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Type 2 Diabetes Mellitus (T2DM) is linked to multiple complications, including cognitive impairment, and the prevalence of memory-related neurodegenerative diseases is higher in T2DM patients. One possible theory is the alteration of the microvascular and macrovascular environment of the blood–brain barrier (BBB). In this study, we employed different approaches, including RT-PCR, functional pharmacokinetic studies using sodium fluorescein (NaFL), and confocal microscopy, to characterize the functional and molecular integrity of the BBB in a T2DM animal model, leptin receptor-deficient mutant mice (Leprdb/db mice). As a result, VCAM-1, ICAM-1, MMP-9, and S100b (BBB-related markers) dysregulation was observed in the Leprdb/db animal model compared to littermate wild-type mice. The brain concentration of sodium fluorescein (NaFL) increased significantly in Leprdb/db untreated mice compared to insulin-treated mice. Therefore, the permeability of NaFL was higher in Leprdb/db control mice than in all remaining groups. Identifying the factors that increase the BBB in Leprdb/db mice will provide a better understanding of the BBB microvasculature and present previously undescribed findings of T2DM-related brain illnesses, filling knowledge gaps in this emerging field of research. [ABSTRACT FROM AUTHOR]

Pathological cardiac remodeling is associated with cardiovascular disease and can lead to heart failure. Nuclear factor-kappa B (NF-κB) is upregulated in the hypertrophic heart. Moreover, the expression of the G-protein-coupled receptor kinase 2 (GRK2) is increased and linked to the progression of heart failure. The inhibitory effects of paroxetine on GRK2 have been established. However, its protective effect on IκBα/NFκB signaling has not been elucidated. This study investigated the cardioprotective effect of paroxetine in an animal model of cardiac hypertrophy (CH), focusing on its effect on GRK2-mediated NF-κB-regulated expression of prohypertrophic and profibrotic genes. Wistar albino rats were administered normal saline, paroxetine, or fluoxetine, followed by isoproterenol to induce CH. The cardioprotective effects of the treatments were determined by assessing cardiac injury, inflammatory biomarker levels, histopathological changes, and hypertrophic and fibrotic genes in cardiomyocytes. Paroxetine pre-treatment significantly decreased the HW/BW ratio (p < 0.001), and the expression of prohypertrophic and profibrotic genes Troponin-I (p < 0.001), BNP (p < 0.01), ANP (p < 0.001), hydroxyproline (p < 0.05), TGF-β1 (p < 0.05), and αSMA (p < 0.01) as well as inflammatory markers. It also markedly decreased pIκBα, NFκB(p105) subunit expression (p < 0.05) and phosphorylation. The findings suggest that paroxetine prevents pathological cardiac remodeling by inhibiting the GRK2-mediated IκBα/NF-κB signaling pathway. [ABSTRACT FROM AUTHOR]

Introduction and aims Junctional epidermolysis bullosa (JEB) is a devastating hereditary skin disorder characterized by severe fragility of skin and mucus membrane, emerges as blistering with little or no trauma. The major classification of JEB includes JEB intermediate and JEB severe; the latter is the most common subtype, which is the result of loss of function mutations in the skin basement membrane protein laminin-332. Lacking this protein impairs basement membrane adhesion of epidermal keratinocytes and leads to deregulated epidermal–dermal crosstalk. The skin dermis is composed of multiple fibroblast subpopulations with different functions in skin homeostasis and repair; however, their specific involvement in skin diseases is largely unknown. Here, we investigate how fibroblast subpopulations, namely papillary and reticular fibroblasts, impact JEB disease progression and skin regeneration. Methods Three-dimensional (3D) organotypic (OT) combining healthy (control) and diseased (JEB) keratinocytes cells with distinct fibroblast subpopulations from different anatomical sites were developed and analysed by immunohistochemistry and RNA-Seq. Results JEB OTs with papillary fibroblasts showed increased epidermal growth, invagination of basal layer and deregulated differentiation compared with reticular fibroblasts. RNA-Seq analysis revealed altered activity in key signalling including immune, cytokine, gap junction, mitogen-activated protein kinase and remodelling of keratinization and ECM proteins, particularly in JEB OT cultured with papillary fibroblast. JEB OTs and cells also demonstrated variation in cell proliferation and activation of transforming growth factor-β1 and immune cell signalling. Currently, we are targeting identified signalling pathways in our two-dimensional and 3D cell culture models and identifying druggable targets, before testing selected candidates in our JEB mouse disease model for further preclinical validation. Conclusions By dissecting the pathological signalling interactions between keratinocytes and distinct fibroblast subpopulations, our goal is to identify new therapeutic target to improve the skin health and regeneration in patients with JEB. [ABSTRACT FROM AUTHOR]

