Motility Response Research Articles

An artificial intelligence platform provides an accurate interpretation of esophageal motility from Functional Lumen Imaging Probe Panometry studies.

Functional lumen imaging probe (FLIP) Panometry is performed at the time of sedated endoscopy and evaluates esophageal motility in response to distension. This study aimed to develop and test an automated artificial intelligence (AI) platform that could interpret FLIP Panometry studies. The study cohort included 678 consecutive patients and 35 asymptomatic controls that completed FLIP Panometry during endoscopy and high-resolution manometry (HRM). "True" study labels for model training and testing were assigned by experienced esophagologists per a hierarchical classification scheme. The supervised, deep learning, AI model generated FLIP Panometry heatmaps from raw FLIP data and based on convolutional neural networks assigned esophageal motility labels using a two-stage prediction model. Model performance was tested on a 15% held-out test set (n=103); the remainder of the studies were utilized for model training (n=610). "True" FLIP labels across the entire cohort included 190 (27%) "normal," 265 (37%) "not normal/not achalasia," and 258 (36%) "achalasia." On the test set, both the Normal/Not normal and the achalasia/not achalasia models achieved an accuracy of 89% (with 89%/88% recall, 90%/89% precision, respectively). Of 28 patients with achalasia (per HRM) in the test set, 0 were predicted as "normal" and 93% as "achalasia" by the AI model. An AI platform provided accurate interpretation of FLIP Panometry esophageal motility studies from a single center compared with the impression of experienced FLIP Panometry interpreters. This platform may provide useful clinical decision support for esophageal motility diagnosis from FLIP Panometry studies performed at the time of endoscopy.

The lymphatic endothelium-derived follistatin: activin A axis regulates neutrophil motility in response to Pseudomonas aeruginosa.

The lymphatic system plays an active role during infection, however the role of lymphatic-neutrophil interactions in host-defense responses is not well understood. During infection with pathogens such as Pseudomonas aeruginosa, Staphylococcus aureus and Yersinia pestis, neutrophils traffic from sites of infection through the lymphatic vasculature, to draining lymph nodes to interact with resident lymphocytes. This process is poorly understood, in part, due to the lack of in vitro models of the lymphatic system. Here we use a 3D microscale lymphatic vessel model to examine neutrophil-lymphatic cell interactions during host defense responses to pathogens. In previous work, we have shown that follistatin is secreted at high concentrations by lymphatic endothelial cells during inflammation. Follistatin inhibits activin A, a member of the TGF-β superfamily, and, together, these molecules form a signaling pathway that plays a role in regulating both innate and adaptive immune responses. Although follistatin and activin A are constitutively produced in the pituitary, gonads and skin, their major source in the serum and their effects on neutrophils are poorly understood. Here we report a microfluidic model that includes both blood and lymphatic endothelial vessels, and neutrophils to investigate neutrophil-lymphatic trafficking during infection with P. aeruginosa. We found that lymphatic endothelial cells produce secreted factors that increase neutrophil migration toward P. aeruginosa, and are a significant source of both follistatin and activin A during Pseudomonas infection. We determined that follistatin produced by lymphatic endothelial cells inhibits activin A, resulting in increased neutrophil migration. These data suggest that the follistatin:activin A ratio influences neutrophil trafficking during infection with higher ratios increasing neutrophil migration.

Search and response mechanisms in active soft matter

Efficient migration in complex environments is crucial for biological systems that search for a target across various length scales. Examples include T-cell locomotion, algae's viscotaxis, bacterial chemotaxis in complex environments, and mammalian sperm cell migration within the female reproductive tract. These microswimmers must search and respond to biophysical and/or biochemical cues from the environment to regulate their motion. This regulation of motion is essential, as the searcher needs to perform various tasks at different regions or carry out a multiphasic search to locate targets. Therefore, identifying the relevant signals present in the microenvironment, and their regulatory effect on the searcher's motion, is key to understanding the search mechanism. As a model system, we study bull sperm migration using microfluidics and apply concepts from soft matter and statistical physics to understand how sperm regulates its motion in a complex environment. I will present a quantitative yet overarching description of the regulation of sperm motility in response to the biophysical/biochemical cues within the female reproductive tract. We develop a holistic view that shows sperm motility can be categorized into multiphasic motions, including progressive, hyperactive, and circular. I will further discuss the outcomes of such interactions between sperm, biochemical stimulants, and biophysical cues, including the rheological properties of mucus and the architecture of the microenvironment. Leveraging this understanding, I will demonstrate new platforms that can detect and, with exquisite precision, isolate cells with high quality to improve assisted reproductive technologies. The approaches and tools we have developed can be further applied to decipher the search strategies utilized by other microorganisms, and study interactions between colloids and polymers under dynamic and non-equilibrium conditions.

