Duct Epithelium Research Articles

Role of Jak3 in cross talk between Intestinal and biliary epithelial cells

BackgroundPrimary biliary cholangitis (PBC) is one of the most common chronic inflammatory condition of the liver affecting 1 in 1000 women over the age of 40. The exact etiology and pathogenetic mechanisms of PBC is unknown and currently, there is no cure to it. PBC progressively affects the intrahepatic and extrahepatic biliary tree leading to bile duct strictures, cholestasis, and hepatic fibrosis. A better understanding of the initial pathologic events is essential to develop appropriate intervention. Recent studies suggest gut‐liver communication has greater implications in hepatic health where the gut microbiome and the gut barrier functions are implicated in bacterial translocation and bile acid metabolism. However, the mechanistic role of intestinal mucosal epithelial cells in gut‐dysbiosis‐mediated PBC are less understood. Janus kinase 3 (Jak3) is a non‐receptor protein kinase that plays essential role in hematopoietic and intestinal epithelial (IEC) differentiation. Previously, we reported that loss of Jak3 exacerbates symptoms in murine model of DSS‐induced colitis through compromised intestinal differentiation. In this work, we investigated the tissue‐specific role of Jak3 in hepatic canalicular remodeling and cholestasis during intestinal chronic low‐grade inflammation (CLGI).MethodsWe used a biliary epithelial cell model and global and IEC Jak3 KO (Jak3−/−) mice and their littermate control and compared the effects of Jak3 on gut dysbiosis and hepatic canalicular remodeling and cholangitis. We also investigated the status of hepatic TLR signaling and TGFb1 pathways using a combination of flow cytometry, confocal microscopy, and western analysis techniques.ResultsOur results show that Jak3 regulates both mucosal epithelial and biliary epithelial (BEC) tolerance through the regulation of TLR where Jak3 impacted BEC remodeling and canalicular structure formation. Studies using hematoxylin and eosin‐stained liver sections demonstrated marked infiltration of lympho‐plasmacytic cells in portal tracts surrounding interlobular bile ducts. An interlobular bile duct showed distorted lumen, and cellular detachment from the basement membrane with infiltration of several lymphoid cells into the cellular layer in both global Jak3‐KO and IEC‐Jak3‐KO mice. Immunohistochemical (IHC) analysis of liver of Jak3−/− and IEC‐Jak3−/−and their littermate control mice using monoclonal antibodies to CD4 and CD8 showed that CD8‐positive cells were predominant along with CD4‐positive cells that were distributed mainly at the peribiliary areas of the portal veins with some CD8 cells in the sinusoidal spaces. CD8 T‐cells and mononuclear cell were also seen attached to bile duct epithelium indicating invasion of these cells into the biliary epithelial layer. A differential flowcytometric assay showed an increased influx of monocytes in Jak3 KO mice compared to control. Moreover, KO mice showed an increased hepatic expression of TLR‐2 and TLR‐4 and a higher IL‐6, IL‐17 and Tnf‐α as compared to the control mice. Together, these results showed that loss of Jak3 expression leads to compromise biliary tolerance coupled with increased extravasation of the circulating monocytes into the liver. In addition, we also found an increased Th17 effector function and simultaneous suppression of Treg cell proliferation in the absence of Jak3, when compared to its control mice. Collectively, these results indicate that Jak3 plays a major role in hepatic canalicular remodeling and cholestasis during intestinal CLGI.

Novel Palliative Chemotherapy for Cholangiocarcinoma

The term cholangiocarcinoma (CC) refers to all tumors arising from bile duct epithelium. CCs are characterized by their rarity, difficulty in diagnosis, and overall poor prognosis. This leads to a paucity of data from which to define the natural history and optimal treatment regimens. Currently, surgical resection remains the only potentially curative treatment, but many patients develop recurrence. In addition, a limited number of patients can be candidates for curative resection at diagnosis. Therefore, chemotherapy is inevitable choice for the treatment of advanced CC. Gemcitabine plus cisplatin (GP) is considered a standard option for advanced biliary cancer. A randomized phase III trial (ABC-02 trial) showed the superiority of gemcitabine plus cisplatin over gemcitabine alone. Treatment with nab-paclitaxel plus gemcitabine-cisplatin prolonged median progression-free survival and overall survival vs. those reported for historical controls treated with gemcitabine-cisplatin alone in a phase II study of 60 patients with locally advanced unresectable or metastatic biliary tract cancer. Recent data of the ABC-06 trial has provided slight evidence for the use of second-line chemotherapy after progression on cisplatin plus gemcitabine combination. Other active regimens, that could be considered in patients who include have disease progression while receiving GP and who retain an adequate performance status, includes capecitabine plus cisplatin, liposomal irinotecan plus leucovorin-modulated fluorouracil and a fluoropyrimidine alone. We herein review recent published data regarding the use of palliative chemotherapies in CC patients, with a particular focus on novel cytotoxic agents.

