Growth Niche Research Articles

Photoperiods and light differentially influence growth and potential niches of phycocyanin‐ and phycoerythrin‐rich picocyanobacteria

AbstractStrains from the picocyanobacteria genus Synechococcus are currently found across a wide range of photoperiods and photosynthetically active radiation. Future scenarios now forecast range expansions of marine Synechococcus into new photic regimes. We found that strains of temperate, coastal phycocyanin‐rich and phycoerythrin‐rich Synechococcus grew fastest under moderate photosynthetically active radiation, and a 24‐h photoperiod, despite a cumulative diel photon dose equivalent to conditions where growth was slower, under higher light and shorter photoperiods. Under optimal conditions, a phycoerythrin‐rich Synechococcus strain achieved a highest recorded cyanobacterial chlorophyll‐specific exponential growth rate (μ) of 4.5 d−1. Two phycoerythrin‐rich strains demonstrated wider ability to modulate light capture capacity, whereas two phycocyanin‐rich strains showed less change in light capture across increasing cumulative diel photon dose. All four coastal strains showed a decrease of effective absorption cross‐section for photosystem II photochemistry, vs. increasing cumulative diel photosynthetically active radiation doses. Within each strain, μ showed consistent, saturating responses to increasing cumulative diel photosystem II electron flux, with more variations in responses of μ to cumulative photosynthetically usable radiation. As photoperiod opportunists, coastal picocyanobacteria show potential to expand into longer photic regimes as higher latitudes warm.

The multifaceted roles of phosphoethanolamine-modified lipopolysaccharides: from stress response and virulence to cationic antimicrobial resistance.

SUMMARYLipopolysaccharides (LPS) are an integral part of the outer membrane of Gram-negative bacteria and play essential structural and functional roles in maintaining membrane integrity as well as in stress response and virulence. LPS comprises a membrane-anchored lipid A group, a sugar-based core region, and an O-antigen formed by repeating oligosaccharide units. 3-Deoxy-D-manno-octulosonic acid-lipid A (Kdo2-lipid A) is the minimum LPS component required for bacterial survival. While LPS modifications are not essential, they play multifaceted roles in stress response and host-pathogen interactions. Gram-negative bacteria encode several distinct LPS-modifying phosphoethanolamine transferases (PET) that add phosphoethanolamine (pEtN) to lipid A or the core region of LPS. The pet genes differ in their genomic locations, regulation mechanisms, and modification targets of the encoded enzyme, consistent with their various roles in different growth niches and under varied stress conditions. The discovery of mobile colistin resistance genes, which represent lipid A-modifying pet genes that are encoded on mobile elements and associated with resistance to the last-resort antibiotic colistin, has led to substantial interest in PETs and pEtN-modified LPS over the last decade. Here, we will review the current knowledge of the functional diversity of pEtN-based LPS modifications, including possible roles in niche-specific fitness advantages and resistance to host-produced antimicrobial peptides, and discuss how the genetic and structural diversities of PETs may impact their function. An improved understanding of the PET group will further enhance our comprehension of the stress response and virulence of Gram-negative bacteria and help contextualize host-pathogen interactions.

The MADS-box gene XAANTAL1 participates in Arabidopsis thaliana primary root growth and columella stem cell patterns in response to ROS, via direct regulation of PEROXIDASE 28 and RETINOBLASTOMA RELATED.

The balance between cell growth, proliferation and differentiation emerges from gene regulatory networks coupled to various signal transduction pathways, including reactive oxygen species (ROS) and transcription factors (TFs), enabling developmental responses to environmental cues. The Arabidopsis thaliana's primary root has become a valuable system for unraveling such networks. Recently, the role of TFs that mediate the ROS's inhibition of primary root growth has begun to be characterized. This study demonstrates that the MADS-box transcription factor XAANTAL1 (XAL1) is an essential regulator of hydrogen peroxide (H2O2) in primary root growth and root stem cell niche identity. Interestingly, our findings suggest that XAL1 acts as a positive regulator of H2O2 concentration in the root meristem by directly regulating genes involved in oxidative stress response, such as PEROXIDASE 28 (PER28). Moreover, we found that XAL1 is necessary for the H2O2-induced inhibition of primary root growth through the negative regulation of peroxidase and catalase activities. Furthermore, XAL1, in conjunction with RETINOBLASTOMA-RELATED (RBR), is essential for positively regulating the differentiation of columella stem cells and for participating in primary root growth inhibition in response to oxidative stress induced by H2O2 treatment.

