Cells Type Research Articles

Epithelial Responses in Radiation-Induced Lung Injury (RILI) Allow Chronic Inflammation and Fibrogenesis.

Radiation models, such as whole thorax lung irradiation (WTLI) or partial-body irradiation (PBI) with bone-marrow sparing, have shown that affected lung tissue displays a continual progression of injury, often for months after the initial insult. Undoubtably, a variety of resident and infiltrating cell types either contribute to or fail to resolve this type of progressive injury, which in lung tissue, often develops into lethal and irreversible radiation-induced pulmonary fibrosis (RIPF), indicating a failure of the lung to return to a homeostatic state. Resident pulmonary epithelium, which are present at the time of irradiation and persist long after the initial insult, play a key role in the maintenance of homeostatic conditions in the lung and have often been described as contributing to the progression of radiation-induced lung injury (RILI). In this study, we took an unbiased approach through RNA sequencing to determine the in vivo response of the lung epithelium in the progression of RIPF. In our methodology, we isolated CD326+ epithelium from the lungs of 12.5 Gy WTLI C57BL/6J female mice (aged 8-10 weeks and sacrificed at regular intervals) and compared irradiated and non-irradiated CD326+ cells and whole lung tissue. We subsequently verified our findings by qPCR and immunohistochemistry. Transcripts associated with epithelial regulation of immune responses and fibroblast activation were significantly reduced in irradiated animals at 4 weeks postirradiation. Additionally, alveolar type-2 epithelial cells (AEC2) appeared to be significantly reduced in number at 4 weeks and thereafter based on the diminished expression of pro-surfactant protein C (pro-SPC). This change is associated with a reduction of Cd200 and cyclooxygenase 2 (COX2), which are expressed within the CD326 populations of cells and function to suppress macrophage and fibroblast activation under steady-state conditions, respectively. These data indicate that either preventing epithelial cell loss that occurs after irradiation or replacing important mediators of immune and fibroblast activity produced by the epithelium are potentially important strategies for preventing or treating this unique injury.

Intestinal epithelial cell-specific deletion of cytokine-inducible SH2- containing protein alleviates experimental colitis in ageing mice.

The incidence of inflammatory bowel disease (IBD) in the elderly has increased in recent years. However, the mechanisms underlying the ageing-related IBD susceptibility remain elusive. Cytokine-inducible SH2-containing protein (CISH) involved in regulating metabolism, the expansion of intestinal tuft cells and type-2 innate lymphoid cells, and ageing-related airway inflammation. Here, we investigated the role of CISH in ageing-related colitis susceptibility. CISH and phosphorylated signal transducer and activator of transcription-3 (p-STAT3) levels were evaluated in the colons of ageing mice and older ulcerative colitis (UC) patients. Mice with intestinal epithelial cell-specific knockout of Cish (CishΔIEC) and Cish-floxed mice were administered dextran sodium sulfate (DSS) or trinitrobenzene sulfonic acid (TNBS) to induce colitis. Colonic tissues were analysed in quantitative real-time polymerase chain reaction, immunoblotting, immunohistochemical, and histological staining experiments. Differentially expressed genes from colonic epithelia were analysed by RNA-sequencing. Ageing increased the severity of DSS-induced colitis and the expression of colonic epithelial CISH in mice. CishΔIEC prevented against the DSS or TNBS-induced colitis in middle-aged mice but not, in the young mice. RNA-sequencing analysis revealed that CishΔIEC significantly suppressed DSS-induced oxidative stress and proinflammatory responses. During ageing CCD841 cell model, knockdown of CISH decreased ageing-induced oxidative stress and proinflammatory responses, whereas these effects were compromised by knocking down or inhibiting STAT3. The increase in CISH expression was higher in the colonic mucosa of older patients with UC than that in healthy controls. CISH might be a proinflammatory regulator in ageing; therefore, targeted therapy against CISH may provide a novel strategy for treating ageing-related IBD.

