterns, nutrition, and exposure to EDCs

Functional Insights Into the Role of gppA in (p)ppGpp Metabolism of Vibrio cholerae.

The stringent response, an adaptive response to nutrient limitation and exposure to xenobiotics in bacteria, is mediated by two intracellular signaling molecules, pppGpp and ppGpp, together called (p)ppGpp. The cellular level of (p)ppGpp in bacterial cells is controlled by the Rel/Spo family of proteins. In the cholera pathogen, Vibrio cholerae, (p)ppGpp metabolism is regulated by the products of at least three genes relA, spoT, and relV. In this study, we identify and characterize the function of the guanosine-5′-triphosphate 3′-diphosphate pyrophosphatase A (GppA) encoding gene gppA of V. cholerae. Genomic analysis indicates that the gppA locus is conserved in vibrios and organized as a bicistronic operon along with the rhlB gene. We engineered the genome of V. cholerae to develop different mutants devoid of GppA and/or other phosphate metabolic enzymes. Our findings indicate that in V. cholerae, GppA plays an important role in the conversion of pppGpp to ppGpp during amino acid deprivation but not during glucose starvation. Quantitative analyses of the gppA transcript level reveal its differential expression pattern at different growth phases and starvation conditions. It has been observed that the GppA deficiency during amino acid starvation condition could be complemented by overexpressing the exopolyphosphatase coding gene ppx of V. cholerae. By deletion analysis, we further demonstrate that the amino and carboxy terminal sequences flanking the Ppx-GppA motif of the GppA protein of V. cholerae are also important for its enzymatic function.

Culture Conditions Affect Chemical-Induced Developmental Toxicity In Vitro: The Case of Folic Acid, Methionine and Methotrexate in the Neural Embryonic Stem Cell Test.

In vitro tests are increasingly applied in chemical hazard assessment. Basic culture conditions may affect the outcome of in vitro tests and should be optimised to reduce false predictions. The neural embryonic stem cell test (ESTn) can predict early neurodevelopmental effects of chemicals, as it mimics the differentiation of stem cells towards the neuroectodermal lineage. Normal early neural differentiation depends crucially on folic acid (FA) and methionine (MET), both elements of the one-carbon (1C) cycle. The aim of this study was to assess the concentration-dependent influence of FA and MET on neural differentiation in the ESTn, and its consequences for assay sensitivity to methotrexate (MTX), a compound that interferes with the 1C cycle. Neural differentiation was inhibited below 0.007 mM and above 0.22 mM FA, while both stem cell viability (< 0.097 mM, > 1.52 mM) and neural differentiation (< 0.181 mM, > 1.35 mM) were affected when changing MET concentrations. A 10-day exposure to 13 nM MTX inhibited neural differentiation, especially in FA- and MET-deficient conditions. However, a 24-hour exposure to 39 nM MTX decreased neural cell and neural crest cell differentiation markers only when the concentration of FA in the medium was three times the standard concentration, which was expected to have a protective effect against MTX. These results show the importance of nutrient concentrations, exposure scenarios and timing of read-outs for cell differentiation and compound sensitivity in the ESTn. Caution should be taken when interpreting results from a single in vitro test, especially when extrapolating to effects on complex morphogenetic processes, like neural tube development.

Simulation of Bottom Phytocenosis in the Crimean Coastal Zone

Purpose. The work is aimed at simulating production processes in the bottom phytocenosis typical of the rocky soils in the Crimean coastal zone. The macroalgae uptake of the main nutrients (phosphates, ammonium and nitrates) and release of oxygen and dissolved compounds are planned to be estimated. Methods and Results. The study was based on the balance model of the macroalgae growth and interaction with the environment. The model empirical parameters were determined from the published data of the laboratory experiments. The bottom phytocenosis in the model was represented by seven algae species: Cystoseira barbata, Ulva lactuca, Ceramium tenuicorne, Cladophora glomerata, Polysiphonia nigrescens, Phyllophora truncata, Enteromorpha prolifera. The main simulation results were: annual estimates of the released and absorbed substances, seasonal dynamics of the macroalgae biomass and evaluation of the bottom phytocenosis response to the changing environmental conditions. The rate of the production processes in the model was influenced both by the external (water temperature, insolation and concentration of nutrients in water) and internal (intracellular quotas of nitrogen and phosphorus, and species-specific parameters determining efficiency of nutrient assimilation by the algae tissues) factors. According to the simulation results, the ratio of nitrogen to phosphorus in the algae tissues was significantly higher than that in the seawater and varied from 30 to 60 for different species depending on water temperature, light exposure and concentration of nutrients. Conclusions. During the periods of intensive macroalgae growth (from April to September), the rate of nitrogen and phosphorus uptake can increase by a factor of 1.5 in comparison with the winter period. Maximum contribution to water purification from the surplus nutrients was made by the green algae: Ulva lactuca, Cladophora glomerata and Enteromorpha prolifera. At that Cladophora glomerata has shown the best adaptive abilities (minimal decrease in the growth rate) under a sharp drop of nutrients concentration in water.