Exosomes and Tumor Progression: Our Current Knowledge

Exosomes are tiny vesicles that cells secrete into the extracellular environment. They are crucial in cellular communication and have wide-ranging physiological and pathological ramifications. Cargo sorting, MVB development and maturation, MVB transport, and MVB fusion with the plasma membrane are the four essential steps in exosome biogenesis. The high heterogeneity of exosomes is due to the fact that each process is modulated by the competition or coordination of multiple mechanisms, resulting in the sorting of diverse compositions of molecular cargos into different subpopulations of exosomes. In cancer, exosomes have been shown to play a crucial role in tumor growth, metastasis, and pre-metastatic niche formation. In this mini-review, we briefly compile what we know about exosomes at present, including how they are made, what they carry, and how they promote tumor growth. Exosomes' potential as diagnostic and prognostic biomarkers is discussed. We also take a look at the research that hasn't been done and the challenges that have been overlooked

Lymph node stromal cells vary in susceptibility to infection but can support the intracellular growth of Listeria monocytogenes.

Lymph node stromal cells (LNSC) are an often overlooked component of the immune system, but play a crucial role in maintaining tissue homeostasis and orchestrating immune responses. Our understanding of the functions these cells serve in the context of bacterial infections remains limited. We previously showed that Listeria monocytogenes, a facultative intracellular foodborne bacterial pathogen, must replicate within an as-yet-unidentified cell type in the mesenteric lymph node (MLN) to spread systemically. Here, we show that L. monocytogenes could invade, escape from the vacuole, replicate exponentially, and induce a type I IFN response in the cytosol of two LNSC populations infected in vitro, fibroblastic reticular cells (FRC) and blood endothelial cells (BEC). Infected FRC and BEC also produced a significant chemokine and pro-inflammatory cytokine response after in vitro infection. Flow cytometric analysis confirmed that GFP+ L. monocytogenes were associated with a small percentage of MLN stromal cells in vivo following foodborne infection of mice. Using fluorescent microscopy, we showed that these cell-associated bacteria were intracellular L. monocytogenes and the number of infected FRC and BEC changed over the course of a three-day infection in mice. Ex vivo culturing of these infected LNSC populations revealed viable, replicating bacteria that grew on agar plates. These results highlight the unexplored potential of FRC and BEC to serve as suitable growth niches for L. monocytogenes during foodborne infection and to contribute to the pro-inflammatory environment within the MLN that promotes clearance of listeriosis.

Synthesis of polymerizable quaternary ammonium bromides and their efficacy against pathogenic and food spoilage bacteria