Immune Phenotypic Characterization of a TRAIL-Knockout Mouse

The TNF-superfamily member TRAIL is known to mediate selective apoptosis in tumor cells suggesting this protein as a potential antitumor drug target. However, initial successful pr-clinical results could not be translated into the clinic. Reasons for the ineffectiveness of TRAIL-targeting in tumor therapies could include acquired TRAIL resistance. A tumor cell acquires TRAIL resistance, for example, by upregulation of antiapoptotic proteins. In addition, TRAIL can also influence the immune system and thus, tumor growth. We were able to show in our previous work that TRAIL-/- mice show improved survival in a mouse model of pancreatic carcinoma. Therefore, in this study we aimed to immunologically characterize the TRAIL-/- mice. We observed no significant differences in the distribution of CD3+, CD4+, CD8+ T-cells, Tregs, and central memory CD4+ and CD8+ cells. However, we provide evidence for relevant differences in the distribution of effector memory T-cells and CD8+CD122+ cells but also in dendritic cells. Our findings suggest that T-lymphocytes of TRAIL-/- mice proliferate at a lower rate, and that the administration of recombinant TRAIL significantly increases their proliferation, while regulatory T-cells (Tregs) from TRAIL-/- mice are less suppressive. Regarding the dendritic cells, we found more type-2 conventional dendritic cells (DC2s) in the TRAIL-/- mice. For the first time (to the best of our knowledge), we provide a comprehensive characterization of the immunological landscape of TRAIL-deficient mice. This will establish an experimental basis for future investigations of TRAIL-mediated immunology.

Study of frontal and ethmoid sinus of sinonasal complex along with olfactory fossa: anatomical considerations for endoscopic sinus surgery.

The Functional endoscopic sinus surgery through transnasal approach is a common modality of treatment for disorders of the nasal cavity, paranasal air sinuses as well as cranial cavity. The olfactory fossa (OF) is located along the superior aspect of cribriform plate which varies in shape and depth. This variable measurement of the depth of OF is mostly responsible for greater risk of intracranial infiltration during endoscopic procedures in and around the nasal cavity. The morphology of frontal and ethmoid sinus (ES) vary from simple to complex. This cadaveric study is planned to improve the ability of the otolaryngologist, radiologist to understand the possible morphological variations and plan steps of less invasive "precision surgery" to have a safe and complication free procedures. A total of 37 human head regions were included in the study. For classification of OF, Modified Kero's classification was used. The size, shape and cells of frontal and ES were noted. We found, type II (60.8%) OF was more common followed by type I (29.7%) than type III (9.5%). The shape of frontal sinus was comma shaped (55.4%) followed by oval (18.9%) than irregular (16.2%). Most common two cells type of ES was seen in 50.0% of both anterior and posterior ES. Out of 74 ES, 8.1% of Onodi cells and 14.9% of agger nasi cells were seen.

Systemic mastocytosis with an associated hematological neoplasm (acute myeloid leukaemia with RUNX1::RUNX1T1 fusion) – A case study

Systemic mastocytosis (SM) is a rare blood disorder that can affect multiple body organs. There are some patients of SM with coexpression of hematological neoplasm designated in the World Health Organization classification as SM with an associated hematological neoplasm (SM-AHN). A 13-year-old female was referred for immunophenotyping since blasts were reported in the bone marrow aspirate. On performing flow cytometry, abnormal myeloid blasts (>20%) were obtained on flow cytometry suggestive of acute myeloid leukemia with aberrant expression of CD19. Furthermore, there were increased mast cells abnormally expressing CD25. The aspiration slides reviewed showed atypical mast cells Type I and Type II. Molecular analysis revealed RUNX1::RUNX1T1 fusion as well as KIT D816V mutation. Karyotyping revealed a three-way translocation involving chromosomes 8, 12, and 21 consistent with RUNX1::RUNX1T1 fusion. Fluorescence in situ hybridization (FISH) performed using a dual color dual fusion probe for RUNX1::RUNX1T1 showed atypical abnormal pattern consistent with RUNX1::RUNX1T1 fusion. This rare diagnosis could be reached as a result of a strong correlation between morphology, immunophenotyping, cytogenetics, and molecular analysis, leading to a better treatment plan for the patient.

