Nutrient Fluctuations Research Articles

Seasonal and long-term variations of nutrients in Liaodong Bay, China: Influencing factors and ecological effects

Nutrients are essential for marine primary productivity and have a critical role in maintaining the structure and function of marginal. Variations in nutrient levels in sea ecosystems can influence ecological disturbances significantly. The Liaodong Bay (LDB) is a semi-enclosed marginal sea in northern China. It has experienced severe eutrophication since the 1990s, leading to considerable environmental challenges. Understanding of seasonal and long-term nutrient dynamics in the LDB is limited. We examined seasonal datasets collected in May (spring), August (summer), November (autumn), and March (winter) of 2019, and analyzed long-term trends through historical records spanning multiple decades. Nutrients accumulated during autumn/winter but were depleted during spring/summer. A low concentration of dissolved inorganic phosphate led to an increased nitrogen ratio exceeding the Redfield ratio (>16) during winter, spring, and summer, driven by phytoplankton growth. In late-autumn, nutrient concentrations increased, with ratios approaching the Redfield ratio. Phosphorus limitation prevailed in spring, summer, and winter, while silicon limitation dominated in autumn. Dissolved inorganic nitrogen and nitrogen ratios in the LDB increased sharply since the 1980s, peaking before declining after 2013. Dissolved silica and silicon ratios decreased steadily, stabilizing in recent years. These trends imply a shift from nitrogen-to-phosphorus limitation, influenced by riverine inputs and atmospheric deposition. These nutrient fluctuations may have significant ecological effects, including dinoflagellate abundance, algal blooms, and jellyfish blooms. Our analyses highlight the complexity of nutrient dynamics and positive impact of local nutrient-reduction policies implemented in recent years in improving the environmental quality of the LDB.

Hydrological-driven changes in the phytoplankton community structure under nutrient stress in island river ecosystems

While the spatiotemporal dynamics of phytoplankton community structures in river ecosystems have been extensively studied, their primary driving factors remain elusive, particularly at the watershed scale. This research examined the phytoplankton community structure across three watersheds of an island and concurrently analyzed its response to water level fluctuations (WLFs). The phytoplankton density was notably higher at elevated water levels, registering an increase of 1.29 times in Nandu River, 1.34 times in Changhua River, and 1.28 times in Wanquan River. Notably, the middle reaches of the Changhua River recorded the highest phytoplankton density (4.46 × 108 cells/L). Broadly speaking, Cyanophyta, Chlorophyta, and Bacillariophyta remained the predominant phytoplankton across varied water levels. Dominant phytoplankton species varied with water levels; however, the most prevalent species consistently belonged to the Cyanophyta group, primarily Microcystis or Merismopedia, with dominance ranging between 0.17 to 0.38 (low water level) and 0.18 to 0.32 (high water level). Enhanced phytoplankton diversity and richness were observed at higher water levels, correlating with increased concentrations of NH4+-N, NO3−-N, and PO43− in the river water. Consequently, nutrient fluctuations driven by hydrodynamics significantly influence the phytoplankton community structure in island river ecosystems.

Optimal resource allocation in micro-organisms under periodic nutrient fluctuations

Although microorganisms often live in dynamic environments, most studies, both experimental and theoretical, are carried out under static conditions. In this work, we investigate the issue of optimal resource allocation in bacteria growing in periodic environments. We consider a dynamic model describing the microbial metabolism under varying conditions, involving a control variable quantifying the protein precursors allocation. Our objective is to determine the optimal strategies maximizing the long-term growth of cells under a piecewise-constant periodic environment. Firstly, we perform a theoretical analysis of the resulting optimal control problem (OCP), based on the application the Pontryagin’s Maximum Principle (PMP). We determine that the structure of the optimal control must be bang–bang, with possibly some singular arcs corresponding to optimal equilibria of the system. If the control presents singular arcs, then these can only be reached and left through chattering arcs. We also use a direct optimization method, implemented in the BOCOP software, to solve the studied OCP. Our study reveals that the optimal solution over a large time horizon is related to the one over a single period of the varying environment with periodic constraints. Moreover, we observe that the maximal average growth rate attainable under periodic conditions can be higher than the one under a constant environment. We further extend our analysis to conduct a qualitative comparison between the predictions from our model and some recent biological experiments on E. coli. This analysis particularly highlights the mechanisms of action of the ppGpp signaling molecule, thus providing relevant explanations of the experimental observations. In conclusion, our study corroborates previous research indicating that this molecule plays a crucial role in the regulation of resource allocation of protein precursors in E. coli.