Despite rigorous cleaning and sanitization protocols, food manufacturing facilities can harbor persistent microorganisms that can cause post processing contamination leading to food safety and food spoilage issues. Of particular concern are ‘growth niches’, regions within a manufacturing facility more likely to accumulate water and debris because they are outside the zones of routine cleaning and sanitization. Such growth niches present a challenge in controlling post-processing microbial contamination which can affect the safety and shelf life of foods and beverages. In recent years, researchers have explored using antimicrobial and nonfouling coatings to control microbial persistence and cross contamination. A challenge in identifying effective coatings is the limited research on efficacy of antimicrobial monomers (suitable for polymerization into antimicrobial coatings) on microorganisms relevant in food safety and spoilage. In this work, we synthesize and characterize the efficacy of five antimicrobial quaternary ammonium bromide (QAB) monomers against pathogenic and food spoilage organisms. The QAB monomers are end-capped with dihydroxy moieties suitable for subsequent polymerization into antimicrobial polyurethanes and polyesters. N,N-bis(2-hydroxyethyl)-N-methyl-N-R-1-ammonium bromides (R= -octan {C8QAB}, -decan {C10QAB}, -dodecan {C12QAB}, -hexadecan {C16QAB}, -octadecan {C18QAB}) were synthesized and characterized using spectral techniques (Fourier transform infrared spectroscopy and nuclear magnetic resonance spectroscopy). Minimum inhibitory concentrations were determined against relevant food safety and food spoilage bacteria Listeria monocytogenes, Salmonella Typhimurium, Escherichia coli, and Pseudomonas poae. Sixteen and eighteen carbon length QAB monomers inhibited growth of L. monocytogenes and P. poae at a concentration of 3 ppm, while sixteen carbon length QAB monomerinhibited growth of Salmonella Typhimurium at 26 ppm and against Escherichia coli at 13 ppm. Quaternary ammonium compounds capped with polymerizable end groups such as those reported here offer an opportunity to reduce persistence and cross-contamination of food pathogens and food spoilage bacteria with particular relevance in growth niches outside of routine cleaning and sanitization regimens.

The Chlamydia effector CpoS modulates the inclusion microenvironment and restricts the interferon response by acting on Rab35.

The obligate intracellular bacterium Chlamydia trachomatis inserts a family of inclusion membrane (Inc) proteins into the membrane of its vacuole (the inclusion). The Inc CpoS is a critical suppressor of host cellular immune surveillance, but the underlying mechanism remained elusive. By complementing a cpoS mutant with various natural orthologs and variants of CpoS, we linked distinct molecular interactions of CpoS to distinct functions. Unexpectedly, we found CpoS to be essential for the formation of inclusion membrane microdomains that control the spatial organization of multiple Incs involved in signaling and modulation of the host cellular cytoskeleton. While the function of CpoS in microdomains was uncoupled from its role in the suppression of host cellular defenses, we found the ability of CpoS to interact with Rab GTPases to be required not only for the manipulation of membrane trafficking, such as to mediate transport of ceramide-derived lipids (sphingolipids) to the inclusion, but also for the inhibition of Stimulator of interferon genes (STING)-dependent type I interferon responses. Indeed, depletion of Rab35 phenocopied the exacerbated interferon responses observed during infection with CpoS-deficient mutants. Overall, our findings highlight the role of Inc-Inc interactions in shaping the inclusion microenvironment and the modulation of membrane trafficking as a pathogenic immune evasion strategy. IMPORTANCE Chlamydia trachomatis is a prevalent bacterial pathogen that causes blinding ocular scarring and urogenital infections that can lead to infertility and pregnancy complications. Because Chlamydia can only grow within its host cell, boosting the intrinsic defenses of human cells may represent a novel strategy to fight pathogen replication and survival. Hence, CpoS, a Chlamydia protein known to block host cellular defenses, or processes regulated by CpoS, could provide new opportunities for therapeutic intervention. By revealing CpoS as a multifunctional virulence factor and by linking its ability to block host cellular immune signaling to the modulation of membrane trafficking, the present work may provide a foundation for such rationale targeting and advances our understanding of how intracellular bacteria can shape and protect their growth niche.

Does deterministic coexistence theory matter in a finite world?