Pharmacist Participation in Diabetes-Related Distress Assessment and Role of Self Management Among Type-2 Diabetes Mellitus Patients in a Tertiary Care Hospital

According to the World Health Organization (WHO), diabetes is a state of chronic hyperglycemia resulting from decrease in insulin engenderment (type 1) (or) decremented insulin uptake by cells (type 2) leading to multitude of complications ranging from diseases of the diminutive vessels of kidney and retina, peripheral neuropathy and coronary artery disease. Diabetes distress is defined as patient concerns about disease management, support, emotional burden and access to care, is an important condition distinct from depression. It is a part of diabetes and it is a non-psychiatric disease. Diabetes related distress (DRD) refers to the emotional and behavioural changes caused by diabetes. A p-value of less than 0.05 is considered significant. Our study shows the scores of specific diet does not show significant correlation in DDS-17 (r=-0.101) whereas a significant correlation was found in SDSCA (0.206). Similarly, the scores in exercise does not show a significant correlation in DDS-17 (r=0.03) but shows a significant correlation in SDSCA (0.666). It reveals Patients tend to have high and low diabetes distress depending upon their diabetes self-care specifically related to diet intake. Moreover women are more prone to experience diabetes distress. This was confirmed by using the (DDS-17) and (SDSCA). Age, smoking, alcohol, consumption, emotional burden, physical inactivity was found to contribute to the development of diabetes related distress in the study subjects.

Insights into the pathogenesis of allergic disease from dedicator of cytokinesis 8 deficiency.

Clinical observations and mechanistic studies in dedicator of cytokinesis 8 (DOCK8)-deficient patients and mice have revealed multiple mechanisms that could contribute to their unusually prevalent and severe allergic disease manifestations. Physical interactions of DOCK8 with STAT3 in B cells and T cells may contribute to increased IgE isotype switching or defective immune synapse formation that decreases T-cell receptor signal strength. A newly discovered TFH13 cell type promotes the development of life-threatening allergy via production of IL-13 and is increased in DOCK8 deficiency. Cytoskeletal derangements and cytothripsis, which were previously shown to account for the increased susceptibility to viral skin infection in DOCK8 deficiency, can lead to interplay between myeloid cells and T cells to ultimately increase production of IL-4, IL-5, and IL-13. Finally, theeffects on type-2 innate lymphoid cells may also contribute to allergic disease.

Carnosine increases insulin-stimulated glucose uptake and reduces methylglyoxal-modified proteins in type-2 diabetic human skeletal muscle cells

Type-2 diabetes (T2D) is characterised by a dysregulation of metabolism, including skeletal muscle insulin resistance, mitochondrial dysfunction, and oxidative stress. Reactive species, such as methylglyoxal (MGO) and 4-hydroxynonenal (4-HNE), positively associate with T2D disease severity and can directly interfere with insulin signalling and glucose uptake in skeletal muscle by modifying cellular proteins. The multifunctional dipeptide carnosine, and its rate-limiting precursor β-alanine, have recently been shown to improve glycaemic control in humans and rodents with diabetes. However, the precise mechanisms are unclear and research in human skeletal muscle is limited. Herein, we present novel findings in primary human T2D and lean healthy control (LHC) skeletal muscle cells. Cells were differentiated to myotubes, and treated with 10 mM carnosine, 10 mM β-alanine, or control for 4-days. T2D cells had reduced ATP-linked and maximal respiration compared with LHC cells (p = 0.016 and p = 0.005). Treatment with 10 mM carnosine significantly increased insulin-stimulated glucose uptake in T2D cells (p = 0.047); with no effect in LHC cells. Insulin-stimulation increased MGO-modified proteins in T2D cells by 47%; treatment with carnosine attenuated this increase to 9.7% (p = 0.011). There was no effect treatment on cell viability or expression of other proteins. These findings suggest that the beneficial effects of carnosine on glycaemic control may be explained by its scavenging actions in human skeletal muscle.

Conventional dendritic cells type 1 are strongly enriched, quiescent and relatively tolerogenic in local inflammatory arthritis.