Tau acts as a selective barrier along microtubules to regulate multi-motor cargo transport.

Organelles and vesicular cargoes are transported by teams of kinesin and dynein motors along microtubules. Tau is a neuronal MAP that stabilizes microtubules within the axon. While single-molecule studies show that tau differentially regulates the motility of kinesins and dynein in vitro, less is known about its role in modulating the trafficking of endogenous cargoes transported by their native teams of motors. To dissect tau's role in regulating transport, we isolate endocytic organelles from cells and reconstitute their motility along tau-decorated microtubules in vitro. We asked how the sets of motors transporting a cargo determine its motility characteristics and response to tau. Here, we found that early phagosomes (EPs) undergo unidirectional retrograde transport while late phagosomes (LPs) move bidirectionally. Correspondingly, different numbers and combinations of kinesins-1, -2, -3, and dynein are bound to EPs and LPs. Previous studies showed that tau preferentially inhibits kinesin motors, which biases the transport of LPs towards the microtubule minus-end. Recently, we showed that tau strongly inhibits long-range, dynein-mediated EP motility. Dynein is the dominant motor in EP transport, and tau strongly impedes forces generated by teams of multiple dynein motors and accelerates dynein unbinding under load. These results are surprising, as previous single-molecule studies showed that dynein is less sensitive to tau compared to kinesin. Our results show that specific cargoes differentially respond to tau, where dynein-complexes on EPs are more sensitive to tau inhibition than those on LPs, suggesting that pathological changes in tau associated with Alzheimer's disease and other tauopathies will also have disparate impact on cargoes. We will next compare the effects of pathogenic tau mutations on the transport of different cargoes in iPSC-derived neurons with the aim of identifying the pathways most disrupted in neurodegenerative disease.

A Mixture of Chemicals Found in Human Amniotic Fluid Disrupts Brain Gene Expression and Behavior in Xenopus laevis.

Thyroid hormones (TH) are essential for normal brain development, influencing neural cell differentiation, migration, and synaptogenesis. Multiple endocrine-disrupting chemicals (EDCs) are found in the environment, raising concern for their potential effects on TH signaling and the consequences on neurodevelopment and behavior. While most research on EDCs investigates the effects of individual chemicals, human health may be adversely affected by a mixture of chemicals. The potential consequences of EDC exposure on human health are far-reaching and include problems with immune function, reproductive health, and neurological development. We hypothesized that embryonic exposure to a mixture of chemicals (containing phenols, phthalates, pesticides, heavy metals, and perfluorinated, polychlorinated, and polybrominated compounds) identified as commonly found in the human amniotic fluid could lead to altered brain development. We assessed its effect on TH signaling and neurodevelopment in an amphibian model (Xenopus laevis) highly sensitive to thyroid disruption. Fertilized eggs were exposed for eight days to either TH (thyroxine, T4 10 nM) or the amniotic mixture (at the actual concentration) until reaching stage NF47, where we analyzed gene expression in the brains of exposed tadpoles using both RT-qPCR and RNA sequencing. The results indicate that whilst some overlap on TH-dependent genes exists, T4 and the mixture have different gene signatures. Immunohistochemistry showed increased proliferation in the brains of T4-treated animals, whereas no difference was observed for the amniotic mixture. Further, we demonstrated diminished tadpoles' motility in response to T4 and mixture exposure. As the individual chemicals composing the mixture are considered safe, these results highlight the importance of examining the effects of mixtures to improve risk assessment.

