Increase In Number Of Cells Research Articles

ANATOMICAL AND HISTOLOGICAL ASPECTS OF THE EVOLUTIONARY MORPHOLOGY OF THE SPINAL NODES OF VERTEBRATES ANIMALIUM

Using anatomical, histological, neurohistological and morphological research methods, the histomorphology of spinal cord nodes in a comparative anatomical series of vertebrates: bony fishes, amphibians, reptiles, birds and mammals, which differ in their motor activity and their place of existence in a certain environment, was clarified in the work. It has been established that in the process of phylogenesis, a certain structural and morphofunctional reorganization of the spinal nodes occurs. They differ in shape and size. Adaptation to various living conditions of animals was formed on the basis of changes in the density and size of neurocytes, an increase in the total number of gliocytes and perineuralneglia cells, and polymorphism in the degree of chromatophilia.
 Thus, according to neurohistological studies, it has been noted that the neurons of the spinal cord nodes of cold-blooded animals (pond frog, pond lizard) are characterized by a functional degree of relative polymorphism – chromatophilia.
 Impregnation of spinal cord nodes with silver nitrate in all studied animals revealed different intensity of staining of nerve cells (light, light-dark, dark), which is related to the specifics of species and age-related neuromorphology, the morpho-functional state of the nervous system and the type of higher nervous activity.
 An important issue of modern neuromorphology of animals is the study of spinal cord nodes, which play an important role as primary centers on the way to transmit sensory information from receptors to the central nervous system, providing appropriate reactions to the action of certain factors. The scientific article uses material that is a fragment of the research work of the adjacent departments "Development, morphology and histochemistry of animal organs in normal and pathological conditions", state registration number – 0120U100796.
 The obtained results of the research have an important general biological significance, which complements and expands the idea of certain regularities of spinal nodes, which relate to their structural organization and comparative characteristics at the cellular and tissue levels in vertebrate animals of various species.

Expansion of ventral foregut is linked to changes in the enhancer landscape for organ-specific differentiation

Cell proliferation is fundamental for almost all stages of development and differentiation that require an increase in cell number. Although cell cycle phase has been associated with differentiation, the actual process of proliferation has not been considered as having a specific role. Here we exploit human embryonic stem cell-derived endodermal progenitors that we find are an in vitro model for the ventral foregut. These cells exhibit expansion-dependent increases in differentiation efficiency to pancreatic progenitors that are linked to organ-specific enhancer priming at the level of chromatin accessibility and the decommissioning of lineage-inappropriate enhancers. Our findings suggest that cell proliferation in embryonic development is about more than tissue expansion; it is required to ensure equilibration of gene regulatory networks allowing cells to become primed for future differentiation. Expansion of lineage-specific intermediates may therefore be an important step in achieving high-fidelity in vitro differentiation.

Lung injury and oxidative stress induced by inhaled chlorine in mice is associated with proinflammatory activation of macrophages and altered bioenergetics

Chlorine (Cl2) gas is a highly toxic and oxidizing irritant that causes life-threatening lung injuries. Herein, we investigated the impact of Cl2-induced injury and oxidative stress on lung macrophage phenotype and function. Spontaneously breathing male C57BL/6J mice were exposed to air or Cl2 (300 ppm, 25 min) in a whole-body exposure chamber. Bronchoalveolar lavage (BAL) fluid and cells, and lung tissue were collected 24 h later and analyzed for markers of injury, oxidative stress and macrophage activation. Exposure of mice to Cl2 resulted in increases in numbers of BAL cells and levels of IgM, total protein, and fibrinogen, indicating alveolar epithelial barrier dysfunction and inflammation. BAL levels of inflammatory proteins including surfactant protein (SP)-D, soluble receptor for glycation end product (sRAGE) and matrix metalloproteinase (MMP)-9 were also increased. Cl2 inhalation resulted in upregulation of phospho-histone H2A.X, a marker of double-strand DNA breaks in the bronchiolar epithelium and alveolar cells; oxidative stress proteins, heme oxygenase (HO)-1 and catalase were also upregulated. Flow cytometric analysis of BAL cells revealed increases in proinflammatory macrophages following Cl2 exposure, whereas numbers of resident and antiinflammatory macrophages were not altered. This was associated with increases in numbers of macrophages expressing cyclooxygenase (COX)-2 and inducible nitric oxide synthase (iNOS), markers of proinflammatory activation, with no effect on mannose receptor (MR) or Ym-1 expression, markers of antiinflammatory activation. Metabolic analysis of lung cells showed increases in glycolytic activity following Cl2 exposure in line with proinflammatory macrophage activation. Mechanistic understanding of Cl2-induced injury will be useful in the identification of efficacious countermeasures for mitigating morbidity and mortality of this highly toxic gas.