Research studies have indicated that the comorbidity burden of mood disorders and obesity is reasonably high. Insulin signaling has been shown to modulate multiple physiological functions in the brain, indicating its association with neuropsychiatric diseases, including mood disorders. Leptin is a hormone responsible for regulating body weight and insulin homeostasis. Previous studies on db/db mice (a mouse model that carries a spontaneous genetic mutation in leptin receptor Leprdb) have shown that they exhibit inflammation as well as neurobehavioral traits associated with mood. Therefore, targeting inflammatory pathways such as TNF-α may be an effective strategy in the treatment of obesity-linked mood disorders. The objective of this study was to investigate the effect of long-term administration of etanercept (a TNF-α blocker) on anxiety and depressive-like behaviors in db/db mice. This was performed using light/dark box, forced swim, and open field tests with lean littermate wild type (WT) mice serving as a control group. Using flow cytometry in peripheral blood, we further examined the molecular effects of etanercept on NF-κB p65, TNF-α, IL-17A, and TLR-4 expressing CD4+, CD8+, and CD14+ cells in the peripheral blood. Our data show that peripheral administration of etanercept decreased these cells in db/db mice. Furthermore, our results indicated that peripheral administration of etanercept reduced anxiety and depressive-like behaviors. Therefore, targeting TNF-α signaling might be an effective strategy for modulating obesity-associated depression and anxiety. [ABSTRACT FROM AUTHOR]

Intra‐synovial tendon injuries are a common orthopedic problem with limited treatment options. The synovium is a specialized connective tissue forming the inner encapsulating lining of diarthrodial joints and intra‐synovial tendons. It contains multipotent mesenchymal stromal cells that render it a viable source of progenitors for tendon repair. This study evaluated the effects of autologous implantation of cells derived from normal synovium (synovial membrane cells [SMCs]) in augmenting repair in an ovine model of intra‐synovial tendon injury. For this purpose, synovial biopsies were taken from the right digital flexor tendon sheath following creation of a defect to the lateral deep digital flexor tendon. Mononuclear cells were isolated by partial enzymatic digestion and assessed for MSC characteristics. Cell tracking and tendon repair were assessed by implanting 5 × 106 cells into the digital flexor tendon sheath under ultrasound guidance with the effects evaluated using magnetic resonance imaging and histopathology. Synovial biopsies yielded an average 4.0 × 105 ± 2.7 × 105 SMCs that exhibited a fibroblastic morphology, variable osteogenic, and adipogenic responses but were ubiquitously strongly chondrogenic. SMCs displayed high expression of CD29 with CD271NEGATIVE and MHC‐IILOW cell‐surface marker profiles, and variable expression of CD73, CD90, CD105, CD166, and MHC‐I. Implanted SMCs demonstrated engraftment within the synovium, though a lack of repair of the tendon lesion over 24 weeks was observed. We conclude healthy synovium is a viable source of multipotent cells, but that the heterogeneity of synovium underlies the variability between different SMC populations, which while capable of engraftment and persistence within the synovium exhibit limited capacity of influencing tendon repair. © 2019 The Authors. Journal of Orthopaedic Research® published by Wiley Periodicals, Inc. on behalf of Orthopaedic Research Society J Orthop Res 38:128–138, 2020 [ABSTRACT FROM AUTHOR]

The axon initial segment (AIS), the site of action potential initiation in neurons, is a critical determinant of neuronal excitability. Growing evidence indicates that appropriate recruitment of the AIS macrocomplex is essential for synchronized firing. However, disruption of the AIS structure is linked to the etiology of multiple disorders, including autism spectrum disorder (ASD), a condition characterized by deficits in social communication, stereotyped behaviors, and very limited interests. To date, a complete understanding of the molecular components that underlie the AIS in ASD has remained elusive. In this research, we examined the AIS structure in a BTBR T+Itpr3tf/J mouse model (BTBR), a valid model that exhibits behavioral, electrical, and molecular features of autism, and compared this to the C57BL/6J wild-type control mouse. Using Western blot studies and high-resolution confocal microscopy in the prefrontal frontal cortex (PFC), our data indicate disrupted expression of different isoforms of the voltage-gated sodium channels (NaV) at the AIS, whereas other components of AIS such as ankyrin-G and fibroblast growth factor 14 (FGF14) and contactin-associated protein 1 (Caspr) in BTBR were comparable to those in wild-type control mice. A Western blot assay showed that BTBR mice exhibited a marked increase in different sodium channel isoforms in the PFC compared to wild-type mice. Our results provide potential evidence for previously undescribed mechanisms that may play a role in the pathogenesis of autistic-like phenotypes in BTBR mice. [ABSTRACT FROM AUTHOR]

