Preclinical Models of Atopic Dermatitis Suitable for Mechanistic and Therapeutic Investigations

Anish R Maskey,1 Xian Mo,1,2 Xiu-Min Li1,3,4 1Department of Pathology, Microbiology and Immunology, New York Medical College, Valhalla, NY, 10595, USA; 2The Department of Allergy and Clinical Immunology, Guangzhou Institute of Respiratory Health, Guangzhou, People’s Republic of China; 3Department of Otolaryngology, New York Medical College, Valhalla, NY, 10595, USA; 4Department of Dermatology, New York Medical College, Valhalla, NY, 10595, USACorrespondence: Xiu-Min Li, Department of Pathology, Microbiology, & Immunology, New York Medical College, Valhalla, New York, 10595, USA, Tel +1-914-595-1497, Email [email protected]: Atopic dermatitis (AD) is a complex immune-mediated abnormality of the skin characterized by impaired barrier function, eczematous dermatitis, chronic pruritus and itch. The immunological response in AD is mediated by a Th2-dominated immune response in the early acute phase followed by a Th1/ Th2 mixed immune response in the chronic phase. AD is the first step of the “atopic march” that progresses into food allergy, allergic rhinitis, and asthma. Different models are indispensable for studying AD pathogenesis and for designing pre-clinical studies for therapeutic discovery. They reflect the characteristic morphological features of typical human AD with regard to epidermal thickening, hyperkeratosis, acanthosis, and spongiosis and help understand the immunopathogenesis of the disease with respect to IgE levels and cellular infiltration of eosinophils, mast cells, and lymphocytes. Although it is difficult to replicate all human AD clinical features in a model, several AD in vivo models comprising spontaneous, induced, transgenic, and humanized and in vitro models, including 2D, co-culture, and 3D, have been described previously. However, several questions remain regarding whether these models satisfactorily reflect the complexity of human AD. Therefore, this review comprehensively highlights the diversity of currently available models and provides insights into the selection of suitable models based on research questions. It also summarizes the diverse mechanisms associated with each model, which may be valuable for better study design to test new therapeutic options.Keywords: atopic dermatitis, eczema, in vitro models, in vivo models, Th2 cytokine, microbiom

Celecoxib exerts protective effects in the vascular endothelium via COX-2-independent activation of AMPK-CREB-Nrf2 signalling

Although concern remains about the athero-thrombotic risk posed by cyclo-oxygenase (COX)-2-selective inhibitors, recent data implicates rofecoxib, while celecoxib appears equivalent to NSAIDs naproxen and ibuprofen. We investigated the hypothesis that celecoxib activates AMP kinase (AMPK) signalling to enhance vascular endothelial protection. In human arterial and venous endothelial cells (EC), and in contrast to ibuprofen and naproxen, celecoxib induced the protective protein heme oxygenase-1 (HO-1). Celecoxib derivative 2,5-dimethyl-celecoxib (DMC) which lacks COX-2 inhibition also upregulated HO-1, implicating a COX-2-independent mechanism. Celecoxib activated AMPKα(Thr172) and CREB-1(Ser133) phosphorylation leading to Nrf2 nuclear translocation. Importantly, these responses were not reproduced by ibuprofen or naproxen, while AMPKα silencing abrogated celecoxib-mediated CREB and Nrf2 activation. Moreover, celecoxib induced H-ferritin via the same pathway, and increased HO-1 and H-ferritin in the aortic endothelium of mice fed celecoxib (1000 ppm) or control chow. Functionally, celecoxib inhibited TNF-α-induced NF-κB p65(Ser536) phosphorylation by activating AMPK. This attenuated VCAM-1 upregulation via induction of HO-1, a response reproduced by DMC but not ibuprofen or naproxen. Similarly, celecoxib prevented IL-1β-mediated induction of IL-6. Celecoxib enhances vascular protection via AMPK-CREB-Nrf2 signalling, a mechanism which may mitigate cardiovascular risk in patients prescribed celecoxib. Understanding NSAID heterogeneity and COX-2-independent signalling will ultimately lead to safer anti-inflammatory drugs

