Production Of Signaling Molecules Research Articles

Ostarine attenuates pyocyanin in Pseudomonas aeruginosa by interfering with quorum sensing systems.

Antimicrobial resistance has been an increasingly serious threat to global public health. Anti-virulence strategies are being developed to manage antibiotic resistance because they apply a lower selective pressure for antimicrobial-resistant pathogens than that created using traditional bactericides. We aimed to discover novel small molecules that can reduce the production of virulence factors in Pseudomonas aeruginosa and determine the mechanism of action underlying these effects. A clinical compound library was screened, and ostarine was identified as a potential anti-virulence agent. The effects of ostarine were studied via antimicrobial susceptibility testing, bacterial growth assays, pyocyanin quantitation assays, transcriptomic analysis, quorum sensing signal molecule quantification, and real-time PCR assays. Ostarine treatment significantly decreased the synthesis of pyocyanin without any bactericidal action. Besides, ostarine treatment did not affect the relative growth rate and cell morphology of bacteria. Treatment with ostarine interfered with quorum sensing by decreasing the transcription of genes associated with quorum sensing systems and the production of signalling molecules. The inhibition of ostarine on pyocyanin production and gene expression can be alleviated when signalling molecules were supplemented externally. Overall, ostarine may act as a novel anti-virulence agent that can attenuate P. aeruginosa pyocyanin by interfering with quorum sensing systems.

The Phenotypic Responses of Vascular Smooth Muscle Cells Exposed to Mechanical Cues.

During the development of atherosclerosis and other vascular diseases, vascular smooth muscle cells (SMCs) located in the intima and media of blood vessels shift from a contractile state towards other phenotypes that differ substantially from differentiated SMCs. In addition, these cells acquire new functions, such as the production of alternative extracellular matrix (ECM) proteins and signal molecules. A similar shift in cell phenotype is observed when SMCs are removed from their native environment and placed in a culture, presumably due to the absence of the physiological signals that maintain and regulate the SMC phenotype in the vasculature. The far majority of studies describing SMC functions have been performed under standard culture conditions in which cells adhere to a rigid and static plastic plate. While these studies have contributed to discovering key molecular pathways regulating SMCs, they have a significant limitation: the ECM microenvironment and the mechanical forces transmitted through the matrix to SMCs are generally not considered. Here, we review and discuss the recent literature on how the mechanical forces and derived biochemical signals have been shown to modulate the vascular SMC phenotype and provide new perspectives about their importance.

Saccharomyces cerevisiae Requires CFF1 To Produce 4-Hydroxy-5-Methylfuran-3(2H)-One, a Mimic of the Bacterial Quorum-Sensing Autoinducer AI-2.

Quorum sensing is a process of cell-to-cell communication that bacteria use to orchestrate collective behaviors. Quorum sensing depends on the production, release, and detection of extracellular signal molecules called autoinducers (AIs) that accumulate with increasing cell density. While most AIs are species specific, the AI called AI-2 is produced and detected by diverse bacterial species, and it mediates interspecies communication. We recently reported that mammalian cells produce an AI-2 mimic that can be detected by bacteria through the AI-2 receptor LuxP, potentially expanding the role of the AI-2 system to interdomain communication. Here, we describe a second molecule capable of interdomain signaling through LuxP, 4-hydroxy-5-methylfuran-3(2H)-one (MHF), that is produced by the yeast Saccharomyces cerevisiae Screening the S. cerevisiae deletion collection revealed Cff1p, a protein with no known role, to be required for MHF production. Cff1p is proposed to be an enzyme, with structural similarity to sugar isomerases and epimerases, and substitution at the putative catalytic residue eliminated MHF production in S. cerevisiae Sequence analysis uncovered Cff1p homologs in many species, primarily bacterial and fungal, but also viral, archaeal, and higher eukaryotic. Cff1p homologs from organisms from all domains can complement a cff1ΔS. cerevisiae mutant and restore MHF production. In all cases tested, the identified catalytic residue is conserved and required for MHF to be produced. These findings increase the scope of possibilities for interdomain interactions via AI-2 and AI-2 mimics, highlighting the breadth of molecules and organisms that could participate in quorum sensing.IMPORTANCE Quorum sensing is a cell-to-cell communication process that bacteria use to monitor local population density. Quorum sensing relies on extracellular signal molecules called autoinducers (AIs). One AI called AI-2 is broadly made by bacteria and used for interspecies communication. Here, we describe a eukaryotic AI-2 mimic, 4-hydroxy-5-methylfuran-3(2H)-one, (MHF), that is made by the yeast Saccharomyces cerevisiae, and we identify the Cff1p protein as essential for MHF production. Hundreds of viral, archaeal, bacterial, and eukaryotic organisms possess Cff1p homologs. This finding, combined with our results showing that homologs from all domains can replace S. cerevisiae Cff1p, suggests that like AI-2, MHF is widely produced. Our results expand the breadth of organisms that may participate in quorum-sensing-mediated interactions.