Glucocorticoid-sensitive period of corticotroph development-Implications for mechanisms of early life stress.

Corticotrophs are intermediaries in the hypothalamic-pituitary-adrenal (HPA) axis, which plays a crucial role in stress response in vertebrates. The HPA axis displays an intricate mode of negative feedback regulation, whereby the peripheral effector, cortisol inhibits the secretion of its upstream regulator, adrenocorticotropic hormone (ACTH) from proopiomelanocortin (POMC)-expressing cells in the pituitary. While the feedback regulation of the HPA axis is well characterized in the adult organism, the effect of feedback regulation on the development of corticotrophs is poorly understood. Here, we studied the effect of glucocorticoids on the development of POMC-expressing cells in the zebrafish pituitary. The development of POMC cells showed a steady increase in numbers between 2-6 days post fertilization. Inhibition of endogenous glucocorticoid synthesis resulted in an increase in POMC cell number due to reduced developmental feedback inhibition of cortisol on POMC cells. Conversely, addition of exogenous dexamethasone at a critical developmental window led to a decrease in corticotroph cell number, mimicking greater feedback control due to increased cortisol levels. Finally, developmental dysregulation of ACTH levels resulted in impaired anxiety-like and stress-coping behaviours. Hence, we identified a sensitive developmental window for the effect of glucocorticoids on corticotrophs and demonstrate the downstream effect on stress-responsive behaviour.

Crashworthiness and optimization of bionic sandwich cores under out-of-plane compression

In this paper, novel bionic sandwich cores are proposed based on the cross section of seagull feather shaft to provide an effective protective mechanism against impact. The relative density of the bionic core is firstly defined, which is closely related to cell configuration and geometric parameters. Through finite element simulations supported by experiments using 3D printed specimens, the novel bionic core is found to be superior in crashworthiness as compared to conventional honeycomb core in terms of specific energy absorption and mean force under out-of-plane compression. To decipher the underlying mechanisms for the enhanced performances, a theoretical model for the bionic core is derived based on the simplified super folding element (SSFE) theory. The effects of wall thickness, number of cells and amplitude of sinusoidal beam on relative density and crashworthiness are explored. It is revealed that the increase in wall thickness and number of cells led to improvements of load bearing capacity. An optimal structure is subsequently obtained based on the complex proportional assessment (COPRAS) and response surface method (RSM). Compared to conventional honeycomb core, the optimal peak force of the bionic core is decreased by 11.53%. Through the combination of experiment, simulation and theoretical analysis, this study provides a novel idea for the optimization of the crashworthiness of the bionic core under out-of-plane compression.

Subacute and low dose of tributyltin exposure leads to brown adipose abnormalities in male rats

Tributyltin (TBT) is an obesogenic endocrine disrupting chemical (EDC) linked with several metabolic complications. Brown adipose tissue (BAT) is the principal site for thermogenesis, making it a potential target for obesity management and metabolic disease. However, few studies have evaluated TBT effect on BAT function. In this investigation, we assessed whether subacute (15 days) and low dose of TBT exposure (100 ng/kg/day) results in abnormal BAT morphophysiology in adult male rats. Body temperature, BAT morphology, inflammation, oxidative stress, collagen deposition and BAT metabolic gene expression markers were assessed in room temperature (Room, ∼24 ºC) and after cold tolerance test (Cold, ∼4 ºC) conditions. A reduction in body temperature was observed in both Room and Cold conditions in TBT rats, suggesting abnormal BAT thermogenic function. Changes in BAT morphology were observed in TBT rats, with an increase in BAT lipid accumulation, an increase in BAT unilocular adipocyte number and a decrease in BAT multilocular adipocyte number in Room condition. All these parameters were opposite in Cold condition TBT rats, leading to a borderline increase in BAT UCP1 protein expression. An increase in BAT mast cell number was observed in TBT rats in Room condition. An increase in ED1 protein expression (macrophage marker) was observed in TBT rats in Cold condition. Oxidative stress and collagen deposition increased in both Room and Cold conditions in TBT rats. TBT exposure caused a borderline increase in BAT COL1A1 protein expression in Cold condition. Further, strong negative correlations were observed between body temperature and BAT lipid accumulation, and BAT lipid accumulation and multilocular adipocyte number. Thus, these data suggest that TBT exposure impaired BAT morphophysiology through impacts on lipid accumulation, inflammation, fibrosis and oxidative stress in male rats.