Contrasting Dynamics of Intracellular and Extracellular Antibiotic Resistance Genes in Response to Nutrient Variations in Aquatic Environments.

The propagation of antibiotic resistance in environments, particularly aquatic environments that serve as primary pathways for antibiotic resistance genes (ARGs), poses significant health risks. The impact of nutrients, as key determinants of bacterial growth and metabolism, on the propagation of ARGs, particularly extracellular ARGs (eARGs), remains poorly understood. In this study, we collected microorganisms from the Yangtze River and established a series of microcosms to investigate how variations in nutrient levels and delivery frequency affect the relative abundance of intracellular ARGs (iARGs) and eARGs in bacterial communities. Our results show that the relative abundance of 7 out of 11 representative eARGs in water exceeds that of iARGs, while 8 iARGs dominate in biofilms. Notably, iARGs and eARGs consistently exhibited opposite responses to nutrient variation. When nutrient levels increased, iARGs in the water also increased, with the polluted group (COD = 333.3 mg/L, COD:N:P = 100:3:0.6, m/m) and the eutrophic group (COD = 100 mg/L, COD:N:P = 100:25:5, m/m) showing 1.2 and 3.2 times higher levels than the normal group (COD = 100 mg/L, COD:N:P = 100:10:2, m/m), respectively. In contrast, eARGs decreased by 6.7% and 8.4% in these groups. On the other hand, in biofilms, higher nutrient levels led to an increase in eARGs by 1.5 and 1.7 times, while iARGs decreased by 17.5% and 50.1% in the polluted and eutrophic groups compared to the normal group. Moreover, while increasing the frequency of nutrient delivery (from 1 time/10 d to 20 times/10 d) generally did not favor iARGs in either water or biofilm, it selectively enhanced eARGs in both. To further understand these dynamics, we developed an ARGs-nutrient model by integrating the Lotka-Volterra and Monod equations. The results highlight the complex interplay of bacterial growth, nutrient availability, and mechanisms such as horizontal gene transfer and secretion influencing ARGs' propagation, driving the opposite trend between these two forms of ARGs. This contrasting response between iARGs and eARGs contributes to a dynamic balance that stabilizes bacterial resistance levels amid nutrient fluctuations. This study offers helpful implications regarding the persistence of bacterial resistance in the environment.

The Driving Mechanism of Phytoplankton Resource Utilization Efficiency Variation on the Occurrence Risk of Cyanobacterial Blooms.

Algae are highly sensitive to environmental factors, especially nutrient fluctuations; excessive nutrients can lead to the proliferation of specific algae species, resulting in dominance. In this study, we aimed to reevaluate changes in algal dominance from the perspective of resource utilization efficiency (RUE). We established 80 monitoring sites across different water systems, collecting water and phytoplankton samples. Using canonical correspondence analysis (CCA) and a generalized additive model (GAM), we analyzed the correlation between phytoplankton RUE and nutrient concentrations, quantifying the corresponding relationship between algal dominance and RUE. Our results indicate a significant negative correlation between the RUE of total phosphorus (TP) and total nitrogen (TN) concentration, but a positive correlation with N:P. The RUE of TN was negatively correlated with TN concentration and N:P. We constructed GAMs with interaction terms and confirmed a nonlinear relationship between algal dominance and RUE. When the RUE of TN was low, a positive correlation was observed, while a negative correlation was observed otherwise. These findings reveal the ecological adaptability of algal communities and provide valuable insights for predicting the risk of algal bloom outbreaks.