The availability of intracellular, stabilized β-catenin, a transcription factor coactivator, is tightly regulated; β-catenin is translocated into the nucleus in response to Wnt ligand binding to its cell membrane receptors. Here we show that Wnt signal activation in mammalian cells activates intracellular mobilization of connexin 43 (Cx43), which belongs to a gap junction protein family, a new target protein in response to extracellular Wnt signal activation. Transmission electron microscopy showed that the nuclear localization of Cx43 was increased by 8- to 10-fold in Wnt5A- and 9B-treated cells compared with controls; this Wnt-induced increase was negated in the cells where Cx43 and β-catenin were knocked down using shRNA. There was a significant (P < 0.001) and concomitant depletion of the cell membrane and cytosolic signal of Cx43 in Wnt-treated cells with an increase in the nuclear signal for Cx43; this was more obvious in cells where β-catenin was knocked down using shRNA. Conversely, Cx43 knockdown resulted in increased β-catenin in the nucleus in the absence of Wnt activation. Coimmunoprecipitation of Cx43 and β-catenin proteins with a casein kinase (CKIδ) antibody showed that Cx43 interacts with β-catenin and may form part of the so-called destruction complex. Functionally, Wnt activation increased the rate of wound reepithelization in rat skin in vivo. [ABSTRACT FROM AUTHOR]

Titanium (Ti) is widely used as a biomaterial for endosseous implants due to its relatively inert surface oxide layer that enables implanted devices the ability of assembling tissue reparative components that culminate in osseointegration. Topographic modifications in the form of micro- and nanoscaled structures significantly promote osseointegration and enhance the osteogenic differentiation of adult mesenchymal stromal cells (MSCs). While the biological mechanisms central to the differential responses of tissues and cells to Ti surface modifications remain unknown, adhesion and morphological adaptation are amongst the earliest events at the cell-biomaterial interface that are highly influenced by surface topography and profoundly impact the regulation of stem cell fate determination. This study correlated the effects of Ti topographic modifications on adhesion and morphological adaptation of human MSCs with phenotypic change. The results showed that modified Ti topographies precluded the adhesion of a subset of MSCs while incurring distinct morphological constraints on adherent cells. These effects anomalously corresponded with a differential expression of stem cell pluripotency and Wnt signalling-associated markers on both modified surfaces while additionally differing between hydrophobic and hydrophilic surface modifications—though extent of osteogenic differentiation induced by both modified topographies yielded similarly significant higher levels of cellular mineralisation in contrast to polished Ti. These results suggest that in the absence of deposited proteins and soluble factors, both modified topographies incur the selective adhesion of a subpopulation of progenitors with relatively higher cytoskeletal plasticity. While the presence of deposited proteins and soluble factors does not significantly affect adherence of cells, nanotopographic modifications enhance expression of pluripotency markers in proliferative conditions, which are conversely overridden by both modified topographies in osteogenic inductive conditions. Further deciphering the mechanisms underlying cellular selectivity and Ti topographic responsiveness will improve our understanding of stem cell heterogeneity and advance the potential of MSCs in regenerative medicine. [ABSTRACT FROM AUTHOR]

The clinical potential of multipotent mesenchymal stem cells (MSCs) has led to the essential development of analytical tools such as antibodies against membrane-bound proteins for the immunophenotypic characterization of human and rodent cells. Such tools are frequently lacking for emerging large animal models like the sheep that have greater relevance for the study of human musculoskeletal diseases. The present study identified a set of commercial nonspecies specific monoclonal antibodies for the immunophenotypic characterization of ovine MSCs. A protocol combining the less destructive proteolytic activity of accutase and EDTA was initially developed for the detachment of cells from plastic with minimum loss of cell surface antigens. A range of commercially available antibodies against human or rodent MSC antigens were then tested in single and multistain-based assays for their cross-reactivity to bone marrow derived ovine MSCs. Antibody clones cross-reactive to ovine CD73 (96.9% ± 5.9), CD90 (99.6% ± 0.3), CD105 (99.1 ± 1.5), CD271 (97.7 ± 2.0), and MHC1 (94.0% ± 7.2) antigens were identified using previously reported CD29, CD44, and CD166 as positive controls. Multistaining analysis indicated the colocalization of these antigens on MSCs. Furthermore, antibody clones identified to cross-react against white blood cell antigens exhibited either negative (CD117 (0.1% ± 0.1)) or low (MHCII (10.5% ± 16.0); CD31 (14.6% ± 4.2), and CD45 (39.4% ± 31.8)) cross-reactivity with ovine MSCs. The validation of these antibody clones to sheep MSC antigens is essential for studies utilizing this large animal model for stem cell-based therapies. © 2016 International Society for Advancement of Cytometry [ABSTRACT FROM AUTHOR]

