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Comparative oncology is an understudied field of science. We are far from understanding the key mechanisms behind Peto's paradox, i.e., understanding how long-lived and large animals are not subject to a higher cancer burden despite the longer exposure time to mutations and the larger number of cells exposed.In this work, we investigated the scientific evidence on such mechanisms through a systematic mini-review of the literature about the relation of longevity and/or large body mass with physiological, genetic, or environmental traits among mammalian species. More than forty thousand articles were retrieved from three repositories, and 383 of them were screened using an active-learning-based tool. Of those, 36 articles on longevity and 37 on body mass were selected for the review. Such articles were examined focusing on: number and type of species considered, statistical methods used, traits investigated, and observed relationship with longevity and/or body mass. Where applicable, the traits investigated were matched with one or more hallmarks of cancer.We obtained a list of potential candidate traits to explain Peto's paradox related to replicative immortality, cell senescence, genome instability and mutations, proliferative signaling, growth suppression evasion, and cell resistance to death.Our investigation suggests that different strategies have been followed to prevent cancer in large and long-lived species. The large number of papers retrieved emphasizes that more studies can be launched in the future, using more efficient analytical approaches to comprehensively evaluate the convergent biological mechanisms essential for acquiring longevity and large body mass without increasing cancer risk.

The current paper covers an experimental study, mathematical modelling, and design optimization of the laser treatment process of denim fabrics. Laser fading is used in the finishing phase of garment production, unfortunately, it decreases fabric durability. Denim fabric is known as a cotton fabric but nowadays it is blended with other fibres to improve fabric properties. For this reason, the main purpose of this study was to test multicomponent fabric longevity after laser treatment. The knowledge from this study is used by garment production company. The tear force, abrasion resistance, and colour difference are considered as performance characteristics of laser treated denim fabrics subjected to maximization. To achieve the goals, novel, mathematical models were developed. In the preliminary tests, the design space was determined. The full factorial design of experiments was utilized in the experimental study and the values of the performance characteristics are measured. Based on the results of the experimental study the artificial neural network and Haar wavelet based mathematical models were developed to predict the behaviour of the tear force, abrasion resistance, and colour difference. The artificial intelligence-based multicriteria design optimization procedure is developed.

Laser fading technology is used to give a unique worn look to a fabric. This finishing technique is environmentally friendly compared to conventional methods because it reduces the use of harmful chemicals and large amounts of water. A carbon dioxide (CO2) laser with a wavelength of 10.6 µm was used in this study. In bulk production, fixed manufacturing parameters help to reduce production preparation time. Thus, two combinations of laser power and speed of the laser cutter head (14 W and 230 mm/s; 16 W and 350 mm/s) were used to determine how universal the fixed laser parameters are for fading five different types of multicomponent twill and satin weave denim fabrics, which contain cotton, elastane, polyester and viscose. Physico-mechanical properties (tear, tensile properties and abrasion resistance) were tested to evaluate the effect of the selected laser parameters on fabric strength properties. Microscopical analysis was performed to assess the effect of laser fading on the yarn and fibre morphology of denim fabrics.

The effect of nanoclay additive on polyamide film oxygen permeability is investigated from the perspective of possible use as a laminate component for low-cost food packaging material. Montmorillonite nanoclay was melt-mixed in an industrial grade polyamide by twin-screw extrusion and the mixture was hot-pressed to a ~50 µm thick film. The film with 10 wt.% of nanoclay loading showed a 17 % decrease in the oxygen transmission rate (OTR), as compared to the pristine polyamide film (72 and 87 cm3/m2∙24 h, respectively). Despite the relatively high loading of the filler the obtained OTR exceeds that of the food packaging preferred upper limit of 10 cm3/m2∙24 h. XRD measurements confirmed the near-complete exfoliation of the nanoclay platelets. The platelets were found to be at an average angle of 9.5 degrees relative to the film’s surface plane. To comply with the requirements for food packaging, this angle needs to be decreased down to 0.4 degrees. To achieve this, different film-making methods enabling better control over the filler particles’ orientation need to be explored. Nanoclay addition increased the films’ yield strength (23 % for 10 wt.% film) and stiffness, while not affecting the films’ optical appearance.

The circular economy is emerging as green technology solution for polymer and composite industries. However, the use of circular economy as an industrial practice is still a global challenge. In this article, polypropylene-cotton hybrid composite was developed using different amounts of cotton fibre waste (0, 10, 30, 40 wt%). Scanning electron microscope (SEM), tribometer, Rockwell hardness tester and binocular microscope were used for investigations of composite surface, hardness and coefficient of friction (COF). The mean coefficient of friction values was 0.64, 0.75, 0.88 and 0.94 for pure propylene, 10, 30 and 40% of cotton reinforced composites, respectively. The scanning electron microscopy characterization of hybrid composite revealed the voids, porosity and asperities due to random fibres orientation. The Rockwell hardness value of composites was increased due to rise of fibre fraction. Based on the COF values, hardness and surface characterization, polypropylene-cotton reinforced hybrid composite could be used functionally for thermal and sound applications.