The Sinorhizobium meliloti NspS-MbaA system affects biofilm formation, exopolysaccharide production and motility in response to specific polyamines.

We previously showed that specific polyamines (PAs) present in the extracellular environment markedly affect extracellular polysaccharide (EPS) production, biofilm formation and motility in Sinorhizobium meliloti Rm8530. We hypothesized that extracellular PA signals were sensed and transduced by the NspS and MbaA proteins, respectively, which are homologs of the PA-sensing, c-di-GMP modulating NspS-MbaA proteins described in Vibrio cholerae. Here we show that the decrease in biofilm formation and EPS production in the quorum-sensing (QS)-deficient S. meliloti wild-type strain 1021 in cultures containing putrescine or spermine did not occur in a 1021 nspS mutant (1021 nspS). The transcriptional expression of nspS in strain 1021 was significantly increased in cultures containing either of these polyamines, but not by exogenous cadaverine, 1,3-diaminopropane (DAP), spermidine (Spd) or norspermidine (NSpd). Cell aggregation in liquid cultures did not differ markedly between strain 1021 and 1021 nspS in the presence or absence of PAs. The S. meliloti QS-proficient Rm8530 wild-type and nspS mutant (Rm8530 nspS) produced similar levels of biofilm under control conditions and 3.2- and 2.2-fold more biofilm, respectively, in cultures with NSpd, but these changes did not correlate with EPS production. Cells of Rm8530 nspS aggregated from two- to several-fold more than the wild-type in cultures without PAs or in those containing Spm. NSpd, Spd and DAP differently affected swimming and swarming motility in strains 1021 and Rm8530 and their respective nspS mutants. nspS transcription in strain Rm8530 was greatly reduced by exogenous Spm. Bioinformatic analysis revealed similar secondary structures and functional domains in the MbaA proteins of S. meliloti and V. cholerae, while their NspS proteins differed in some residues implicated in polyamine recognition in the latter species. NspS-MbaA homologs occur in a small subset of soil and aquatic bacterial species that commonly interact with eukaryotes. We speculate that the S. meliloti NspS-MbaA system modulates biofilm formation, EPS production and motility in response to environmental or host plant-produced PAs.

Evaluation of two laboratory model methods for diarrheal irritable bowel syndrome

BackgroundDiarrheal irritable bowel syndrome (IBS-D) is a common chronic functional gastrointestinal disorder, and the underlying pathogenic mechanism is still unclear. Animal models that mimic the pathological state of IBS-D patients were constructed to provide a reference for later drug research and model development.MethodsThe IBS-D model was induced using restraint stress and chemical stimulation (rhubarb), and rats were divided into normal control group (NC), chemically stimulated group (CS), and restraint stress group (RS). Visceral motility responses to Colorectal Balloon Dilation (CRD) were measured by Abdominal Withdrawal Reflex (AWR); evaluation of faecal properties and water content; determination of colonic tissue tight junction (TJ) mRNA expression by RT-PCR; measurement of inflammatory cytokines by ELISA; and intestinal flora and short chain fatty acids.ResultsCompared to NC group, CS and RS group rats showed increased intestinal sensitivity and Bristol stool score, significant diarrheal symptoms and weight loss. Mucin 2, ZO-1, OCLN, CLDN4 mRNA expression was reduced and the intestinal mucosal barrier function was diminished. In addition, the levels of inflammatory factors IL-1β, IL-6, IL-8, IL-10 and TNF-α increased, the abundance and diversity of intestinal flora decreased, the content of beneficial bacteria such as Bifidobacteria decreased, and SCFAs such as acetic acid, propionic acid and butyric acid decreased to different degrees. Although, no significant difference was observed for any molecular and inflammatory marker, but compared to CS group, RS group had less water in the stool, higher visceral sensitivity, and higher relative abundance of beneficial intestinal bacteria such as Actinobacteria.ConclusionIn conclusion, restraint stress combined with chemical stimulation can mimic the pathological state of diarrhoea symptoms, visceral hypersensitivity, reduced intestinal mucosal barrier permeability, immune regulatory dysfunction and dysbiosis in IBS-D patients. However, herbs with antibacterial effects such as rhubarb and senna, for example, are not suitable as the first choice for chemical stimulation, as they may lead to a decrease in harmful bacteria and an increase in beneficial bacteria in the intestinal fraction and do not perfectly mimic the imbalanced state of intestinal flora in IBS-D patients, while restraint stress may be a key factor in modelling.