This paper describes the mechanistic details of an electrochemical method to control the withdrawal of a liquid metal alloy, eutectic gallium indium (EGaIn), from microfluidic channels. EGaIn is one of several alloys of gallium that are liquid at room temperature and form a thin (nm scale) surface oxide that stabilizes the shape of the metal in microchannels. Applying a reductive potential to the metal removes the oxide in the presence of electrolyte and induces capillary behavior; we call this behavior 'recapillarity' because of the importance of electrochemical reduction to the process. Recapillarity can repeatably toggle on and off capillary behavior by applying voltage, which is useful for controlling the withdrawal of metal from microchannels. This paper explores the mechanism of withdrawal and identifies the applied current as the key factor dictating the withdrawal velocity. Experimental observations suggest that this current may be necessary to reduce the oxide on the leading interface of the metal as well as the oxide sandwiched between the wall of the microchannel and the bulk liquid metal. The ability to control the shape and position of a metal using an applied voltage may prove useful for shape reconfigurable electronics, optics, transient circuits, and microfluidic components. [ABSTRACT FROM AUTHOR]

Contamination of water due to bromate is a severe health hazard. The aim of the present study was to remove bromate from water using a crosslinked polystyrene based strongly basic anion exchange resin De-Acidite FF-IP. Batch experiments were performed to study the influence of various experimental parameters such as effect of pH, contact time, temperature, and effect of competing anions on bromate removal by De-Acidite FF-IP resin. At optimum parameters, the removal rate of bromate was very fast and 90% removal took place in 5 min and equilibrium was established within 10 min. The presence of competitive anions reduced the bromate adsorption in the order of Cl− > F− > CO [ABSTRACT FROM AUTHOR]

Aflatoxins AFB1, AFB2, AFG1, and AFG2 are toxic secondary metabolites produced by Aspergillus flavus and Aspergillus parasiticus and posses a potential threat to food safety. In the present work, liquid-liquid extraction and ultraperformance LC- MS/ MS method has been applied for the determination of four naturally occurring aflatoxins AFB1, AFB2, AFG1, and AFG2 in nonalcoholic beer. Aflatoxins extraction from nonalcoholic beer was carried out using liquid-liquid extraction procedure. The effects of solvent-types were studied to obtain maximum recovery of the target analytes with minimum contamination. Among different solvents, the aflatoxins extraction was best achieved using ethyl acetate. The obtained recoveries were ranged from 85 to 96% with good quality parameters: LOD values between 0.001 and 0.003 ng/mL, linearity of the calibration curve ( r2 > 0.999), and repeatability (run-to-run) and reproducibility (day-to-day) precisions with RSDs lower than 5% ( n = 5) achieved at 0.50 ng/mL concentration. The optimized liquid-liquid extraction in combination with ultraperformance LC- MS/ MS was applied successfully to the analysis of AFB1, AFB2, AFG1, and AFG2 aflatoxins in 11 nonalcoholic beers and were detected up to 15.31 ng/L in some of the samples. [ABSTRACT FROM AUTHOR]

Beclin-1 has a central role in the regulation of autophagy. Barrett's esophagus (BE) is associated with a significantly increased risk for the development of esophageal adenocarcinoma (EAC). In the current study, we evaluated the role of Beclin-1 and autophagy in the EAC. Biopsies obtained from patients with BE and EAC, tissues from a rat model of BE and EAC, and esophageal cell lines were evaluated for the expression of Beclin-1 by immunohistochemistry, immunoblotting, or RT-PCR. Since reflux of bile acids is important in EAC, we also evaluated the effect of exposure to deoxycholic acid (DCA) on autophagy and Beclin-1 expression. Beclin-1 expression was high in squamous epithelium and nondysplastic BE, whereas its expression was low in dysplastic BE and EAC. The same pattern of expression was observed in rat tissues and in esophageal cell lines. Normal esophageal epithelium and HET-1A cells (derived from normal squamous epithelium) show high levels of Beclin-1, but lower levels of Beclin-1 were found in BE and EAC cell lines (CP-A, CP-C, and OE33). Acute exposure to DCA led to increased Beclin-1 expression and increased autophagy as evaluated by electron microscopy and counting percentage of GFP-LC3- positive BE cells with punctate pattern. In contrast, chronic exposure to DCA did not result in the alteration of Beclin-1 levels or autophagy. In summary, these data suggest that autophagy is initially activated in response to bile acids, but chronic exposure to bile acids leads to decreased Beclin-1 expression and autophagy resistance. [ABSTRACT FROM AUTHOR]