Characteristics and serviceability of denim fabrics have undergone major changes. Nowadays denim is commonly used for casual wear. Durability and comfort are important parameters for consumers when choosing a denim garment. Therefore, in this study, abrasion resistance, tear and tensile properties of core–spun yarns and air permeability of denim fabrics with different weft yarns per centimetre and fibre content were analysed. The test results showed that weft yarns per centimetre influences fabric air permeability negatively but abrasion resistance increases. Higher weft yarns per centimetre influences fabric air permeability negatively but abrasion resistance increases. Polyester, elastane, modal, viscose and Lycra T400 were used in the core of weft yarn to analyse the impact of those fibres on the durability and comfort properties. Elastane is used to add stretchability to the fabric, which provides comfort to the wearer. The higher the elastomeric fibre content in the fabric, the greater is its elasticity; however, the tensile properties of the woven fabric decrease. The tear strength of the fabric was increased by the presence of the polyester fibre in the core.

In this study, propolis additives provide antibacterial and antifungal effects that prolong the product’s shelf life. The aim of the study is to obtain homogeneous fiber membranes of polyvinyl alcohol and propolis by the electrospinning method and to evaluate their suitability for food packaging. Three propolis extracts are used in the study—water, ethyl alcohol, and glycerin-based. The membranes’ morphology and fiber diameter distribution, tensile deformation, air permeability, thermogravimetric analysis, differential scanning calorimetry, Fourier-transform infrared spectroscopy, and microbiological tests (Listeria monocytogenes, Salmonella enteritidis, Escherichia coli) were analyzed for electrospun samples. The results of the study show that propolis extracts are incorporated into membranes and the additive provides an antimicrobial effect with the contact surface. The obtained membranes are breathable: gas exchange can be controlled by using a material of appropriate thickness (air permeability coefficient is 0.046 and 0.276 mm/s). The mechanical properties of membranes are affected by moisture, but tensile strength can be improved with thermal post-processing at 100 °C. The propolis-containing fibers’ diameters are from 293 ± 8 to 664 ± 11 nm. Depending on membranes’ demonstrated properties, it can be concluded that the composites have the potential to increase the shelf life of fresh fruits and berries.

Abstract Congenital intrahepatic portosystemic venous shunts are rare vascular malformations which are incidentally discovered on imaging or once hepatic encephalopathy becomes clinically apparent. Surgical ligation and endovascular embolization are potential treatments.

Due to the high prevalence of infectious diseases and their concurrent outbreaks, there is a high interest in developing novel materials with antimicrobial properties. Antibacterial and antiviral properties of a range of metal-based nanoparticles (NPs) are a promising means to fight airborne diseases caused by viruses and bacteria. The aim of this study was to test antimicrobial metals and metal-based nanoparticles efficacy against three viruses, namely influenza A virus (H1N1; A/WSN/1933) and coronaviruses TGEV and SARS-CoV-2; and two bacteria, Escherichia coli and Staphylococcus aureus. The efficacy of ZnO, CuO, and Ag NPs and their respective metal salts, i.e., ZnSO4, CuSO4, and AgNO3, was evaluated in suspensions, and the compounds with the highest antiviral efficacy were chosen for incorporation into fibers of cellulose acetate (CA), using electrospinning to produce filter materials for face masks. Among the tested compounds, CuSO4 demonstrated the highest efficacy against influenza A virus and SARS-CoV-2 (1 h IC50 1.395 mg/L and 0.45 mg/L, respectively), followed by Zn salt and Ag salt. Therefore, Cu compounds were selected for incorporation into CA fibers to produce antiviral and antibacterial filter materials for face masks. CA fibers comprising CuSO4 decreased SARS-CoV-2 titer by 0.38 logarithms and influenza A virus titer by 1.08 logarithms after 5 min of contact; after 1 h of contact, SARS-COV-2 virus was completely inactivated. Developed CuO- and CuSO4-based filter materials also efficiently inactivated the bacteria Escherichia coli and Staphylococcus aureus. The metal NPs and respective metal salts were potent antibacterial and antiviral compounds that were successfully incorporated into the filter materials of face masks. New antibacterial and antiviral materials developed and characterized in this study are crucial in the context of the ongoing SARS-CoV-2 pandemic and beyond.

Fungi contain a significant amount of chitin in their cell walls presenting an attractive source for this commercially significant material. In this study, chitin was extracted from eight different mushroom species native to Estonia. Significant differences in molar mass, chitin content, and fibre size were observed while the degree of acetylation was mostly similar.