Survival prediction on intrahepatic cholangiocarcinoma with histomorphological analysis on the whole slide images

Intrahepatic cholangiocarcinoma (ICC) is cancer that originates from the liver's secondary ductal epithelium or branch. Due to the lack of early-stage clinical symptoms and very high mortality, the 5-year postoperative survival rate is only about 35%. A critical step to improve patients' survival is accurately predicting their survival status and giving appropriate treatment. The tumor microenvironment of ICC is the immediate environment on which the tumor cell growth depends. The differentiation of tumor glands, the stroma status, and the tumor-infiltrating lymphocytes in such environments are strictly related to the tumor progress. It is crucial to develop a computerized system for characterizing the tumor environment. This work aims to develop the quantitative histomorphological features that describe lymphocyte density distribution at the cell level and the different components at the tumor's tissue level in H&E-stained whole slide images (WSIs). The goal is to explore whether these features could stratify patients' survival. This study comprised of 127 patients diagnosed with ICC after surgery, where 78 cases were randomly chosen as the modeling set, and the rest of the 49 cases were testing set. Deep learning-based models were developed for tissue segmentation and lymphocyte detection in the WSIs. A total of 107-dimensional features, including different type of graph features on the WSIs were extracted by exploring the histomorphological patterns of these identified tumor tissue and lymphocytes. The top 3 discriminative features were chosen with the mRMR algorithm via 5-fold cross-validation to predict the patient's survival. The model's performance was evaluated on the independent testing set, which achieved an AUC of 0.6818 and the log-rank test p-value of 0.03. The Cox multivariable test was used to control the TNM staging, γ-Glutamytransferase, and the Peritumoral Glisson's Sheath Invasion. It showed that our model could independently predict survival risk with a p-value of 0.048 and HR (95% confidence interval) of 2.90 (1.01–8.32). These results indicated that the composition in tissue-level and global arrangement of lymphocytes in the cell-level could distinguish ICC patients' survival risk.

PENDIDIKAN KESEHATAN DETEKSI DINI KANKER PAYUDARA DENGAN MELAKUKAN ‘SADARI’ DI TP PKK DESA RAMBAH TENGAH HILIR KECAMATAN RAMBAH KABUPATEN ROKAN HULU

Breast cancer is a malignancy in breast tissue that can originate from the ductal epithelium or its lobules. Based on Pathological Based Registration in Indonesia, Breast cancer ranks first with a relative frequency of 18.6%. In Indonesia, more than 80% of cases are found to be in an advanced stage, where treatment is difficult. Therefore, it is necessary to understand the prevention efforts as the first step in preventing breast cancer before entering the final phase which is difficult to treat. Breast self-examination or SADARI until now is a way of early detection of breast cancer which is quite effective. SADARI is easy to do and can be applied to all ages, both teenagers and adult women. The purpose of this activity is to disseminate health education through BSE in TP PKK Desa Rambah Tengah Hilir Village, Rambah District for early detection of breast cancer in order to increase public knowledge in prevention and early treatment. The result of this service is the increasing knowledge of women of childbearing age in making efforts for breast cancer self-detection. In the end, it is hoped that activities like this can be carried out in other villages in Rambah District to further increase public knowledge in detecting breast cancer which is the most cancer in women.

Effects of Lipolysis-Stimulated Lipoprotein Receptor on Tight Junctions of Pancreatic Ductal Epithelial Cells in Hypertriglyceridemic Acute Pancreatitis.