EVALUATION OF THE TOLERANCE THRESHOLD OF THE MARINE BENTHIC DIATOM PLEUROSIGMA AESTUARII (BRéB. IN KüTZ.) W. SMITH, 1853 (BACILLARIOPHYTA) UNDER THE IMPACT OF COPPER (II) IONS

Copper compounds accumulate in marine bottom sediments as the result of human activity and, being highly toxic, affect microphytobenthos. Evaluation of the tolerance ranges of benthic diatoms to copper pollution is important for biotesting and assessment of coastal marine environment. This work is aimed to reveal the dynamics of growth and cell death of marine benthic diatom Pleurosigma aestuarii (Bréb. in Kütz.) W. Smith, 1853 under the impact of a wide range of copper (II) ions concentrations over the course of a 10-day experiment in clonal culture, as well as to identify a tolerance threshold for this species, critical for its survival when exposed to the toxicant. The study has focused on assessment of the changes in the proportion (%) of alive cells, absolute cell number and specific growth rate of the culture at different exposure durations and concentrations of Cu2+ ions (32-1024 μg/L) and was meant to evaluate the applicability of this species as a new test object for ecotoxicology. In the control culture and in the cultures exposed to Cu2+ in the range of concentrations 32-256 μg/L, the proportion of alive cells did not change over the course of the experiment (95-99 %). At Cu2+ concentration 320 μg/L, the proportion of alive cells decreased to 23 % on the 3rd day and to 10 % on the 5th day, which provided a basis to consider this value as a threshold one for P. aestuarii survival. At Cu2+ concentrations 384 μg/L and higher, up to the maximum one (1024 μg/L), drastic inhibition of the culture was recorded as early as on the 1st day, and on the days 3-5, all the cells died. The increase in the absolute cell number in the concentration range 32- 256 μg/L were consistent with the dose-response sigmoid model. Over the timespan of days 1-7, the cell number increased by 3-5 times, reaching its maximum, and then it decreased by 10-12 % by the 10th day. At Cu2+ concentrations 320 μg/L and higher, the increase in the cell number was strongly suppressed since the 1st day. Within the 32-256 μg/L range, the test culture is characterized by positive specific cell growth rate for the period up to 7 days; at the threshold concentration 320 µg/L and higher, this test parameter becomes negative. Thus, P. aestuarii should be recommended as a new appropriate test object both for toxicological experiments and for monitoring of coastal marine environment affected by technogenic pollution.

Evaluation and Performance of Three-Phase Inverter Using Multiple Bidirectional Choppers Intended for 1.5-kVdc PV Systems

This article focuses on the chopper-cell number of a novel three-phase inverter for utility-scale photovoltaic (PV) systems where multiple cascaded bidirectional chopper cells and a three-phase line-frequency transformer with a three-legged core are used. The inverter per phase is composed of a main converter, which is equivalent to the conventional bidirectional chopper, and an auxiliary converter, which is composed of multiple cascaded chopper cells. Although the inverter performance can be improved by increasing the chopper-cell number because of increased switching frequency and reduced voltage steps, the increased cell number may result in increased converter loss and cost. However, no paper has evaluated the chopper-cell number of the inverter to the best of the authors’ knowledge. Further, no paper has carried out experimental verification of the inverter during the power faults. This article evaluates the inverter using two cells per phase (two-cell inverter) with the one using three cells per phase (three-cell inverter) under the same equivalent switching frequency in terms of converter loss, efficiency, and steady/transient state performance. The numerical analysis shows that the two-cell inverter shows superiority over the three-cell inverter in converter efficiency. Further, the experiments using a 1.5-kW downscaled model verify the inverter performance under the normal and fault conditions.

