Epithelial Alterations Research Articles

Conditional gene regulation models demonstrate a pro-proliferative role for growth hormone receptor in prostate cancer.

Humans with inactivating mutations in growth hormone receptor (GHR) have lower rates of cancer, including prostate cancer. Similarly, mice with inactivating Ghr mutations are protected from prostatic intraepithelial neoplasia in the C3(1)/TAg prostate cancer model. However, gaps in clinical relevance in those models persist. The current study addresses these gaps and the ongoing role of Ghr in prostate cancer using loss-of-function and gain-of-function models. Conditional Ghr inactivation was achieved in the C3(1)/TAg model by employing a tamoxifen-inducible Cre and a prostate-specific Cre. In parallel, a transgenic GH antagonist was also used. Pathology, proliferation, and gene expression of 6-month old mouse prostates were assessed. Analysis of The Cancer Genome Atlas data was conducted to identify GHR overexpression in a subset of human prostate cancers. Ghr overexpression was modeled in PTEN-P2 and TRAMP-C2 mouse prostate cancer cells using stable transfectants. The growth, proliferation, and gene expression effects of Ghr overexpression was assessed in vitro and in vivo. Loss-of-function for Ghr globally or in prostatic epithelial cells reduced proliferation and stratification of the prostatic epithelium in the C3(1)/TAg model. Genes and gene sets involved in the immune system and tumorigenesis, for example, were dysregulated upon global Ghr disruption. Analysis of The Cancer Genome Atlas revealed higher GHR expression in human prostate cancers with ERG-fusion genes or ETV1-fusion genes. Modeling the GHR overexpression observed in these human prostate cancers by overexpressing Ghr in mouse prostate cancer cells with mutant Pten or T-antigen driver genes increased proliferation of prostate cancer cells in vitro and in vivo. Ghr overexpression regulated the expression of multiple genes oppositely to Ghr loss-of-function models. Loss-of-function and gain-of-function Ghr models, including prostatic epithelial cell specific alterations in Ghr, altered proliferation, and gene expression. These data suggest that changes in GHR activity in human prostatic epithelial cells play a role in proliferation and gene regulation in prostate cancer, suggesting the potential for disrupting GH signaling, for example by the FDA approved GH antagonist pegvisomant, may be beneficial in treating prostate cancer.

Apoptotic biliary epithelial cells and gut dysbiosis in the induction of murine primary biliary cholangitis

Primary biliary cholangitis (PBC) is a female-predominant liver autoimmune disease characterized by the specific immune-mediated destruction of the intrahepatic small bile duct. Although apoptosis of biliary epithelial cells (BECs) and alterations in gut microbiota are observed in patients with PBC, it is still unclear whether these events happen in the early stage and cause the breakdown of tolerance in PBC. In this study, we examined the early events in the loss of tolerance in our well-defined 2-OA-OVA-induced murine autoimmune cholangitis (AIC) model. We report herein that apoptosis of BECs was notable in the early stage of murine AIC. An altered gut microbiota, in particular, an increased percentage of gram-positive Firmicutes in AIC mice was also observed. BECs in AIC mice expressed adhesion molecule ICAM-1, cytokines/chemokines TNF-α, CCL2, CXCL9, CXCL10, and toll-like receptor (TLR) 2. Moreover, BECs treated with TLR2 ligand had elevated apoptosis and CXCL10 production. These data collectively suggest a new mechanism of tolerance breakdown in AIC. Altered gut microbiota induces apoptosis of BECs through TLR2 signaling. BECs secrete chemokines to recruit CD8 T cells to damage BECs further.

Effect of gluten on the digestive tract and fat body of Telodeinopus aoutii (Diplopoda).