Development of a Broad-Spectrum Antiviral Agent with Activity Against Herpesvirus Replication and Gene Expression

Purpose: To evaluate the broad-spectrum antiviral activity of peptide H9 (H9) in vitro in order to gain insight into its underlying molecular mechanisms.Method: Antiviral activity against Herpes simplex virus type 1 (HSV-1) was determined using thiazolyl blue (MTT) assay. Polymerase Chain Reaction (PCR) was employed to assay H9 antiviral activity against human cytomegalovirus (HCMV) and Epstein-Barr virus (EBV). The inhibitory effect of H9 on the replication of these viral genes including early genes was assayed by real time-Ppolymerase chain reaction (RT-PCR) and Western blot.Results: H9 possessed significant inhibitory effect on the four different herpesviruses with 50 % inhibitory concentration (IC50) of 1.21 ng/mL (HSV-1). AD169 infection was strongly inhibited with an EC50 value of 0.46 ng/ml. The anti-herpesviral activity of H9 was dose-dependent. The peptide acted primarily during the early stage of infection by detection of the early genes.Conclusion: The results demonstrate that H9 can inhibit the infection of HSV-1, EBV and HCMV. Furthermore, H9 has a broad-spectrum anti-herpesviral effect in vitro based on targeted killing of infected cells expressing genes.Keywords: Antagonist, Trapping receptor/ligand, Broad-spectrum, Anti-herpesvirus, H9 peptide, Gene expressio

Planar and van der Waals heterostructures for vertical tunnelling single electron transistors

Despite a rich choice of two-dimensional materials, which exists these days, heterostructures, both vertical (van der Waals) and in-plane, offer an unprecedented control over the properties and functionalities of the resulted structures. Thus, planar heterostructures allow p-n junctions between different two-dimensional semiconductors and graphene nanoribbons with well-defined edges; and vertical heterostructures resulted in the observation of superconductivity in purely carbon-based systems and realisation of vertical tunnelling transistors. Here we demonstrate simultaneous use of in-plane and van der Waals heterostructures to build vertical single electron tunnelling transistors. We grow graphene quantum dots inside the matrix of hexagonal boron nitride, which allows a dramatic reduction of the number of localised states along the perimeter of the quantum dots. The use of hexagonal boron nitride tunnel barriers as contacts to the graphene quantum dots make our transistors reproducible and not dependent on the localised states, opening even larger flexibility when designing future devices

Large-Eddy Simulation of Flow and Pollutant Transport in Urban Street Canyons with Ground Heating

Our study employed large-eddy simulation (LES) based on a one-equation subgrid-scale model to investigate the flow field and pollutant dispersion characteristics inside urban street canyons. Unstable thermal stratification was produced by heating the ground of the street canyon. Using the Boussinesq approximation, thermal buoyancy forces were taken into account in both the Navier–Stokes equations and the transport equation for subgrid-scale turbulent kinetic energy (TKE). The LESs were validated against experimental data obtained in wind-tunnel studies before the model was applied to study the detailed turbulence, temperature, and pollutant dispersion characteristics in the street canyon of aspect ratio 1. The effects of different Richardson numbers (Ri) were investigated. The ground heating significantly enhanced mean flow, turbulence, and pollutant flux inside the street canyon, but weakened the shear at the roof level. The mean flow was observed to be no longer isolated from the free stream and fresh air could be entrained into the street canyon at the roof-level leeward corner. Weighed against higher temperature, the ground heating facilitated pollutant removal from the street canyon.Singapore-MIT Alliance for Research and Technology. Center for Environmental Sensing and Monitorin

Assessment of Neurodevelopmental Outcomes in Children With Congenital Heart Disease Using Magnetic Resonance Imaging (MRI): Focus on Brain Volume as a Predictor of Neurodevelopmental Abnormalities