Plastic rearrangement of basal forebrain parvalbumin-immunoreactive neurons in the kainite model of epilepsy.

Temporal lobe epilepsy (TLE) is the most prevalent form of epilepsy, through the neuronal mechanisms of this syndrome remain elusive. In addition to the temporal lobe structures, it was found that the basal forebrain cholinergic cells are also involved in epileptogenesis. However, little is known about the involvement of the basal forebrain GABAergic neurons in epilepsy; despite this, they largely project to the temporal lobe and are crucial for the regulation of the hippocampal circuitry. In this study, we assessed epilepsy-induced changes in parvalbumin (PARV) immunoreactive neurons of the medial septum (MS) and of the magnocellular preoptic nucleus (MCPO) using the kainic acid (KA) model in rats. In addition, we estimated the respective changes in the cholinergic varicosities in the MS, where we observed a significant reduction in the PARV cell number (12849 ± 2715 vs. 9372 ± 1336, p = .029) and density (16.2 ± 2.62 vs. 10.5 ± 1.00 per .001 mm3, p =.001), and an increase in the density of cholinergic varicosities (47.9 ± 11.1 vs. 69.4 ± 17.8 per 30,000 µm2, p =.036) in KA-treated animals. In the MCPO, these animals showed a significant increase in somatic volume (827.9 ± 235.2 µm3 vs. 469.9 ± 79.6 µm3, p = .012) and total cell number (2268.6 ± 707.1 vs. 1362.4 ± 262.0, p =.028). These results show that the basal forebrain GABAergic cell populations undergo numerical and morphological changes in epileptic animals, which may contribute to an increased vulnerability of brain circuits to epilepsy and epilepsy-related functional impairments.

Functionalization of 3D printed polymeric bioresorbable stents with a dual cell-adhesive peptidic platform combining RGDS and YIGSR sequences.

Biomimetic surface modification with cell-adhesive peptides is a promising approach to improve endothelialization of current bioresorbable stents (BRS). Among them, RGDS and YIGSR sequences have been reported to mediate adhesion and migration of endothelial cells (ECs) while preventing platelet activation. This work presents the functionalization of novel 3D-printed poly-L-lactic acid (PLLA) and poly(L-lactic-co-ε-caprolactone) (PLCL) BRS with linear RGDS and YIGSR sequences, as well as a dual platform (PF) containing both motifs within a single biomolecule. Functionalized surfaces were characterized in terms of static contact angle, biomolecule distribution under confocal fluorescence microscopy and peptide quantification via detachment from the surface, showing a biomolecule density in the range of 0.5 to 3.5 nmol cm-2. Biological evaluation comprised a cell adhesion test on functionalized films with ECs and a blood perfusion assay on functionalized stents to assess ECs response and device hemocompatibility, respectively. Cell adhesion assays evidenced significantly increased cell number and spreading onto functionalized films with respect to control samples. Regarding stents' hemocompatibility, platelet adhesion onto PLCL stents was severely decreased with respect to PLLA. In addition, functionalization with RGDS, YIGSR and the PF rendered BRS stents displaying even further reduced platelet adhesion. In conclusion, the combination of intrinsically less prothrombogenic materials such as PLCL and its functionalization with EC-discriminating adhesive biomolecules paves the way for a new generation of BRS based on accelerated re-endothelialization approaches.