Adult specimens or larvae of invertebrates used as food for vertebrates are often maintained close to gluten so they might become vectors for cereal proteins. However, the tissues and internal organs can respond differently in animals with different feeding habits. The midgut epithelium might be a first and sufficient barrier preventing uptake and effects of gluten on the whole body, while the fat body is the main organ that accumulates different xenobiotics. Good models for such research are animals that do not feed on gluten-rich products in their natural environment. The project's goal was to investigate alterations in the midgut epithelium and fat body of the herbivorous millipede Telodeinopus aoutii (Diplopoda) and analyze cell death processes activated by gluten. It enabled us to determine whether changes were intensified or reversed by adaptive mechanisms. Adult specimens were divided into control and experimental animals fed with mushrooms supplemented with gluten and analyzed using transmission electron microscopy, flow cytometry, and confocal microscopy. Two organs were isolated for the qualitative and quantitative analysis: the midgut and the fat body. Our study of the herbivorous T. aoutii which does not naturally feed on gluten containing diet showed that continuous and prolonged gluten feeding activates repair processes that inhibit the processes of cell death (apoptosis and necrosis) and induce an increase in cell viability.

A role for age-associated alterations in esophageal epithelium in eosinophilic esophagitis-associated fibrosis.

Subepithelial fibrosis occurs in a subset of eosinophilic esophagitis (EoE) patients and is associated with esophageal stricture. While mechanisms driving EoE fibrosis remain incompletely understood, findings from experimental systems support roles for epithelial-fibroblast crosstalk in this type of tissue remodeling. The current paradigm presents EoE as a progressive fibrostenotic disease in which aged patients develop fibrosis as a function of disease chronicity. In the current study we provide evidence that altered epithelial biology in the aging esophagus may also contribute to EoE-associated fibrosis. We find that induction of EoE inflammation in young and aged mice using the MC903/Ovalbumin protocol for the same time period results in increased lamina propria thickness uniquely in aged animals. Additionally, epithelial cells from aged mice less efficiently limit fibroblast contractility in collagen plug contraction assays compared to those from their young counterparts. Finally, to identify potential mechanisms through which aged esophageal epithelial cells may stimulate fibrotic remodeling, we perform cytokine array experiments in young and aged mice. These studies are significant as identification of age-associated factors that contribute to fibrotic remodeling may aid in the design of strategies toward early detection, prevention, and therapy of fibrostenotic EoE.

Impairment mechanism of nasal mucosa after radiotherapy for nasopharyngeal carcinoma.

The nasal mucosa, which performs the crucial functions of filtering, humidifying and temperature regulation, is one of the most vulnerable areas of nasopharyngeal carcinoma (NPC) patients after radiotherapy (RT). Following RT, NPC patients experience a series of pathological changes in the nasal mucosa, ultimately leading to physiological dysfunction of the nasal epithelium. This article systematically reviews the clinical and pathological manifestations of RT-related nasal damage in NPC patients and summarizes the potential mechanism of damage to the human nasal epithelium by RT. Finally, we outline the current mechanistic models of nasal epithelial alterations after RT in NPC patients and provide additional information to extend the in-depth study on the impairment mechanisms of the nasal mucosa resulting from RT. We also describe the relationship between structural and functional alterations in the nasal mucosa after RT to help mitigate and treat this damage and provide insights informing future clinical and fundamental investigations.

Autophagy in age‐related macular degeneration

AbstractAge‐related macular degeneration (AMD) is the leading cause of visual impairment in the aging population with limited understanding of its pathogenesis and a lack of effective treatment. The progression of AMD is initially characterized by atrophic alterations in the retinal pigment epithelium, as well as the formation of lysosomal lipofuscin and extracellular drusen deposits. Cellular damage caused by chronic oxidative stress, protein aggregation and inflammatory processes may lead to geographic atrophy and/or choroidal neovascularization and fibrosis. The role of macroautophagy/autophagy in AMD pathology is steadily emerging. Selective and secretory autophagy and their role in drusen biogenesis, senescence‐associated secretory phenotype, inflammation and epithelial‐mesenchymal transition in the pathogenesis of AMD are discussed.