Metabolomics and physiological analyses validates previous findings on the mechanism of response to wounding stress of different intensities in broccoli

The mechanism of response of plant to wounding stress is a complex process that physiologically modifies the wounded tissue for protection. In this study, untargeted metabolomics and physiological analyses were performed to validate the molecular mechanism of response to wounding stress of two intensities (0.04 and 1.85 m2 kg−1) in broccoli florets and shreds, respectively. The results showed that 97 and 1220 differentially expressed metabolites could be identified in broccoli subjected to the Florets vs. Control and Shreds vs. Control experiments, respectively. The Kyoto Encyclopedia Genes and Genomes pathway analyses revealed that these metabolites were mainly involved in aminoacyl-tRNA, amino acid, and secondary metabolite biosynthesis; purine metabolism; and plant signal molecule production. This study validated that wounding stress induced plant signal molecule production. Activation of jasmonic acid biosynthesis and H2O2 production were more susceptible to wounding stress of higher intensities, whereas induction of salicylic acid biosynthesis and O2− production were more susceptible to wounding stress of lower intensities. Furthermore, wounding stress also activated glucosinolate and phenylpropanoid biosynthesis by regulating the levels of the precursors, including L-leucine, phenylalanine, tyrosine, valine, isoleucine, tryptophan, methionine, and phenylalanine. Wounding stress induced phenylpropanoid biosynthesis and the antioxidant system by upregulating the corresponding critical enzyme activity and gene expression, contributing greatly to the enhancement of phenolic compound levels, free radical scavenging ability, and resistance to wounding in broccoli.

Emergence of multistability and strongly asymmetric collective modes in two quorum sensing coupled identical ring oscillators.

The simplest ring oscillator is made from three strongly nonlinear elements repressing each other unidirectionally, resulting in the emergence of a limit cycle. A popular implementation of this scheme uses repressor genes in bacteria, creating the synthetic genetic oscillator known as the Repressilator. We consider the main collective modes produced when two identical Repressilators are mean-field-coupled via the quorum-sensing mechanism. In-phase and anti-phase oscillations of the coupled oscillators emerge from two Andronov-Hopf bifurcations of the homogeneous steady state. Using the rate of the repressor's production and the value of coupling strength as the bifurcation parameters, we performed one-parameter continuations of limit cycles and two-parameter continuations of their bifurcations to show how bifurcations of the in-phase and anti-phase oscillations influence the dynamical behaviors for this system. Pitchfork bifurcation of the unstable in-phase cycle leads to the creation of novel inhomogeneous limit cycles with very different amplitudes, in contrast to the well-known asymmetrical limit cycles arising from oscillation death. The Neimark-Sacker bifurcation of the anti-phase cycle determines the border of an island in two-parameter space containing almost all the interesting regimes including the set of resonant limit cycles, the area with stable inhomogeneous cycle, and very large areas with chaotic regimes resulting from torus destruction and period doubling of resonant cycles and inhomogeneous cycles. We discuss the structure of the chaos skeleton to show the role of inhomogeneous cycles in its formation. Many regions of multistability and transitions between regimes are presented. These results provide new insights into the coupling-dependent mechanisms of multistability and collective regime symmetry breaking in populations of identical multidimensional oscillators.