Who's talking to whom: microbiome-enteric nervous system interactions in early life.

The enteric nervous system (ENS) is the intrinsic nervous system of the gastrointestinal tract (GI) and regulates important GI functions, including motility, nutrient uptake, and immune response. The development of the ENS begins during early organogenesis and continues to develop once feeding begins, with ongoing plasticity into adulthood. There has been increasing recognition that the intestinal microbiota and ENS interact during critical periods, with implications for normal development and potential disease pathogenesis. In this review, we focus on insights from mouse and zebrafish model systems to compare and contrast how each model can serve in elucidating the bidirectional communication between the ENS and the microbiome. At the end of this review, we further outline implications for human disease and highlight research innovations that can lead the field forward.

Sex differences in the central regulation of colorectal motility in response to noxious stimuli.

Distinct sex differences in the prevalence and symptoms of abnormal bowel habits in patients with irritable bowel syndrome (IBS) have been reported. We have elucidated the sex differences in the regulation of colorectal motility via the central nervous system. Noxious stimuli in the colorectum of anesthetized male rats enhance colorectal motility by activating monoaminergic neurons in descending pain inhibitory pathways from the brainstem to the lumbosacral spinal cord. These monoaminergic neurons release serotonin and dopamine into the lumbosacral spinal cord, resulting in the increment of colorectal motility. In female rats, in contrast, noxious stimuli in the colorectum have no effect on colorectal motility. We clarified that GABAergic inhibition in the lumbosacral spinal cord masks the enhancement of colorectal motility induced by monoamines in female animals. Considering that IBS patients often show visceral hypersensitivity and hyperalgesia, our studies suggest that differences in the descending neurons that respond to painful stimuli are involved in various sex differences in abnormal bowel habits.

A novel integrated experimental and computational approach to unravel fibroblast motility in response to chemical gradients in 3D collagen matrices.

Fibroblasts play an essential role in tissue repair and regeneration as they migrate to wounded areas to secrete and remodel the extracellular matrix. Fibroblasts recognize chemical substances such as growth factors, which enhance their motility towards the wounded tissues through chemotaxis. Although several studies have characterized single-cell fibroblast motility before, the migration patterns of fibroblasts in response to external factors have not been fully explored in 3D environments. We present a study that combines experimental and computational efforts to characterize the effect of chemical stimuli on the invasion of 3D collagen matrices by fibroblasts. Experimentally, we used microfluidic devices to create chemical gradients using collagen matrices of distinct densities. We evaluated how cell migration patterns were affected by the presence of growth factors and the mechanical properties of the matrix. Based on these results, we present a discrete-based computational model to simulate cell motility, which we calibrated through the quantitative comparison of experimental and computational data via Bayesian optimization. By combining these approaches, we predict that fibroblasts respond to both the presence of chemical factors and their spatial location. Furthermore, our results show that the presence of these chemical gradients could be reproduced by our computational model through increases in the magnitude of cell-generated forces and enhanced cell directionality. Although these model predictions require further experimental validation, we propose that our framework can be applied as a tool that takes advantage of experimental data to guide the calibration of models and predict which mechanisms at the cellular level may justify the experimental findings. Consequently, these new insights may also guide the design of new experiments, tailored to validate the variables of interest identified by the model.