Abstract IA029: Noncoding RNAs and DNA methylation alterations as liquid biopsy biomarkers in colorectal cancer

Abstract Colorectal cancer (CRC), a leading cause of cancer-related death worldwide, evolves as a result of the stepwise accumulation of a series of genetic and epigenetic alterations in the normal colonic epithelium, leading to the development of colorectal adenomas and invasive adenocarcinomas. Although genetic alterations have a major role in a subset of CRCs, the pathophysiological contribution of epigenetic aberrations in this malignancy has attracted considerable attention. Data from the past couple of decades has unequivocally illustrated that epigenetic marks are important molecular hallmarks of cancer, as they occur very early in disease pathogenesis, involve virtually all key cancer-associated pathways and, most importantly, can be exploited as clinically relevant disease biomarkers for diagnosis, prognostication and prediction of treatment response. In this regard, various types of molecular biomarkers have been explored as potential options for early cancer detection, disease prognosis and predictive biomarkers that can assist in providing the probability of therapeutic response to specific therapies or disease relapse. While this list of molecular markers continues to grow, in the context of early-detection markers or markers for disease monitoring when the neoplastic tissues has been surgically removed, the majority of the focus has been on the development of non-invasive liquid biopsy markers that can be easily assayed in bodily fluids including blood, urine and stool. In contrast, majority of the prognostic and predictive biomarkers have heavily relied on the analysis of surgically resected tissues in patients before the implementation of adjuvant chemotherapy. Among various categories of molecular assays, the major categories of analytes involve analysis of cell-free tumor DNA (ctDNA) for mutational analysis of specific gene(s) and the interrogation of epigenetic DNA methylation patterns, as well as the analysis of various transcriptomic markers including expression of mRNAs and various categories of small non-coding RNAs (ncRNAs) that have been recognized to play a central role in CRC pathogenesis. While there is a lot of interest in developing these cell-free DNA and RNA markers, most of these markers are still in relatively early stages of development, but with intriguing leads and promise for their future development. While the analysis of mutational and methylation analysis is quite specific, the challenge remains the lack of sensitivity of these assays due to the limited quantities of ctDNA present in systemic circulation. In contrast, analysis of cell-free RNAs is quite robust, their tissue-of-origin and specificity remains debatable. More recent data suggests that interrogation of some of these markers within tumor-derived exosomes or extracellular vesicles in blood might offer a adequate balance between appropriate sensitivity and specificity – which is much needed for a given biomarker before its adaption in the clinic. This presentation will provide a summary of the role of DNA methylation alterations and non-coding RNAs as potential diagnostic, prognostic and predictive biomarkers in CRC, as we usher into the era of precision oncology. Citation Format: Ajay Goel. Noncoding RNAs and DNA methylation alterations as liquid biopsy biomarkers in colorectal cancer [abstract]. In: Proceedings of the AACR Special Conference on Colorectal Cancer; 2022 Oct 1-4; Portland, OR. Philadelphia (PA): AACR; Cancer Res 2022;82(23 Suppl_1):Abstract nr IA029.

Central serous chorioretinopathy complicated by neovascularization following panretinal photocoagulation for diabetic retinopathy

AbstractPurpose: To report a case of chronic central serous chorioretinopathy (CSCR) complicated by neovascularization following panretinal photocoagulation (PRP) for a proliferative diabetic retinopathy (DR).Methods: A 51‐year‐old man, with a medical history of diabetes mellitus and chronic CSCR presented to our department for alteration of central vision of the left eye (LE) 6 months after bilateral PRP for proliferative DR. Complete ophthalmic examination with best corrected visual acuity (BCVA) measurement, fundus photography, fundus autofluorescence (FAF), swept source optical coherence tomography (SS‐OCT), swept source optical coherence tomography angiography (SS‐OCTA) and indocyanine green angiography (ICG‐A) were performed.Results: BCVA was 20/100 in the right eye (RE) and 20/400 in the LE. Anterior segment examination was unremarkable except for a cataract in the RE. Fundoscopic examination of the LE showed multiple areas of retinal pigment epithelium (RPE) alteration and a temporal pigment epithelium detachment (PED). FAF showed multiple hypoautofluorescent lesions associated with gravitational tracks. SS‐OCT revealed a serous retinal detachment, a flat irregular PED with a double layer sign overlying a thickened choroid and dilated choroidal vessels. Mid‐phase ICGA revealed 2 discrete areas of hyperfluorescence. SS‐OCTA confirmed the presence of 2 well‐defined choroidal neovascular membranes. The patient was diagnosed with Pachychoroid Neovasculopathy (PNV). After 2 intravitreal bevacizumab injections, SS‐OCT showed complete resolution of the subretinal fluid whereas SS‐OCTA visualized a mature “dead‐tree” shaped membrane.Conclusions: This is an unusual case of chronic CSCR progressing to PNV following PRP for RD. The disruption of an active RPE pump mechanism or release of endogenous stress hormones after PRP have been suggested by some authors as possible causes of this progression. Further studies are needed to elucidate the exact pathogenesis.