Objective We investigated the effects of lipolysis-stimulated lipoprotein receptor (LSR) on the tight junctions (TJs) of pancreatic ductal epithelial cells (PDECs) in hypertriglyceridemic acute pancreatitis (HTGAP). Methods Sprague-Dawley rats were fed standard rat chow or a high-fat diet and injected with sodium taurocholate to obtain normal and HTGAP rats, respectively. Serum triglyceride (TG) levels, pathological changes, TJ proteins in the pancreas, and TJ ultrastructure of PDECs were assessed. LSR overexpression (OE) and knockdown (KD) HPDE6-C7 models were designed and cultured in a high-fat environment. Protein levels were quantified by Western blotting. Cell monolayer permeability was detected using FITC-Dextran. Results Serum TG concentration and pancreatic scores were higher in the HTGAP group than in the normal group. Among the TJ proteins, LSR protein expression was significantly lower in the HTGAP group than in the acute pancreatitis (AP) group. Tricellulin (TRIC) expression in the pancreatic ductal epithelia was higher in the HTGAP group than in the AP group. The HTGAP group had lower TJ protein levels, wider intercellular space, and widespread cellular necrosis with disappearance of cell junction structures. In the cell study, TJ proteins were downregulated and the cellular barrier was impaired by palmitic acid (PA), which was reversed by LSR-OE, whereas LSR-KD downregulated the TJ proteins and aggravated PA-induced cellular barrier impairment. Conclusions Hypertriglyceridemia downregulates the TJ proteins in PDECs, which may impair the pancreatic ductal mucosal barrier function. LSR regulation can change the effects of HTG on cellular barrier function by upregulating the TJ proteins.

Digital image analysis of intraepithelial B-lymphocytes to assess lymphoepithelial lesions in salivary glands of Sjögren's syndrome patients.

Salivary glands of primary SS (pSS) patients characteristically harbour periductal infiltrates, in which lymphoepithelial lesions (LELs) can develop. LELs are composed of hyperplastic ductal epithelium with infiltrating lymphocytes and may assist in the challenging diagnostic process of pSS. As manual identification of LELs remains difficult, we aimed to identify LELs by using an objective digital image analysis (DIA) algorithm that detects intraepithelial lymphocytes. A virtual triple-staining technique developed for this study was used to count intraepithelial lymphocytes in consecutive slides stained for CD3 (T-lymphocytes), high-molecular-weight cytokeratin (hmwCK) (striated ducts) and CD20 (B-lymphocytes) in labial and parotid gland biopsies in a diagnostic cohort of 109 sicca patients. Patients were classified as having pSS or non-SS according to the ACR-EULAR classification criteria. T-lymphocytes were detected in almost all analysed ducts of pSS and non-SS sicca patients, whereas intraepithelial B-lymphocytes were present in 59-68% of labial and parotid gland biopsies of pSS patients, against only 2-3% of patients classified as non-SS. Intraepithelial B-lymphocytes were found in almost all striated ducts with hyperplasia (LELs). Remarkably, ∼25% of analysed striated ducts without hyperplasia of pSS patients also contained B-lymphocytes (precursor-LELs). Furthermore, presence of intraepithelial B-lymphocytes was associated with clinical parameters of pSS (i.e. serology). The presence of intraepithelial B-lymphocytes in salivary gland biopsies of sicca patients is a clear indicator of pSS and can be used as an objective alternative to LEL scoring. Therefore, identification of B-lymphocyte-containing ducts should be added to the diagnostic histopathological work-up of patients suspected of pSS.

Ibuprofen and diclofenac differentially affect cell viability, apoptosis and morphology changes of human cholangiocarcinoma cell lines

ObjectivesCholangiocarcinoma is a malignant biliary epithelial duct neoplasm caused by chronic inflammation after liver fluke infection. It is a major public health concern in the Greater Mekong sub-region in northeast Thailand. Herein, the effects of the non-steroidal anti-inflammatory drugs (NSAIDs) ibuprofen and diclofenac on the cell proliferation activity of the human cholangiocarcinoma cell lines KKU-M139 and KKU-213B were studied. MethodsCell viability was assessed with MTT assays. Inverted phase-contrast light microscopy, scanning electron microscopy and transmission electron microscopy were used to investigate the cells’ morphological alterations. Caspase 3/7 and Annexin V/PI were detected with a multimode microplate reader. ResultsIbuprofen and diclofenac decreased viability in both cell lines, and ibuprofen-treated cells exhibited reversible cell injury. In both KKU-M139 and KKU-213B cell lines, the diclofenac-treated cells had the greatest injury. The cells exhibited features of irreversible cell injury. In addition, caspase 3/7 and Annexin V/PI detection revealed early cell apoptotic characteristics. ConclusionThese findings suggest that NSAIDs may potentially suppress cell viability. Ibuprofen and diclofenac both induced morphological changes and apoptosis.