Contemporary studies of species coexistence are underpinned by deterministic models that assume that competing species have continuous (i.e., noninteger) densities, live in infinitely large landscapes, and coexist over infinite time horizons. By contrast, in nature, species are composed of discrete individuals subject to demographic stochasticity and occur in habitats of finite size where extinctions occur in finite time. One consequence of these discrepancies is that metrics of species' coexistence derived from deterministic theory may be unreliable predictors of the duration of species coexistence in nature. These coexistence metrics include invasion growth rates and niche and fitness differences, which are now commonly applied in theoretical and empirical studies of species coexistence. In this study, we tested the efficacy of deterministic coexistence metrics on the duration of species coexistence in a finite world. We introduce new theoretical and computational methods to estimate coexistence times in stochastic counterparts of classic deterministic models of competition. Importantly, we parameterized this model using experimental field data for 90 pairwise combinations of 18 species of annual plants, allowing us to derive biologically informed estimates of coexistence times for a natural system. Strikingly, we found that for species expected to deterministically coexist, community sizes containing only 10 individuals had predicted coexistence times of more than 1000 years. We also found that invasion growth rates explained 60% of the variation in intrinsic coexistence times, reinforcing their general usefulness in studies of coexistence. However, only by integrating information on both invasion growth rates and species' equilibrium population sizes could most (>99%) of the variation in species coexistence times be explained. This integration was achieved with demographically uncoupled single-species models solely determined by the invasion growth rates and equilibrium population sizes. Moreover, because of a complex relationship between niche overlap/fitness differences and equilibrium population sizes, increasing niche overlap and increasing fitness differences did not always result in decreasing coexistence times, as deterministic theory would predict. Nevertheless, our results tend to support the informed use of deterministic theory for understanding the duration of species' coexistence while highlighting the need to incorporate information on species' equilibrium population sizes in addition to invasion growth rates.

Coupled myovascular expansion directs cardiac growth and regeneration.

Heart regeneration requires multiple cell types to enable cardiomyocyte (CM) proliferation. How these cells interact to create growth niches is unclear. Here, we profile proliferation kinetics of cardiac endothelial cells (CECs) and CMs in the neonatal mouse heart and find that they are spatiotemporally coupled. We show that coupled myovascular expansion during cardiac growth or regeneration is dependent upon VEGF-VEGFR2 signaling, as genetic deletion of Vegfr2 from CECs or inhibition of VEGFA abrogates both CEC and CM proliferation. Repair of cryoinjury displays poor spatial coupling of CEC and CM proliferation. Boosting CEC density after cryoinjury with virus encoding Vegfa enhances regeneration. Using Mendelian randomization, we demonstrate that circulating VEGFA levels are positively linked with human myocardial mass, suggesting that Vegfa can stimulate human cardiac growth. Our work demonstrates the importance of coupled CEC and CM expansion and reveals a myovascular niche that may be therapeutically targeted for heart regeneration.

Advances in Nonfouling and Antimicrobial Coatings: Perspectives for the Food Industry

Microbial cross-contamination represents a significant burden to public health and food wasted due to microbial spoilage. Despite established protocols for cleaning and sanitizing surfaces and advances in antimicrobial and nonfouling coatings research, the persistence of pathogenic and spoilage microorganisms remains a challenge in food manufacturing. Specifically, the design and application of antimicrobial and nonfouling coatings in nonfood contact regions of food manufacturing facilities remain an area of research need as these regions are known microbial growth niches. In this review, we survey the state-of-the-art antimicrobial and nonfouling technologies compatible with polyurethane and epoxy coatings suitable for Zone 2/3 regions, such as drain ditches and equipment framework. The properties of polyurethane and epoxy coatings are discussed, followed by a detailed survey of antimicrobial and nonfouling compounds suitable for incorporation into polyurethane and epoxy polymerization chemistries. Challenges and opportunities in the application of antimicrobial and nonfouling coatings to Zones 2/3 are discussed, with an emphasis placed on the design of effective application studies.

Abstract P1122: Coupled Myovascular Expansion Directs Cardiac Growth And Regeneration

Background: Heart regeneration requires multiple cell types to enable cardiomyocyte (CM) proliferation. How these cells interact to create growth niches is unclear. Here we profile proliferation kinetics of cardiac endothelial cells (CECs) and CMs in the neonatal mouse heart and find that they are spatiotemporally coupled. Methods and Results: We show that coupled myovascular expansion during cardiac growth or regeneration is dependent upon VEGF-VEGFR2 signaling, as genetic deletion of Vegfr2 from CECs or inhibition of VEGFA abrogates both CEC and CM proliferation. Repair of cryoinjury displays poor spatial coupling of CEC and CM proliferation. Boosting CEC density after cryoinjury with virus encoding Vegfa enhances regeneration. Using Mendelian randomization, we demonstrate that circulating VEGFA levels are positively linked with human myocardial mass, suggesting that Vegfa can stimulate human cardiac growth. Conclusions: Our work demonstrates the importance of coupled CEC and CM expansion and reveals a myovascular niche that may be therapeutically targeted for heart regeneration.