Microglial activation and retinal pigment epithelium alteration in a model of focal LED‐induced phototoxicity. Neuroprotection afforded by bFGF and minocycline

AbstractPurpose: To study the microglial reaction and degeneration of the retinal pigment epithelium (RPE) and its protection with basic fibroblast growth factor (bFGF) administered alone or combined with minocycline (MC) in a model of focal light‐emitting diode (LED)‐induced phototoxicity (LIP) in mice.Methods: In dark‐adapted adult C57 mice (20–25 g), the left eye was dilated and exposed for 45 seconds to a blue LED (400 nm; 500 lux) placed 2 mm perpendicular to the corneal apex (n = 71). Mice were treated with bFGF (0.5 μg) administered alone or in combination with MC (45 mg/kg). bFGF was administered in a single intravitreal injection just after LIP and the MC was daily injected intraperitoneally, starting the day before LIP (n = 10–13 per group). Vehicle or naïve groups were used as controls (n = 6–10 each group). Retinas were immunodetected with anti‐arrestin and anti‐Iba1 antibodies to study cone outer segments (a+OS) and microglial cells (Iba1+cells) and RPEs were immunodetected with anti‐ZO1 to study RPE cells, at 3 and 7 days after LIP. The number of cones and microglial cells was studied within a pre‐fixed area (PFA) of 0.9 mm diameter at the centre of the lesion.Results: Treatment with bFGF alone or in combination with MC was effective in protecting a+OS within the PFA compared to vehicles at 7 days after LIP (p = 0.034 and p = 0.001, respectively). The combined treatment of bFGF and MC was effective in reducing the number of Iba1+cells within the PFA in the outer plexiform layer at 7 days after LIP (p = 0.0003). However, the treatments had no restorative effect on RPE cell survival or morphological integrity at 7 days after LIP (p = 0.6762).Conclusions: Administration of bFGF alone or in combination with MC increased a+OS survival and reduces Iba1+cell activation but does not reduce RPE damage.

Biomarkers predicting central serous chorioretinopathy episode persistence.

The purpose of this study was to identify biomarkers at presentation that are associated with a persistent central serous chorioretinopathy (CSC) episode. The prospective study included 35 patients with an acute CSC episode. Potential clinical and imaging biomarkers were evaluated at baseline and 3 months from the episode onset. As biomarkers age, sex, steroid use, episode recurrence, central retinal thickness (CRT), macular volume (MV), choroidal thickness (CT), pigment epithelial detachment (PED) height, and width, number of retinal hyperreflective foci (HF), leakage pattern, and area of retinal pigment epithelial (RPE) alterations were investigated. At 3 months from the CSC episode onset, spontaneous resolution occurred in 19 patients, while 16 patients had a persistent CSC episode. The group of patients with a persistent episode was statistically significantly associated with female sex (p = 0.032), older age (p = 0.015), wider PED (p = 0.005), and higher number of HF (p = 0.02). Moreover, this group of patients had a significant association with thinner choroid and diffuse RPE alterations as a pair (p = 0.008). Older and female CSC patients with wider PED, increased number of HF, thinner choroid, and diffuse RPE alterations at presentation are inclined to episode persistence and could benefit from earlier treatment.

A Linkage between Angiogenesis and Inflammation in Neovascular Age-Related Macular Degeneration.

