Physiological Manipulations Research Articles

Epigenetic and epitranscriptomic regulation during oncogenic γ-herpesvirus infection.

Oncogenic gamma herpesviruses, including Epstein-Barr Virus (EBV) and Kaposi's Sarcoma-associated Herpesvirus (KSHV), are opportunistic cancer-causing viruses and induces oncogenesis through complex mechanisms, which involves manipulation of cellular physiology as well as epigenetic and epitranscriptomic reprogramming. In this review, we describe the intricate processes by which these viruses interact with the epigenetic machinery, leading to alterations in DNA methylation, histone modifications, and the involvement of non-coding RNAs. The key viral proteins such as EBNA1 and LMP1 encoded by EBV; LANA and vGPCR encoded by KSHV; play pivotal roles in these modifications by interacting with host factors, and dysregulating signaling pathways. The resultant reprogramming can lead to activation of oncogenes, silencing of tumor suppressor genes, and evasion of the immune response, which ultimately contributes to the oncogenic potential of these viruses. Furthermore, in this review, we explore current therapeutic strategies targeting these epigenetic alterations and discuss future directions for research and treatment. Through this comprehensive examination of the epigenetic and epitranscriptomic reprogramming mechanisms employed by oncogenic gamma herpesviruses, we aim to provide valuable insights into potential avenues for novel therapeutic interventions.

Trypanosoma cruzi reprograms mitochondrial metabolism within the anterior midgut of its vector Rhodnius prolixus during the early stages of infection

BackgroundTrypanosoma cruzi is transmitted to humans by hematophagous bugs belonging to the Triatominae subfamily. Its intra-vectorial cycle is complex and occurs exclusively in the insect's midgut. Dissecting the elements involved in the cross-talk between the parasite and its vector within the digestive tract should provide novel targets for interrupting the parasitic life cycle and affecting vectorial competence. These interactions are shaped by the strategies that parasites use to infect and exploit their hosts, and the host's responses that are designed to detect and eliminate parasites. The objective of the current study is to characterize the impact of T. cruzi establishment within its vector on the dynamics of its midgut.MethodsIn this study, we evaluated the impact of T. cruzi infection on protein expression within the anterior midgut of the model insect Rhodnius prolixus at 6 and 24 h post-infection (hpi) using high-throughput quantitative proteomics.ResultsShortly after its ingestion, the parasite modulates the proteome of the digestive epithelium by upregulating 218 proteins and negatively affecting the expression of 11 proteins involved in a wide array of cellular functions, many of which are pivotal due to their instrumental roles in cellular metabolism and homeostasis. This swift response underscores the intricate manipulation of the vector's cellular machinery by the parasite. Moreover, a more in-depth analysis of proteins immediately induced by the parasite reveals a pronounced predominance of mitochondrial proteins, thereby altering the sub-proteomic landscape of this organelle. This includes various complexes of the respiratory chain involved in ATP generation. In addition to mitochondrial metabolic dysregulation, a significant number of detoxifying proteins, such as antioxidant enzymes and P450 cytochromes, were immediately induced by the parasite, highlighting a stress response.ConclusionsThis study is the first to illustrate the response of the digestive epithelium upon contact with T. cruzi, as well as the alteration of mitochondrial sub-proteome by the parasite. This manipulation of the vector's physiology is attributable to the cascade activation of a signaling pathway by the parasite. Understanding the elements of this response, as well as its triggers, could be the foundation for innovative strategies to control the transmission of American trypanosomiasis, such as the development of targeted interventions aimed at disrupting parasite proliferation and transmission within the triatomine vector.Graphical