Nutrient-toxic element mixtures and the early postnatal gut microbiome in a United States longitudinal birth cohort

BackgroundThe infant microbiome contributes to health status across the lifespan, but environmental factors affecting microbial communities are poorly understood, particularly when toxic and essential elements interact. ObjectiveWe aimed to identify the associations between a spectrum of other early-postnatal nutrient or toxic elemental exposures measured and the infant gut microbiome. MethodsOur analysis included 179 six-week-old infants from the New Hampshire Birth Cohort Study. Eleven elements were measured in infant toenail clippings. The gut microbiome was assessed using 16S rRNA V4-V5 hypervariable region targeted sequencing. Multivariable zero-inflated logistic normal regression (MZILN) was used to model the association between element concentrations and taxon relative abundance. To explore interactive and nonlinear associations between the exposures and specific taxa we employed Bayesian Kernel Machine Regression (BKMR). Effect modification by delivery mode, feeding mode, peripartum antibiotic exposure, and infant sex was assessed with stratified models. ResultsWe found a negative association between arsenic and microbial diversity in the full population that was accentuated among infants exposed to peripartum antibiotics. Arsenic, cadmium, copper, iron, lead, manganese, nickel, selenium, tin, and zinc were each associated with differences in at least one taxon in the full study population, with most of the related taxa belonging to the Bacteroides and Lactobacillales. In stratified analyses, mercury, in addition to the other elements, was associated with specific taxa. Bifidobacterium, which associated negatively with zinc in MZILN and BKMR models, had a quadratic association with arsenic concentrations. These associations varied with the concentration of the other element. ConclusionsEarly postnatal toxic and nutrient elemental exposures are associated with differences in the infant microbiome. Further research is needed to clarify the whether these alterations are a biomarker of exposure or if they have implications for child and lifelong health.

Nutritional prevention of colorectal cancer.

The preventability estimate for colorectal cancer (CRC) is approximately 50%, highlighting the huge potential for altering modifiable lifestyle factors (including diet and body fatness) in order to reduce risk of this common malignancy. There is strong evidence that dietary factors (including intake of wholegrains, fibre, red and processed meat and alcohol) affect CRC risk. The lack of positive intervention trials and limited mechanistic understanding likely explain limited public health impact of epidemiological observations, to date. An alternative strategy for nutritional prevention of CRC is use of supplements that provide higher individual nutrient exposure than obtained through the diet (chemoprevention). There are positive data for calcium and/or vitamin D and the n-3 fatty acid EPA from polyp prevention trials using colorectal adenoma as a CRC risk biomarker. Although CRC is an obesity-related malignancy, there remains a paucity of observational data supporting intentional weight loss for CRC risk reduction. Some types of obesity surgeries (Roux-en-Y gastric bypass) might actually increase subsequent CRC risk due to alteration of local intestinal factors. There is intense interest in nutritional therapy of patients after diagnosis of CRC, in order to impact on recurrence and overall survival (now often termed cancer interception). In conclusion, nutritional prevention of CRC continues to hold much promise. Increased mechanistic understanding of the role of individual nutrients (linked to intestinal microbiota), as well as a precision medicine approach to CRC chemoprevention and interception based on both tumour and host factors, should enable translation of nutritional interventions into effective CRC risk reduction measures.

Melanization in Cryptococcus neoformans Requires Complex Regulation.

The fungal human pathogen Cryptococcus neoformans undergoes melanization in response to nutrient starvation and exposure to exogenous melanin precursors. Melanization protects the fungus against host defense mechanisms such as oxidative damage and other environmental stressors (e.g., heat/cold stress, antimicrobial compounds, ionizing radiation). Conversely, the melanization process generates cytotoxic intermediates, and melanized cells are potentially susceptible to overheating and to certain melanin-binding drugs. Despite the importance of melanin in C. neoformans biology, the signaling mechanisms regulating its synthesis are poorly understood. The recent report by D. Lee, E.-H. Jang, M. Lee, S.-W. Kim, et al. [mBio 10(5):e02267-19, 2019, https://doi.org/10.1128/mBio.02267-19] provides new insights into how C. neoformans regulates melanization. The authors identified a core melanin regulatory network consisting of transcription factors and kinases required for melanization under low-nutrient conditions. The redundant and epistatic connections of this melanin-regulating network demonstrate that C. neoformans melanization is complex and carefully regulated at multiple levels. Such complex regulation reflects the multiple functions of melanin in C. neoformans biology.

Simultaneous exploration of nutrients and pollutants in human milk and their impact on preterm infant growth: An integrative cross-platform approach

Early nutritional management including fortified human breastmilk is currently recommended to fulfil the energy demands and counterbalance risks associated to preterm birth. However, little is known about the potential adverse effects of exposure to persistent organic pollutants (POPs) carried in human milk on preterm infant growth. We conducted a pilot study proving the application of an integrative analytical approach based on mass spectrometry (MS) coupled to advanced statistical models, favouring the comprehensive molecular profiling to support the identification of multiple biomarkers. We applied this workflow in the frame of a preterm infants’ cohort to explore environmental determinants of growth. The combination of high resolution gas and liquid chromatography MS platforms generated a large molecular profile, including 102 pollutants and nutrients (targeted analysis) and 784 metabolites (non-targeted analysis). Data analysis consisted in a preliminary examination of associations between the signatures of POPs and the normalised growth of preterm infants, using multivariate linear regression adjusting for known confounding variables. A second analysis aimed to identify multidimensional biomarkers using a multiblock algorithm allowing the integration of multiple datasets in the growth model of preterm infants. The preliminary results did not suggest an impairment of preterm growth associated to the milk concentrations of POPs. The multiblock approach however revealed complex interrelated molecular networks of POPs, lipids, metabolites and amino acids in breastmilk associated to preterm infant growth, supporting the high potential of biomarkers exploration of this proposed workflow. Whereas the present study intended to identify simultaneously pollutant and nutrient exposure profiles associated to early preterm infant growth, this workflow may be easily adapted and applied to other matrices (e.g. serum) and research settings, favouring the functional exploration of environmental determinants of complex and multifactorial diseases.