Age-related macular degeneration (AMD) is the leading cause of visual impairment in the aging population with a limited understanding of its pathogenesis and the number of patients are all the time increasing. AMD is classified into two main forms: dry and neovascular AMD (nAMD). Dry AMD is the most prevalent form (80-90%) of AMD cases. Neovascular AMD (10-20% of AMD cases) is treated with monthly or more sparsely given intravitreal anti-vascular endothelial growth factor inhibitors, but unfortunately, not all patients respond to the current treatments. A clinical hallmark of nAMD is choroidal neovascularization. The progression of AMD is initially characterized by atrophic alterations in the retinal pigment epithelium, as well as the formation of lysosomal lipofuscin and extracellular drusen deposits. Cellular damage caused by chronic oxidative stress, protein aggregation and inflammatory processes may lead to advanced geographic atrophy and/or choroidal neovascularization and fibrosis. Currently, it is not fully known why different AMD phenotypes develop. In this review, we connect angiogenesis and inflammatory regulators in the development of nAMD and discuss therapy challenges and hopes.

Metagenomic insights into the rumen epithelial integrity responses to the vitamin B1 supplement under high-concentrate diets treatments.

Subacute ruminal acidosis (SARA) becomes the most common nutritional metabolic disease in high-yielding dairy cows and later fatting beef cattle because of the increasing consumption of high-concentrate diets in modern feeding patterns. Our previous research found a certain piece of evidence that adding 180 mg thiamine/kg DMI could increase the rumen pH and regulate the structure of the rumen microbial community in vivo. However, there is still limited experimental data on the effects of SARA on thiamine status, the damage to the structure of rumen epithelial cells, and the underlying mechanism of the epithelium alterations. For this purpose, a total of 18 Angus bulls (average 22.0-months-old) with an average live weight of 567.6 ± 27.4 kg were randomly allocated into a control treatment (CON), a high-concentrate diet treatment (HC), and a high-concentrate diet with the vitamin B1 supplement treatment (HCB). All bulls were conducted with a 7-day adjustment period followed by a 60-day-long main feeding procedure. Results indicated that ADFI and ADG significantly decreased in the HC treatment compared with CON (P < 0.05), while significantly increased after the VB1 supplement (P < 0.05). Besides, ruminal acetate content was significantly downregulated while propionate was significantly upregulated under the HC treatment compared with CON (P < 0.05); however, these alterations showed a completely inverse regulatory effect on the VB1 supplement compared with HC (P < 0.05). These changes causatively induced a significant decrease in the A/P ratio in the HC treatment compared with CON and HCB treatments (P < 0.05). Bacterial communities in the HC treatment could be separated from those in CON through PCoA axes 1 and 2. Meanwhile, the VB1 supplement significantly altered the bacterial communities compared with the HC treatment, except for HCB-3. Furthermore, the HC treatment significantly upregulated the expression of JNK, Bax, Caspase-8, Caspase-3, Caspase-9, and Cyt-C compared with CON, while significantly downregulated the expression of Bcl-2. The VB1 supplement showed a complete converse gene expression compared with HC. In conclusion, the VB1 supplement could effectively attenuate the alterations that occurred when exposed to high-concentrate diets, and help promote production performance through increased fermentability.

Lipopolysaccharide induced intestinal epithelial injury: a novel organoids-based model for sepsis in vitro.

Advances in organoid culture technology have provided a greater understanding of disease pathogenesis, which has been rarely studied in sepsis before. We aim to establish a suitable organoids-based intestinal injury model for sepsis. Stable passaged organoids were constructed and pre-treated with lipopolysaccharide (LPS) to mimic sepsis-induced intestinal injury. The LPS-induced sepsis model was used as a reference. We used quantitative real-time polymerase chain reaction to evaluate the RNA levels of inflammatory factors and antimicrobial peptides. Enzyme-linked immunosorbent assay was used to evaluate the protein levels, hematoxylin and eosin staining was used to evaluate the pathology of the small intestine of mice, and immunohistochemistry and immunofluorescence were used to evaluate the intestinal epithelial barrier function. Perkin Elmer Operetta™ was used to obtain high-resolution images of three-dimensional organoids. An LPS concentration >150 μg/mL after 24 h was identified to cause organoid growth restriction. The fluorescence intensity of zonula occludens-1 and occludins at LPS concentrations >100 μg/mL decreased significantly after 24 h. After LPS stimulation for 8 h, the RNA expression levels of interleukin (IL)-1α, tumor necrosis factor alpha, granulocyte-macrophage colony-stimulating factor, IL-6, and regenerating islet-derived protein 3 alpha, beta, and gamma increased. These results resembled those of intestinal epithelial layer alterations in a mouse sepsis model. For IL-10, the RNA expression level increased only when the LPS level >200 μg/mL for 24 h. This study provides the primary intestinal in vitro model to study the effects of LPS-induced intestinal injury resembling sepsis. This model provides a platform for immune associated mechanism exploration and effective drug screening.