Electrospun polymer nanofiber materials have been studied as basic materials for various applications. Depending on the intended use of the fibers, their morphology can be adjusted by changing the technological parameters, the properties of the spinning solutions, and the combinations of composition. The aim of the research was to evaluate the effect of electrode type, spinning parameters, polymer molecular weight, and solution concentration on membranes morphology. The main priority was to obtain the smallest possible fiber diameters and homogeneous electrospun membranes. As a result, five electrode types were selected, the lowest PVA solution concentration for stable spinning process was detected, spinning parameters for homogenous fibers were obtained, and the morphology of electrospun fiber membranes was analyzed. Viscosity, conductivity, pH, and density were evaluated for PVA polymers with five different molecular weights (30–145 kDa) and three concentration solutions (6, 8, and 10 wt.%). The membrane defects and fiber diameters were compared as a function of molecular weight and electrode type. The minimum concentration of PVA in the solution allowed more additives to be added to the solution, resulting in thinner diameters and a higher concentration of the additive in the membranes. The molecular weight, concentration, and electrode significantly affected the fiber diameters and the overall quality of the membrane.

Novel composite self-disinfecting films of polylactic acid (PLA) filled with nanosized particles of double sodium–copper(II) paratungstate B Na2Cu3(CuOH)2[W12O40(OH)2]·32H2O (POM) were developed. The solvent casting (POM/PLA film) and solvent-free melt extrusion methods (Extr. POM/PLA film) were applied for film preparation. The copper (II) ion release to water from both types of the films after 10 days at different temperatures demonstrated that the PLA matrix acts as a diffusion barrier, and the resulting concentration of released copper in water at room temperature remained low, at 0.79% for POM/PLA film and 0.51% for Extr. POM/PLA film. The POM-containing films reveals a significant inhibitory effect against E. coli ATCC 25922 in the agar diffusion test. The numbers of CFUs in washes of the films after incubation for 24 h were found to be 3.6 log CFU mL–1 (POM/PLA film) and 4.1 log CFU mL–1 (Extr. POM/PLA film). The films combine the antibacterial properties of POM and a bio-based polymer matrix, which makes them a prospective coating material for applications in hospital indoor environments. Excellent thermal stability of POM gives a technological advantage for industrial manufacturing to allow the processing of novel composite material in the solvent free (molten) state.

We have applied a novel and integrative analysis framework for next-generation sequencing (NGS) data to 503 human subjects provided by the Religious Orders Study and Memory and Aging Project (ROSMAP) to examine changes in transcriptomic organization and common variants in association with late-onset Alzheimer’s disease (LOAD). Our framework identified seven reproducible, co-regulated modules after quality control (QC), clinical segregation, preservation filtering, and functional ontology analysis. These modules were specifically enriched in several innate and adaptive immune system processes, the synaptic vesicle cycle, and Hippo signaling. Topological and functional erosion of these modules due to shedding of genes and loss of in-module connectivity was diagnostic of disease progression. Perturbation analysis revealed that only 1% of eQTLs overlapped genes participating in these co-regulated modules. Common variants nevertheless identified components of the immune systems like human leukocyte antigen (HLA) complex and microtubule-associated protein tau (MAPT) regions in association with LOAD. Our results implicate microglial function, adaptive immune response, and the structural degeneration of neurons as contributors to the transcriptional deregulation observed along with common genetic variants in the progression of LOAD.

Endovascular retrieval of a foreign body is becoming an increasingly common procedure in the management of complications resulting from more frequent endovascular procedures. Many procedures are performed on a regular basis in assessment of vascular anatomy, endovascular-guided therapy, and catheter placement. This case report depicts a complication of a chemoport placement resulting in a foreign body. Evaluation of the foreign body raised attention to aberrant anatomy, a persistent left-sided superior vena cava. We further discuss briefly the embryology behind a persistent left-sided superior vena cava, technical errors leading to the foreign body, and assessing the nature of the foreign body through different imaging modalities. This is followed by the subsequent endovascular retrieval by Interventional Radiology and a literature review and individual case assessment of endovascular foreign body retrieval. We discuss considerations for practice based upon our literature review. Keywords: Superior vena cava, Persistent, Endovascular, Chemoport, Interventional radiology, Aberrant anatomy