The Role of Surgical Approaches in the Multi-Modal Management of Adult Craniopharyngiomas.

Craniopharyngiomas are rare, benign primary brain tumors that arise from remnants of the craniopharyngeal duct epithelium within the sellar and suprasellar region. Despite their benign biology, they may cause significant morbidity, secondary to involvement of nearby eloquent neural structures, such as the pituitary gland, hypothalamus, and optic apparatus. Historically, aggressive surgical resection was the treatment goal to minimize risk of tumor recurrence via open transcranial midline, anterolateral, and lateral approaches, but could lead to clinical sequela of visual, endocrine, and hypothalamic dysfunction. However, recent advances in the endoscopic endonasal approach over the last decade have mostly supplanted transcranial surgery as the optimal surgical approach for these tumors. With viable options for adjuvant radiation therapy, targeted medical treatment, and alternative minimally invasive surgical approaches, the management paradigm for craniopharyngiomas has shifted from aggressive open resection to more minimally invasive but maximally safe resection, emphasizing quality of life issues, particularly in regards to visual, endocrine, and hypothalamic function. This review provides an update on current multi-modal approaches for craniopharyngiomas, highlighting the modern surgical treatment paradigm for this disease entity.

COVID-19–Related Chronic Bilateral Dacryoadenitis

Pathological features of ophthalmic aftereffects of COVID-19 are important for new insight in treating patients. To examine the expression of SARS-CoV-2 nucleocapsid protein and angiotensin-converting enzyme 2 (ACE2) in lacrimal gland tissues of a patient with COVID-19 and a patient without COVID-19. In this retrospective case-control study, the case of a 35-year-old woman with positive test results for SARS-CoV-2 who had had lacrimal gland enlargements for 6 months was analyzed. A 43-year-old woman without COVID-19 who had idiopathic chronic bilateral dacryoadenitis served as a negative control. Histopathology and immunohistochemistry with anti-SARS-CoV-2 nucleocapsid protein and ACE2 in the lacrimal glands. Both patients were Japanese women aged 35 years (case) and 43 years (control). Histopathologic findings in the patient with COVID-19 demonstrated marked inflammatory cell infiltration, lymphoid follicles, and germinal center formation in the lacrimal gland. The inflammation was mainly made up of lymphocytes and plasma cells with several polymorphonuclear leukocytes, where the lacrimal glands were atrophic. Of note, a number of lacrimal gland ducts markedly contained eosinophilic materials in the lumens, which indicated glandular damage. Immunoreactivity for SARS-CoV-2 nucleocapsid protein was noted in the inflammatory cells around the lacrimal gland ductal epithelia. In addition, strong ACE2 expression was noted in the lacrimal gland. In the patient without COVID-19, marked inflammation was noted in the lacrimal gland; however, there were no eosinophilic material deposits in the ductal lumens. SARS-CoV-2 nucleocapsid protein immunoreactivity was not observed, whereas ACE2 was expressed in the lacrimal glands. In this case-control study, expression of ACE2 indicated that the lacrimal gland could be a target organ for SARS-CoV-2 to adhere to. Chronic bilateral dacryoadenitis in the patient with COVID-19 showed SARS-CoV-2-positive inflammatory cells with glandular damage, which might be a COVID-19-associated ophthalmic aftereffect.