Paternal influence on pregnancy: Setting the precedence

According to sociologist, Sylvia patriarchy is “a system of social structures and practices in which men dominate, oppress, and exploit women.” Derived from the Greek word patriarkhēs, patriarchy means “the rule of the father.” American sociologist Allan Johnson further mentions that patriarchy is a kind of society in which even though men and women participate, but is male-privileged, maledominated, male-identified, and male-centered. Apart from the sociologist’s point of view of society, it seems that developmental programs in biology also have a patriarchal trend. At least, that’s what is reported from several studies recently. Lopez-Tello et al., reported a paternal insulinlike growth factor 2 (Igf2)-related selective trafficking of maternal resources toward the fetus for its development. Metabolic demands for the fetus are enhanced during pregnancy. It is, therefore, observed that there is a relatively greater shunting of metabolic fuels toward the fetus to increase nutrient availability, more than it is perhaps allowed by the maternal system. The placenta assists in this shuttle by promoting insulin resistance, which leads to glucose intolerance in the mother. This glucose is now made available to the fetal compartments. The paternal Igf2 gene undergoes genomic imprinting, and only the copy inherited from the father is active. Paternally active Igf2 expressed in placental endocrine cells directs the maternal lipids and carbohydrates toward the fetus, an excellent example of paternal manipulation of maternal physiology and metabolic patriarchy. Further, studies have shown that fathers fed with a high-fat diet reduce the pregnancy success rate because of the reduction of their sperm motility. Excess intake of processed food, highsugar diets, or fructose has consequences on offspring’s cardiovascular and metabolic diseases. Males fed a low protein diet in mice results in glucose intolerance, metabolic and cardiovascular dysfunctions, and altered patterns of bone mineralization in pups. We, therefore, can conclude a robust paternal influence on pregnancy and fetal outcome. Pregnancy was once thought of as an exclusively maternal affair. Increasing research in this arena seems to have smashed that stereotype with paternal influence seems to play a significant role in shaping the birth process. It is high time that Sylvia patriarchal understanding is redefined with the paternal share of responsibility beyond just dominance and an authoritarian role.

One-Trial Appetitive Learning Tasks for Drug Targeting.

One-trial appetitive learning developed from one-trial passive avoidance learning as a standard test of retrograde amnesia. It consists of one learning trial followed by a retention test, in which physiological manipulations are presented. As in passive avoidance learning, food- or waterdeprived rats or mice finding food or water inside an enclosure are vulnerable to the retrograde amnesia produced by electroconvulsive shock treatment or the injection of various drugs. In one-trial taste or odor learning conducted in rats, birds, snails, bees, and fruit flies, there is an association between a food item or odorant and contextual stimuli or the unconditioned stimulus of Pavlovian conditioning. The odor-related task in bees was sensitive to protein synthesis inhibition as well as cholinergic receptor blockade, both analogous to results found on the passive avoidance response in rodents, while the task in fruit flies was sensitive to genetic modifications and aging, as seen in the passive avoidance response of genetically modified and aged rodents. These results provide converging evidence of interspecies similarities underlying the neurochemical basis of learning.

The Neurochemical Anatomy of Runway Acquisition and Extinction

Abstract: A review is presented as to the neurochemical basis of the straight runway task, usually consisting of an acquisition phase followed by an extinction phase. During the acquisition of the appetitive runway task, running speeds from the start box to the goal box progressively increase over trials and then decrease when the reward is withheld. Runway extinction is susceptible to lesions of the limbic system, including the medial frontal cortex, the hippocampus, the septum, the amygdala, and the dorsomedial thalamus. When specific neurotransmitter systems are examined, extinction was delayed when noradrenaline transmission was impeded, perhaps involving noradrenergic projections to the hippocampus and neocortex. Extinction was likewise delayed after either facilitation or blocking of dopamine transmission, a result implicating an inverted U-shaped function caused by dopamine’s role in behavioral activation or reward processes. Extinction was also delayed by indirect GABAA receptor agonists injected during acquisition, explained by druginduced disinhibitory tendencies. This simple paradigm may provide information about the effects of a physiological manipulation on both cognition and emotion.

Holistic transcriptional responses of Cordyceps militaris to different culture temperatures

Cordyceps militaris is a medicinal entomopathogenic fungus containing valuable biometabolites for pharmaceutical applications. Its genetic inheritance and environmental factors play a crucial role in the production of biomass enriched with cordycepin. While temperature is a crucial controlled parameter for fungal cultivation, its impacts on growth and metabolite biosynthesis remains poorly characterized. This study aimed to investigate the metabolic responses and cordycepin production of C. militaris strain TBRC6039 under various temperature conditions through transcriptome analysis. Among 9599 expressed genes, 576 genes were significantly differentially expressed at culture temperatures of 15 and 25 °C. The changes in the transcriptional responses induced by these temperatures were found in several metabolisms involved in nutrient assimilation and energy source, including amino acids metabolism (e.g., glycine, serine and threonine metabolism) and lipid metabolism (e.g., biosynthesis of unsaturated fatty acids and steroid biosynthesis). At the lower temperature (15 °C), the biosynthetic pathways of lipids, specifically ergosterol and squalene, were the target for maintaining membrane function by transcriptional upregulation. Our study revealed the responsive mechanisms of C. militaris in acclimatization to temperature conditions that provide an insight on physiological manipulation for the production of metabolites by C. militaris.