Apoptosis resistance is a hallmark of cancer cells. Typically, bile acids induce apoptosis. However during gastrointestinal (GI) tumorigenesis the cancer cells develop resistance to bile acid-induced cell death. To understand how bile acids induce apoptosis resistance we first need to identify the molecular pathways that initiate apoptosis in response to bile acid exposure. In this study we examined the mechanism of deoxycholic acid (DCA)-induced apoptosis, specifically the role of Na+/H+ exchanger (NHE) and Na+ influx in esophageal cells. In vitro studies revealed that the exposure of esophageal cells (JH-EsoAd1, CP-A) to DCA (0.2 mM -0.5 mM) caused lysosomal membrane perturbation and transient cytoplasmic acidification. Fluorescence microscopy in conjunction with atomic absorption spectrophotometry demonstrated that this effect on lysosomes correlated with influx of Na+, subsequent loss of intracellular K+, an increase of Ca2+ and apoptosis. However, ethylisopropyl-amiloride (EIPA), a selective inhibitor of NHE, prevented Na+, K+ and Ca2+ changes and caspase 3/7 activation induced by DCA. Ouabain and amphotericin B, two drugs that increase intracellular Na+ levels, induced similar changes as DCA (ion imbalance, caspase3/7 activation). On the contrary, DCA-induced cell death was inhibited by medium with low a Na+ concentrations. In the same experiments, we exposed rat ileum ex-vivo to DCA with or without EIPA. Severe tissue damage and caspase-3 activation was observed after DCA treatment, but EIPA almost fully prevented this response. In summary, NHE-mediated Na+ influx is a critical step leading to DCA-induced apoptosis. Cells tolerate acidification but evade DCA-induced apoptosis if NHE is inhibited. Our data suggests that suppression of NHE by endogenous or exogenous inhibitors may lead to apoptosis resistance during GI tumorigenesis. [ABSTRACT FROM AUTHOR]

Liquid metals composed of gallium stay within networks of microfluidic channels due to the presence of a thin surface oxide that forms spontaneously on the surface of the metal. On page 671 M. R. Khan, C. Trlica, and M. D. Dickey use “recapillarity” to reduce the surface oxide and thereby turn on capillary behavior locally in a manner that withdraws the metal from the channels and solves the maze. [ABSTRACT FROM AUTHOR]

Abstract Autism spectrum disorder (ASD) is a neurodevelopmental disorder whose symptoms include communication deficits, a lack of social skills, and stereotyped repetitive behaviors. We used BTBR T+ Itpr3tf/J (BTBR) mice, a model that demonstrates most of the core behavioral features of ASD, such as decreased sociability and high levels of repetitive behaviors. Currently, there is no treatment available that is able to improve most of the ASD disorder symptoms; thus, finding novel therapies is immediately required. Stat3 inhibitors are potential targets in the treatment of several immune disorders. The aim of the present study was to investigate the effects of S3I-201, a selective Stat3 inhibitor, to determine its potential mechanism in BTBR mice. In this study, we first examined the effects of S3I-201 on repetitive behavior and marble burying. We also examined the treatment of S3I-201 on Th1 (IFN-γ and T-bet), Th17 (IL-17A, RORγt, Stat3, IL-21, and IL-22), and T regulatory (Treg, Foxp3 and Helios) production in spleen CD4+ T cells. We further assessed Th1, Th17, and Treg mRNA and protein expression levels in brain tissues. S3I-201 treatment in BTBR mice significantly prevents marble burying and repetitive behavior. Furthermore, S3I-201 administration causes a considerable decrease in IFN-γ, T-bet, IL-17A, RORγt, Stat3, IL-21, and IL-22 levels, and increases in Foxp3 and Helios production CD4+ T cells in BTBR mice. Additionally, S3I-201 treatment also significantly decreases Th1 and Th17 levels, and increases Treg mRNA and protein expression levels. Therefore, these results suggest that S3I-201 could be considered as a therapeutic option for ASD. Highlights • S3I-201 prevented repetitive behavior in a BTBR animal model of autism • S3I-201 treatment in BTBR mice increased the number of T regulatory cells • S3I-201 attenuated neuroimmune dysfunction by decreasing proinflammatory cytokine levels • S3I-201 downregulated IL-17A/RORγt signaling in BTBR mice • S3I-201 could be used as a therapeutic treatment for autism [ABSTRACT FROM AUTHOR]

Pectin based quaternary amino anion exchanger (Pc-QAE) was prepared using simple crosslinking polymerization method. This anion exchanger was characterized by X-ray diffraction (XRD), scanning electron microscopy (SEM) and Fourier transform infrared spectroscopy (FTIR). Pc-QAE was applied for the removal of phosphate anion from the aqueous solution. The adsorption process which was pH dependent showed maximum adsorption of phosphate anions at pH 7. Pc-QAE showed good monolayer adsorption capacity for phosphate anions which demonstrated its good capability towards Langmuir isotherm model. Moreover, the adsorption was evaluated thermodynamically and the negative value of Gibbs free energy (−1.791 KJ/mol) revealed the spontaneity of adsorption process. The value of ΔH° and ΔS° were found to be 15.28 and 49.48 KJ/mol, respectively representing the endothermic nature and enhancement in degree of freedom due to the adsorption process. [ABSTRACT FROM AUTHOR]