Compensability of an enhanced incidence of spermatozoa with cytoplasmic droplets in boar semen for use in artificial insemination: a single cell approach

This single cell study aimed to clarify whether an elevated incidence of sperm with a retained cytoplasmic droplet (CD) can be compensated by a higher sperm number in boar semen doses to maintain fertility. Cluster analysis of motile spermatozoa (ten boars) revealed that spermatozoa with a CD are underrepresented in the fast, linearly moving sperm cohort compared to morphologically normal sperm. Nonetheless, the response to the motility stimulator procaine was barely affected in spermatozoa with distal CD (Cramer’s V = 0.14), but moderately affected in sperm with proximal CD (V = 0.28). Viability was lower in sperm with distal CD (p < 0.05) but not with proximal CD compared to normal sperm during 168 h storage of extended semen samples (n = 11) and subsequent thermic stress. Morphologically normal sperm from normospermic samples (n = 10) or samples with a high incidence (≥ 15%) of sperm with CD (n = 9) had similar motility patterns and responses to procaine. The origin of morphologically normal sperm had no effect on sperm viability (p > 0.05; n = 26). In conclusion, a moderately enhanced prevalence of sperm with CD seems to be compensable by an increase in sperm numbers in boar semen doses.

CAMPYLOBACTER: A BRIEF REVIEW OF ITS CAUSES, DIAGNOSTIC APPROACHES AND PREVENTION

The most frequent bacterial which is cause of gastroenteritis in humans is Campylobacter. According to Centers for Disease Control and Prevention (CDC) reports around 1.3 million cases of Campylobacter infection in the US occurs per year. This review was designed with the aim to discuss in detail the root causing agents of Campylobacter infection, its diagnostic and prevention methods. Campylobacteriosis starts with the attachment of the pathogenic bacteria to intestinal cells, followed by colonization and, lastly, penetration of the cells following ingestion by the host. Consumption of raw milk, undercooked poultry, and contaminated water have all been associated with Campylobacter infection. Intestinal mucosal Campylobacter toxins proliferate, necrotizing the intestinal villi. A toxin known as cytolethal distending toxins (CDT) damages DNA by acting as a deoxyribonuclease (DNase). Old aged patients and immunocompromised patients are more at risk of morbidity, mortality, and long-term sickness. In addition to additional virulence and survival factors, this review gathers information on motility, chemotaxis, adhesion, invasion, multidrug resistance, and stress response variables. It has been found that mishandling of raw poultry and consumption of undercooked poultry are the major risk factors for human campylobectriosis. Various preventive measures can be adopted to decrease the transmission of the pathogens and the subsequent disease such as the vaccination of the poultry, the health surveillance and the precise food hygiene all over the entire production chain.

Spatial self-segregation of pioneer cyanobacterial species drives microbiome organization in biocrusts

Microbial communities are typically characterized by some degree of self-organization. In biological soil crust (biocrust) communities, vertical organization of resident populations at the mm scale is driven by organismal adaptations to physicochemical microniches. However, the extent of horizontal organization and its driving processes are unknown. Using a combination of observational and genetic mapping, we provide evidence for a highly defined, horizontal self-organization (patchiness) at the mm to cm scale in a successionally early biocrust community dominated by the pioneer cyanobacteria, Microcoleus vaginatus (Microcoleaceae) and Parifilum sp. (Coleofasciculaceae). Experiments with representative isolates of each species demonstrate that the phenomenon is driven by active spatial segregation based on cross-species sensing through the exometabolome acted upon with motility responses. Further, we show that both species share the ability to enrich for specialized cyanospheres of heterotrophic bacteria at smaller scales, and that these cyanospheres are characterized by compositional host-specificity, thus expanding the reach of spatial patchiness beyond primary producers. Our results highlight the importance of specific microbial interactions in the emergence of microbiome compositional architecture and the enhancement of microbial diversity.

Spermatozoa HSP90b expression correlates with ROS generation and altered motility in response to methyl parathion treatment in vitro

Objectives: Widely used organophosphorus pesticides, methyl parathion (MePa), alter the reproductive functions in various animals and humans by induction of oxidative stress on augmented release of reactive oxygen species (ROS). MePa affects semen quality by inducing DNA damage through spermatogenic stages. Several heat shock proteins (HSPs) are expressed in response to environmental stressors particularly the redox-active ones for regulation of protein turnover. Since oxidative stress and sperm motility are implicated in MePa toxicity, studying the expression of HSP90b will unravel the mechanism behind its noxiousness. Material and Methods: Spermatozoa isolated from healthy donors were subjected to various concentrations of MePa (50, 250, 500, and 750 μM) for studying its effect on sperm motility, ROS generation, sperm chromatin integrity, and expression of stress responsive molecular chaperone HSP90b. Results: In vitro exposure of MePa at concentrations ≥500 μM results in a decline in sperm motility and an increased generation of ROS, DNA damage, and HSP90b expression. Conclusion: ROS-mediated modulation of HSP90b expression may affect the structural integrity of client proteins and oxidative injury to membrane lipid, along with DNA integrity resulting in declined sperm motility in response to MePa.