ACE2 and TMPRSS2 immunolocalization and oral manifestations of COVID-19.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) entry into the host cells depends on the expression of angiotensin-converting enzyme 2 (ACE2) and transmembrane protease serine 2 (TMPRSS2). We investigated the distribution of ACE2- and TMPRSS2-expressing cells in various oral tissues to identify the underlying mechanism of oral manifestations in patients with coronavirus disease 2019. We analyzed the expression patterns of ACE2 and TMPRSS2 in the oral mucosa (tongue, palate, and buccal mucosa), trigeminal ganglion, vessels, and salivary glands of 9 Sprague-Dawley rats using immunohistochemistry and immunofluorescence. ACE2 and TMPRSS2 were strongly expressed in the intermediate layer of the squamous epithelia of tongue papillae and buccal mucosa. ACE2- and TMPRSS2-positive cells were observed in the taste buds of the tongue. Additionally, ACE2 and TMPRSS2 were co-expressed in the ductal epithelium and acinar cells of salivary glands. Furthermore, both ACE2 and TMPRSS2 were stained in the neuronal cell body of trigeminal ganglia, but not in Schwann cells. Moreover, ACE2 and TMPRSS2 were expressed in capillaries, but not in venules/arterioles. SARS-CoV-2 can spread the suprabasal area of squamous epithelia of the oral mucosa, invades taste bud, trigeminal nerve, parotid gland, and microvessel, resulting in oral manifestations.

Mitigation of portal fibrosis and cholestatic liver disease in ANKS6-deficient livers by macrophage depletion.

Congenital hepatic fibrosis (CHF) is a developmental liver disease that is caused by mutations in genes that encode ciliary proteins and is characterized by bile duct dysplasia and portal fibrosis. Recent work has demonstrated that mutations in ANKS6 can cause CHF due to its role in bile duct development. Here, we report a novel ANKS6 mutation, which was identified in an infant presenting with neonatal jaundice due to underlying biliary abnormalities and liver fibrosis. Molecular analysis revealed that ANKS6 liver pathology is associated with the infiltration of inflammatory macrophages to the periportal fibrotic tissue and ductal epithelium. To further investigate the role of macrophages in CHF pathophysiology, we generated a novel liver-specific Anks6 knockout mouse model. The mutant mice develop biliary abnormalities and rapidly progressing periportal fibrosis reminiscent of human CHF. The development of portal fibrosis in Anks6 KO mice coincided with the accumulation of inflammatory monocytes and macrophages in the mutant liver. Gene expression and flow cytometric analysis demonstrated the preponderance of M1- over M2-like macrophages at the onset of fibrosis. A critical role for macrophages in promoting peribiliary fibrosis was demonstrated by depleting the macrophages with clodronate liposomes which effectively reduced inflammatory gene expression and fibrosis, and ameliorated tissue histology and biliary function in Anks6 KO livers. Together, this study demonstrates that macrophages play an important role in the initiation of liver fibrosis in ANKS6-deficient livers and their therapeutic elimination may provide an avenue to mitigate CHF in patients.

Hyperresponsive cytosolic DNA-sensing pathway in monocytes from primary Sjögren's syndrome.

ObjectivesCytosolic DNA-sensing pathway stimulation prompts type I IFN (IFN-I) production, but its role in systemic IFN-I pathway activation in primary SS (pSS) is poorly studied. Here we investigate the responsiveness of pSS monocytes and plasmacytoid dendritic cells (pDCs) to stimulator of interferon genes (STING) activation in relation to systemic IFN-I pathway activation and compare this with SLE.MethodsExpression of DNA-sensing receptors cGAS, IFI16, ZBP-1 and DDX41, signalling molecules STING, TBK1 and IRF3, positive and negative STING regulators, and IFN-I-stimulated genes MxA, IFI44, IFI44L, IFIT1 and IFIT3 was analysed in whole blood, CD14+ monocytes, pDCs, and salivary glands by RT-PCR, monocyte RNA sequencing data, flow cytometry and immunohistochemical staining. Peripheral blood mononuclear cells (PBMCs) from pSS, SLE and healthy controls (HCs) were stimulated with STING agonist 2′3′-cGAMP. STING phosphorylation (pSTING) and intracellular IFNα were evaluated using flow cytometry.ResultsSTING activation induced a significantly higher proportion of IFNα-producing monocytes, but not pDCs, in both IFN-low and IFN-high pSS compared with HC PBMCs. Additionally, a trend towards more pSTING+ monocytes was observed in pSS and SLE, most pronounced in IFN-high patients. Positive STING regulators TRIM38, TRIM56, USP18 and SENP7 were significantly higher expression in pSS than HC monocytes, while the dual-function STING regulator RNF26 was downregulated in pSS monocytes. STING was expressed in mononuclear infiltrates and ductal epithelium in pSS salivary glands. STING stimulation induced pSTING and IFNα in pSS and SLE pDCs.ConclusionpSS monocytes and pDCs are hyperresponsive to stimulation of the STING pathway, which was not restricted to patients with IFN-I pathway activation.