Dendritic cells (DC) are crucial for initiating and shaping immune responses. So far, little is known about the functional specialization of human DC subsets in (local) inflammatory conditions. We profiled conventional (c)DC1, cDC2 and monocytes based on phenotype, transcriptome and function from a local inflammatory site, namely synovial fluid (SF) from patients suffering from a chronic inflammatory condition, Juvenile Idiopathic Arthritis (JIA) as well as patients with rheumatoid arthritis (RA). Paired PB and SF samples from 32 JIA and 4 RA patients were collected for mononuclear cell isolation. Flow cytometry was done for definition of antigen presenting cell (APC) subsets. Cell sorting was done on the FACSAria II or III. RNA sequencing was done on SF APC subsets. Proliferation assays were done on co-cultures after CD3 magnetic activated cell sorting (MACS). APC Toll-like receptor (TLR) stimulation was done using Pam3CSK4, Poly(I:C), LPS, CpG-A and R848. Cytokine production was measured by Luminex. cDC1, a relatively small DC subset in blood, are strongly enriched in SF, and showed a quiescent immune signature without a clear inflammatory profile, low expression of pathogen recognition receptors (PRRs), chemokine and cytokine receptors, and poor induction of T cell proliferation and cytokine production, but selective production of IFNλ upon polyinosinic:polycytidylic acid exposure. In stark contrast, cDC2 and monocytes from the same environment, showed a pro-inflammatory transcriptional profile, high levels of (spontaneous) pro-inflammatory cytokine production, and strong induction of T cell proliferation and cytokine production, including IL-17. Although the cDC2 and monocytes showed an overlapping transcriptional core profile, there were clear differences in the transcriptional landscape and functional features, indicating that these cell types retain their lineage identity in chronic inflammatory conditions. Our findings suggest that at the site of inflammation, there is specific functional programming of human DCs, especially cDC2. In contrast, the enriched cDC1 remain relatively quiescent and seemingly unchanged under inflammatory conditions, pointing to a potentially more regulatory role.

Light Microscopy and Ultrastructure of Body Wall in Leech Haemadipsa zeylanica.

Leeches are widely distributed worldwide in a diversity of habitats, such as freshwater, seas, desert, and oases. Since literature does not have an elaborate study on histology of leeches, thus emphasis is given to study the histology. The aim of this study is to examine the histology of free-living leeches Haemadipsa zeylanica under light microscopy and extended to ultrastructural study under transmission electron microscopy. Tissue was fixed in 10% formalin, dehydrated with alcohol, infiltrated and embedded in wax, and thin section was cut using a rotary microtome. Sections were stained with hematoxylin and eosin. The epithelial layer of cells in H. zeylanica had a thin layer of cuticles covering it. In light microscopy, the epidermis has three types of epidermal cells, namely, glandular cells (Type I, Type II, and Type III), supporting cells (Type IV), and pigment cells (Type V). When the three glandular cells of the body wall of H. zeylanica epidermis were examined under an electron microscope, three types of distinct electron-dense granules were identified. Type I cells with granules, Type II cells with course granules close to the dermis were identified as pear-shaped secretory cells, and Type III cells with electron-dense granules.

Reprogramming Cancer Cells to Antigen-presenting Cells.

Cancer cells evade the immune system by downregulating antigen presentation. Although immune checkpoint inhibitors (ICI) and adoptive T-cell therapies revolutionized cancer treatment, their efficacy relies on the intrinsic immunogenicity of tumor cells and antigen presentation by dendritic cells. Here, we describe a protocol to directly reprogram murine and human cancer cells into tumor-antigen-presenting cells (tumor-APCs), using the type 1 conventional dendritic cell (cDC1) transcription factors PU.1, IRF8, and BATF3 delivered by a lentiviral vector. Tumor-APCs acquire a cDC1 cell-like phenotype, transcriptional and epigenetic programs, and function within nine days (Zimmermannova et al., 2023). Tumor-APCs express the hematopoietic marker CD45 and acquire the antigen presentation complexes MHC class I and II as well as co-stimulatory molecules required for antigen presentation to T cells, but do not express high levels of negative immune checkpoint regulators. Enriched tumor-APCs present antigens to Naïve CD8+ and CD4+ T cells, are targeted by activated cytotoxic T lymphocytes, and elicit anti-tumor responses in vivo. The tumor-APC reprogramming protocol described here provides a simple and robust method to revert tumor evasion mechanisms by increasing antigen presentation in cancer cells. This platform has the potential to prime antigen-specific T-cell expansion, which can be leveraged for developing new cancer vaccines, neoantigen discovery, and expansion of tumor-infiltrating lymphocytes. Key features • This protocol describes the generation of antigen-presenting cells from cancer cells by direct reprogramming using lineage-instructive transcription factors of conventional dendritic cells type I. • Verification of reprogramming efficiency by flow cytometry and functional assessment of tumor-APCs by antigen presentation assays.

Continuous Measurement of Reactive Oxygen Species Formation in Bacteria-infected Bone Marrow-derived Macrophages Using a Fluorescence Plate Reader.