In this research work we report Gum-ghatti supported ZnSe-WO 3 nano-hetero-assembly for solar powered degradation of endocrine disruptor Bisphenol S (BPA). The photocatalyst was characterized by Scanning electron microscopy (SEM), High resolution transmission electron microscopy (HRTEM), Small area electron diffraction (SAED), X-Ray diffraction (XRD), Fourier transform infra red spectroscopy (FTIR), Photoluminescence (PL), Energy dispersive X-ray (EDX), UV–vis spectrophotometry and Brauner Emmet Teller surface area analyzer (BET). We achieve a Z-scheme photocatalyst (ZnSe-WO 3 ) with a higher charge flow and visible absorption. Gum ghatti acts as a superadsorbent and a sink for charge carriers. The removal of BPA has been studied under three experimental protocols where 99.5% removal was achieved by symbiose of photocatalysis-adsorption-ozonation in just 45 min hetero-assembly has a high surface area, stability and reduced carrier recombination. The results have been analyzed by scavenger effect, mass spectrometry, kinetics and total organic carbon (TOC) analysis. 49.4% of TOC was removed and COD was reduced to 16.7% after 2 h in symbiotic condition. From the band edges and scavenger effect it was inferred that superoxide radical anions are major attacking species. The work paves way for designing of novel photocatalysts with increasing biogenic quotient and higher efficiency. [ABSTRACT FROM AUTHOR]

Multiple sclerosis (MS) is an autoimmune disease characterized by inflammatory infiltration in association with demyelination in the central nervous system. Among the factors involved in the immunological mechanisms of MS, Th1, Th17, and Th22 cells play a critical role. In the present study, we investigated the role of CA-074, a potent Cathepsin B inhibitor, in MS progression, using the SJL/J mouse model of experimental autoimmune encephalomyelitis (EAE). Following induction of EAE, mice were administered CA-074 (10 mg/kg) intraperitoneally each day, beginning on day 14 and continuing until day 28, and were evaluated for clinical signs. We further investigated the effect of CA-074 on Th1 (T-bet/STAT4), Th17 (IL-17A/RORγT), Th22 (TNF-α/IL-22), and regulatory T (Treg/Foxp3) cells in the spleen, using flow cytometry. We also analyzed the effect of CA-074 on T-bet, IL-17A, RORγT, IL-22, and mRNA and protein levels using RT-PCR and western blot analysis for brain tissues. Cathepsin B expression were also assessed by western blot in the brain tissues. The severity of clinical scores decreased significantly in CA-074-treated mice compared with that in EAE control mice. Moreover, the percentage of CD4+T-bet+, CXCR5+T-bet+, CD4+STAT4+, CD4+IL-17A+, CXCR5+IL-17A+, CD4+RORγT+, CCR6+RORγT+, CD4+TNF-α+, CD4+IL-22+, and CCR6+IL-22+ cells decreased while CD25+Foxp3+ increased in CA-074-treated EAE mice as compared to vehicle-treated EAE mice. Further, CA-074-treated EAE mice had downregulated Cathepsin B protein expression which was associated with decreased T-bet, IL-17A, RORγT, and IL-22 mRNA/protein expression. These results suggest that Cathepsin B could be a novel therapeutic candidate against for the treatment of MS. • We investigated the possible neuroprotective effects of Cathepsin B inhibitor. • CA-074 ameliorates disease severity by relapsing-remitting in EAE. • CA-074 reduces Th1 transcription factor signaling. • CA-074 downregulated Th17/Th22 signaling in EAE mice. • CA-074 could be used as a novel candidate for MS treatment. [ABSTRACT FROM AUTHOR]