The aim of this study was to investigate the electrochemical behaviour of aqueous electrolytes on thin-layer (20 µm) nanoporous carbide-derived carbon (CDC) composite fibrous directly electrospun electrodes without further carbonisation. There have been previously investigated fibrous electrodes, which are produced by applying different post-treatment processes, however this makes the production of fibrous electrodes more expensive, complex and time consuming. Furthermore, in the present study high specific capacitance was achieved with directly electrospun nanoporous CDC-based fibrous electrodes in different neutral aqueous electrolytes. The benefit of fibrous electrodes is the advanced mechanical properties compared to the existing commercial electrode technologies based on pressure-rolled or slurry-cast powder mix electrodes. Such improved mechanical properties are preferred in more demanding applications, such as in the space industry. Electrospinning technology also allows for larger electrode production capacities without increased production costs. In addition to the influence of aqueous electrolyte chemical composition, the salt concentration effects and cycle stability with respect to organic electrolytes are investigated. Cyclic voltammetry (CV) measurements on electrospun electrodes showed the highest capacitance for asymmetrical cells with an aqueous 1 M NaNO3-H2O electrolyte. High CV capacitance was correlated with constant current charge–discharge (CC) data, for which a specific capacitance of 191 F g−1 for the positively charged electrode and 311 F g−1 for the negatively charged electrode was achieved. The investigation of electrolyte salt concentration on fibrous electrodes revealed the typical capacitance dependence on ionic conductivity with a peak capacitance at medium concentration levels. The cycle-life measurements of selected two-electrode test cells with aqueous and non-aqueous electrolytes revealed good stability of the electrospun electrodes.

This study focuses on the electrochemical behavior of thin-layer fibrous carbide-derived carbon (CDC) electrospun electrodes in commercial and research and development stage organic-solvent and ionic liquid (IL) based electrolytes. The majority of earlier published works stated various electrolytes with asymmetric cells of powder-based pressure-rolled (PTFE), or slurry-cast electrodes, were significantly different from the presented CDC-based fibrous spun electrodes. The benefits of the fibrous structure are relatively low thickness (20 µm), flexibility and mechanical durability. Thin-layered durable electrode materials are gaining more interest and importance in mechanically more demanding applications such as the space industry and in wearable devices, and need to achieve a targeted balance between mechanical, electrical and electrochemical properties. The existing commercial electrode technologies lack compatibility in such applications due to their limited mechanical properties and high cost. The test results showed that the widest potential window dU ≤ 3.5 V was achieved in 1.5 M 1-ethyl-3-methylimidazoliumbis(trifluoromethyl-sulfonyl)imide (EMIm-TFSI) solution in acetonitrile (ACN). Gravimetric capacitance reached 105.6 F g−1 for the positively charged electrode. Cycle-life results revealed stable material capacitance and resistance over 3000 cycles.

Biological aging is a complex process dependent on the interplay of cell autonomous and tissue contextual changes which occur in response to cumulative molecular stress and manifest through adaptive transcriptional reprogramming. Here we describe a transcription factor (TF) meta-analysis of gene expression datasets accrued from 18 tissue sites collected at different biological ages and from 7 different in-vitro aging models. In-vitro aging platforms included replicative senescence and an energy restriction model in quiescence (ERiQ), in which ATP was transiently reduced. TF motifs in promoter regions of trimmed sets of target genes were scanned using JASPAR and TRANSFAC. TF signatures established a global mapping of agglomerating motifs with distinct clusters when ranked hierarchically. Remarkably, the ERiQ profile was shared with the majority of in-vivo aged tissues. Fitting motifs in a minimalistic protein-protein network allowed to probe for connectivity to distinct stress sensors. The DNA damage sensors ATM and ATR linked to the subnetwork associated with senescence. By contrast, the energy sensors PTEN and AMPK connected to the nodes in the ERiQ subnetwork. These data suggest that metabolic dysfunction may be linked to transcriptional patterns characteristic of many aged tissues and distinct from cumulative DNA damage associated with senescence.

Quiescence is the prevailing state of many cell types under homeostatic conditions. Yet, surprisingly little is known about how quiescent cells respond to energetic and metabolic challenges. To better understand compensatory responses of quiescent cells to metabolic stress, we established, in human primary dermal fibroblasts, an experimental ‘energy restriction’ model. Quiescence was achieved by short-term culture in serum-deprived media and ATP supply restricted using a combination of glucose transport inhibitors and mitochondrial uncouplers. In aggregate, these measures led to markedly reduced intracellular ATP levels while not compromising cell viability over the observation period of 48 h. Analysis of the transcription factor landscape induced by this treatment revealed alterations in several signal transduction nodes beyond the expected biosynthetic adaptations. These included increased abundance of NF-κB regulated transcription factors and altered transcription factor subsets regulated by Akt and p53. The observed changes in gene regulation and corresponding alterations in key signaling nodes are likely to contribute to cell survival at intracellular ATP concentrations substantially below those achieved by growth factor deprivation alone. This experimental model provides a benchmark for the investigation of cell survival pathways and related molecular targets that are associated with restricted energy supply associated with biological aging and metabolic diseases.