MANAGEMENT OF SUSPECT GRANULOMATOUS EPULIS OF ANTERIOR MANDIBULAR TEETH WITH GINGIVECTOMY

ABSTRACT Gingival enlargement is a manifestation of hypertrophy (increase in cell size) and hyperplasia (increase in cell number). Epulis is a gingival hyperplasia (tumor-like) that originates from periodontal connective tissue. One type of epulis that often occurs is granulomatous epulis. Granulomatous epulis is a type of epulis that occurs from a granulomatous tissue reaction due to chronic irritation due to calculus, root residues, or carious edges. This case report discusses the management of suspect epulis granulomatous in mandibular anterior teeth. A 45-year-old female patient complained of swollen lower front gums and frequent bleeding when brushing her teeth. Intraoral examination showed that there was gingival enlargement in the form of papules with a size of 6 mm x 8 mm, reddish in color, smooth surface, soft consistency, well demarcated, painless but bleeding on palpation in the tooth area 41. OHI score 4.9 (moderate), BOP ( Bleeding in probing) was positive, and the probing depth of the mesiolabial, midlabial, and distolabial surfaces i.e. for tooth 41 was 5-5-5. The diagnosis in this case led to suspect epulis granulomatous on tooth 41. The treatment plan was KIE (Communication, Information and Education), initial therapy in the form of scaling and root planing, gingivectomy and then control. The results of the treatment showed that the gingival contour was physiologically formed, there was no swelling, negative palpation, negative BOP, and decreased pocket depth from the mesiolabial, midlabial, and distolabial surfaces of tooth 41 to 2-2-3.Keywords: epulis, granulomatous epulis, gingivectomy, gingival enlargement

Dendritic Cell-Based Vaccines Recruit Neutrophils to the Local Draining Lymph Nodes to Prime Natural Killer Cell Responses

Dendritic cell (DC)-based cancer vaccines are a form of immunotherapy that activates the innate and adaptive immune systems to combat cancers. Neutrophils contribute to cancer biology and have the potential to be exploited by immunotherapeutic platforms to enhance anti-tumor immune responses. We previously showed that DC vaccines elicit the expansion of mouse interferon (IFN)γ-producing mature natural killer (NK) cells to elevate anti-tumor responses. Here, we demonstrate the rapid recruitment of neutrophils to the draining lymph nodes of DC-vaccinated mice. This was accompanied by an increase in the total number of NK cells producing IFNγ and expressing CD107a, a marker of degranulation that demonstrates NK cell functional activity. Furthermore, the depletion of neutrophils in DC-immunized mice resulted in decreased numbers of NK cells in draining lymph nodes compared to the controls. Interestingly, the increased number of IFNγ- and CD107a-expressing NK cells in DC-immunized mice was not detected in mice depleted of neutrophils. Further investigations showed that DC vaccines induced IFNγ- and TNFα-producing CD8+ T cells that also expressed CD107a, but depletion of neutrophils did not have any impact on the CD8+ T cell population. Our findings suggest that neutrophil-mediated anti-tumor immunity induced by a DC vaccine platform could be targeted to provide innovative strategies to enhance its clinical efficacy.

Anatomical Assessment of Skin Separation in Date Palm (Phoenix dactylifera L. var. Mejhoul) Fruit during Maturation and Ripening Stages

Skin separation (puffiness) is a critical physiological disorder that significantly reduces the date palm (Phoenix dactylifera L.) fruit’s visual appearance and quality. The objectives of this study were to analyze fruit anatomy in skin-separated and normal date palm (var. Mejhoul) fruit and to assess the microclimatic conditions (temperature and relative humidity) during the fruit developmental stages and their association with skin separation. Fruit anatomy analysis was carried at different growth stages (Kimri-Khalal, Khalal, Rutab and Tamr) for two growing seasons. In addition, microclimatic conditions (specifically, temperature, and relative humidity) as well as soil moisture content were assessed across the study period 2020 and 2021. The anatomical analysis of the date palm fruit revealed that the exocarp or skin (cuticle, epidermis, hypodermis) thickness were quite similar across the developmental stages and over the study period (2020, 2021). Conversely, significantly more sclereid cells were found in skin-separated fruit (compared to normal fruit) at the Tamr stage. At that stage (Tamr), the percentage increase in sclereid cell number in skin-separated fruit ranged from 34–183%, sclereid clusters number 23–92%, cluster area 37–206%, and cluster perimeter 25–64%, as compared to normal fruit. In addition, sclereid cells at skin-separated areas of the fruit were clustered together, forming a chain of aggregates underneath the fruit skin, and were closer (42–50%, than normal) to the cuticle, leading to the partial separation of the exocarp from the fruit mesocarp. Although the weather data were similar across the growing seasons (temperatures, 33–34 °C; relative humidity, 45–46%), skin separation occurred in 14–15% of total fruit in 2020 and 30–34% in 2021. Therefore, we believe that skin separation disorder is not as heavily weather dependent as it seems to have been thought. In conclusion, changes in microclimate conditions were not the conclusive component for inducing the skin separation physiological disorder in date palm fruit. Further studies are required to identify the main factors that stimulate the development of sclereid clusters aggregates and the potential of agricultural practices to reduce skin separation.