Macrophages are at the center of innate immunity and are the main target cells of the intracellular pathogen Salmonella enterica serovar Typhi. The production of reactive oxygen and nitrogen species (ROS/RNS) is the host's early response to invading microbes, as oxidative stress is highly toxic for bacteria. Adequate ROS/RNS production in infected macrophages is critical for the clearance of intracellular pathogens; this is achieved by several enzymes, including inducible NADPH phagocyte oxidase (NOX) and nitric oxide synthase (iNOS), respectively. The pro-inflammatory cytokine interferon gamma (IFNγ), primarily produced by activated natural killer cells and T-helper cells type 1, is a potent inducer of iNOS. Therefore, it is crucial for infection control through oxidative microbicidal activity. To characterize the early oxidative stress response via ROS formation, which is critical for the reduction of Salmonella proliferation within macrophages, we established an in vitro model of murine macrophages infected with Salmonella enterica serovar Typhimurium (S.tm). This serovar induces a systemic infection in mice that is frequently used as a model for typhoid fever, which, in human subjects, is caused by Salmonella Typhi. We generated bone marrow-derived macrophages (BMDM) from C57BL/6N wildtype mice using macrophage colony-stimulating factor (M-CSF) stimulation for six days. Thereafter, we infected BMDM withS. tm for one hour. Shortly before infection, cells were stained with CellROXTM Deep Red reagent. In its reduced form, CellROXTM is non-fluorescent. As a result of oxidation by ROS, this reagent exhibits strong fluorescence and persists within the cells. Subsequently, changes as a result of the oxidative stress response can be measured with a TECAN Spark microplate reader over time. We designed this protocol to measure oxidative stress in macrophages through the course of an infection with an intracellular bacterium. The protocol has several advantages over established techniques. First, it allows to continuously monitor and quantify ROS production in living cells from the very start of the infection to the final clearance of the intracellular pathogen. Second, this protocol enables efficient ROS detection without stressing the cells by detaching or staining procedures. Graphical abstract.

Aplastic anemia severity and IL-6 and IL-8 blood levels.

Aplastic anemia is a rare, fatal bone marrow disorder that is presumed to be an autoimmune-mediated illness that actively destroys haematopoietic cells through a T helper type-1 cell response. Different cell types in the bone marrow and peripheral circulation produce chemokines, such as interleukin-6 (IL-6) and interleukin-8 (IL-8). The myelopoiesis that is profoundly impaired in aplastic anemia may be inhibited by these two, as critical and powerful inhibitors. Therefore, it is conceivable that their ongoing overproduction may contribute to aplastic anemia. We performed a quantitative enzyme-linked immunosorbent assay on the peripheral blood plasma to reveal the levels of IL-6 and IL-8 and their correlation to aplastic anaemia. A total of 80 cases of aplastic anemia were included in this study, diagnosed according to the criteria laid down by the International Agranulocytosis and Aplastic Anemia study group. A total of 10 healthy individuals served as controls in this study. With the help of a commercial ELISA kit and the instructions from the kit's maker, the levels of IL-6 and IL-8 were measured in a quantitative way. Mean serum IL-6 and IL-8 levels in cases were 283.28±220.27 and 122.56±97.79 pg/ml, respectively, as compared to 7.52±1.43 and 3.42±1.73 pg/ml levels in controls. Statistically, mean IL-6, as well as IL-8 levels, were significantly higher in aplastic anemia patients than in controls (p< 0.001). Levels of interleukins were also assessed in relation to the severity of the disease. Patients with very severe aplastic anaemia had significantly higher mean IL-6 and IL-8 levels (516.71±36.73 and 220.50±23.45 pg/ml, respectively), followed by severe aplastic anaemia (198.84±150.39 and 89.82±77.18 pg/ml, respectively) and non-severe aplastic anaemia (26.71±33.40 and 10.29±2.63 pg/ml, respectively) (p<0.001). Blood serum levels of IL-6 and IL-8 were increased in aplastic anemia and showed a correlation with the severity of the disease. Hence, IL-6 and IL-8 may play an important role in the immune-mediated pathophysiology of aplastic anemia and their increasing levels are giving alarming signals for timely implementation of the appropriate treatment regimen to stop further progression of the disease. Additional studies are required in order to further investigate the exact involvement and role of IL-6 and IL-8 in aplastic anemia.