Quorum Sensing System Affects the Plant Growth Promotion Traits of Serratia fonticola GS2.

Quorum sensing (QS) enables bacteria to organize gene expression programs, thereby coordinating collective behaviors. It involves the production, release, and population-wide detection of extracellular signaling molecules. The cellular processes regulated by QS in bacteria are diverse and may be used in mutualistic coordination or in response to changing environmental conditions. Here, we focused on the influence of the QS-dependent genes of our model bacterial strain Serratia fonticola GS2 on potential plant growth promoting (PGP) activities including indole-3-acetic acid (IAA) production, 1-aminocyclopropane-1-carboxylate (ACC) deaminase activity, and biofilm formation. Based on genomic and phenotypic experimental data we identified and investigated the function of QS genes in the genome of the model strain. Our gene deletion study confirmed the biological functionality of the QS auto-inducer (gloI) and receptor (gloR) on potential PGP activities of GS2. A transcriptomic approach was also undertaken to understand the role of QS genes in regulation of genes primarily involved in PGP activities (IAA, ACC deaminase activity, and biofilm formation). Both transcriptomic and phenotypic data revealed that the QS-deletion mutants had considerably less PGP activities, as compared to the wild type. In addition, in vivo plant experiments showed that plants treated with GS2 had significantly higher growth rates than plants treated with the QS-deletion mutants. Overall, our results showed how QS-dependent genes regulate the potential PGP activities of GS2. This information may be helpful in understanding the relationship between QS-dependent genes and the PGP activity of bacteria, which aid in the production of practical bio-fertilizers for plant growth promotion.

Characterisation of a mural cell network in the murine pituitary gland

The anterior and intermediate lobes of the pituitary are composed of endocrine cells, as well as vasculature and supporting cells, such as folliculostellate cells. Folliculostellate cells form a network with several postulated roles in the pituitary, including production of paracrine signalling molecules and cytokines, coordination of endocrine cell hormone release, phagocytosis, and structural support. Folliculostellate cells in rats are characterised by expression of S100B protein, and in humans by glial fibrillary acid protein. However, there is evidence for another network of supporting cells in the anterior pituitary that has properties of mural cells, such as vascular smooth muscle cells and pericytes. The present study aims to characterise the distribution of cells that express the mural cell marker platelet derived growth factor receptor beta (PDGFRβ) in the mouse pituitary and establish whether these cells are folliculostellate. By immunohistochemical localisation, we determine that approximately 80% of PDGFRβ+ cells in the mouse pituitary have a non-perivascular location and 20% are pericytes. Investigation of gene expression in a magnetic cell sorted population of PDGFRβ+ cells shows that, despite a mostly non-perivascular location, this population is enriched for mural cell markers but not enriched for rat or human folliculostellate cell markers. This is confirmed by immunohistochemistry. The present study concludes that a mural cell network is present throughout the anterior pituitary of the mouse and that this population does not express well-characterised human or rat folliculostellate cell markers.

Short Communication:Transgender Women on Feminizing Hormone Therapy Demonstrate a Distinct Rectal Mucosal Transcriptome from Cisgender Men.

Among transgender women (TGW), the effects of feminizing hormone therapy use on rectal mucosal (RM) HIV transmission are largely unknown. In this small pilot study, we compared the RM transcriptome in TGW utilizing feminizing hormone therapy with a group of cisgender men who have sex with men (MSM) engaging in condomless receptive anal intercourse (AI) and to a group of cisgender men who had never engaged in AI. There were 498 differentially expressed genes (DEGs) in TGW compared with men who had never engaged in AI, and 154 DEGs compared with the MSM. Among TGW, a unique RM transcriptome was identified that implicated pathways critical for mucosal immune responses, including upregulation of genes that mediate immune cell activation and the production of cytokines and other immune signaling molecules. Furthermore, gene set enrichment analyses identified immune signatures that implicated enrichment of proinflammatory immunological pathways in TGW, specifically involving interferon-α, interleukin-6, and tumor necrosis factor-α signaling, whereas metabolic pathways were shown to be enriched among the cisgender male groups. These findings suggest that TGW have a distinct RM immune environment influenced by the use of feminizing hormones, and consequently, there is an urgent need for further investigation into the immunological effects of gender-affirming hormone therapy and its potential impact on HIV mucosal transmission risk for transgender individuals.