Anp32a Promotes Neuronal Regeneration after Spinal Cord Injury of Zebrafish Embryos.

After spinal cord injury (SCI) in mammals, neuronal regeneration is limited; in contrast, such regeneration occurs quickly in zebrafish. Member A of the acidic nuclear phosphoprotein 32 (ANP32a) family is involved in neuronal development, but its function is controversial, and its involvement in zebrafish SCI remains unknown. To determine the role of zebrafish ANP32a in the neuronal regeneration of SCI embryos, we microinjected ANP32a mRNA into embryos from zebrafish transgenic line Tg(mnx1:GFP) prior to SCI. Compared to control SCI embryos, the results showed that the regeneration of spinal cord and resumption of swimming capability were promoted by the overexpression of ANP32a mRNA but reduced by its knockdown. We next combined fluorescence-activated cell sorting with immunochemical staining of anti-GFAP and immunofluorescence staining against anti-PH3 on Tg(gfap:GFP) SCI embryos. The results showed that ANP32a promoted the proliferation and cell number of radial glial cells at the injury epicenter at 24 h post-injury (hpi). Moreover, when we applied BrdU labeling to SCI embryos derived from crossing the Tg(gfap:GFP) and Tg(mnx1:TagRFP) lines, we found that both radial glial cells and motor neurons had proliferated, along with their increased cell numbers in Anp32a-overexpression SCI-embryos. On this basis, we conclude that ANP32a plays a positive role in the regeneration of zebrafish SCI embryos.

Sodium Butyrate Supplementation Modulates Neuroinflammatory Response Aggravated by Antibiotic Treatment in a Mouse Model of Binge-like Ethanol Drinking.

Growing evidence supports the pivotal role of the bidirectional interplay between the gut microbiota and the central nervous system during the progression of alcohol use disorder (AUD). In our previous study, supplementation with sodium butyrate (SB) in C57BL/6J mice prevented increased ethanol consumption in a binge-like drinking paradigm (DID) as a result of treatment with a non-absorbable antibiotic cocktail (ABX). In this study, we tested the hypothesis that SB protection against enhanced ABX-induced ethanol consumption in mice is partially due to modulation of neuroinflammatory responses. Pro- and anti-inflammatory cytokines, as well as changes in microglia and astrocytes were analyzed in hippocampus tissues from ABX-, SB-, ABX+SB-treated mice subjected to 4-week DID. We found that ethanol without or with ABX treatment increased mRNA levels of key brain cytokines (MCP-1, TNF-α, IL-1β, IL-6 and IL-10) while SB supplementation prevented these changes. Additionally, SB supplementation prevented changes in microglia, i.e., increase in Iba-1 positive cell number and morphology, and in astrocytes, i.e., decrease in GFAP-positive cell number, induced by combination of ethanol and ABX treatments. Our results suggest that gut microbiota metabolites can influence drinking behavior by modulation of neuroinflammation, highlighting the potential for microbiome-targeting strategies for treatment or prevention of AUD.

The Issue of Monocyte Activation in ASD: Troubles with Translation.

Autism spectrum disorder (ASD) prevalence has increased year on year for the past two decades and currently affects 1 in 44 individuals in the US. An increasing number of studies have pointed to increased immune activation as both an etiological agent and also involved in the ongoing pathological process of ASD. Both adaptive and innate immune responses have been implicated. Evidence of innate dysregulation has so far included increased production of innate inflammatory cytokines, increased cell numbers, and altered activation in monocytes in the blood and microglia in the brain. Suggesting an orchestrated innate immune response may be involved in ASD. Hughes et al. (2022) recently assessed transcriptome differences that could underlie altered activation of monocytes using next-generation bulk-RNA sequencing on isolated CD14+ monocytes at baseline and after activation with different Toll-like receptor agonists. Circulating CD14+ monocyte from children with autistic disorder (AD) and children diagnosed with perverse developmental disorder not otherwise specified (PDD-NOS) were found to differ in a number of activation pathways after gene enrichment analysis compared to typically developing children. There was an overall upregulation in translational machinery in both neurodevelopmental disorder groups, whereas typically developing children were downregulated, indicating an issue with monocyte activation. Several identified differentially expressed genes in monocytes were also identified as ASD at-risk genes, according to the Simons Foundation Autism Research Initiative (SFARI), and genes involved in inflammatory bowel diseases. This work implicates altered monocyte activation with a lack of regulation as a potential mechanistic issue in ASD. Future work is warranted to evaluate how monocyte regulatory mechanisms differ in ASD individuals.