Globally, cardiovascular disease (CVD) continues to be the primary cause of morbidity and mortality. The risk of cardiovascular disease is markedly increased in individuals with type 2 diabetes mellitus (T2DM), making managing cardiovascular health a top priority. Initially developed for their glucose-lowering properties, sodium-glucose cotransporter 2 (SGLT2) inhibitors have emerged as a transformative class of pharmaceuticals with profound cardiovascular benefits that extend far beyond glycemic control. One of the most striking findings is the substantial reduction in major adverse cardiovascular events (MACE), including myocardial infarction, stroke, and cardiovascular mortality, observed in clinical trials evaluating SGLT2 inhibitors. These extraordinary cardioprotective effects are demonstrated by landmark trials such as EMPA-REG OUTCOME, CANVAS, and DECLARE-TIMI 58, which are discussed in detail. In addition, SGLT2 inhibitors have demonstrated positive outcomes in heart failure (HF) with reduced ejection fraction, which has led to their incorporation into HF treatment guidelines. SGLT2 inhibitors offer renoprotection by delaying the progression of diabetic kidney disease, reducing albuminuria, preserving glomerular filtration rates, and their immediate cardiovascular benefits. We investigate the potential mechanisms underlying these renal benefits, focusing on the role of hemodynamic alterations and intraglomerular pressure reduction. In addition, SGLT2 inhibitors have a distinct diuretic effect that can contribute to volume reduction and symptom alleviation in patients with heart failure (HF). This diuretic action, distinct from conventional diuretics, warrants additional research to optimize their use in T2DM and HF patients. The risk of euglycemic diabetic ketoacidosis, genital mycobacterial infections, and bone fractures are also discussed. Understanding these issues is essential for making educated clinical decisions. In conclusion, SGLT2 inhibitors have transcended their initial function as anti-diabetic agents to become essential components of cardiovascular and renal protection strategies in T2DM patients. Their diverse benefits, which include cardioprotection, renoprotection, and the potential for HF management, highlight their potential to transform cardiovascular medicine. Optimizing the use of SGLT2 inhibitors in clinical practice bears the promise of improved cardiovascular outcomes for patients with T2DM and beyond as we navigate this changing landscape., Competing Interests: The authors have declared that no competing interests exist., (Copyright © 2023, Fatima et al.)

Background The coronavirus disease (COVID-19) virus has caused millions of deaths. It is difficult to differentiate between pure viral COVID-19 pneumonia and secondary infection. Clinicians often use procalcitonin (PCT) to decide on empiric antibiotic therapy. Methodology We performed a retrospective study of patients admitted with COVID-19 between January 1st, 2020, and June 30th, 2020. Patient demographics, clinical findings, and laboratory findings with a focus on PCT levels were recorded. Coinfection was considered if clinicians ordered a septic workup (urine, blood, and respiratory cultures) or if the physicians started or escalated antimicrobial therapy. PCT levels on the day of culture and daily for the next three days were recorded. Significant PCT change was defined as a decrease in PCT levels of >50% from the initial elevated PCT level. Results In total, 143 (59.8%) patients had one secondary infection. These included pulmonary infections (118, 49.4%), blood infections (99, 41.4%), and urine infections (64, 26.8%). Many patients had more than one documented positive culture: respiratory system and blood together in 80 (33.4%) patients, sputum and urine in 55 (23.1%) patients, and urine and blood in 46 (19.2%) patients. Out of the 143 patients with a positive culture, PCT was abnormal on the day of positive culture in 93 (65.5%), while PCT was abnormal in 64 out of 96 on the day of negative culture (66.7%) (p = 0.89). Individual analysis for PCT levels of respiratory cultures showed out of 118 positive sputum cultures, 86 (72%) had abnormal PCT on the day of culture. PCT in positive versus negative cultures was not significantly different, with median PCT (interquartile range, IQR) of 1.66 (6.61) versus 1.03 (2.23) (p = 0.172). For blood cultures, out of 99 positive blood cultures, 73 (73%) had abnormal PCT levels on the day of the culture. PCT in positive versus negative cultures was significantly elevated, with a median of 1.61 (5.97) vs. 0.65 (1.77) (p < 0.001). For urine, out of 64 positive cultures, 41 (64.1%) had abnormal PCT levels on the day of the culture. PCT in positive versus negative cultures was not significantly different, with a median of 0.71 (2.92) vs. 0.93 (4.71) (p = 0.551). To observe the change in PCT after culture, PCT values for the next three days after culture were analyzed. We found that patients with positive cultures had higher PCT levels than those with negative cultures. There was no significant improvement over the following three days. Patients with abnormal PCT on the day of the suspected infection had a longer length of stay in the hospital, with a median (IQR) of 23.9 days (3.16) vs. 16.9 days (2.18) (p = 0.021). Conclusions Secondary coinfections in patients with COVID-19 infections are not associated with PCT elevation on the day of suspected secondary infection. However, most patients with bacteremia had a significant elevation of PCT on the day of bacteremia before collection and reporting of positive culture. Patients with abnormal PCT levels on the day of suspected infection had a longer hospital stay than patients with normal PCT levels. Subsequent testing of PCT in patients showed no significant improvement in PCT., Competing Interests: The authors have declared that no competing interests exist., (Copyright © 2022, Nadeem et al.)