Evaluation of the Effect of Dermatochalasis and Upper Eyelid Blepharoplasty Surgery on Corneal Epithelial Thickness Alterations.

The aim of this study was to assess the effect of different grades of dermatochalasis (DC) and upper eyelid blepharoplasty (UEB) surgery on corneal epithelial thickness (CET), objectively using anterior segment-optical coherence tomography (AS-OCT) measurements. 90 eyes of patients with DC were divided into three groups according to the severity of the DC. Forty-one eyes of age and sex-matched patients without DC were randomly selected as the control group. The study did not include patients with more than 2 D of spherical refractive error and more than 1.5 D of astigmatism, a history of previous eyelid surgery, ocular surface disease, contact lens use, and ophthalmic eye drop use. CET measurements of all the patients were performed with an AS-OCT (RTVue-XR, Optovue Inc., USA). There were statistically significant differences in the CET of the superior, superonasal, superotemporal, inferotemporal, and temporal sectors between the DC and control groups (p = 0.001, p = 0.02, p = 0.03, p = 0.02, p = 0.04, respectively). While there were no differences in CET among the sectors of the control group, there was a difference in CET among some sectors of the DC group. When the DC group was subdivided by severity, there was no difference between subgroups for CET across all sectors both preoperatively and postoperatively. In the measurements made 6months after UEB surgery, the CET in all sectors increased statistically significantly compared to those measured in the preoperative period. This study revealed that DC reshaped the corneal epithelium and UEB surgery restored this remodeling 6months after surgery. This journal requires that authors assign a level of evidence to each article. For a full description of these Evidence-Based Medicine ratings, please refer to the Table of Contents or the online Instructions to Authors www.springer.com/00266 .

Structural and functional changes in rat uterus induced by neonatal androgenization.

Fetal or neonatal androgen exposure has a programming effect on ovarian function inducing a polycystic ovarian syndrome-like condition. Its effects on uterine structure and function are poorly studied. The aim of this work was to characterize the temporal course of changes in the rat uterine structure induced by neonatal exposure to aromatizable or not aromatizable androgens. Rats were daily treated with testosterone, dihydrotestosterone or vehicle during follicle assembly period (postnatal days 1 to 5). Uterine histoarchitecture, hormonal milieu, endometrial stromal collagen and capillary density were analyzed at prepubertal, pubertal and adult ages. Our data shows that neonatal androgen exposure induces early and long-lasting deleterious effects on uterine development, including altered adenogenesis and superficial epithelial alterations and suggest a role for altered serum estradiol levels in the maintenance and worsening of the situation. Our results suggest that alterations of the neonatal androgenic environment on the uterus could be responsible for alterations in the processes of implantation and maintenance of the embryo in women with polycystic ovary syndrome.

Comprehensive single cell analysis of pandemic influenza A virus infection in the human airways uncovers cell-type specific host transcriptional signatures relevant for disease progression and pathogenesis.