Expression and Crystallization of HDAC6 Tandem Catalytic Domains.

Histone deacetylase 6 (HDAC6) is an atypical lysine deacetylase with tandem catalytic domains and an ubiquitin-binding zinc finger domain. HDAC6 is involved in various biological processes, such as cell motility or stress responses, and has been implicated in pathologies ranging from cancer to neurodegeneration. Due to this broad range of functions, there has been considerable interest in developing HDAC6-specific small molecule inhibitors, several of which are already available. The crystal structure of the tandem catalytic domains of zebrafish HDAC6 has revealed an arrangement with twofold symmetry and extensive surface interaction between the catalytic domains. Further dissection of the biochemical properties of HDAC6 and the development of novel inhibitors will benefit from being able to routinely express high-quality protein. We present here our optimized protocol for expression and crystallization of the zebrafish tandem catalytic domains.

Contribution of the EnvZ/OmpR two-component system to growth, virulence and stress tolerance of colistin-resistant Aeromonas hydrophila.

Aeromonas hydrophila is an important zoonotic pathogen responsible for septicemia, diarrhea and gastroenteritis, and has attracted considerable attention. The EnvZ/OmpR two-component system (TCS) mediates environmental stress responses in gram-negative bacteria. We investigated the role of the TCS in A. hydrophila by comparing the characteristics of the parental (23-C-23), EnvZ/OmpR knockout (23-C-23:ΔEnvZ/OmpR), and complemented strains (23-C-23:CΔEnvZ/OmpR). Under non-stress conditions, the 23-C-23:ΔEnvZ/OmpR strain showed a significant decrease in growth rate compared to that of 23-C-23. Transcriptome and metabonomic analysis indicated that many metabolic pathways were remarkably affected in the ΔEnvZ/OmpR strain, including the TCA cycle and arginine biosynthesis. In addition, the virulence of the ΔEnvZ/OmpR strain was attenuated in a Kunming mouse model. The ΔEnvZ/OmpR strain exhibited notably reduced tolerance to environmental stresses, including high temperature, different pH conditions, oxidative stress, and high osmotic stress. The downregulated expression of genes related to cell metabolism, motility, and virulence in the ΔEnvZ/OmpR mutant strain was further validated by real-time quantitative PCR. Consequently, our data suggest that the EnvZ/OmpR TCS is required for growth, motility, virulence, and stress response in A. hydrophila, which has significant implications in the development of novel antibacterial and vaccine therapies targeting EnvZ/OmpR against A. hydrophila.

Dynamic in Situ Confinement Triggers Ligand-Free Neuropeptide Receptor Signaling.

Membrane receptor clustering is fundamental to cell–cell communication; however, the physiological function of receptor clustering in cell signaling remains enigmatic. Here, we developed a dynamic platform to induce cluster formation of neuropeptide Y2 hormone receptors (Y2R) in situ by a chelator nanotool. The multivalent interaction enabled a dynamic exchange of histidine-tagged Y2R within the clusters. Fast Y2R enrichment in clustered areas triggered ligand-independent signaling as determined by an increase in cytosolic calcium and cell migration. Notably, the calcium and motility response to ligand-induced activation was amplified in preclustered cells, suggesting a key role of receptor clustering in sensitizing the dose response to lower ligand concentrations. Ligand-independent versus ligand-induced signaling differed in the binding of arrestin-3 as a downstream effector, which was recruited to the clusters only in the presence of the ligand. This approach allows in situ receptor clustering, raising the possibility to explore different receptor activation modalities.