Nasal mucoepidermoid carcinoma after radiotherapy: Case report

Abstract Introduction Mucoepidermoid carcinoma (MEC) is a tumor originated from the epithelium of the glandular excretory ducts and has highly variable biological potential. It is the most prevalent cancer of the salivary glands. The present report aims to describe a case of nasal mucoepidermoid carcinoma that developed after adjuvant radiotherapy (RT) treatment of a recurrent pituitary macroadenoma. Case Report Male patient, 62 years old, presented with recurrent nasal epistaxis on the right, associated with intense pulsatile headache, visual analogical scale (VAS) 10/10, with improvement only with the use of opioids and morphine. After undergoing oncological screening and study by imaging exams, the presence of an expansive seal lesion with suprasellar extension was seen, involving the medial wall of the cavernous segment of the right carotid artery and the anterior cerebral artery, as well as the presence of a new expansive lesion in the right nasal cavity, with ethmoid bone invasion superiorly and medial orbit wall invasion laterally, compressing the ipsilateral optic nerve canal. Discussion Sinonasal neoplasms represent a small portion of all malignancies of the upper aerodigestive tract, accounting for < 5% of these neoplasms. The development of MEC involves risk factors such as occupational issues, history of trauma and surgery involving the nasal area, and radiation exposure, as in previous RT. Conclusion Mucoepidermoid carcinoma is an uncommon neoplasia and can be associated with RT treatment, as used in cases of recurrent pituitary macroadenoma. In general, surgical resection to obtain free margins of neoplastic tissue is the aimed treatment, seeking better prognosis.

Delineation of biliary epithelial cell dynamics in maternal liver during pregnancy.

In pregnant mice, the maternal liver expands drastically during gestation, which is believed to be essential to accommodate various metabolic demands caused by physiological changes and fetal growth. Although hepatocyte proliferation and hypertrophy have been reported, little is known about the dynamics of biliary epithelial cells (BECs), which comprise the bile duct epithelium in the liver. Here, we show that BECs transiently proliferate during the early stage of gestation. Lineage tracing revealed that BEC progeny were retained in the bile duct epithelium and did not differentiate into hepatocytes, indicating BEC self-replication during pregnancy. RNA-sequencing analysis of BECs identified their early pregnancy-signature transcriptomes, which highlighted Yes-associated protein (YAP) signaling-related genes. Nuclear accumulation of YAP was enhanced in BECs during pregnancy but was barely detectable in hepatocytes. In addition, the pharmacological inhibition of YAP attenuated BEC proliferation and liver weight gain during pregnancy. Our results delineate the proliferation and transcriptomic dynamics of BECs during pregnancy and suggest the relevance of YAP-mediated signals.

Specific features of the pathology of the respiratory system in SARS-CoV-2 (Coronaviridae: Coronavirinae: Betacoronavirus: Sarbecovirus) infected Syrian hamsters (Mesocricetus auratus)

Verification of histological changes in respiratory system using Syrian (golden) hamsters (Mesocricetus auratus) as experimental model is an important task for preclinical studies of drugs intended for prevention and treatment of the novel coronavirus infection COVID-19.The aim of this work was to study pathological changes of pulmonary tissue in SARS-CoV-2 (Coronaviridae: Coronavirinae: Betacoronavirus; Sarbecovirus) experimental infection in Syrian hamsters. Male Syrian hamsters weighting 80-100 g were infected by intranasal administration of culture SARS-CoV-2 at dose 4 × 104 TCID50/ml (TCID is tissue culture infectious dose). Animals were euthanatized on 3, 7 and 14 days after infection, with gravimetric registration. The viral load in lungs was measured using the polymerase chain reaction (PCR). Right lung and trachea tissues were stained with hematoxylin-eosin and according to Mallory. The highest viral replicative activity in lungs was determined 3 days after the infection. After 7 days, on a background of the decrease of the viral load in lungs, a pathologically significant increase of the organ's gravimetric parameters was observed. Within 3 to 14 days post-infection, the lung histologic pattern had been showing the development of inflammation with a succession of infiltrative-proliferative, edematousmacrophagal and fibroblastic changes. It was found that initial changes in respiratory epithelium can proceed without paranecrotic interstitial inflammation, while in the formation of multiple lung parenchyma lesions, damage to the epithelium of bronchioles and acinar ducts can be secondary. The appearance of epithelioid large-cell metaplastic epithelium, forming pseudoacinar structures, was noted as a pathomorphological feature specific to SARS-CoV-2 infection in Syrian hamsters. As a result of the study, the specific features of the pathology of the respiratory system in SARSCoV-2 infected Syrian hamsters were described. These findings are of practical importance as reference data that can be used for preclinical studies to assess the effectiveness of vaccines and potential drugs.