Исследование вагинального микробиома женщин с привычным невынашиванием беременности, а также спектра выделяемых ими газовых сигнальных молекул

Введение. Проблема невынашивания беременности в настоящее время не теряет своей актуальности. Частота данной патологии по оценкам различных авторов составляет от 2 до 5% в общей популяции. Как известно, инфекционный фактор играет весьма значимую роль в генезе привычного невынашивания. Вопрос об этиологической роли инфекции в привычном невынашивании широко дискутируется в литературе. Цель исследования - анализ микробиома влагалища здоровых женщин и женщин с привычным невынашиванием беременности, а также спектра выделяемых лактобациллами газовых сигнальных молекул, играющих важную роль в поддержании здоровой жизнедеятельности организма. Методика. Забор материала из влагалища производили стерильным тампоном на полистироловой палочке с площади 1 см2 и в течение 2 ч доставляли в бактериологическую лабораторию. Для выделения факультативно анаэробных и аэробных бактерий использовали питательные среды: Эндо агар для энтеробактерий, маннит-солевой агар (М118) для стафилококков, микрококков, для выявления лецитиназной активности - агар Бэрда-Паркера и так далее. Культивирование проводили при температуре 37 °С в течение 24-48 ч. Количество колоний выражали в lg КОЕ/мл. Продукцию газовых сигнальных молекул (H2, O2, N2, CO, CH4, CO2, NO, H2S) определяли с помощью метода газовой хроматографии на приборе Хроматэк-кристалл 5000.2. Количество выделенных газов измеряли в ppm (млн-1) . Результаты. Спектр основного микробиома влагалища здоровых женщин в возрасте 19-23 лет представлен бактериями нормальной микробиоты родов Lactobacillus, Enterococcus, Bifidobacterium, Peptococcus, Peptostreptococcus, Bacillus, Staphylococcus (эпидермальные штаммы). Условно-патогенные грибы рода Candida, бактероиды, золотистый стафилококк, стрептококки, микрококки, вейлонеллы, гарднереллы и актиномицеты выделялись в редких случаях. Различные штаммы лактобацилл выделялась у 91% здоровых женщин. Среди газовых молекул, продуцируемых лактобациллами, преобладали: СО2, СО и NO. Установлены дисбиотические нарушения микробиома влагалища у небеременных женщин с привычным невынашиванием беременности в анамнезе. Также у данной группы пациенток отмечено снижение продукции газовых сигнальных молекул: СО и NO, и повышение продукции H2S и CH4. Выводы. Полученные результаты свидетельствуют об угнетении иммунного ответа у данной категории больных и необходимости коррекции дисбиотических нарушений еще на этапе прегравидарной подготовки.Background. The issue of habitual miscarriage is very important at the present time. Prevalence of this pathology is approximately 2-5%. Infection considerably contributes to the pathogenesis of habitual miscarriage. The etiology of infection has been widely discussed in the literature. Aim. To study the vaginal microbiome of healthy women and women with habitual miscarriage as well as the spectrum of gaseous signal molecules released by lactobacillus, which is essential for maintaining healthy vital activity. Method. Samples of vaginal material were collected with a sterile swab on a polystyrene stick from an area of 1 cm2; the samples were delivered to the bacteriological laboratory within 2 hours. Facultative anaerobic and aerobic bacteria were isolated using the following nutrient media: Endo agar for enterobacteria; mannitol-salt agar (M118) for staphylococci and micrococci; Baird-Parker agar for detection of lecithinase activity, etc. Cultivation was performed at 37°C for 24-48 hours. The number of colonies was expressed in l g CFU/ml. Production of gaseous signaling molecules (H2, O2, N2, CO, CH4, CO2, NO, and H2S) was determined using a Chromatech Crystal 5000.2 gas chromatograph. The amount of gases was measured in ppm. Results. The major vaginal microbiome spectrum of healthy women (19-23 years) was represented by normal microbiota bacteria of the genera Lactobacillus, Enterococcus, Bifidobacterium, Peptococcus, Peptostreptococcus, Bacillus, and Staphylococcus (epidermal strains). Opportunistic fungi of the genera Candida, Bacteroides, Staphylococcus aureus, Streptococci, Micrococci, Weylonella, Gardnerella, and Actinomycetes were found rarely. Lactobacillus strains were isolated from 91% of healthy women. Lactobacilli produced most of CO2, CO, and NO molecules. Dysbiotic vaginal microbiome was found in non-pregnant women with a history of habitual miscarriage. These patients also had decreased production of CO and NO, and increased production of H2S and CH4. Conclusion. The study demonstrated suppression of the immune response in this category of patients. Dysbiosis requires correction prior to pregnancy.