Late-onset Alzheimer’s disease (LOAD) and age are significantly correlated such that one-third of Americans beyond 85 years of age are afflicted. We have designed and implemented a pilot study that combines systems biology approaches with traditional next-generation sequencing (NGS) analysis techniques to identify relevant regulatory pathways, infer functional relationships and confirm the dysregulation of these biological pathways in LOAD. Our study design is a most comprehensive systems approach combining co-expression network modeling derived from RNA-seq data, rigorous quality control (QC) standards, functional ontology, and expression quantitative trait loci (eQTL) derived from whole exome (WES) single nucleotide variant (SNV) genotype data. Our initial results reveal several statistically significant, biologically relevant genes involved in sphingolipid metabolism. To validate these findings, we performed a gene set enrichment analysis (GSEA). The GSEA revealed the sphingolipid metabolism pathway and regulation of autophagy in association with LOAD cases. In the execution of this study, we have successfully tested an integrative approach to identify both novel and known LOAD drivers in order to develop a broader and more detailed picture of the highly complex transcriptional and regulatory landscape of age-related dementia.

Cellular responses to energy stress involve activation of pro-survival signaling nodes, compensation in regulatory pathways and adaptations in organelle function. Specifically, energy restriction in quiescent cells (ERiQ) through energetic perturbations causes adaptive changes in response to reduced ATP, NAD+ and NADP levels in a regulatory network spanned by AKT, NF-κB, p53 and mTOR. Based on the experimental ERiQ platform, we have constructed a minimalistic theoretical model consisting of feedback motifs that enable investigation of stress-signaling pathways. The computer simulations reveal responses to acute energetic perturbations, promoting cellular survival and recovery to homeostasis. We speculated that the very same stress mechanisms are activated during aging in post-mitotic cells. To test this hypothesis, we modified the model to be deficient in protein damage clearance and demonstrate the formation of energy stress. Contrasting the network’s pro-survival role in acute energetic challenges, conflicting responses in aging disrupt mitochondrial maintenance and contribute to a lockstep progression of decline when chronically activated. The model was analyzed by a local sensitivity analysis with respect to lifespan and makes predictions consistent with inhibitory and gain-of-function experiments in aging.

After spinal cord injury (SCI), re-establishing cellular homeostasis is critical to optimize functional recovery. Central to that response is PERK signaling, which ultimately initiates a pro-apoptotic response if cellular homeostasis cannot be restored. Oligodendrocyte (OL) loss and white matter damage drive functional consequences and determine recovery potential after thoracic contusive SCI. We examined acute (<48 h post-SCI) and chronic (6 weeks post-SCI) effects of conditionally deleting Perk from OLs prior to SCI. While Perk transcript is expressed in many types of cells in the adult spinal cord, its levels are disproportionately high in OL lineage cells. Deletion of OL-Perk prior to SCI resulted in: (1) enhanced acute phosphorylation of eIF2α, a major PERK substrate and the critical mediator of the integrated stress response (ISR), (2) enhanced acute expression of the downstream ISR genes Atf4, Ddit3/Chop, and Tnfrsf10b/Dr5, (3) reduced acute OL lineage-specific Olig2 mRNA, but not neuronal or astrocytic mRNAs, (4) chronically decreased OL content in the spared white matter at the injury epicenter, (5) impaired hindlimb locomotor recovery, and (6) reduced chronic epicenter white matter sparing. Cultured primary OL precursor cells with reduced PERK expression and activated ER stress response showed: (1) unaffected phosphorylation of eIF2α, (2) enhanced ISR gene induction, and (3) increased cytotoxicity. Therefore, OL-Perk deficiency exacerbates ISR signaling and potentiates white matter damage after SCI. The latter effect is likely mediated by increased loss of Perk -/- OLs., (© 2024 Wiley Periodicals LLC.)

Protein kinase R (PKR)-like endoplasmic reticulum kinase (PERK) is a major signal transducer of the endoplasmic reticulum stress response (ERSR) pathway. Outcomes of PERK activation range from abrogating ER stress to induction of cell death, dependent on its level, duration, and cellular context. Current data demonstrate that after mouse spinal cord injury (SCI), acute inhibition of PERK (0-72 h) with the small molecule inhibitor GSK2656157 reduced ERSR while improving white matter sparing and hindlimb locomotion recovery. GSK2656157-treated mice showed increased numbers of oligodendrocytes at the injury epicenter. Moreover, GSK2656157 protected cultured primary mouse oligodendrocyte precursor cells from ER stress-induced cytotoxicity. These findings suggest that in the context of SCI, excessive acute activation of PERK contributes to functionally relevant white matter damage. Pharmacological inhibition of PERK is a potential strategy to protect central nervous system (CNS) white matter following acute injuries, including SCI.