The respiratory epithelium constitutes the first line of defense against invading respiratory pathogens, such as the 2009 pandemic strain of influenza A virus (IAV, H1N1pdm09), and plays a crucial role in the host antiviral response to infection. Despite its importance, however, it remains unknown how individual cell types within the respiratory epithelium respond to IAV infection or how the latter may influence IAV disease progression and pathogenesis. Here, we used single cell RNA sequencing (scRNA-seq) to dissect the host response to IAV infection in its natural target cells. scRNA-seq was performed on human airway epithelial cell (hAEC) cultures infected with either wild-type pandemic IAV (WT) or with a mutant version of IAV (NS1R38A) that induced a robust innate immune response. We then characterized both the host and viral transcriptomes of more than 19,000 single cells across the 5 major cell types populating the human respiratory epithelium. For all cell types, we observed a wide spectrum of viral burden among single infected cells and a disparate host response between infected and bystander populations. Interestingly, we also identified multiple key differences in the host response to IAV among individual cell types, including high levels of pro-inflammatory cytokines and chemokines in secretory and basal cells and an important role for luminal cells in sensing and restricting incoming virus. Multiple infected cell types were shown to upregulate interferons (IFN), with type III IFNs clearly dominating the antiviral response. Transcriptional changes in genes related to cell differentiation, cell migration, and tissue repair were also identified. Strikingly, we also detected a shift in viral host cell tropism from non-ciliated cells to ciliated cells at later stages of infection and observed major changes in the cellular composition. Microscopic analysis of both WT and NS1R38A virus-infected hAECs at various stages of IAV infection revealed that the transcriptional changes we observed at 18 hpi were likely driving the downstream histopathological alterations in the airway epithelium. To our knowledge, this is the first study to provide a comprehensive analysis of the cell type-specific host antiviral response to influenza virus infection in its natural target cells – namely, the human respiratory epithelium.

Use of Personalized Biomarkers in Metastatic Colorectal Cancer and the Impact of AI.

Simple SummaryColorectal cancer is one of the most frequent cancers worldwide, with a high incidence and mortality. Although many treatment options are available for metastatic disease, patient survival is still limited. The molecular classification of colorectal cancer proposed in 2015 has helped us to better understand colorectal cancer and realize a more effective implementation of therapeutic sequences. It has also been observed that the existing mutational landscape is closely correlated with the epigenetics of colorectal cancer. The identification of prognostic and predictive biomarkers in this context becomes a necessity closely related to therapeutics, and artificial intelligence can be used to discover new biomarkers.Colorectal cancer is a major cause of cancer-related death worldwide and is correlated with genetic and epigenetic alterations in the colonic epithelium. Genetic changes play a major role in the pathophysiology of colorectal cancer through the development of gene mutations, but recent research has shown an important role for epigenetic alterations. In this review, we try to describe the current knowledge about epigenetic alterations, including DNA methylation and histone modifications, as well as the role of non-coding RNAs as epigenetic regulators and the prognostic and predictive biomarkers in metastatic colorectal disease that can allow increases in the effectiveness of treatments. Additionally, the intestinal microbiota’s composition can be an important biomarker for the response to strategies based on the immunotherapy of CRC. The identification of biomarkers in mCRC can be enhanced by developing artificial intelligence programs. We present the actual models that implement AI technology as a bridge connecting ncRNAs with tumors and conducted some experiments to improve the quality of the model used as well as the speed of the model that provides answers to users. In order to carry out this task, we implemented six algorithms: the naive Bayes classifier, the random forest classifier, the decision tree classifier, gradient boosted trees, logistic regression and SVM.

Behavioural, physiological and molecular responses of the Antarctic fairy shrimp Branchinecta gaini (Daday, 1910) to polystyrene nanoplastics

Plastic pollution represents an emerging environmental issue in terrestrial Antarctica, especially in the Antarctic Peninsula and Maritime Antarctica, which have been recently recognized as hot spots for plastic litter. In these regions, freshwater (FW) environments such as lakes host isolated ecosystems and species that can be severely affected by increasing environmental and anthropogenic stressors, which include plastics that are still overlooked. In this study, we investigated for the first time the impact of nanoplastics on adults of the fairy shrimp Branchinecta gaini (Order Anostraca) populating Antarctic FW ecosystems, using surface charged polystyrene nanoparticles (PS NPs) as a proxy. Short-term acute toxicity (48 h) was investigated by exposing adults to carboxyl (-COOH, 60 nm) and amino-modified (-NH2, 50 nm) PS NPs at 1 and 5 μg mL−1. Biodisposition of PS NPs and lethal and sub-lethal effects (i.e., swimming, moulting, histology, gene expression) were assessed. Behaviour of PS NPs in Antarctic FW media was monitored through 48 h of exposure showing that both PS NPs kept their nanoscale size in the Antarctic FW media. Survival of fairy shrimp adults over short-term exposure was not affected, on the other hand an increase in moulting rate and alterations in the gut epithelium were observed upon exposure to both PS NPs. Significant alterations at the behavioural (ventilation rate) and molecular (up-regulation of Hsp70mit, Hsp83, Sod, P450) levels were related to PS NP surface charge and associated with PS-NH2 exposure only. Nanoplastics could represent a threat for Antarctic FW biodiversity and the Antarctic fairy shrimp could be a valuable model for assessing their impact on such remote and pristine aquatic ecosystems.