Identification of Quorum Sensing Activators and Inhibitors in The Marine Sponge Sarcotragus spinosulus.

Marine sponges, a well-documented prolific source of natural products, harbor highly diverse microbial communities. Their extracts were previously shown to contain quorum sensing (QS) signal molecules of the N-acyl homoserine lactone (AHL) type, known to orchestrate bacterial gene regulation. Some bacteria and eukaryotic organisms are known to produce molecules that can interfere with QS signaling, thus affecting microbial genetic regulation and function. In the present study, we established the production of both QS signal molecules as well as QS inhibitory (QSI) molecules in the sponge species Sarcotragus spinosulus. A total of eighteen saturated acyl chain AHLs were identified along with six unsaturated acyl chain AHLs. Bioassay-guided purification led to the isolation of two brominated metabolites with QSI activity. The structures of these compounds were elucidated by comparative spectral analysis of 1HNMR and HR-MS data and were identified as 3-bromo-4-methoxyphenethylamine (1) and 5,6-dibromo-N,N-dimethyltryptamine (2). The QSI activity of compounds 1 and 2 was evaluated using reporter gene assays for long- and short-chain AHL signals (Escherichia coli pSB1075 and E. coli pSB401, respectively). QSI activity was further confirmed by measuring dose-dependent inhibition of proteolytic activity and pyocyanin production in Pseudomonas aeruginosa PAO1. The obtained results show the coexistence of QS and QSI in S. spinosulus, a complex signal network that may mediate the orchestrated function of the microbiome within the sponge holobiont.

The Importance of Cancer-Associated Fibroblasts in the Pathogenesis of Head and Neck Cancers.

Despite progress in anticancer therapies, head and neck squamous cell carcinoma (HNSCC) has still a low survival rate. Recent studies have shown that tumour stroma may play an important role in the pathogenesis of this malignant disease. Fibroblasts are a major component of the tumour microenvironment and may significantly influence HNSCC progression as indicated by the contribution they make to important hallmarks of cancer, such as inflammation, non-restricted growth, angiogenesis, invasion, metastasis, and therapy resistance. It is well known that tumour cells can confer a cancer-associated fibroblast (CAF) phenotype that supports the growth and dissemination of cancer cells. CAFs can stimulate cancer progression through cell-cell contacts and communication, remodelling of extracellular matrix, and production of many signal molecules and matrix metalloproteinases. Consequently, genetic changes in epithelial cells are probably not the only factor that drives HNSCC carcinogenesis. Non-genetic changes in the tumour stroma can also be significantly involved. Stress-induced signals can induce a multicellular program, creating a field of tissue that is predisposed to malignant transformation. The “field cancerization” concept represents a process of active evolution of intercellular interactions and feedback loops between tumour and stromal cells. This model paves the way to study cancer from a new perspective and identify new therapeutic targets. In this review, we discuss current knowledge about CAFs, such as their cellular origin, phenotypical plasticity and functional heterogeneity, and stress their contribution to HNSCC progression. This article was supported by the project AZV 16-29835A. The authors declare they have no potential conflicts of interest concerning drugs, products, or services used in the study. The Editorial Board declares that the manuscript met the ICMJE recommendation for biomedical papers. Submitted: 18. 6. 2019 Accepted: 9. 9. 2019.