Zooplankton‐derived dissolved organic matter composition and its bioavailability to natural prokaryotic communities

AbstractZooplankton grazing onphytoplankton promotes the release of particulate and dissolved organic matter (DOM) into the water column and therefore plays a key role in organic matter cycling in aquatic systems. Prokaryotes are the main DOM consumers in the ocean by actively remineralizing and transforming it, contributing to its molecular diversification. To explore the molecular composition of zooplankton‐derived DOM and its bioavailability to natural prokaryotic communities, the DOM generated by a mixed zooplankton community in the coastal Atlantic off Spain was used as substrate for a natural prokaryotic community and monitored over a ~ 5‐d incubation experiment. The molecular composition of solid‐phase extracted DOM was characterized via Fourier‐transform ion cyclotron resonance mass spectrometry. After ~ 4 d in the zooplankton‐derived DOM amended incubation, the prokaryotic community demonstrated a 17‐fold exponential increase in cell number. The prokaryotic growth resulted in a reduction in bulk dissolved organic carbon concentration and the zooplankton‐derived DOM was considerably transformed at molecular and bulk elemental levels over the incubation period. The C : N ratio (calculated from the obtained molecular formulae) increased while the functional diversity decreased over the incubation time. In addition, molecular indices pointed to a reduced bioavailability of DOM at the end of the experiment. These findings show that zooplankton excreta are a source of labile organic matter that is quickly metabolized by the prokaryotic community. Therefore, a fraction of carbon is shunted from transfer to secondary consumers similarly to the viral shunt, suggesting that the zooplankton–prokaryotic interactions play an important role in the ocean's carbon cycle.

Macroglial activation over time in an experimental glaucoma model

AbstractGlaucoma is a neurodegenerative retinal disease characterized by irreversible loss of retinal ganglion cells (RGCs), leading to visual loss. Increased intraocular pressure (IOP) is the only risk factor for which treatment is available. However, other factors such as glial activation and dysfunction can also induce RGC death in glaucoma.Astrocytes and Müller glia (macroglial cells) play important roles in neuronal activity by providing physical and metabolic support to neurons. In glaucoma, when tissue damage occurs, macroglial cells undergo reactive gliosis, to defend the nerve tissue against damage and attempt to maintain its homeostasis. However, alterations in the function of these cells could cause damage and even neuronal death. In reactive macrogliosis, cells undergo complex biochemical and functional remodelling and exhibit morphological changes characterized by cell body thickening, increased number and length of cell processes, increased cell number and up‐regulation of cytoskeletal components such as gliofibrillary acidic protein (GFAP), which is considered one of the main markers of this process and may act as antigen‐presenting cells by expressing major histocompatibility complex class II (MHC‐II). Reactivation of macroglia can initially be beneficial, as it increases their metabolic activity, increases the expression of antioxidant defence, and restores ion, water and neurotransmitter balance. However, if macroglia become chronic, it is detrimental by directly or indirectly damaging the tissue and preventing its repair. Astrocytes may act in concert with microglia during the inflammatory process, so inflammatory mediators produced by astrocytes can chronically activate microglial cells, contributing to neuronal death, and similarly, inflammatory mediators released by microglia can chronically activate astrocytes.One of the most widely used models to elevate IOP is the unilateral laser‐induced ocular hypertension (OHT) model. Studies in this model have analysed the temporal pattern of RGC death and microglial cell activation at time points: 1, 3, 5, 8 and 15 days. Molecular changes in proinflammatory and anti‐inflammatory cytokines involved in the neuroinflammatory process, released by glial cells (microglia, astrocytes, and Müller cells) at the same time points mentioned above, have also been identified.Given the close relationship between the activation of macroglia and microglia and their involvement in the death of RGCs in this pathology, the present work analysed how the activation of retinal macroglial cells occurred at different time points (1, 3, 5, 8 and 15 days) after the induction of OHT in an experimental model of laser‐induced OHT in mice. For this purpose, we have analysed the GFAP‐labelled retinal area, the intensity of GFAP labelling, and the expression of MHC‐II in both OHT eyes and normotensive contralateral eyes compared to naïve eyes.In this study, in both OHT and contralateral eyes, macroglial cells showed morphological changes as well as in GFAP and MHC‐II expression at all time points analysed, being slightly more intense at 3–5 days, coinciding with the pattern of microglial activation previously studied, demonstrating this fact, a bilateral inflammatory process maintained over time in which microglia, astrocytes and Müller glia are involved.