Effects of storm events on nutrient characteristics in a stratified drinking water reservoir: Behavior, transmission pathways and management strategy

Storm events result in nutrient fluctuations and deterioration of reservoir water supply quality. Understanding of nutrient dynamics (e.g., concentration, composition, loads and transport pathways) and adoption of effective management strategies are critical for safeguarding water quality. A comprehensive monitoring was conducted for three storm events during the rainy season in 2023. Results showed nitrogen (N) and phosphorus (P) dynamics demonstrate a significant response to hydrological process. Rainfall resulted in the highest event mean concentrations (EMCs) of total nitrogen (TN), nitrate nitrogen (NO3−-N), ammonia nitrogen (NH4+-N), total phosphorus (TP), and particulate phosphorus (PP) in the runoff being 1.97, 2.15, 2.30, 44.17, and 62.38 times higher than those observed in baseflow. On average, NO3−-N/PP accounted for 82 %/96 % of N/P exports. Hysteresis analyses reveal that NH4+-N and PP were mainly transported by surface runoff from over-land sources, whereas TN and NO3−-N were primarily delivered by subsurface runoff. Additionally, nutrient concentrations were significantly higher in the intrusive layer in reservoir compared to the pre-storm period, which gradually decreased from the tail to the head as particulate sedimentation and water column mixing occurred. Water-lifting-aerators (WLAs) were employed to alter the reservoir thermal stratification regime via artificial mixing to affect the intrusive layer of storm runoff. Comparison of the intrusive layer for three storms reveals that WLAs triggers the storm runoff to form an underflow via increasing the reservoir bottom water temperature above that the runoff, ensuring that water quality at the intake position remains unaffected by inflows. These findings serve as a reference for the response of reservoir eutrophication levels to storm events and present practical engineering experience for enhancing water quality safety during the rainy season.

Increasing and fluctuating resource availability enhances invasional meltdown.

Exotic plant invaders can promote others via direct or indirect facilitation, known as "invasional meltdown." Increased soil nutrients can also promote invaders by increasing their competitive impacts, but how this might affect meltdown is unknown. In a mesocosm experiment, we evaluated how eight exotic plant species and eight Eurasian native species responded individually to increasing densities of the invasive plant Conyza canadensis, while varying the supply and fluctuations of nutrients. We found that increasing density of C. canadensis intensified competitive suppression of natives but intensified facilitation of other exotics. Higher and fluctuating nutrients exacerbated the competitive effects on natives and facilitative effects on exotics. Overall, these results show a pronounced advantage of exotics over native target species with increased relative density of C. canadensis under high nutrient availability and fluctuation. We integrate these results with the observation that exotic species commonly drive increases in soil resources to suggest the Resource-driven Invasional Meltdown and Inhibition of Natives hypothesis in which biotic acceleration of resource availability promotes other exotic species over native species, leading to invasional meltdown.

Microbial ecology of northern Gulf of Mexico estuarine waters.