The role of leptin in primary Sjögren syndrome: a clinical and histopathologicalassessment study

The underlying pathogenetic mechanisms of Sjögren syndrome (SjS) have not been elucidated yet [1]. Leptin is a glycosylated peptide structurally similar to some cytokines [2]. Leptin has proinflammatory effects via various mechanisms. It has been previously reported that serum leptin levels increase in some autoimmune diseases and that leptin levels are also associated with disease activity [3, 4]. In addition, leptin and its receptor in the salivary glands stimulate epithelial proliferation and modulate the local immune system with their autocrine effects [5]. In this study, we assessed the presence of leptin in minor salivary glands (MSG) of the patients with and without SjS. Additionally, we evaluated the association between leptin staining intensity and disease activity among patients with SjS. We included patients who underwent MSG biopsy with the suspicion of SjS between 2013 and 2014. Among these patients, those who met the 2012 American College of Rheumatology Sjögren's syndrome classification criteria (6) and had at least 50 mononuclear cell infiltrates in a 4 mm2 minor salivary gland section were classified as the SjS group. The rest of the patients who underwent MSG biopsy but did not meet the histopathological and/or clinical criteria for SjS were considered the control group. Leptin staining was assessed by immunohistochemistry (Bio-Rad AbDSerotec, Oxford, UK) in the stroma, acinar and ductal epithelium, and staining intensities were graded by a semiquantitative method; no staining (score 0), mild (score 1), moderate (score 2), and diffuse (score 3). Definitions of leptin staining intensities were described as mild; focal staining of few cells in one focus, moderate; mild staining in more than one focus, diffuse; diffuse staining in all assessed sections (Figure 1). The sum of leptin staining in all areas was defined as total leptin staining. The SjS group was divided into 3 groups according to the focus scores (FS) (FS1: FS = 1, FS2: FS = 2, FS3: FS ≥ 3). In addition, patients' disease activities were assessed by EULAR Sjogren's syndrome disease activity index (ESSDAI) [7]. Visual (histograms, probability plots) and analytical methods (Kolmogorov-Smirnov test) were used to analyze the variables' distribution and select the test method. Descriptive analyses were presented as mean or median according to their distribution patterns. A chi-square test was used for univariate analyses to identify variables associated with leptin staining and clinical features. Patients' disease activities were assessed by ESSDAI [9]. The correlations between ESSDAI scores and leptin staining intensities were analyzed using Spearman's test. A p value of 66%, in FS3 group: 100%) areas were higher in the patients with FS3. There was a positive correlation between ESSDAI and leptin staining scores in the acinar area (p < 0.05; r = 0.406) regardless of focus scores. However, the correlations between disease activity and leptin staining in ductal areas (p: 0.1; r: 0.334), in stromal areas (p: 0.1; r: 0.184), and total leptin (p: 0.2; r: 0.268) staining was not significant. We identified that majority of the patients in both groups had leptin staining in acinar (92% and 84%, respectively), ductal (94% and 84%, respectively), and stromal areas (96% and 100%, respectively) of their MSGs. Patients with higher FSs had significantly more intense leptin staining patterns. The possible explanations for the similar staining pattern in patients with an FS of ≤2 was the insufficient number of patients (β error) or absolute indifference. In contrast, a study by Erbasan F et al. argued that leptin has no role in primary SjS due to similar leptin staining patterns among patients with SjS and healthy controls. However, they did not compare patients according to the ESSDAI scores [8]. Thus, we still need more robust data to identify whether leptin has an inciting or prognostic role in pSS. Limitations of our study were the lack of some clinical characteristics and metabolic parameters which may affect leptin metabolism and lack of leptin receptor staining patterns in MSG biopsies, which together with leptin staining could better define the role of leptin. Lack of power analysis was another limitation. In conclusion, leptin staining properties were similar in both SjS and non-SjS groups. Additionally, the only clinical significance of leptin density in the MSG was in the acinar region. The authors have no financial or competing interests. All authors declare no conflict of interest We received funding support from Kartal Dr. Lutfi Kirdar City Hospital research assistance department. Authors contributed to writing this study and have approved the final version. Ethical approval: This article does not contain any studies with human participants performed by any of the authors. The study protocol was approved by the Local Ethics Committee of Dr Lutfi Kirdar Kartal City Hospital.