Karrikins-induced accumulation of Salvianolic acid B is regulated by Jasmonic acid and hydrogen peroxide in Salvia miltiorrhiza

The influence of smoke-isolated karrikins (KAR1) on the production of endogenous signal molecules and secondary metabolite Salvianolic acid B (SAB) in Salvia miltiorrhiza has been studied. These results indicated that KAR1 induced increase of the accumulation of jasmonic acid (JA) and hydrogen peroxide (H2O2) as well as SAB production. The contents of JA and H2O2 were increased by treatment of KAR1, suggesting that production of JA and H2O2 was early biochemical reaction of S. miltiorrhiza responding to KAR1 in this experiment. KAR1-stimulated generation of JA and H2O2 was inhibited by JA inhibitors, which indicated that KAR1-induced accumulation of JA was located in the upstream of H2O2 biosynthesis. Treatments with JA-specific scavengers (salicylhydroxamic acid (SHAM) and n-propylgallate (PrGall)) and H2O2 biosynthesis inhibitors (catalase (CAT) and diphenylene iodonium (DPI)) inhibited KAR1-induced accumulation of SAB, which suggested that JA and H2O2 played signal roles in KAR1-stimulated accumulation of SAB. CAT/DPI could inhibit KAR1-induced promotion of SAB induced by exogenous JA. In addition, application of H2O2 could reverse KAR1-induced inhibitory effects of SHAM, PrGall toward SAB production. Our results indicated that JA mediated the KAR1-induced increase of SAB dependent on the generation of H2O2 in S. miltiorrhiza.

The Possible Role of the Nitroso-Sulfide Signaling Pathway in the Vasomotoric Effect of Garlic Juice.

The beneficial cardiovascular effects of garlic have been reported in numerous studies. The major bioactive properties of garlic are related to organic sulfides. This study aimed to investigate whether garlic juice works exclusively due to its sulfur compounds or rather via the formation of new products of the nitroso-sulfide signaling pathway. Changes in isometric tension were measured on the precontracted aortic rings of adult normotensive Wistar rats. We evaluated NO-donor (S-nitrosoglutathione, GSNO)-induced vasorelaxation and compare it with effects of hydrogen sulfide (H2S)/GSNO and garlic/GSNO. Incubation with garlic juice increased the maximal GSNO-induced relaxation and markedly changed the character of the relaxant response. Although incubation with an H2S donor enhanced the maximal vasorelaxant response of GSNO, neither the absolute nor the relative relaxation changed over time. The mixture of GSNO with an H2S donor evoked a response similar to GSNO-induced relaxation after incubation with garlic juice. This relaxation of the H2S and GSNO mixture was soluble guanylyl cyclase (sGC) dependent, partially reduced by HNO scavenger and it was adenosine triphosphate-sensitive potassium channels (KATP) independent. In this study, we demonstrate for the first time the suggestion that H2S itself is probably not the crucial bioactive compound of garlic juice but rather potentiates the production of new signaling molecules during the GSNO-H2S interaction.

Emergence of colony-level quorum sensing due to expansion based on simple dynamics

The growth of bacterial colonies is described in terms of a simple mathematical model that incorporates aspects of nutrient acquisition and quorum sensing signal molecule production. Quorum sensing describes cell communication mechanism that allows bacterial cells to control colony’s behaviour in relation to the population density. The consecutive relation of cell transition and quorum sensing signal molecule production is explored using a continuum representation. This paper demonstrates a simple interaction model for non-motile bacteria based on existing mathematical descriptions. The model considers the population of bacteria as consisting of down-regulated and up-regulated bacteria in which the signal molecules being produced at much faster rate by the up-regulated bacteria. The finding highlights the existence of fold bifurcation phenomenon on the fraction of up-regulated bacteria that behaves as an “on-off” quorum sensing switch in response to the effective diffusion constant.