UHRF1-mediated ferroptosis promotes pulmonary fibrosis via epigenetic repression of GPX4 and FSP1 genes

Pulmonary fibrosis (PF), as an end-stage clinical phenotype of interstitial lung diseases (ILDs), is frequently initiated after alveolar injury, in which ferroptosis has been identified as a critical event aggravating the pathophysiological progression of this disease. Here in, a comprehensive analysis of two mouse models of pulmonary fibrosis developed in our lab demonstrated that lung damage-induced ferroptosis of alveolar epithelial Type2 cells (AEC2) significantly accumulates during the development of pulmonary fibrosis while ferroptosis suppressor genes GPX4 and FSP1 are dramatically inactivated. Mechanistically, upregulation of de novo methylation regulator Uhrf1 sensitively elevates CpG site methylation levels in promoters of both GPX4 and FSP1 genes and induces the epigenetic repression of both genes, subsequently leading to ferroptosis in chemically interfered AEC2 cells. Meanwhile, specific inhibition of UHRF1 highly arrests the ferroptosis formation and blocks the progression of pulmonary fibrosis in both of our research models. This study first, to our knowledge, identified the involvement of Uhrf1 in mediating the ferroptosis of chemically injured AEC2s via de novo promoter-specific methylation of both GPX4 and FSP1 genes, which consequently accelerates the process of pulmonary fibrosis. The above findings also strongly suggested Uhrf1 as a novel potential target in the treatment of pulmonary fibrosis.

Long COVID outcomes in an asthmatic cohort and its implications for asthma control

Long COVID outcomes in an asthmatic cohort and its implications for asthma control

Childhood asthma is associated with development of type 1 diabetes and inflammatory bowel diseases: a Danish nationwide registry study

Asthma and autoimmune disorders might be affected by opposing immune mechanisms, T helper cells type 2 (Th2) and T helper cells type 1 (Th1) immunity, respectively. Knowledge on comorbidity can increase understanding of the underlying etiologies. We aim to examine the association between childhood asthma and subsequent risk of type 1 diabetes (T1D) and inflammatory bowel diseases (IBD) in Danish children. Children of Danish origin born during 1991–1996 were included and childhood asthma, defined as a minimum of two collected prescriptions of inhalation corticosteroids age 5–7 years, was linked to hospitalisations with either T1D or IBD after age 8. Associations between childhood asthma and incidence of T1D and IBD were analysed using sex- and year stratified Cox regression. A total of 366,200 children were included in the study, 4.9% had asthma, which increased the risk of both T1D and IBD, hazard ratios of 1.32 (1.08–1.61) and 1.27 (1.09–1.48). In this large nationwide Danish study, we found that children with asthma have increased risk of developing immune diseases T1D and IBD. This contradicts the Th1 vs Th2 paradigm and points towards shared disease mechanisms and risk factors.

Metformin enhances the antitumor activity of oncolytic herpes simplex virus HF10 (canerpaturev) in a pancreatic cell cancer subcutaneous model

Oncolytic virus (OV) therapy is a promising cancer immunotherapy, especially for cold tumors by inducing the direct lysis of cancer cells and initiation of potent antitumor response. Canerpaturev (C-REV) is an attenuated oncolytic herpes simplex virus-1, which demonstrated a potent antitumor effect in various preclinical models when used either alone or combined. Metformin is a commonly prescribed antidiabetic drug that demonstrated a potent immune modulator effect and antitumor response. We combined C-REV with metformin in a low immunogenic bilateral murine tumor model to enhance C-REV’s antitumor efficacy. In vitro, metformin does not enhance the C-REV cell cytotoxic effect. However, in in vivo model, intratumoral administration of C-REV with the systemic administration of metformin led to synergistic antitumor effect on both sides of tumor and prolonged survival. Moreover, combination therapy increased the effector CD44+ CD8+ PD1- subset and decreased the proportion of terminally-differentiated CD103+ KLRG-1+ T-regulatory cells on both sides of tumor. Interestingly, combination therapy efficiently modulates conventional dendritic cells type-1 (cDC1) on tumors, and tumor-drained lymph nodes. Our findings suggest that combination of C-REV and metformin enhances systemic antitumor immunity. This study may provide insights into the mechanism of action of OV therapy plus metformin combination against various tumor models.