The availability of intracellular, stabilized β-catenin, a transcription factor coactivator, is tightly regulated; β-catenin is translocated into the nucleus in response to Wnt ligand binding to its cell membrane receptors. Here we show that Wnt signal activation in mammalian cells activates intracellular mobilization of connexin 43 (Cx43), which belongs to a gap junction protein family, a new target protein in response to extracellular Wnt signal activation. Transmission electron microscopy showed that the nuclear localization of Cx43 was increased by 8- to 10-fold in Wnt5A- and 9B-treated cells compared with controls; this Wnt-induced increase was negated in the cells where Cx43 and β-catenin were knocked down using shRNA. There was a significant ( P < 0.001) and concomitant depletion of the cell membrane and cytosolic signal of Cx43 in Wnt-treated cells with an increase in the nuclear signal for Cx43; this was more obvious in cells where β-catenin was knocked down using shRNA. Conversely, Cx43 knockdown resulted in increased β-catenin in the nucleus in the absence of Wnt activation. Coimmunoprecipitation of Cx43 and β-catenin proteins with a casein kinase (CKIδ) antibody showed that Cx43 interacts with β-catenin and may form part of the so-called destruction complex. Functionally, Wnt activation increased the rate of wound reepithelization in rat skin in vivo.

Activated carbon was prepared from date pits via chemical activation with H3PO4. The effects of activating agent concentration and activation temperature on the yield and surface area were studied. The optimal activated carbon was prepared at 450 °C using 55 % H3PO4. The prepared activated carbon was characterized by Fourier transform infrared spectroscopy, scanning electron microscopy, thermogravimetric-differential thermal analysis, and Brunauer, Emmett, and Teller (BET) surface area. The prepared date pit-based activated carbon (DAC) was used for the removal of bromate (BrO3 (-)). The concentration of BrO3 (-) was determined by ultra-performance liquid chromatography-mass tandem spectrometry (UPLC-MS/MS). The experimental equilibrium data for BrO3 (-) adsorption onto DAC was well fitted to the Langmuir isotherm model and showed maximum monolayer adsorption capacity of 25.64 mg g(-1). The adsorption kinetics of BrO3 (-) adsorption was very well represented by the pseudo-first-order equation. The analytical application of DAC for the analysis of real water samples was studied with very promising results.

Eutectic gallium indium (EGaIn) is a promising liquid metal for a variety of electrical and optical applications that take advantage of its soft and fluid properties. The presence of a rapidly forming oxide skin on the surface of the metal causes it to stick to many surfaces, which limits the ability to easily reconfigure its shape on demand. This paper shows that water can provide an interfacial slip layer between EGaIn and other surfaces, which allows the metal to flow smoothly through capillaries and across surfaces without sticking. Rheological and surface characterization shows that the presence of water also changes the chemical composition of the oxide skin and weakens its mechanical strength, although not enough to allow the metal to flow freely in microchannels without the slip layer. The slip layer provides new opportunities to control and actuate liquid metal plugs in microchannels-including the use of continuous electrowetting-enabling new possibilities for shape reconfigurable electronics, sensors, actuators, and antennas.

Background: Radiotracers techniques are offering a unique way to diagnosis deep tissue infection in its early stages. The radiotracers including radio-antibiotics have shown promising results in the early diagnosis of infection and its discrimination from infectious foci but wide ranges of microorganisms still poses threats., Objectives: Synthesis of Technetium-99m ((99m)Tc)-temafloxacin (TMC) complex for the localization of in vivo Streptococci pneumoniae infection in the early stages., Material and Methods: The (99m)Tc-TMC complex was prepared by adding 50 µg of stannous chloride (SnCl2) with 37 MBq (0.5 mL) of sodium pertechnetate (Na(99m)TcO4-) at a pH of 5.2. Then 1 mg of the TMC was added to the mixture followed by incubation at room temperature for 10 min. The same procedure was repeated by changing the amount of the SnCl2 from 50 to 250 µg along with the TMC from 2 to 5 mg and Na(99m)TcO4- from 74 to 185 MBq. In higher concentrations of cysteine the stability of the (99m)Tc-TMC complex was evaluated. In vitro Streptococci pneumoniae uptake was investigated to validate the accuracy and preciseness of the (99m)Tc-TMC complex. In vivo uptake of the (99m)Tc-TMC complex was evaluated in ten (10) normal male Wistar rats (MWR) (140-160 g) divided into two groups (I and II)., Results: Maximum stability of 98.00 ± 0.34% at 30 min after reconstitution was observed by mixing 2.5 mg of TMC, 100 µg of SnCl2 with 74 MBq of the Na(99m)TcO4-. The stability of the complex remained 90% up to 4 hours. In serum the complex showed stability up to 16 hours. A saturated in vitro binding was noted with live Streptococci pneumoniae. In the infected region (left thigh) of the MWR, almost five times higher uptake was observed as compared to the inflamed and normal muscles., Conclusions: The above results confirm the suitability of the (99m)Tc-TMC complex as a potential Streptococci pneumoniae infection localizing agent.