Acetylation of lysine 7 of AhyI affects the biological function in Aeromonas hydrophila

Acyl-homoserine-lactone synthase (AhyI) of Aeromonas hydrophila can produce quorum sensing (QS) auto-inducer 1 (AI-1) type signal molecule, which plays important roles in various biological phenomenons such as biofilm formation, hemolysin production and motility. Previous research revealed that the AhyI of A. hydrophila has acetylation modification on lysine 7 site, but its intrinsic biological function is still largely unknown. To study the effect of AhyI protein and its acetylation modification on the physiological traits of A. hydrophila, the site-directed mutagenesis strains including ΔahyI::ahyI-K7Q and ΔahyI::ahyI-K7R were made in this study. The mutation at K7 site of lysine acetylation in AhyI protein decreased the protease production, but the lysine acetylations do not affect the biofilm formation and hemolysin production. To further study the effect of lysine acetylation on AI-1 signal molecule production, the acyl-homoserine lactones (AHLs) extraction and bioluminescence quantification were performed. Compared with the rescue strain, the acetylation on K7 of AhyI resulted in a decreased level of AHLs and bioluminescence production. It indicated that the lysine acetylation modification on the AhyI protein can regulate the production of signalling molecules. Overall, the obtained data in this study provide a theoretical basis for further understanding the role of lysine acetylation of AhyI protein and lay a foundation to systematically study the regulatory mechanism of QS.

Injectable supramolecular polymer-nanoparticle hydrogels enhance human mesenchymal stem cell delivery.

Stem cell therapies have emerged as promising treatments for injuries and diseases in regenerative medicine. Yet, delivering stem cells therapeutically can be complicated by invasive administration techniques, heterogeneity in the injection media, and/or poor cell retention at the injection site. Despite these issues, traditional administration protocols using bolus injections in a saline solution or surgical implants of cell‐laden hydrogels have highlighted the promise of cell administration as a treatment strategy. To address these limitations, we have designed an injectable polymer–nanoparticle (PNP) hydrogel platform exploiting multivalent, noncovalent interactions between modified biopolymers and biodegradable nanoparticles for encapsulation and delivery of human mesenchymal stem cells (hMSCs). hMSC‐based therapies have shown promise due to their broad differentiation capacities and production of therapeutic paracrine signaling molecules. In this work, the fundamental hydrogel mechanical properties that enhance hMSC delivery processes are elucidated using basic in vitro models. Further, in vivo studies in immunocompetent mice reveal that PNP hydrogels enhance hMSC retention at the injection site and retain administered hMSCs locally for upwards of 2 weeks. Through both in vitro and in vivo experiments, we demonstrate a novel scalable, synthetic, and biodegradable hydrogel system that overcomes current limitations and enables effective cell delivery.

Biomaterial Surface Hydrophobicity-Mediated Serum Protein Adsorption and Immune Responses.

The nature of the protein corona forming on biomaterial surfaces can affect the performance of implanted devices. This study investigated the role of surface chemistry and wettability on human serum-derived protein corona formation on biomaterial surfaces and the subsequent effects on the cellular innate immune response. Plasma polymerization, a substrate-independent technique, was employed to create nanothin coatings with four specific chemical functionalities and a spectrum of surface charges and wettability. The amount and type of protein adsorbed was strongly influenced by surface chemistry and wettability but did not show any dependence on surface charge. An enhanced adsorption of the dysopsonin albumin was observed on hydrophilic carboxyl surfaces while high opsonin IgG2 adsorption was seen on hydrophobic hydrocarbon surfaces. This in turn led to a distinct immune response from macrophages; hydrophilic surfaces drove greater expression of anti-inflammatory cytokines by macrophages, whilst surface hydrophobicity caused increased production of proinflammatory signaling molecules. These findings map out a unique relationship between surface chemistry, hydrophobicity, protein corona formation, and subsequent cellular innate immune responses; the potential outcomes of these studies may be employed to tailor biomaterial surface modifications, to modulate serum protein adsorption and to achieve the desirable innate immune response to implanted biomaterials and devices.