Traumatic spinal cord injury (SCI) is a devastating neurological condition that involves both primary and secondary tissue loss. Various cytotoxic events including hypoxia, hemorrhage and blood lysis, bioenergetic failure, oxidative stress, endoplasmic reticulum (ER) stress, and neuroinflammation contribute to secondary injury. The HIF prolyl hydroxylase domain (PHD/EGLN) family of proteins are iron-dependent, oxygen-sensing enzymes that regulate the stability of hypoxia inducible factor-1α (HIF-1α) and also mediate oxidative stress caused by free iron liberated from the lysis of blood. PHD inhibition improves outcome after experimental intracerebral hemorrhage (ICH) by reducing activating transcription factor 4 (ATF4)-driven neuronal death. As the ATF4-CHOP (CCAAT-enhancer-binding protein homologous protein) pathway plays a role in the pathogenesis of contusive SCI, we examined the effects of PHD inhibition in a mouse model of moderate T9 contusive SCI in which white matter damage is the primary driver of locomotor dysfunction. Pharmacological inhibition of PHDs using adaptaquin (AQ) moderately lowers acute induction of Atf4 and Chop mRNAs and prevents the acute decline of oligodendrocyte (OL) lineage mRNAs, but does not improve long-term recovery of hindlimb locomotion or increase chronic white matter sparing. Conditional genetic ablation of all three PHD isoenzymes in OLs did not affect Atf4, Chop or OL mRNAs expression levels, locomotor recovery, and white matter sparing after SCI. Hence, PHDs may not be suitable targets to improve outcomes in traumatic CNS pathologies that involve acute white matter injury., Competing Interests: The authors have declared that no competing interests exist.

One of the difficulties in identifying novel therapeutic strategies to manage central nervous system (CNS) trauma is the need for behavioral assays to assess chronic functional recovery. In vitro assays and/or acute behavioral assessments cannot accurately predict long-term functional outcome. Using data from 13 independent T9 moderate contusive spinal cord injury (SCI) studies, we asked whether the ratio of acute (24-72 h post-injury) changes in the levels of neuron-, oligodendrocyte-, astrocyte-specific and/or endoplasmic reticulum stress response (ERSR) messenger ribonucleic acids (mRNAs) could predict the extent of chronic functional recovery. Increased levels of neuron, oligodendrocyte, and astrocyte mRNAs all correlated with enhanced Basso Mouse Scale (BMS) scores. Reduced levels of the ERSR mRNAs Atf4 and Chop correlate with improved chronic locomotor function. Neither neural or ERSR mRNAs were predictive for chronic recovery across all behavioral changes. The ratio of oligodendrocyte/ERSR mRNAs, however, did predict "improved," "no change," or "worse" functional recovery. Neuronal/ERSR mRNA ratios predicted functional improvement, but could not distinguish between worse or no change outcomes. Astrocyte/ERSR mRNA ratios were not predictive. This approach can be used to confirm biological action of injected drugs in vivo and to optimize dose and therapeutic window. It may prove useful in cervical and lumbar SCI and in other traumatic CNS injuries such as traumatic brain injury and stroke, where prevention of neuronal loss is paramount to functional recovery. Although the current analysis was directed toward ERSR whose activity was targeted in all but one study, acute mRNA markers for other pathophysiological cascades may be as predictive of chronic recovery when those cascades are targeted for neuroprotection.

The endoplasmic reticulum stress response (ERSR) is activated in various neurodegenerative diseases and/or after CNS traumatic injuries. The ERSR is comprised of three major arms, PERK, IRE-1, and activating transcription factor-6, with the latter two contributing to the unfolded protein response (UPR). PERK activity overlaps with the integrated stress response (ISR) kinases, PKR, HRI, and GCN2 which all signal through, eukaryotic initiation factor 2α, ATF4, and CHOP. All initially attempt to restore endoplasmic reticulum (ER) homeostasis, but if ER stress is unresolved, ATF4/CHOP-mediated cell death is initiated. Here, we investigate the contribution of the inositol-requiring protein-1α-X-box binding protein-1 (XBP1)-mediated UPR signaling pathway to the pathogenesis of spinal cord injury (SCI). We demonstrate that deletion of Xbp1 caused an exacerbated ATF4/CHOP signaling in cultured mouse oligodendrocyte (OL) progenitor cells and enhanced their sensitivity to ER stress. Similar effects were also observed with the Xbp1 pathway inhibitor toyocamycin. Furthermore, OL lineage-specific loss of Xbp1 resulted in enhanced ISR in mice that underwent moderate contusive SCI at the T9 level. Consistently, post-injury recovery of hindlimb locomotion and white matter sparing were reduced in OL Xbp1-deficient mice, which correlated with chronically decreased relative density of OPCs and OLs at the injury epicenter at 6 weeks post-SCI. We conclude that the IRE1-XBP1-mediated UPR signaling pathway contributes to restoration of ER homeostasis in OLs and is necessary for enhanced white matter sparing and functional recovery post-SCI., (© 2020 Wiley Periodicals LLC.)