Estuarine and coastal ecosystems are of high economic and ecological importance, owing to their diverse communities and the disproportionate role they play in carbon cycling, particularly in carbon sequestration. Organisms inhabiting these environments must overcome strong natural fluctuations in salinity, nutrients, and turbidity, as well as numerous climate change-induced disturbances such as land loss, sea level rise, and, in some locations, increasingly severe tropical cyclones that threaten to disrupt future ecosystem health. The northern Gulf of Mexico (nGoM) along the Louisiana coast contains dozens of estuaries, including the Mississippi-Atchafalaya River outflow, which dramatically influence the region due to their vast upstream watershed. Nevertheless, the microbiology of these estuaries and surrounding coastal environments has received little attention. To improve our understanding of microbial ecology in the understudied coastal nGoM, we conducted a 16S rRNA gene amplicon survey at eight sites and multiple time points along the Louisiana coast and one inland swamp spanning freshwater to high brackish salinities, totaling 47 duplicated Sterivex (0.2-2.7 µm) and prefilter (>2.7 µm) samples. We cataloged over 13,000 Amplicon Sequence ariants (ASVs) from common freshwater and marine clades such as SAR11 (Alphaproteobacteria), Synechococcus (Cyanobacteria), and acI and Candidatus Actinomarina (Actinobacteria). We observed correlations with freshwater or marine habitats in many organisms and characterized a group of taxa with specialized distributions across brackish water sites, supporting the hypothesis of an endogenous brackish-water community. Additionally, we observed brackish-water associations for several aquatic clades typically considered marine or freshwater taxa, such as SAR11 subclade II, SAR324, and the acI Actinobacteria. The data presented here expand the geographic coverage of microbial ecology in estuarine communities, help delineate the native and transitory members of these environments, and provide critical aquatic microbiological baseline data for coastal and estuarine sites in the nGoM.IMPORTANCEEstuarine and coastal waters are diverse ecosystems influenced by tidal fluxes, interconnected wetlands, and river outflows, which are of high economic and ecological importance. Microorganisms play a pivotal role in estuaries as "first responders" and ecosystem architects, yet despite their ecological importance, they remain underrepresented in microbial studies compared to open ocean environments. This leads to substantial knowledge gaps that are important for understanding global biogeochemical cycling and making decisions about conservation and management strategies in these environments. Our study makes key contributions to the microbial ecology of estuarine and coastal habitats in the northern Gulf of Mexico. Our microbial community data support the concept of a globally distributed, core brackish microbiome and emphasize previously underrecognized brackish-water taxa. Given the projected worsening of land loss, oil spills, and natural disasters in this region, our results will serve as important baseline data for researchers investigating the microbial communities found across estuaries.

Integrating Omics Approaches for Climate-Resilient Crops: A Comprehensive Review

Climate change is a looming threat to global agriculture, impacting temperature, rainfall patterns, pest dynamics, and soil quality. These challenges transcend numerous crops vital for global food security. Drought, soil acidity, and nutrient fluctuations induced by climate change impede crop productivity, necessitating the development of breeding strategies to produce climate-resilient varieties. This review delves into the integration of omics approaches, including genomics, transcriptomics, proteomics, and metabolomics, to bolster breeding programs across diverse crops. By harnessing high-throughput technologies, researchers gain insights into the genetic and molecular mechanisms underlying traits such as stress resistance, yield, and disease tolerance. The cultivation of elite cultivars with enhanced stress tolerance is paramount for sustainable agriculture in the face of climate change. Various breeding approaches, encompassing functional genomics and mutagenomics, are explored alongside the application of genome editing tools like CRISPR/Cas9 and TALEN for targeted trait enhancement. Through the integration of multi-omics data, novel genetic targets are unearthed, facilitating the development of crop varieties resilient to climate-induced stressors beyond maize. This review underscores the significance of multi-omics in crop breeding and highlights strides made toward climate-resilient crop production. Understanding crop responses to abiotic stresses induced by climate change is imperative for the development of resilient varieties. The integration of modern genetics into classical breeding methods aims to cultivate stress-resistant cultivars, mitigating food security risks. Multi-omics approaches play pivotal roles in unraveling crop performance and stress tolerance mechanisms under diverse environmental conditions. Moving forward, the integration of multi-omics approaches will pinpoint candidate genes and pathways, enabling precision breeding to enhance crop performance amidst changing climates. These endeavors will propel the advancement of climate-resilient crops, safeguarding global food security in the face of climate change's challenges.