130 Skin-associated side effects in patients with chronic rhinosinusitis with nasal polyposis following treatment with dupilumab

Chronic rhinosinusitis with nasal polyposis (CRSwNP) is a typical type-2 inflammation involving T-helper type-2 cells and impairing quality of life due to nasal obstruction, discharge and reduced sense of smell. Recently, the anti-IL4Ra antibody dupilumab was approved for CRSwNP and while dermatologic side effects in patients treated with dupilumab for atopic dermatitis are frequently observed, there is limited knowledge about these effects in patients with CRSwNP. We aimed to investigate frequency and characteristics of dermatologic side effects following initiation of dupilumab treatment in a cohort of Austrian CRSwNP patients.

Xenobiotic-sensing nuclear receptors as targets for phthalates-induced lung injury and antagonism of lycopene

Phthalates are extensively used in the production of plastics products and have been verified to induce lung injury. Lycopene (LYC) has proved an effective preventive and can be utilized to prevent phthalates-induced toxicity. However, the role of phthalate in pathogenesis of lung injury remain poorly researched, and little work has been devoted whether LYC could alleviate phthalate-induced lung toxicity via modulating nuclear xenobiotic receptors (NXRs) response. Here, di (2-ethylhexyl) phthalate (DEHP) is used as the representative of phthalates for further studies on toxicity of phthalates and the antagonistic role of LYC in phthalates-induced lung injury. We found that DEHP exposure caused alveoli destruction and alveolar epithelial cells type II damage. Mechanistically, DEHP exposure increased nuclear accumulation of aryl hydrocarbon receptor (AHR) and its downstream genes level, including cytochrome P450-dependent monooxygenase (CYP) 1A1 and CYP1B1. Constitutive androstane receptor (CAR) and their downstream gene level, including CYP2E1 are also increased after phthalates exposure. Significantly, LYC supplementation relieves lung injury from DEHP exposure by inhibiting the activation of NXRs. We confirm that NXRs plays a key role in phthalates-induced lung injury. Our study showed that LYC may have a positive role in alleviating the toxicity effects of phthalates, which provides an effective strategy for revising phthalates-induced injury.

IoMT-Based Automated Diagnosis of Autoimmune Diseases Using MultiStage Classification Scheme for Sustainable Smart Cities

The resolution of complex medical diagnoses using pattern recognition requires an artificial neural network-based expert system to automate autoimmune disease diagnosis in blood samples. This process is done using image-based computer-aided diagnosis (CAD) to reduce errors in the diagnosis process. This paper describes a Multistage Classification Scheme (MSCS), which uses antinuclear antibody (ANA) tests to identify and classify the existence of autoantibodies in the blood serum that bind to antigens found in the nuclei of mammalian cells. The MSCS classified HEp-2 cells into three stages by using Binary Tree (BT), Artificial Neural Network (ANN), and Support Vector Machine (SVM) as basic blocks. The Indirect Immunofluorescence (IIF) technique is used in the ANA test with Human Epithelial type-2 (HEp-2) cells as substrates. The efficiency of the proposed methodology is assessed using the dataset of ICPR 2016. The intermediate cells (IMC) and positive cells (PC) were separated in Stage 1 prior to preprocessing based on their total strength, and special preprocessing is applied to intermediate cells for improved output, and positive cells are subjected to mild preprocessing. The mean class accuracy (MCA) was 84.9% for intermediate cells and 95.8% for positive cells, although the carefully picked 24 features and SVM classifier were applied. ANN showed better performance by adjusting the weights using the SCGBP algorithm. So, the MCA is 88.4% and 97.1% for intermediate and positive cells, respectively. BT had an MCA of 95.3% for intermediate and 98.6% for positive. In Stage 2, the meta learners BT2, ANN2, and SVM2 were trained for an augmented feature set (24 + 3 results from base learners). Therefore, the performance of BT2, ANN2, and SV M2 was increased by 1.8%, 4.5%, and 4.1% as compared to Stage 1. In Stage 3, the final prediction was performed by majority voting among the results of the three meta learners to achieve 99.1% MCA. The proposed algorithm can be embedded into a CAD framework built for the ANA examination. The proposed model will improve operational efficiency, decrease medical expenses, expand accessibility to healthcare, and improve patient safety in the sector, enabling enterprises to lower unplanned downtime, develop new products or services, increase operational effectiveness, and enhance risk management.