Late-Stage Functionalization of Living Organisms: Rethinking Selectivity in Biology.

With unlimited selectivity, full post-translational chemical control of biology would circumvent the dogma of genetic control. The resulting direct manipulation of organisms would enable atomic-level precision in "editing" of function. We argue that a key aspect that is still missing in our ability to do this (at least with a high degree of control) is the selectivity of a given chemical reaction in a living organism. In this Review, we systematize existing illustrative examples of chemical selectivity, as well as identify needed chemical selectivities set in a hierarchy of anatomical complexity: organismo- (selectivity for a given organism over another), tissuo- (selectivity for a given tissue type in a living organism), cellulo- (selectivity for a given cell type in an organism or tissue), and organelloselectivity (selectivity for a given organelle or discrete body within a cell). Finally, we analyze more traditional concepts such as regio-, chemo-, and stereoselective reactions where additionally appropriate. This survey of late-stage biomolecule methods emphasizes, where possible, functional consequences (i.e., biological function). In this way, we explore a concept of late-stage functionalization of living organisms (where "late" is taken to mean at a given state of an organism in time) in which programmed and selective chemical reactions take place in life. By building on precisely analyzed notions (e.g., mechanism and selectivity) we believe that the logic of chemical methodology might ultimately be applied to increasingly complex molecular constructs in biology. This could allow principles developed at the simple, small-molecule level to progress hierarchically even to manipulation of physiology.

Gut metabolite L-lactate supports Campylobacter jejuni population expansion during acute infection.

How the microaerobic pathogen Campylobacter jejuni establishes its niche and expands in the gut lumen during infection is poorly understood. Using 6-wk-old ferrets as a natural disease model, we examined this aspect of C. jejuni pathogenicity. Unlike mice, which require significant genetic or physiological manipulation to become colonized with C. jejuni, ferrets are readily infected without the need to disarm the immune system or alter the gut microbiota. Disease after C. jejuni infection in ferrets reflects closely how human C. jejuni infection proceeds. Rapid growth of C. jejuni and associated intestinal inflammation was observed within 2 to 3 d of infection. We observed pathophysiological changes that were noted by cryptic hyperplasia through the induction of tissue repair systems, accumulation of undifferentiated amplifying cells on the colon surface, and instability of HIF-1α in colonocytes, which indicated increased epithelial oxygenation. Metabolomic analysis demonstrated that lactate levels in colon content were elevated in infected animals. A C. jejuni mutant lacking lctP, which encodes an L-lactate transporter, was significantly decreased for colonization during infection. Lactate also influences adhesion and invasion by C. jejuni to a colon carcinoma cell line (HCT116). The oxygenation required for expression of lactate transporter (lctP) led to identification of a putative thiol-based redox switch regulator (LctR) that may repress lctP transcription under anaerobic conditions. Our work provides better insights into the pathogenicity of C. jejuni.

Potential of computational models in personalized treatment of obstructive sleep apnea: a patient-specific partial 3D finite element study.

The upper airway experiences mechanical loads during breathing. Obstructive sleep apnea is a very common sleep disorder, in which the normal function of the airway is compromised, enabling its collapse. Its treatment remains unsatisfactory with variable efficacy in the case of many surgeries. Finite element models of the upper airway to simulate the effects of various anatomic and physiologic manipulations on its mechanics could be helpful in predicting surgical success. Partial 3D finite element models based on patient-specific CT-scans were undertaken in a pilot study of 5 OSA patients. Upper airway soft tissues including the soft palate, hard palate, tongue, and pharyngeal wall were segmented around the midsagittal plane up to a width of 2.5 cm in the lateral direction. Simulations of surgical interventions such as Uvulopalatopharyngoplasty (UPPP), maxillo-mandibular advancement (MMA), palatal implants, and tongue implants have been performed. Our results showed that maxillo-mandibular advancement (MMA) surgery of 1cm improved the critical closing pressure by at least 212.2%. Following MMA, the best improvement was seen via uvulopalatopharyngoplasty (UPPP), with an improvement of at least 19.12%. Palatal and tongue implants also offered a certain degree of improvement. Further, we observed possible interacting mechanisms that suggested simultaneous implementation of UPPP and tongue stiffening; and palatal and tongue stiffening could be beneficial. Our results suggest that computational modeling is a useful tool for analyzing the influence of anatomic and physiological manipulations on upper airway mechanics. The goal of personalized treatment in the case of OSA could be achieved with the use of computational modeling.