A novel smart fertigation system for irrigation with treated wastewater: Effects on nutrient recovery, crop and soil

Both southern and northern regions of Italy are experiencing reduced precipitation and increased heat waves due to climate change, negatively affecting agricultural sector. Urban wastewater could be a solution to this problem, providing a constant source for irrigation and reducing synthetic fertilizer use. This research presents a two-year field study on using tertiary treated wastewater for processing tomato crop irrigation through an innovative smart fertigation system, designed to supply the exact doses of NPK nutrients considering those already delivered to the plants via the irrigation water. With the aim of studying the effects of irrigation with treated wastewater, three water sources were compared: fresh water added with chemical fertilizer, tertiary treated wastewater added with chemical fertilizer and tertiary treated wastewater without addition of fertilizer. The proposed system was efficient and consistent with the design, it saved considerable amounts of fertilizers, handling nutrient fluctuations in wastewater. Of the three irrigation water types that have been tested, only the one that used tertiary treated wastewater alone without additional fertilizers was not capable of meeting tomato nutritional needs, despite the fact that significant macronutrient savings were achieved. No negative effects on soil or plant physiological performances were observed. Plants irrigated with wastewater showed similar growth and productivity to those irrigated with fresh water and no significant differences in fruit quality were found, highlighting the benefits of wastewater reuse for crop irrigation.

Challenges and opportunities in obesity: the role of adipocytes during tissue fibrosis.

Obesity is a chronic disease that affects the energy balance of the whole body. In addition to increasing fat mass, tissue fibrosis occurred in white adipose tissue in obese condition. Fibrosis is the over-activation of fibroblasts leading to excessive accumulation of extracellular matrix, which could be caused by various factors, including the status of adipocytes. The morphology of adipocytes responds rapidly and dynamically to nutrient fluctuations. Adaptive hypertrophy of normal adipocytes protects peripheral organs from damage from lipotoxicity. However, the biological behavior of hypertrophic adipocytes in chronic obesity is abnormally altered. Adipocytes lead to fibrotic remodeling of the extracellular matrix by inducing unresolved chronic inflammation, persistent hypoxia, and increasing myofibroblast numbers. Moreover, adipocyte-induced fibrosis not only restricts the flexible expansion and contraction of adipose tissue but also initiates the development of various diseases through cellular autonomic and paracrine effects. Regarding anti-fibrotic therapy, dysregulated intracellular signaling and epigenetic changes represent potential candidate targets. Thus, modulation of adipocytes may provide potential therapeutic avenues for reversing pathological fibrosis in adipose tissue and achieving the anti-obesity purpose.

Continuous Flow with Reagent Injection on an Inlaid Microfluidic Platform Applied to Nitrite Determination.

A continuous flow with reagent injection method on a novel inlaid microfluidic platform for nitrite determination has been successfully developed. The significance of the high-frequency monitoring of nutrient fluctuations in marine environments is crucial for understanding our impacts on the ecosystem. Many in-situ systems face limitations in high-frequency data collection and have restricted deployment times due to high reagent consumption. The proposed microfluidic device employs automatic colorimetric absorbance spectrophotometry, using the Griess assay for nitrite determination, with minimal reagent usage. The sensor incorporates 10 solenoid valves, four syringes, two LEDs, four photodiodes, and an inlaid microfluidic technique to facilitate optical measurements of fluid volumes. In this flow system, Taylor-Aris dispersion was simulated for different injection volumes at a constant flow rate, and the results have been experimentally confirmed using red food dye injection into a carrier stream. A series of tests were conducted to determine a suitable injection frequency for the reagent. Following the initial system characterization, seven different standard concentrations ranging from 0.125 to 10 µM nitrite were run through the microfluidic device to acquire a calibration curve. Three different calibrations were performed to optimize plug length, with reagent injection volumes of 4, 20, and 50 µL. A straightforward signal processing method was implemented to mitigate the Schlieren effect caused by differences in refractive indexes between the reagent and standards. The results demonstrate that a sampling frequency of at least 10 samples per hour is achievable using this system. The obtained attenuation coefficients exhibited good agreement with the literature, while the reagent consumption was significantly reduced. The limit of detection for a 20 µL injection volume was determined to be 94 nM from the sample intake, and the limit of quantification was 312 nM. Going forward, the demonstrated system will be packaged in a submersible enclosure to facilitate in-situ colorimetric measurements in marine environments.