The transcription factor BMAL1/ARNTL is a non-redundant component of the clock pathway that regulates circadian oscillations of gene expression. Loss of BMAL1 perturbs organismal homeostasis and usually exacerbates pathological responses to many types of insults by enhancing oxidative stress and inflammation. Surprisingly, we observed improved locomotor recovery and spinal cord white matter sparing in Bmal1 -/- mice after T9 contusive spinal cord injury (SCI). While acute loss of neurons and oligodendrocytes was unaffected, Bmal1 deficiency reduced the chronic loss of oligodendrocytes at the injury epicenter 6 weeks post SCI. At 3 days post-injury (dpi), decreased expression of genes associated with cell proliferation, neuroinflammation and disruption of the blood spinal cord barrier (BSCB) was also observed. Moreover, intraspinal extravasation of fibrinogen and immunoglobulins was decreased acutely at dpi 1 and subacutely at dpi 7. Subacute decrease of hemoglobin deposition was also observed. Finally, subacutely reduced levels of the leukocyte marker CD45 and even greater reduction of the pro-inflammatory macrophage receptor CD36 suggest not only lower numbers of those cells but also their reduced inflammatory potential. These data indicate that Bmal1 deficiency improves SCI outcome, in part by reducing BSCB disruption and hemorrhage decreasing cytotoxic neuroinflammation and attenuating the chronic loss of oligodendrocytes.

Deficient myelination, the spiral wrapping of highly specialized membrane around axons, causes severe neurological disorders. Maturation of oligodendrocyte progenitor cells (OPC) to myelinating oligodendrocytes (OL), the sole providers of central nervous system (CNS) myelin, is tightly regulated and involves extensive morphological changes. Here, we present evidence that autophagy, the targeted isolation of cytoplasm and organelles by the double-membrane autophagosome for lysosomal degradation, is essential for OPC/OL differentiation, survival, and proper myelin development. A marked increase in autophagic activity coincides with OL differentiation, with OL processes having the greatest increase in autophagic flux. Multiple lines of evidence indicate that autophagosomes form in developing myelin sheathes before trafficking from myelin to the OL soma. Mice with conditional OPC/OL-specific deletion of the essential autophagy gene Atg5 beginning on postnatal Day 5 develop a rapid tremor and die around postnatal Day 12. Further analysis revealed apoptotic death of OPCs, reduced differentiation, and reduced myelination. Surviving Atg5 -/- OLs failed to produce proper myelin structure. In vitro, pharmacological inhibition of autophagy in OPC/dorsal root ganglion (DRG) co-cultures blocked myelination, producing OLs surrounded by many short processes. Conversely, autophagy stimulation enhanced myelination. These results implicate autophagy as a key regulator of OPC survival, maturation, and proper myelination. Autophagy may provide an attractive target to promote both OL survival and subsequent myelin repair after injury., (© 2019 Wiley Periodicals, Inc.)

Emerging evidence links changes in the gut microbiome and intestinal barrier function to alterations in CNS function. We examined the role of endotoxin-responsive, cAMP-specific, Pde4 subfamily b (Pde4b) enzyme in gut dysbiosis induced neuro-inflammation and white matter loss following spinal cord injury (SCI). Using a thoracic contusion model in C57Bl/6 wild type female mice, SCI led to significant shifts in the gut bacterial community including an increase in the phylum Proteobacteria, which consists of endotoxin-harboring, gram-negative bacteria. This was accompanied by increased systemic inflammatory marker, soluble CD14, along with markers of the endoplasmic reticulum stress response (ERSR) and inflammation in the SCI epicenter. Deletion of Pde4b reduced epicenter expression of markers for the ERSR and inflammation, at both acute and chronic time points post-SCI. Correspondingly, expression of oligodendrocyte mRNAs increased. Within the injury penumbra, inflammatory protein markers of activated astrocytes (GFAP), macrophage/microglia (CD11b, Iba1), and the proinflammatory mediator Cox2, were decreased in Pde4b -/- mice. The absence of Pde4b improved white matter sparing and recovery of hindlimb locomotion following injury. Importantly, SCI-induced gut dysbiosis, bacterial overgrowth and endotoxemia were also prevented in Pde4b -/- mice. Taken together, these findings indicate that PDE4B plays an important role in the development of acute and chronic inflammatory response and consequent recovery following SCI., (Copyright © 2018. Published by Elsevier Inc.)