Intestinal changes in permeability, tight junction and mucin synthesis in a mouse model of Alzheimer's disease.

Alzheimer's disease (AD) is a neurodegenerative disorder characterized by the accumulation of amyloid‑β (Aβ) in the brain. The gut/brain axis may serve a role in AD pathogenesis. The present study investigated deposition of Aβ in the intestinal epithelium and its potential effects on intestinal barrier function in a transgenic mouse model of AD. To investigate alterations in the structure and functionality of the intestinal mucosal barrier in AD model mice, hematoxylin and eosin staining for Paneth cell count, Alcian blue‑periodic acid Schiff staining for goblet cells, immunohistochemistry and immunofluorescence for mucin (MUC)2 and wheat germ agglutin expression, transmission electron microscopy for mucosal ultrastructure, FITC‑labeled dextran assay for intestinal permeability, quantitative PCR for goblet cell precursor expression and western blot analysis for tight junction proteins, MUC2 and inflammatory cytokine detection were performed. The results showed that AD model mice exhibited excessive Aβ deposition in the intestinal epithelium, which was accompanied by increased intestinal permeability, inflammatory changes and decreased expression of tight junction proteins. These alterations in the intestinal barrier led to an increased proliferation of goblet and Paneth cells and increased mucus synthesis. Dysfunction of gut barrier occurs in AD and may contribute to its etiology. Future therapeutic strategies to reverse AD pathology may involve early manipulation of gut physiology and its microbiota.

Potential of Mortierellaceae for polyunsaturated fatty acids production: mini review.

The health benefits of polyunsaturated fatty acids (PUFAs) have encouraged the search for rich sources of these compounds. However, the supply chain of PUFAs from animals and plants presents environmental concerns, such as water pollution, deforestation, animal exploitation and interference in the trophic chain. In this way, a viable alternative has been found in microbial sources, mainly in single cell oil (SCO) production by yeast and filamentous fungi. Mortierellaceae is a filamentous fungal family world-renowned for PUFA-producing strains. For example, Mortierella alpina can be highlighted due to be industrially applied to produce arachidonic acid (20:4 n6), an important component of infant supplement formulas. Thus, the state of the art of strategies to increase PUFAs production by Mortierellaceae strains is presented in this review. Firstly, we have discussed main phylogenetic and biochemical characteristics of these strains for lipid production. Next, strategies based on physiological manipulation, using different carbon and nitrogen sources, temperature, pH and cultivation methods, which can increase PUFA production by optimizing process parameters are presented. Furthermore, it is possible to use metabolic engineering tools, controlling the supply of NADPH and co-factors, and directing the activity of desaturases and elongase to the target PUFA. Thus, this review aims to discuss the functionality and applicability of each of these strategies, in order to support future research for PUFA production by Mortierellaceae species.

Targeting de novo lipid synthesis induces lipotoxicity and impairs DNA damage repair in glioblastoma mouse models.

Deregulated de novo lipid synthesis (DNLS) is a potential druggable vulnerability in glioblastoma (GBM), a highly lethal and incurable cancer. Yet the molecular mechanisms that determine susceptibility to DNLS-targeted therapies remain unknown, and the lack of brain-penetrant inhibitors of DNLS has prevented their clinical evaluation as GBM therapeutics. Here, we report that YTX-7739, a clinical-stage inhibitor of stearoyl CoA desaturase (SCD), triggers lipotoxicity in patient-derived GBM stem-like cells (GSCs) and inhibits fatty acid desaturation in GSCs orthotopically implanted in mice. When administered as a single agent, or in combination with temozolomide (TMZ), YTX-7739 showed therapeutic efficacy in orthotopic GSC mouse models owing to its lipotoxicity and ability to impair DNA damage repair. Leveraging genetic, pharmacological, and physiological manipulation of key signaling nodes in gliomagenesis complemented with shotgun lipidomics, we show that aberrant MEK/ERK signaling and its repression of the energy sensor AMP-activated protein kinase (AMPK) primarily drive therapeutic vulnerability to SCD and other DNLS inhibitors. Conversely, AMPK activation mitigates lipotoxicity and renders GSCs resistant to the loss of DNLS, both in culture and in vivo, by decreasing the saturation state of phospholipids and diverting toxic lipids into lipid droplets. Together, our findings reveal mechanisms of metabolic plasticity in GSCs and provide a framework for the rational integration of DNLS-targeted GBM therapies.