Ming-Cui Fu,1 Ye Lin,2 Feng Yang,1 Ying Wang,1 Xu-Ming Mo2 1Department of Radiology, Children’s Hospital of Nanjing Medical University, Nanjing, People’s Republic of China; 2Department of Cardiothoracic Surgery, Children’s Hospital of Nanjing Medical University, Nanjing, People’s Republic of ChinaCorrespondence: Xu-Ming Mo, Email [email protected]: This study aims to evaluate the neurodevelopmental outcomes in children with congenital heart disease (CHD) using magnetic resonance imaging (MRI), and to assess the role of brain volume metrics as predictors of neurodevelopmental abnormalities.Methods: In this retrospective cohort study, 160 children with CHD treated at Children’s Hospital of Nanjing Medical University from January 2020 to December 2023 were analyzed. Patients were classified into normal (DQ ≥ 70, n=106) and abnormal neurodevelopment (DQ < 70, n=54) groups based on Developmental Quotient (DQ) scores. MRI scans were used to measure total brain volume, cortical gray matter, deep gray matter, white matter, and cerebrospinal fluid volumes. Neurodevelopmental assessments focused on adaptive behavior, motor skills, language, and personal-social behavior. ROC analysis was performed to determine the predictive value of brain volume metrics for neurodevelopmental abnormalities.Results: Total brain volume in the normal group (341.82 ± 10.43 mL) was significantly higher than in the abnormal group (323.92 ± 10.24 mL) (P < 0.05). Cortical gray matter volume in the normal group (131.47 ± 4.02 mL) was also significantly greater than in the abnormal group (121.63 ± 6.91 mL) (P < 0.05). No significant differences were observed in white matter, deep gray matter, or cerebrospinal fluid volumes. Children in the abnormal group scored significantly lower in all developmental domains (P < 0.05). ROC analysis showed that total brain volume (AUC = 0.968) and cortical gray matter volume (AUC = 0.936) were strong predictors of neurodevelopmental abnormalities (P < 0.001).Conclusion: Total brain volume and cortical gray matter volume, as measured by MRI, are effective predictors of neurodevelopmental abnormalities in children with CHD and can serve as valuable tools for early neurodevelopmental assessment.Keywords: magnetic resonance imaging, MRI, congenital heart disease, children, neurodevelopmental outcome

Interaction of Temperature and Light in the Development of Freezing Tolerance in Plants

Abstract Freezing tolerance is the result of a wide range of physical and biochemical processes, such as the induction of antifreeze proteins, changes in membrane composition, the accumulation of osmoprotectants, and changes in the redox status, which allow plants to function at low temperatures. Even in frost-tolerant species, a certain period of growth at low but nonfreezing temperatures, known as frost or cold hardening, is required for the development of a high level of frost hardiness. It has long been known that frost hardening at low temperature under low light intensity is much less effective than under normal light conditions; it has also been shown that elevated light intensity at normal temperatures may partly replace the cold-hardening period. Earlier results indicated that cold acclimation reflects a response to a chloroplastic redox signal while the effects of excitation pressure extend beyond photosynthetic acclimation, influencing plant morphology and the expression of certain nuclear genes involved in cold acclimation. Recent results have shown that not only are parameters closely linked to the photosynthetic electron transport processes affected by light during hardening at low temperature, but light may also have an influence on the expression level of several other cold-related genes; several cold-acclimation processes can function efficiently only in the presence of light. The present review provides an overview of mechanisms that may explain how light improves the freezing tolerance of plants during the cold-hardening period

Experimental studies of e + e -→ some charmless processes containing K S0 at √s = 3.773 and 3.65 GeV