Extracellular DNA release, quorum sensing, and PrrF1/F2 small RNAs are key players in Pseudomonas aeruginosa tobramycin-enhanced biofilm formation

Biofilms are structured microbial communities that are the leading cause of numerous chronic infections which are difficult to eradicate. Within the lungs of individuals with cystic fibrosis (CF), Pseudomonas aeruginosa causes persistent biofilm infection that is commonly treated with aminoglycoside antibiotics such as tobramycin. However, sublethal concentrations of this aminoglycoside were previously shown to increase biofilm formation by P. aeruginosa, but the underlying adaptive mechanisms still remain elusive. Herein, we combined confocal laser scanning microscope analyses, proteomics profiling, gene expression assays and phenotypic studies to unravel P. aeruginosa potential adaptive mechanisms in response to tobramycin exposure during biofilm growth. Under this condition, we show that the modified biofilm architecture is related at least in part to increased extracellular DNA (eDNA) release, most likely as a result of biofilm cell death. Furthermore, the activity of quorum sensing (QS) systems was increased, leading to higher production of QS signaling molecules. We also demonstrate upon tobramycin exposure an increase in expression of the PrrF small regulatory RNAs, as well as expression of iron uptake systems. Remarkably, biofilm biovolumes and eDNA relative abundances in pqs and prrF mutant strains decrease in the presence of tobramycin. Overall, our findings offer experimental evidences for a potential adaptive mechanism linking PrrF sRNAs, QS signaling, biofilm cell death, eDNA release, and tobramycin-enhanced biofilm formation in P. aeruginosa. These specific adaptive mechanisms should be considered to improve treatment strategies against P. aeruginosa biofilm establishment in CF patients’ lungs.

A deep insight into the toxic mechanism for sulfonamides based on bacterial cell-cell communication

Sulfonamides (SAs), a kind of commonly used antibiotics, have been frequently detected in the environment. Due to their potential threat to the ecological environment, the toxicity of SAs to bacteria have been determined in depth, and the toxic mechanism is found to target dihydropteroate synthase (DHPS). However, other toxic mechanism may also exist for SAs, which is still unclear to us. In this paper, the toxic mechanism of sulfachloropyridazine (SCP, as a representative of SAs) was investigated thoroughly on Aliivibrio fischeri (A. fischeri) from the perspective of quorum sensing (QS). The results reveal that SCP may act on three signaling pathways, i.e., Pathway I, II and III, which triggers, maintains and destroys the homeostasis of A. fischeri respectively. In Pathway I, SCP changes the active form of LitR that regulates the expression of lux-related genes, inducing inhibition on bacterial growth while stimulation on luminescence; in Pathway II, SCP facilitates the production of C6HSL signaling molecules, maintaining the homeostasis in LitR and DHPS proteins; whereas in Pathway III, SCP typically combines with DHPS to inhibit the folate metabolism. This study can provide a deeper and more comprehensive understanding on the toxicity of SAs and help the environmental risk assessment of SAs.

Isolation of novel quorum-sensing active bacteria from microbial mats in Shark Bay Australia.

Quorum sensing is a potent system of genetic control allowing phenotypes to be coordinated across localized communities. In this study, quorum sensing systems in Shark Bay microbial mats were delineated using a targeted approach analyzing whole mat extractions as well as the creation of an isolate library. A library of 165 isolates from different mat types were screened using the AHL biosensor E. coli MT102. Based on sequence identity 30 unique isolates belonging to Proteobacteria, Actinobacteria and Firmicutes were found to activate the AHL biosensor, suggesting AHLs or analogous compounds were potentially present. Several of the isolates have not been shown previously to produce signal molecules, particularly the members of the Actinobacteria and Firmicutes phyla including Virgibacillus, Halobacillius, Microbacterium and Brevibacterium. These active isolates were further screened using thin-layer chromatography (TLC) providing putative identities of AHL molecules present within the mat communities. Nine isolates were capable of producing several spots of varying sizes after TLC separation, suggesting the presence of multiple signalling molecules. This study is the first to delineate AHL-based signalling in the microbial mats of Shark Bay, and suggests quorum sensing may play a role in the ecosphysiological coordination of complex phenotypes across microbial mat communities.