Acute Myeloid Leukemia Cells Functionally Compromise Hematopoietic Stem/Progenitor Cells Inhibiting Normal Hematopoiesis Through the Release of Extracellular Vesicles.

Acute myeloid leukemia (AML) is an aggressive and heterogeneous clonal disorder of hematopoietic stem/progenitor cells (HSPCs). It is not well known how leukemia cells alter hematopoiesis promoting tumor growth and leukemic niche formation. In this study, we investigated how AML deregulates the hematopoietic process of HSPCs through the release of extracellular vesicles (EVs). First, we found that AML cells released a heterogeneous population of EVs containing microRNAs involved in AML pathogenesis. Notably, AML-EVs were able to influence the fate of HSPCs modifying their transcriptome. In fact, gene expression profile of AML-EV-treated HSPCs identified 923 down- and 630 up-regulated genes involved in hematopoiesis/differentiation, inflammatory cytokine production and cell movement. Indeed, most of the down-regulated genes are targeted by AML-EV-derived miRNAs. Furthermore, we demonstrated that AML-EVs were able to affect HSPC phenotype, modifying several biological functions, such as inhibiting cell differentiation and clonogenicity, activating inflammatory cytokine production and compromising cell movement. Indeed, a redistribution of HSPC populations was observed in AML-EV treated cells with a significant increase in the frequency of common myeloid progenitors and a reduction in granulocyte-macrophage progenitors and megakaryocyte-erythroid progenitors. This effect was accompanied by a reduction in HSPC colony formation. AML-EV treatment of HSPCs increased the levels of CCL3, IL-1B and CSF2 cytokines, involved in the inflammatory process and in cell movement, and decreased CXCR4 expression associated with a reduction of SDF-1 mediated-migration. In conclusion, this study demonstrates the existence of a powerful communication between AML cells and HSPCs, mediated by EVs, which suppresses normal hematopoiesis and potentially contributes to create a leukemic niche favorable to neoplastic development.

Genomic discovery and structural dissection of a novel type of polymorphic toxin system in gram-positive bacteria

Bacteria have developed several molecular conflict systems to facilitate kin recognition and non-kin competition to gain advantages in the acquisition of growth niches and of limited resources. One such example is a large class of so-called polymorphic toxin systems (PTSs), which comprise a variety of the toxin proteins secreted via T2SS, T5SS, T6SS, T7SS and many others. These systems are highly divergent in terms of sequence/structure, domain architecture, toxin-immunity association, and organization of the toxin loci, which makes it difficult to identify and characterize novel systems using traditional experimental and bioinformatic strategies. In recent years, we have been developing and utilizing unique genome-mining strategies and pipelines, based on the organizational principles of both domain architectures and genomic loci of PTSs, for an effective and comprehensive discovery of novel PTSs, dissection of their components, and prediction of their structures and functions. In this study, we present our systematic discovery of a new type of PTS (S8-PTS) in several gram-positive bacteria. We show that the S8-PTS contains three components: a peptidase of the S8 family (subtilases), a polymorphic toxin, and an immunity protein. We delineated the typical organization of these polymorphic toxins, in which a N-terminal signal peptide is followed by a potential receptor binding domain, BetaH, and one of 16 toxin domains. We classified each toxin domain by the distinct superfamily to which it belongs, identifying nine BECR ribonucleases, one Restriction Endonuclease, one HNH nuclease, two novel toxin domains homologous to the VOC enzymes, one toxin domain with the Frataxin-like fold, and several other unique toxin families such as Ntox33 and HicA. Accordingly, we identified 20 immunity families and classified them into different classes of folds. Further, we show that the S8-PTS-associated peptidases are analogous to many other processing peptidases found in T5SS, T7SS, T9SS, and many proprotein-processing peptidases, indicating that they function to release the toxin domains during secretion. The S8-PTSs are mostly found in animal and plant-associated bacteria, including many pathogens. We propose S8-PTSs will facilitate the competition of these bacteria with other microbes or contribute to the pathogen-host interactions.