Host Transcriptome Analysis of Spodoptera frugiperda Larvae Parasitized by Microplitis manilae

It has been extensively found that parasitoids manipulate host physiology to benefit the survival and development of their offspring. However, the underlying regulatory mechanisms have not received much attention. To reveal the effects of parasitization of the larval solitary endoparasitoid Microplitis manilae (Hymenoptera: Braconidae) on host Spodoptera frugiperda (Lepidoptera: Noctuidae), one of the most destructive agricultural pests in China, deep-sequencing-based transcriptome analysis was conducted to compare the host gene expression levels after 2 h, 24 h, and 48 h parasitization. A total of 1861, 962, and 108 differentially expressed genes (DEGs) were obtained from the S. frugiperda larvae at 2 h, 24 h, and 48 h post-parasitization, respectively, compared with unparasitized controls. The changes in host gene expressions were most likely caused by the injection of wasp parasitic factors, including PDVs, that were injected along with the eggs during oviposition. Based on the functional annotations in GO and KEGG databases, we revealed that most DEGs were implicated in host metabolism and immunity. Further analysis of the common DEGs in three comparisons between the unparasitized and parasitized groups identified four genes, including one unknown and three prophenoloxidase (PPO) genes. Moreover, 46 and 7 common DEGs involved in host metabolism and immunity were identified at two or three time points after parasitization, respectively. Among these, most DEGs showed increased expressions at 2 h post-wasp parasitization while exhibiting significantly decreased expression levels at 24 h post-parasitization, demonstrating the expression regulations of M. manilae parasitization on host metabolism and immune-related genes. Further qPCR verification in 20 randomly selected DEGs confirmed the accuracy and reproducibility of the gene expression profiles generated from RNA-seq. This study reveals the molecular regulatory network about how host insects respond to wasp parasitism, laying a solid foundation for revealing the physiological manipulation of wasp parasitization on host insects, which facilitates the development of biological control practices for parasitoids.

Next-Generation Microfluidics for Biomedical Research and Healthcare Applications.

Microfluidic systems offer versatile biomedical tools and methods to enhance human convenience and health. Advances in these systems enables next-generation microfluidics that integrates automation, manipulation, and smart readout systems, as well as design and three-dimensional (3D) printing for precise production of microchannels and other microstructures rapidly and with great flexibility. These 3D-printed microfluidic platforms not only control the complex fluid behavior for various biomedical applications, but also serve as microconduits for building 3D tissue constructs-an integral component of advanced drug development, toxicity assessment, and accurate disease modeling. Furthermore, the integration of other emerging technologies, such as advanced microscopy and robotics, enables the spatiotemporal manipulation and high-throughput screening of cell physiology within precisely controlled microenvironments. Notably, the portability and high precision automation capabilities in these integrated systems facilitate rapid experimentation and data acquisition to help deepen our understanding of complex biological systems and their behaviors. While certain challenges, including material compatibility, scaling, and standardization still exist, the integration with artificial intelligence, the Internet of Things, smart materials, and miniaturization holds tremendous promise in reshaping traditional microfluidic approaches. This transformative potential, when integrated with advanced technologies, has the potential to revolutionize biomedical research and healthcare applications, ultimately benefiting human health. This review highlights the advances in the field and emphasizes the critical role of the next generation microfluidic systems in advancing biomedical research, point-of-care diagnostics, and healthcare systems.