Regulation of the transcription factor CdnL promotes adaptation to nutrient stress in Caulobacter.

In response to nutrient deprivation, bacteria activate a conserved stress response pathway called the stringent response (SR). During SR activation in Caulobacter crescentus, SpoT synthesizes the secondary messengers guanosine 5'-diphosphate 3'-diphosphate and guanosine 5'-triphosphate 3'-diphosphate (collectively known as (p)ppGpp), which affect transcription by binding RNA polymerase (RNAP) to down-regulate anabolic genes. (p)ppGpp also impacts the expression of anabolic genes by controlling the levels and activities of their transcriptional regulators. In Caulobacter, a major regulator of anabolic genes is the transcription factor CdnL. If and how CdnL is controlled during the SR and why that might be functionally important are unclear. In this study, we show that CdnL is down-regulated posttranslationally during starvation in a manner dependent on SpoT and the ClpXP protease. Artificial stabilization of CdnL during starvation causes misregulation of ribosomal and metabolic genes. Functionally, we demonstrate that the combined action of SR transcriptional regulators and CdnL clearance allows for rapid adaptation to nutrient repletion. Moreover, cells that are unable to clear CdnL during starvation are outcompeted by wild-type cells when subjected to nutrient fluctuations. We hypothesize that clearance of CdnL during the SR, in conjunction with direct binding of (p)ppGpp and DksA to RNAP, is critical for altering the transcriptome in order to permit cell survival during nutrient stress.

Abstract 3080: Activation of mTORC2/Akt is amplified during glutamine starvation and is mediated by GCN2

Abstract Highly proliferative cells like cancer cells depend on glucose and glutamine to augment flux through metabolic and biosynthetic pathways. The mechanistic target of rapamycin (mTOR) integrates signals from nutrients and other growth signals to maintain metabolic homeostasis. Whereas mTOR complex 1 (mTORC1) is activated by the presence of nutrients to promote anabolic metabolism, mTORC2 is activated by nutrient fluctuations, leading to increased Akt phosphorylation at Ser473. How mTORC2 signaling can be augmented by nutrient starvation remains poorly understood. Here, we found that General Control Nonderepressible kinase 2 (GCN2), an amino acid starvation-sensing kinase, conveys glutamine depletion to mTORC2. mTORC2 enhances Akt phosphorylation and Akt feeds back to mTORC2 to augment its activity during glutamine starvation. Amino acid starvation in vivo also enhances mTORC2/Akt signaling via GCN2. Our findings reveal that manipulating the GCN2/mTORC2/Akt signaling cassette could have therapeutic benefit to more effectively starve cancer cells. Citation Format: Tatiana Hernandez, Nikhil Patel*, Jedrick Magsino, Jay Kavia, Aparna Ragupathi, Alysta Paneque, Tracy G. Anthony, Guy Werlen, Estela Jacinto. Activation of mTORC2/Akt is amplified during glutamine starvation and is mediated by GCN2 [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 3080.

Hepatic nutrient and hormone signaling to mTORC1 instructs the postnatal metabolic zonation of the liver

The metabolic functions of the liver are spatially organized in a phenomenon called zonation, linked to the differential exposure of portal and central hepatocytes to nutrient-rich blood. The mTORC1 signaling pathway controls cellular metabolism in response to nutrients and insulin fluctuations. Here we show that simultaneous genetic activation of nutrient and hormone signaling to mTORC1 in hepatocytes results in impaired establishment of postnatal metabolic and zonal identity of hepatocytes. Mutant hepatocytes fail to upregulate postnatally the expression of Frizzled receptors 1 and 8, and show reduced Wnt/β-catenin activation. This defect, alongside diminished paracrine Wnt2 ligand expression by endothelial cells, underlies impaired postnatal maturation. Impaired zonation is recapitulated in a model of constant supply of nutrients by parenteral nutrition to piglets. Our work shows the role of hepatocyte sensing of fluctuations in nutrients and hormones for triggering a latent metabolic zonation program.