CMV reactivation initiates long-term expansion and differentiation of the NK cell repertoire.

NK cells play an important role in suppression of viral replication and are critical for effective control of persistent infections such as herpesviruses. Cytomegalovirus infection is associated with expansion of 'adaptive-memory' NK cells with a characteristic CD56dimCD16bright NKG2C+ phenotype but the mechanisms by which this population is maintained remain uncertain. We studied NK cell reconstitution in patients undergoing haemopoietic stem cell transplantation and related this to CMV reactivation. NK cells expanded in the early post-transplant period but then remained stable in the absence of viral reactivation. However, CMV reactivation led to a rapid and sustained 10-fold increase in NK cell number. The proportion of NKG2C-expressing cells increases on all NK subsets although the kinetics of expansion peaked at 6 months on immature CD56bright cells whilst continuing to rise on the mature CD56dim pool. Phenotypic maturation was observed by acquisition of CD57 expression. Effective control of viral reactivation was seen when the peripheral NK cell count reached 20,000/ml. These data show that short term CMV reactivation acts to reprogramme hemopoiesis to drive a sustained modulation and expansion of the NK cell pool and reveal further insight into long term regulation of the innate immune repertoire by infectious challenge.

Deletion of Notch1 during Cochlear Maturation Leads to Rapid Supporting Cell Death and Profound Deafness.

The sensory region of the mammalian hearing organ contains two main cell types-hair cells and supporting cells. During development, Notch signaling plays an important role in whether a cell becomes either a hair cell or supporting cell by mediating lateral inhibition. However, once the cell fate decisions have been determined, little is understood about the role Notch plays in cochlear maturation. Here, we report that deletion of Notch1 from the early postnatal mouse cochlea in both male and female animals resulted in profound deafness at 6 weeks of age. Histologic analyses at 6 weeks revealed significant hair cell and supporting cell loss throughout the Notch1-deficient cochlea. Early analyses revealed a reduction in supporting cells in the outer hair cell region between postnatal day (P) 2 and P6, without a comparable increase in outer hair cell number, suggesting a mechanism other than lateral inhibition. Consistent with this, we found apoptotic cells in the outer supporting cell region of the cochlea at P1 and P2, indicating that Notch1 is required for outer supporting cell survival during early cochlear maturation. Interestingly, inner supporting cell types were not lost after Notch1 deletion. Surprisingly, we do not detect outer hair cell loss in Notch1 mutants until after the onset of hearing, around P14, suggesting that hair cell loss is caused by loss of the supporting cells. Together, these results demonstrate that Notch1 is required for supporting cell survival during early maturation and that loss of these cells causes later loss of the hair cells and cochlear dysfunction.SIGNIFICANCE STATEMENT During development, Notch signaling has been shown to be critical in regulating the cell fate choices between hair cells and supporting cells. However, little is known about how Notch functions after those cell fate choices are made. Here, we examine the role of Notch1 in the maturing cochlea. We demonstrate that deletion of Notch1 results in profound deafness by 6 weeks of age. Histologic analyses revealed rapid supporting cell death shortly after Notch1 deletion, followed by eventual loss of the hair cells. These results reveal an unexpected role for Notch in supporting cell survival during cochlear maturation.