LIPID PEROXIDATION AND RETINAL PIGMENT EPITHELIUM: A REVIEW ARTICLE

Reactive oxygen species (ROS) may be damaging to any biological system when their levels exceed those of antioxidants. ROS has both positive and negative effects. An imbalance of antioxidants and reactive oxygen species leads to oxidative stress (OS) (ROS. OS, which is more prevalent in elderly persons, is the cause of retinal diseases. Any damage to the retina that results in a reduction in visual acuity is referred to as retinopathy. Age-related macular degeneration (AMD), DR, and other retinal degenerative illnesses are caused by low antioxidant levels. Due to the presence of polyunsaturated fatty acids and photooxidative damage, the retina is especially vulnerable to OS. In vitro and in vivo antioxidant alterations in the retinal pigment epithelium were also noted. The fundamental mechanism must be understood since either OS causes retinopathies or the sickness creates stressful conditions in tissue or cells. We aimed to summarize the latest relationship between lipid peroxidation and retinopathies in the current review article.

233. ENDOSCOPIC REFERENCE SCORE AND HISTOLOGICAL SCORING SYSTEM ARE COMPLEMENTARY ASSESSMENTS FOR CHARACTERIZING FIBROSTENOTIC OR INFLAMMATORY PHENOTYPES IN EOSINOPHILIC ESOPHAGITIS PATIENTS

Abstract The Histologic Scoring System (HSS) assesses severity/extent of histologic features (eosinophil density [ED]/basal zone hyperplasia [BZH]/eosinophil abscesses [EA]/eosinophil surface layering [SL]/surface epithelial alteration [SEA]/dyskeratotic epithelial cells [DEC]/dilated intercellular spaces [DIS]/lamina propria fibrosis [LPF]) in eosinophilic esophagitis (EoE). The Endoscopic Reference Score (EREFS) assesses endoscopic feature severity (edema/exudates/furrows/rings/strictures). We correlated HSS components with inflammatory/remodeling EREFS components in placebo-treated EoE patients from 2 trials—phase 2 proof-of-concept (POC; NCT02379052), phase 3 TREET part A (Phase3-PartA; NCT03633617). POC was a 12-week trial in adults, and Phase3-PartA was a 24-week trial in adults and adolescents. Proximal/mid/distal esophagus biopsies were collected at baseline/end of treatment (EOT) and scored by HSS for grade (severity)/stage (extent)/components (higher scores = greater severity). Endoscopies were scored by EREFS (higher scores = greater severity) and inflammatory (edema+exudates+furrows) and remodeling (rings+stricture) subscores calculated. Pearson correlation and single co-variate regression model analyses were performed between baseline HSS grade/stage, total/individual components scores and EREFS total/components scores and subscores, and for the change in scores from baseline at EOT (ΔEOT). Moderate-to-strong correlations (r &amp;gt; 0.3) were observed between total EREFS and HSS grade/stage for baseline/ΔEOT in POC and baseline in Phase3-PartA (Table). Moderate correlations were observed between BZH and EREFS remodeling subscore and rings for ΔEOT in Phase3-PartA; DIS grade and EREFS remodeling subscore and strictures for baseline and ΔEOT in POC but not Phase3-PartA; and EREFS inflammation and remodeling subscores and components for SEA and SL at baseline and ΔEOT in POC. Moderate-to-strong correlations were observed between EREFS inflammation subscore and DIS stage and between edema and DIS grade/stage at baseline in both studies. Similar results were observed using regression analyses. Specific components of HSS and EREFS, reflecting inflammatory and remodeling processes, are closely correlated, when examined by Pearson correlations or regression analyses, and can aid identification of fibrostenotic or inflammatory phenotypes in EoE. Correlations of DIS with endoscopic inflammatory and remodeling features reflect the role of barrier dysfunction in maintaining chronic inflammation that ultimately leads to remodeling.