Changes in Thyroid Hormones Serum Profiles During Late Pregnancy and Post Lambing in the Awassi Sheep

Thyroid gland (TG) function and thyroid hormones (TH) activities are vital to sustaining the productive performanceof sheep. Changes in serum TH concentrations are an indirect measure of the changes in thyroid activity andcirculating TH, which are considered indicators of the metabolic and nutritional status of the animals. PregnantAwassi ewes (n=17) were utilized to assess the changes in the serum profiles of thyroid-stimulating hormone(TSH), 3,3’,5-triiodothyronine (T3), and thyroxine (T4) 4 days prior to lambing (D/BL) and at 2,6,10,14,18 and 22weeks post-lambing (W/PL). Our study revealed the TSH serum profile had gradually and significantly increased(p< 0.05) between 4D/BL and 22W/PL. TSH levels recorded their lowest drop at 4D/BL, while also peaking at22W/PL. TSH levels at 22W/PL was two times as high as the TSH levels at 4D/BL. Here we demonstrated that T3concentration didn’t exhibit a particular trend throughout all intervals and that the T3 levels attained their peaklevels at 2W/PL, while these levels dropped significantly at 14W/PL. Furthermore, we exhibited a significantdecrease in T4 level (p < 0.01) at 6W/PL, while a significant increase (p<0.05) in its level was recorded at l0W/PL. T4 levels did not display any specific pattern; apart from its lowest concentration at 6W/PL and its highest at10W/PL, the T4 levels remained almost in parallel, with minor fluctuations throughout all intervals. Analysis ofthe T4:T3 ratio at various intervals didn’t reveal any specific trend. Our study provides vital elements on the serumprofiles of TH in pre-and postpartum periods, which mirrors physiological changes and allows the monitoringand manipulation of thyroid physiology, to improve animal health and production.

Subsets of leg proprioceptors influence leg kinematics but not interleg coordination in Drosophila melanogaster walking.

Legged locomotion in terrestrial animals is often essential for mating and survival, and locomotor behavior must be robust and adaptable to be successful. This adaptability is largely provided by proprioceptors monitoring positions and movements of body parts and providing feedback to other components of locomotor networks. In insects, proprioceptive chordotonal organs span joints and encode parameters of relative movement between segments. Previous studies have used whole-organ ablation, reduced preparations or broad physiological manipulations to impair the function of the femoral chordotonal organ (fCO), which monitors the femur-tibia joint, and have demonstrated its contribution to interleg coordination and walking behavior. The fCO in Drosophila melanogaster comprises groups of neurons that differ in their morphology and encoding properties (club, hook, claw); sub-population-level manipulations of fCO function have not been methodologically accessible. Here, we took advantage of the genetic toolkit available in D. melanogaster to identify sub-populations of fCO neurons and used transient optogenetic inhibition to investigate their roles in locomotor coordination. Our findings demonstrate that optogenetic inhibition of a subset of club and hook neurons replicates the effects of inhibiting the whole fCO; when inhibited alone, however, the individual subset types did not strongly affect spatial aspects of single-leg kinematics. Moreover, fCO subsets seem to play only a minor role in interleg temporal coordination. Thus, the fCO contains functionally distinct subgroups, and this functional classification may differ from those based on anatomy and encoding properties; this should be investigated in future studies of proprioceptors and their involvement in locomotor networks.

319 Reflections at the Halfway Point of a Multi-Institutional Comprehensive Usda Sustainable Agriculture Systems Project: Empowering the US Broiler Industry for Transformation and Sustainability

Abstract We received funding for a USDA SAS project that began in September (2019) that involves 7 institutions and approximately 30 faculty. Overarching goals of the project are to improve the system wide efficiency of nutrient and water use in broiler production through a) a better understanding and manipulation of the host physiology and microbial influence on water and nutrient efficiency, b) implementation of novel algal-based technology, and c) effecting educational-behavioral changes through enhanced education and extension programs. To date: a) We have had group progress report meetings 6-8 times per year, and b) we have supported or involved 19 undergraduate students, 22 grad students, and 6 post-docs on the project. The group collectively has published 15 book chapters, 74 refereed publications, 7 theses/dissertations, and filed 2 patents. We also had a symposium at a national meeting. Despite restrictions due to Covid, we have been able to have students take part in internships at U of A and Cornell. A minor in Sustainable Agriculture Systems has been established at Cornell. A class offered by Cornell (The global food, energy, and water nexus – engaging students from the US, China, and India to Chart a Sustainable Future) enables students to participate in the class and receive credit from their home institution. This class incorporates faculty involved in the SAS grant as well as many international guest speakers who are experts in their field. An area we hope to improve upon is getting our message out to our clientele and general public via several different methods.

Pneumatic equiaxial compression device for mechanical manipulation of epithelial cell packing and physiology.