Autophagy mechanisms are well documented in neurons after spinal cord injury (SCI), but the direct functional role of autophagy in oligodendrocyte (OL) survival in SCI pathogenesis remains unknown. Autophagy is an evolutionary conserved lysosomal-mediated catabolic pathway that ensures degradation of dysfunctional cellular components to maintain homeostasis in response to various forms of stress, including nutrient deprivation, hypoxia, reactive oxygen species, DNA damage, and endoplasmic reticulum (ER) stress. Using pharmacological gain and loss of function and genetic approaches, we investigated the contribution of autophagy in OL survival and its role in the pathogenesis of thoracic contusive SCI in female mice. Although upregulation of Atg5 (an essential autophagy gene) occurs after SCI, autophagy flux is impaired. Purified myelin fractions of contused 8 d post-SCI samples show enriched protein levels of LC3B, ATG5, and BECLIN 1. Data show that, while the nonspecific drugs rapamycin (activates autophagy) and spautin 1 (blocks autophagy) were pharmacologically active on autophagy in vivo , their administration did not alter locomotor recovery after SCI. To directly analyze the role of autophagy, transgenic mice with conditional deletion of Atg5 in OLs were generated. Analysis of hindlimb locomotion demonstrated a significant reduction in locomotor recovery after SCI that correlated with a greater loss in spared white matter. Immunohistochemical analysis demonstrated that deletion of Atg5 from OLs resulted in decreased autophagic flux and was detrimental to OL function after SCI. Thus, our study provides evidence that autophagy is an essential cytoprotective pathway operating in OLs and is required for hindlimb locomotor recovery after thoracic SCI. SIGNIFICANCE STATEMENT This study describes the role of autophagy in oligodendrocyte (OL) survival and pathogenesis after thoracic spinal cord injury (SCI). Modulation of autophagy with available nonselective drugs after thoracic SCI does not affect locomotor recovery despite being pharmacologically active in vivo , indicating significant off-target effects. Using transgenic mice with conditional deletion of Atg5 in OLs, this study definitively identifies autophagy as an essential homeostatic pathway that operates in OLs and exhibits a direct functional role in SCI pathogenesis and recovery. Therefore, this study emphasizes the need to discover novel autophagy-specific drugs that specifically modulate autophagy for further investigation for clinical translation to treat SCI and other CNS pathologies related to OL survival., (Copyright © 2018 the authors 0270-6474/18/385900-13$15.00/0.)

After unresolved endoplasmic reticulum stress, recovery of protein synthesis including increased expression of ribosomal components and translation factors may induce cell death. Using a mouse model of moderate contusive spinal cord injury (SCI) at the T9 level, upregulation of ribosomal biogenesis was observed in the injury epicenter at 24h after trauma. Such upregulation coincided with endoplasmic reticulum stress response as previously reported in this model. It was also accompanied by changes in expression of many other genes associated with translational regulation. Systemic treatment with a pharmacological inhibitor of RNA-Polymerase-1, BMH-21 reduced rRNA transcription in the spinal cord. Moreover, in the injury epicenter, treatment with BMH-21 increased expression of oligodendrocyte-specific transcripts including Mbp and Cldn11 at 3days post injury. Although such findings may suggest at least transient reduction of oligodendrocyte death, locomotor outcome was mostly unaffected except slightly accelerated recovery of hindlimb function at week 2 post-injury. Therefore, at least in mice, RNA-Polymerase-1 does not appear to be a robust target for therapies to protect spinal cord tissue after contusion. However, these findings raise an interesting possibility that altered rate of ribosomal biogenesis contributes to the apparent translational reprogramming after contusive SCI. Such a reprogramming could be a major regulator of SCI-induced gene expression., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Motivation exists to develop tobacco cultivars with reduced nicotine content for the purpose of facilitating compliance with expected tobacco product regulations that could mandate the lowering of nicotine levels per se, or the reduction of carcinogenic alkaloid-derived tobacco specific nitrosamines (TSNAs). A berberine bridge enzyme-like (BBL) gene family was recently characterized for N. tabacum and found to catalyze one of the final steps in pyridine alkaloid synthesis for this species. Because this gene family acts downstream in the nicotine biosynthetic pathway, it may represent an attractive target for genetic strategies with the objective of reducing alkaloid content in field-grown tobacco. In this research, we produced transgenic doubled haploid lines of tobacco cultivar K326 carrying an RNAi construct designed to reduce expression of the BBL gene family. Field-grown transgenic lines carrying functional RNAi constructs exhibited average cured leaf nicotine levels of 0.684%, in comparison to 2.454% for the untransformed control. Since numerous barriers would need to be overcome to commercialize transgenic tobacco cultivars, we subsequently pursued a mutation breeding approach to identify EMS-induced mutations in the three most highly expressed isoforms of the BBL gene family. Field evaluation of individuals possessing different homozygous combinations of truncation mutations in BBLa, BBLb, and BBLc indicated that a range of alkaloid phenotypes could be produced, with the triple homozygous knockout genotype exhibiting greater than a 13-fold reduction in percent total alkaloids. The novel source of genetic variability described here may be useful in future tobacco breeding for varied alkaloid levels.