Bio-impedance method to monitor colon motility response to direct distal colon stimulation in anesthetized pigs

Electrical stimulation has been demonstrated as an alternative approach to alleviate intractable colonic motor disorders, whose effectiveness can be evaluated through colonic motility assessment. Various methods have been proposed to monitor the colonic motility and while each has contributed towards better understanding of colon motility, a significant limitation has been the spatial and temporal low-resolution colon motility data acquisition and analysis. This paper presents the study of employing bio-impedance characterization to monitor colonic motor activity. Direct distal colon stimulation was undertaken in anesthetized pigs to validate the bio-impedance scheme simultaneous with luminal manometry monitoring. The results indicated that the significant decreases of bio-impedance corresponded to strong colonic contraction in response to the electrical stimulation in the distal colon. The magnitude/power of the dominant frequencies of phasic colonic contractions identified at baseline (in the range 2–3 cycles per minute (cpm)) were increased after the stimulation. In addition, positive correlations have been found between bio-impedance and manometry. The proposed bio-impedance-based method can be a viable candidate for monitoring colonic motor pattern with high spatial and temporal resolution. The presented technique can be integrated into a closed-loop therapeutic device in order to optimize its stimulation protocol in real-time.

Biofilm formation during wastewater treatment: Motility and physiological response of aerobic denitrifying bacteria under ammonia stress based on surface plasmon resonance imaging

A bacterial image analysis system based on surface plasmon resonance imaging was established to investigate the effect of bacterial motility on biofilm formation under high ammonia nitrogen at the single-cell level. The results showed that the bacterial mean rotation speed and vertical motility distance decreased with the increasing concentration of ammonia nitrogen. Ammonia nitrogen inhibited the metabolic activity of the bacteria, decreasing bacterial motility. Bacterial motility was negatively correlated with the biofilm-formation ability. The biofilm formation ability of Enterobacter cloacae strain HNR exposed to ammonia nitrogen was enhanced by reducing its movement and promoting EPS secretion. Genes related to the tricarboxylic acid cycle and oxidative phosphorylation were down-regulated, indicating inhibition of microbial energy metabolism. Genes related to bacterial secretion and lipopolysaccharide synthesis were up-regulated, facilitating the formation of biofilms and enabling the bacteria to resist ammonia nitrogen stress. This study provides new insights into the biofilm formation under ammonia stress.

Role of hyperpolarization-activated cyclic nucleotide-gated channel HCN2 in embryonic neural stem cell proliferation and differentiation

Hyperpolarization-activated cyclic nucleotide-gated channels (HCN channels) are involved in spontaneous activity in many electrically active cell types such as cardiomyocytes and neurons. In this study, the role of HCN channels in proliferation and migration of Nestin and Sox2 expressing embryonic neural progenitor cells (NPC) originating from the subventricular zone (SVZ) was examined. Immunostaining and PCR data showed that the HCN2 subtype was highly expressed in these cells. Patch clamp recordings revealed a hyperpolarization-activated current, which was sensitive to inhibitors of HCN channels. Using the fluorescence dye bis-(1,3-dibutylbarbituric acid)-trimethineoxonol (DiBAC(4)(3)) we found that a prompt reduction of the extracellular K+ concentration, or exposing the cells to acute hypoxia, induced an instant hyperpolarization in the whole cell population. Recovery from low K+ induced hyperpolarization after extracellular calcium removal, or by re-oxygenation of hypoxic cells, was sensitive to ZD7288, a HCN channel inhibitor. Treatment of neurosphere cultures from the SVZ with ZD7288 caused a significant and reversible inhibition of neurosphere formation from single cells indicating that proliferation of progenitor cells was reduced. Furthermore, the migration of neuronal cells from neurospheres was considerably retarded in the presence of ZD7288. The results suggest that HCN2 channels are involved in controlling the proliferation of NPC and that HCN2 channel-induced spontaneous electrical activity may trigger the motility response of neurosphere-derived neurons in concert with other ion channels. Furthermore, the response to hypoxia suggests that HCN2 channels may trigger the chemotactic response of NPC to ischemic brain regions seen in many studies.