We measure the observed cross sections for the charmless processes e + e -→K S0 K - K - K + π ++ c.c., K S0 K - π + η+c.c., K S0 K - π + π + π - η+c.c., K S0 K - K - K + π + η+c.c., K S0 K - K - K + π + π 0+c.c., K S0 K - ρ ++c.c. and K S0 K - π + ρ 0+c.c. We also extract upper limits on the branching fractions for ψ(3770) decays into these final states at 90% C.L. Analyzed data samples correspond to 17.3 pb-1 and 6.5 pb-1 integrated luminosities registered, respectively, at √s = 3.773 and 3.65 GeV, with the BES-II detector at the BEPC collider. © 2009 Springer-Verlag / Società Italiana di Fisica.published_or_final_versionSpringer Open Choice, 21 Feb 201

X-exome sequencing of 405 unresolved families identifies seven novel intellectual disability genes

X-linked intellectual disability (XLID) is a clinically and genetically heterogeneous disorder. During the past two decades in excess of 100 X-chromosome ID genes have been identified. Yet, a large number of families mapping to the X-chromosome remained unresolved suggesting that more XLID genes or loci are yet to be identified. Here, we have investigated 405 unresolved families with XLID. We employed massively parallel sequencing of all X-chromosome exons in the index males. The majority of these males were previously tested negative for copy number variations and for mutations in a subset of known XLID genes by Sanger sequencing. In total, 745 X-chromosomal genes were screened. After stringent filtering, a total of 1297 non-recurrent exonic variants remained for prioritization. Co-segregation analysis of potential clinically relevant changes revealed that 80 families (20%) carried pathogenic variants in established XLID genes. In 19 families, we detected likely causative protein truncating and missense variants in 7 novel and validated XLID genes (CLCN4, CNKSR2, FRMPD4, KLHL15, LAS1L, RLIM and USP27X) and potentially deleterious variants in 2 novel candidate XLID genes (CDK16 and TAF1). We show that the CLCN4 and CNKSR2 variants impair protein functions as indicated by electrophysiological studies and altered differentiation of cultured primary neurons from Clcn4−/− mice or after mRNA knock-down. The newly identified and candidate XLID proteins belong to pathways and networks with established roles in cognitive function and intellectual disability in particular. We suggest that systematic sequencing of all X-chromosomal genes in a cohort of patients with genetic evidence for X-chromosome locus involvement may resolve up to 58% of Fragile X-negative cases

Integrated transcriptomic and proteomic analysis of the global response of Wolbachia to doxycycline-induced stress

The bacterium Wolbachia (order Rickettsiales), representing perhaps the most abundant vertically transmitted microbe worldwide, infects arthropods and filarial nematodes. In arthropods, Wolbachia can induce reproductive alterations and interfere with the transmission of several arthropod-borne pathogens. In addition, Wolbachia is an obligate mutualist of the filarial parasites that cause lymphatic filariasis and onchocerciasis in the tropics. Targeting Wolbachia with tetracycline antibiotics leads to sterilisation and ultimately death of adult filariae. However, several weeks of treatment are required, restricting the implementation of this control strategy. To date, the response of Wolbachia to stress has not been investigated, and almost nothing is known about global regulation of gene expression in this organism. We exposed an arthropod Wolbachia strain to doxycycline in vitro, and analysed differential expression by directional RNA-seq and label-free, quantitative proteomics. We found that Wolbachia responded not only by modulating expression of the translation machinery, but also by upregulating nucleotide synthesis and energy metabolism, while downregulating outer membrane proteins. Moreover, Wolbachia increased the expression of a key component of the twin-arginine translocase (tatA) and a phosphate ABC transporter ATPase (PstB); the latter is associated with decreased susceptibility to antimicrobials in free-living bacteria. Finally, the downregulation of 6S RNA during translational inhibition suggests that this small RNA is involved in growth rate control. Despite its highly reduced genome, Wolbachia shows a surprising ability to regulate gene expression during exposure to a potent stressor. Our findings have general relevance for the chemotherapy of obligate intracellular bacteria and the mechanistic basis of persistence in the Rickettsiales