It is well established that mechanical cues, e.g., tensile- compressive- or shear forces, are important co-regulators of cell and tissue physiology. To understand the mechanistic effects these cues have on cells, technologies allowing precise mechanical manipulation of the studied cells are required. As the significance of cell density i.e., packing on cellular behavior is beginning to unravel, we sought to design an equiaxial cell compression device based on our previously published cell stretching system. We focused on improving the suitability for microscopy and the user-friendliness of the system. By introducing a hinge structure to the substrate stretch generating vacuum chamber, we managed to decrease the z-displacement of the cell culture substrate, thus reducing the focal plane drift. The vacuum battery, the mini-incubator, as well as the custom-made vacuum pressure controller make the experimental setup more flexible and portable. Furthermore, we improved the efficiency and repeatability of manufacture of the device by designing a mold that can be used to cast the body of the device. We also compared several different silicone membranes, and chose SILPURAN® due to its best microscopy imaging properties. Here, we show that the device can produce a maximum 8.5% radial pre-strain which leads to a 15% equiaxial areal compression as the pre-strain is released. When tested with epithelial cells, upon compression, we saw a decrease in cell cross-sectional area and an increase in cell layer height. Additionally, before compression the cells had two distinct cell populations with different cross-sectional areas that merged into a more uniform population due to compression. In addition to these morphological changes, we detected an alteration in the nucleo-cytoplasmic distribution of YAP1, suggesting that the cellular packing is enough to induce mechanical signaling in the epithelium.

In vitro characterisation of murine pre-adipose nucleated cells reveals electrophysiological cycles associated with biological clocks.

Adipocytes are energy stores of the body which also play a role in physiological regulation and homeostasis through their endocrine activity. Adipocyte circadian clocks drive rhythms in gene expression, and dysregulation of these circadian rhythms associates with pathological conditions such as diabetes. However, although the role of circadian rhythms in adipose cells and related tissues has been studied from phsyiological and molecular perspectives, they have not yet been explored from an electrical perspective. Research into electro‐chronobiology has revealed that electrical properties have important roles in peripheral clock regulation independently of transcription–translation feedback loops. We have used dielectrophoresis to study electrophysiological rhythms in pre‐adipocytes – representing an adipocyte precursor and nucleated cell‐based model, using serum shocking as the cellular method of clock entrainment. The results revealed significant electrophysiological rhythms, culminating in circadian (ca. 24 hourly) cycles in effective membrane capacitance and radius properties, whereas effective membrane conductance was observed to express ultradian (ca. 14 hourly) rhythms. These data shed new light into pre‐adipocyte electrical behaviour and present a potential target for understanding and manipulation of metabolic physiology.

Childhood Trauma, the HPA Axis and Psychiatric Illnesses: A Targeted Literature Synthesis.

Studies of early life stress (ELS) demonstrate the long-lasting effects of acute and chronic stress on developmental trajectories. Such experiences can become biologically consolidated, creating individual vulnerability to psychological and psychiatric issues later in life. The hippocampus, amygdala, and the medial prefrontal cortex are all important limbic structures involved in the processes that undermine mental health. Hyperarousal of the sympathetic nervous system with sustained allostatic load along the Hypothalamic Pituitary Adrenal (HPA) axis and its connections has been theorized as the basis for adult psychopathology following early childhood trauma. In this review we synthesize current understandings and hypotheses concerning the neurobiological link between childhood trauma, the HPA axis, and adult psychiatric illness. We examine the mechanisms at play in the brain of the developing child and discuss how adverse environmental stimuli may become biologically incorporated into the structure and function of the adult brain via a discussion of the neurosequential model of development, sensitive periods and plasticity. The HPA connections and brain areas implicated in ELS and psychopathology are also explored. In a targeted review of HPA activation in mood and psychotic disorders, cortisol is generally elevated across mood and psychotic disorders. However, in bipolar disorder and psychosis patients with previous early life stress, blunted cortisol responses are found to awakening, psychological stressors and physiological manipulation compared to patients without previous early life stress. These attenuated responses occur in bipolar and psychosis patients on a background of increased cortisol turnover. Although cortisol measures are generally raised in depression, the evidence for a different HPA activation profile in those with early life stress is inconclusive. Further research is needed to explore the stress responses commonalities between bipolar disorder and psychosis in those patients with early life stress.