Assessing the Potential Contribution of Coffee-Based Tourism to the Rwandan Coffee Sector Development. The Case of Nyamasheke District

Coffee farming sector is affected by two main factors; the coffee price at international market goes up and down depending on the current situation. On the other hands the cost of farms’ inputs changes up-down due to inflation and others related factors. These factors have a negative impact on profitability of coffee farming to the extent that some farmers are demotivated. There is therefore a need for an innovative solution to this problem. The coffee-based tourism is regarded as a growth niche tourism market and may come up with solutions to problems the coffee sector is facing. This paper’s main objectives are to assess the potential contribution of Coffee based tourism to the coffee sector, and to identify factors that may promote or hinder the coffee-based tourism development in Nyamasheke District. The study followed a qualitative research methodology. Data were collected through face-to-face interviews with ten Coffee Washing Stations managers, four farmers representing coffee farmers organizations and the District’s Vice Mayor of economic affairs. Respondents were selected purposively because are not only involved in the coffee farming business but are also interested in the development of the coffee sector. The study found that Coffee Based Tourism may contribute to the improvement of coffee production as it may motivate coffee farmers to improve their farming activities, improve the coffee farmers’ livelihood, and improve the coffee sector in Rwanda. The SWOT analysis presents factors that may promote or hinder the coffee-based tourism in that district. Strength and opportunities are the promoting factors, while weakness and treats depict the hindering factors. In addition, this paper concludes by arguing that the success of this new venture would depend heavily on the central government and the local government’s will through policy development and other supports needed to develop this business venture.

The multifaceted roles of the adipose tissue vasculature.

The prevalence of obesity and its associated pathologies continue to increase, which has led to a renewed interest in our major weight-regulating organ, the white adipose tissue. It has become clear that its development, expansion, and physiological function depend on proper crosstalk between each of its cellular constituents, with a central role for the vascular endothelium lining the blood vessels. Although first considered a mere barrier, the endothelium has emerged as a dynamic unit modulating many critical adipose tissue functions. It not only oversees the uptake of all nutrients to be stored in the adipocytes but also provides an important growth niche for adipocyte progenitors and regulates the expandability of the tissue during overfeeding and obesity. In this review, we describe the reciprocal relationship between endothelial cells, adipocytes, and obesity. We present recent studies that support an important role for endothelial cells as central mediators of many of the physiological and pathological functions of the adipose tissue and highlight several unknown aspects of adipose tissue vascular biology. This new perspective could present exciting opportunities to develop new therapeutic approaches against obesity-related pathologies and is thus of great interest in our increasingly obese society.

Hypertrophied human adipocyte spheroids as in vitro model of weight gain and adipose tissue dysfunction.