The PP2A-like phosphatase Ppg1 mediates assembly of the Far complex to balance gluconeogenic outputs and enables adaptation to glucose depletion.

To sustain growth in changing nutrient conditions, cells reorganize outputs of metabolic networks and appropriately reallocate resources. Signaling by reversible protein phosphorylation can control such metabolic adaptations. In contrast to kinases, the functions of phosphatases that enable metabolic adaptation as glucose depletes are poorly studied. Using a Saccharomyces cerevisiae deletion screen, we identified the PP2A-like phosphatase Ppg1 as required for appropriate carbon allocations towards gluconeogenic outputs-trehalose, glycogen, UDP-glucose, UDP-GlcNAc-after glucose depletion. This Ppg1 function is mediated via regulation of the assembly of the Far complex-a multi-subunit complex that tethers to the ER and mitochondrial outer membranes forming localized signaling hubs. The Far complex assembly is Ppg1 catalytic activity-dependent. Ppg1 regulates the phosphorylation status of multiple ser/thr residues on Far11 to enable the proper assembly of the Far complex. The assembled Far complex is required to maintain gluconeogenic outputs after glucose depletion. Glucose in turn regulates Far complex amounts. This Ppg1-mediated Far complex assembly, and Ppg1-Far complex dependent control of gluconeogenic outputs enables adaptive growth under glucose depletion. Our study illustrates how protein dephosphorylation is required for the assembly of a multi-protein scaffold present in localized cytosolic pools, to thereby alter gluconeogenic flux and enable cells to metabolically adapt to nutrient fluctuations.

A miniaturized culture platform for control of the metabolic environment.

The heart is a metabolic "omnivore" and adjusts its energy source depending on the circulating metabolites. Human cardiac organoids, a three-dimensional in vitro model of the heart wall, are a useful tool to study cardiac physiology and pathology. However, cardiac tissue naturally experiences shear stress and nutrient fluctuations via blood flow in vivo, whilst in vitro models are conventionally cultivated in a static medium. This necessitates the regular refreshing of culture media, which creates acute cellular disturbances and large metabolic fluxes. To culture human cardiac organoids in a more physiological manner, we have developed a perfused bioreactor for cultures in a 96-well plate format. The designed bioreactor is easy to fabricate using a common culture plate and a 3D printer. Its open system allows for the use of traditional molecular biology techniques, prevents flow blockage issues, and provides easy access for sampling and cell assays. We hypothesized that a perfused culture would create more stable environment improving cardiac function and maturation. We found that lactate is rapidly produced by human cardiac organoids, resulting in large fluctuations in this metabolite under static culture. Despite this, neither medium perfusion in bioreactor culture nor lactate supplementation improved cardiac function or maturation. In fact, RNA sequencing revealed little change across the transcriptome. This demonstrates that cardiac organoids are robust in response to fluctuating environmental conditions under normal physiological conditions. Together, we provide a framework for establishing an easily accessible perfusion system that can be adapted to a range of miniaturized cell culture systems.

Understanding the autophagic functions in cancer stem cell maintenance and therapy resistance.