Adipocyte enlargement is a key feature of obesity and associated with insulin resistance and metabolic disease The cause and consequences of adipocyte enlargement have remained hard to study in vitro due to a lack of human cell models with representative morphology This paper provides an easily set up spheroid culture method, HUVAS (human unilocular vascularized adipocyte spheroids), for the differentiation and culturing of human adipocytes with a more unilocular morphology We show that providing adipocyte progenitors with a vascular differentiation niche is key for achieving in vitro differentiated adipocytes with large lipid droplets Lipid treatment of the HUVAS spheroids can further adipocyte enlargement and induce cellular dysfunction, mimicking the in vivo effects of weight gain The model will allow a wider research community to perform mechanistic studies of the factors impacting on human adipocyte differentiation and growth, increasing our understanding of how obesity develops and why it has such detrimental consequences on whole body metabolism ABSTRACT: The rise in obesity prevalence has created an urgent need for new and improved methods to study human adipocytes and the pathogenic effects of weight gain in vitro. Despite the proven advantage of culturing adipocyte progenitors as 3D structures, the majority of studies continue to use traditional 2D cultures which result in small, multilocular adipocytes with poor representability. We hypothesized that providing differentiating pre-adipocytes with a vascular growth niche would mimic in vivo adipogenesis and improve the differentiation into unilocular adipocytes. Here we present HUVAS (human unilocular vascularized adipocyte spheroids), a simple, easily applicable culture protocol that allows for the differentiation of human adipocytes with a more unilocular morphology and larger lipid droplets than previous protocols. Moreover, we offer a protocol for inducing adipocyte enlargement in vitro, resulting in larger lipid droplets and development of several key features of adipocyte dysfunction, including altered adipokine secretion, impaired lipolysis and insulin resistance. Taken together, our HUVAS model offers an improved culture system for studying the cellular and molecular mechanisms causing metabolic dysfunction and inflammation in human adipose tissue during weight gain.

Tissue-Engineered Skin Regenerative Units for Epidermal Keratinocytes Expansion and Wound Healing.

Chronic wounds have become an increasing medical and economic problem of aging societies because they are difficult to manage. Tissue engineering provides new perspectives for the clinically applicable skin substitutes. Epidermal keratinocytes play an important role in wound epithelization and construction of tissue-engineered skin substitutes. How to obtain a large number of autologous epidermal keratinocytes in a short time is the main problem that limits the application of tissue-engineered skin and epidermal cell membranes. Developing an appropriate method for reproducing the biological potential of cell–cell interactions and simulating the three-dimensional structure between cells has great significance for epidermal keratinocytes expansion and full-thickness skin regeneration. In this article, we propose the concept of tissue-engineered skin regeneration units (TESRUs) as the smallest unit with complete full-thickness skin regeneration ability. First, autologous dermal fibroblasts are cultured in biodegradable macroporous microcarriers to provide the mesenchyme support. Second, autologous epidermal keratinocytes and autologous melanocytes are incubated with the fibroblasts-loaded microcarriers and expand in vitro. Incorporating the above co-culture method into the macroporous microcarriers is reasonable for maintaining cell–cell interactions in spatial and temporal context and providing a suitable growth niche for epidermal keratinocytes. Moreover, TESRUs are composed of fibroblasts, keratinocytes, and melanocytes and have complete full-thickness skin regeneration ability. We suggest that TESRUs could be a promising strategy to repair full-thickness skin defects for clinical applications if the hypothesis proves to be practical.

Spatio-temporal niche plasticity of a freshwater invader as a harbinger of impact variability.

Invasive alien fishes have detrimental ecological effects on aquatic ecosystems and the services they provide. Impacts from an invasion in a single ecosystem may differ across space and time due to variability in prey availability and environmental conditions. We hypothesize that such variability can be profound, even within a single ecosystem. Stable isotopes analysis (SIA) is commonly used to quantitatively describe the trophic niche of a species. However, spatial and temporal variability in occupied niches are often not incorporated into management strategies and policy options. Here, we used long-term monitoring data to investigate the invasion stage as well as SIA to analyse the trophic niche of the invasive channel catfish Ictalurus punctatus in Lake Kasumigaura (Japan), a long-term ecological research site (LTER), across distant sampling sites and years. We found a significant spatio-temporal variability in relative growth and isotopic niche occupation. Moreover, we defined a new index, the Isotopic Plasticity Index (IPI), which is the ratio between core and total home range of an occupied isotopic niche, to be used as a proxy for the trophic niche stretch or density. We found that this IPI varied considerably, confirming the spatio-temporal variability in trophic niches, suggesting the IPI to be an adequate new isotopic metric. Our results further provide evidence for the existence of variation across invaded landscapes, implying heterogeneous impacts on recipient native communities. Therefore, our work emphasizes the importance of exploring trophic plasticity in feeding ecology and growth as such information enables a better understanding of impacts and can inform the design and implementation of effective management responses.

Sirtuin 3 Downregulation in Mycobacterium tuberculosis-Infected Macrophages Reprograms Mitochondrial Metabolism and Promotes Cell Death.

Mycobacterium tuberculosis induces metabolic reprogramming in macrophages like the Warburg effect. This enhances antimicrobial performance at the expense of increased inflammation, which may promote a pathogen-permissive host environment. Since the NAD+-dependent protein deacetylase Sirtuin 3 (SIRT3) is an important regulator of mitochondrial metabolism and cellular redox homeostasis, we hypothesized that SIRT3 modulation mediates M. tuberculosis-induced metabolic reprogramming. Infection of immortalized and primary murine macrophages resulted in reduced levels of SIRT3 mRNA and protein and perturbation of SIRT3-regulated enzymes in the tricarboxylic acid cycle, electron transport chain, and glycolytic pathway. These changes were associated with increased reactive oxygen species and reduced antioxidant scavenging, thereby triggering mitochondrial stress and macrophage cell death. Relevance to tuberculosis disease in vivo was indicated by greater bacterial burden and immune pathology in M. tuberculosis-infected Sirt3-/- mice. CD11b+ lung leukocytes isolated from infected Sirt3-/- mice showed decreased levels of enzymes involved in central mitochondrial metabolic pathways, along with increased reactive oxygen species. Bacterial burden was also greater in lungs of LysMcreSirt3L2/L2 mice, demonstrating the importance of macrophage-specific SIRT3 after infection. These results support the model of SIRT3 as a major upstream regulatory factor, leading to metabolic reprogramming in macrophages by M. tuberculosisIMPORTANCE Tuberculosis, the disease caused by the bacterium M. tuberculosis, remains one of the top 10 causes of death worldwide. Macrophages, the first cells to encounter M. tuberculosis and critical for defense against infection, are hijacked by M. tuberculosis as a protected growth niche. M. tuberculosis-infected macrophages undergo metabolic reprogramming where key mitochondrial pathways are modulated, but the mechanisms driving this metabolic shift is unknown. Our study demonstrates that M. tuberculosis downregulates Sirtuin 3 (SIRT3), an important regulator of mitochondrial metabolism, leading to SIRT3-dependent transcriptional downregulation of mitochondrial metabolic proteins, which is followed by oxidative stress and macrophage necrosis. This study identifies SIRT3 modulation as a key event in M. tuberculosis-induced metabolic reprograming in macrophages that defend against tuberculosis.

Waste activated sludge stimulates in situ microbial reductive dehalogenation of organohalide-contaminated soil

Due to its enriched organic matter, nutrients and growth cofactors, as well as a diverse range of microorganisms, waste activated sludge (WAS) might be an ideal additive to stimulate organohalide respiration for in situ bioremediation of organohalide-contaminated sites. In this study, we investigated the biostimulation and bioaugmentation impacts of WAS-amendment on the performance and microbiome in tetrachloroethene (PCE) and polychlorinated biphenyls (PCBs) dechlorinating microcosms. Results demonstrated that WAS-amendment increased PCE- and PCBs-dechlorination rate as much as 6.06 and 10.67 folds, respectively. The presence of WAS provided a favorable growth niche for organohalide-respiring bacteria (OHRB), including redox mediation and generation of electron donors and carbon sources. Particularly for the PCE dechlorination, indigenous Geobacter and WAS-derived Dehalococcoides were identified to play key roles in PCE-to-dichloroethene (DCE) and DCE-to-ethene dechlorination, respectively. Similar biostimulation and bioaugmentation effects of WAS-amendment were observed on both PCE- and PCBs-dechlorination in three different soils, i.e., laterite, brown loam and paddy soil. Risk assessment suggested low potential ecological risk of WAS amendment in remediation of organohalide-contaminated soil. Overall, this study provided an economic and efficient strategy to stimulate the organohalide respiration-based bioremediation in field applications.