Complex tumour ecosystem comprising tumour cells and its associated tumour microenvironment (TME) constantly influence the tumoural behaviour and ultimately impact therapy failure, disease progression, recurrence and poor overall survival of patients. Crosstalk between tumour cells and TME amplifies the complexity by creating metabolic changes such as hypoxic environment and nutrient fluctuations. These changes in TME initiate stem cell-like programmes in cancer cells, contribute to tumoural heterogeneity and increase tumour robustness. Recent studies demonstrate the multifaceted role of autophagy in promoting fibroblast production, stemness, cancer cell survival during longer periods of dormancy, eventual growth of metastatic disease and disease resistance. Recent ongoing studies examine autophagy/mitophagy as a powerful survival strategy in response to environmental stress including nutrient deprivation, hypoxia and environmental stress in TME. It prevents irreversible senescence, promotes dormant stem-like state, induces epithelial-mesenchymal transition and increases migratory and invasive potential of tumour cells. The present review discusses various theories and mechanisms behind the autophagy-dependent induction of cancer stem cell (CSC) phenotype. Given the role of autophagic functions in CSC aggressiveness and therapeutic resistance, various mechanisms and studies based on suppressing cellular plasticity by blocking autophagy as a powerful therapeutic strategy to kill tumour cells are discussed.

Diel Patterns in the Composition and Activity of Planktonic Microbes in a Subtropical Bay

Planktonic microbes play a crucial role in the matter transformation and energy transfer of marine ecosystems. Marine microbial communities are profoundly influenced by various environmental factors, and their variations are typically investigated in a time-dependent manner. However, studies conducted on a monthly or annual basis do not offer sufficient temporal resolution to reveal changes that can occur on an hourly basis. We conducted a high-resolution time-series study using amplicon sequencing on seawater samples taken at 2-h intervals over a 3-d period from the subtropical Daya Bay to investigate changes in the microbial community composition and activity. The results showed that 46.8% of the microbial rRNA sequences exhibited circadian rhythms, including phytoplankton (e.g., Bacillariophyta, Cryptophyta, and Dinophyceae), heterotrophic bacteria (e.g., Actinomarinaceae, Flavobacteriaceae, and SAR11_clade), and zooplankton (e.g., Bestiolina and Phyllopharyngea). The genera of Bacillariophyta (e.g., Chaetoceros and Pseudo-nitzschia ) exhibited higher activity than those of Chlorophyta (e.g., Micromonas ). A similar trend was observed for zooplankton, where Choanozoa (e.g., Copepoda ) showed a strong circadian rhythm in abundance whereas Ciliophora (e.g., Pithites ) were more active. Light intensity and tides were identified as the driving factors behind the periodic succession and activity of the heterotrophic bacteria and phytoplankton with the exception of picocyanobacteria, which were mainly influenced by fluctuations in temperature, nutrients, and salinity. These high-resolution time-dependent observations showed that, despite the dynamic coastal environment, the periodic changes in the microbial community and activity were distinct and phylogenetically conserved. The findings may help in the development of ecosystem models for predicting microbial abundance and activity in rapidly changing coastal and estuary environments.

The tonoplast localized protein PtNPF1 participates in the regulation of nitrogen response in diatoms.

Diatoms are a highly successful group of phytoplankton, well adapted also to oligotrophic environments and capable of handling nutrient fluctuations in the ocean, particularly nitrate. The presence of a large vacuole is an important trait contributing to their adaptive features. It confers diatoms the ability to accumulate and store nutrients, such as nitrate, when they are abundant outside and then to reallocate them into the cytosol to meet deficiencies, in a process called luxury uptake. The molecular mechanisms that regulate these nitrate fluxes are still not known in diatoms. In this work, we provide new insights into the function of Phaeodactylum tricornutum NPF1, a putative low-affinity nitrate transporter. To accomplish this, we generated overexpressing strains and CRISPR/Cas9 loss-of-function mutants. Microscopy observations confirmed predictions that PtNPF1 is localized on the vacuole membrane. Furthermore, functional characterizations performed on knock-out mutants revealed a transient growth delay phenotype linked to altered nitrate uptake. Together, these results allowed us to hypothesize that PtNPF1 is presumably involved in modulating intracellular nitrogen fluxes, managing intracellular nutrient availability. This ability might allow diatoms to fine-tune the assimilation, storage and reallocation of nitrate, conferring them a strong advantage in oligotrophic environments.