Mammalian uterine morphogenesis and variations.

In eutherian and marsupial mammals, the site of embryo implantation and gestation is the uterus. Uterine morphologies vary between mammalian species. For example, laboratory mice have a bipartite uterus with two uterine horns and a single cervix, whereas humans have a simplex uterus with a single chamber and single cervix. The precursor tissue of the uterus, oviducts, and upper vagina is the Müllerian duct epithelium and its adjacent mesenchyme. Morphological variation between species is established during embryogenesis by species-specific differences in Müllerian duct fusion at the midline, growth, and differentiation. In humans, alterations in Müllerian duct development can lead to variations in uterine morphology that correlate with increased risks of miscarriage and infertility. Here we review the developmental genetic factors that regulate Müllerian duct development, uterine morphogenesis, and human uterine variation.

Immunohistochemical Analysis in Salivary Duct Carcinoma of the Upper Labial Mucosa: A Rare Case Report

Salivary Duct Carcinoma (SDC) is a rare typically high grade, aggressive malignancy arising from the ductal epithelium of salivary glands characterised by ductal formations and central necrosis having pathomorphological resemblance to ductal breast carcinoma. Parotid and submandibular salivary glands are the most commonly affected while very few cases involving the minor salivary glands of the palate, labial mucosa, floor of mouth have been reported. This high grade malignancy must be treated aggressively by complete local excision with radical neck dissection and postoperative radiation therapy seems to offer maximum benefit for the patients. This article highlights a case of a 80-year-old male patient having an intra oral ulcer over a swelling which was since 5 years on the left side of upper lip with left submandibular lymph nodes enlargement is reported. An immunohistochemical analysis of the biopsy specimen was carried out which concluded a SDC. The swelling was surgically excised followed by postoperative radiotherapy and adjuvant chemotherapy. The patient was on a regular follow-up for two and a half years without any local recurrence or distant metastasis.

Multidimensional Fluorescence Imaging of Embryonic and Postnatal Mammary Gland Development.

Multidimensional fluorescence imaging represents a powerful approach for studying the dynamic cellular processes underpinning the development, function, and maintenance of the mammary gland. Here, we describe key multidimensional imaging strategies that enable visualization of mammary branching morphogenesis and epithelial cell fate dynamics during postnatal and embryonic mammary gland development. These include 4-dimensional intravital microscopy and ex vivo imaging of embryonic mammary cultures, in addition to methods that facilitate 3-dimensional imaging of the ductal epithelium at single-cell resolution within its native stroma. Collectively, these approaches provide a window into mammary developmental dynamics, and the perturbations underlying tissue dysfunction and disease.

Combined OPCML and AXL Expression as a Prognostic Marker and OPCML Enhances AXL Inhibitor in Cholangiocarcinoma.

Cholangiocarcinoma (CCA) is a type of liver cancer originating from bile duct epithelium which has an unfavorable prognosis. Therefore, novel prognostic markers and effective therapeutic regimens are required. Opioid-binding protein/cell adhesion molecule-like (OPCML) is a tumor-suppressor protein that suppresses CCA cell proliferation via AXL receptor tyrosine kinase/signal transducer and activator of transcription 3 (AXL/STAT3) inactivation. However, this association in clinical samples remains unknown. We aimed to determine OPCML and AXL expression and investigate their association with clinicopathological features in patients with CCA. In addition, we also addressed whether OPCML enhanced the sensitivity of CCA cells to AXL inhibitor R428 in vitro. The expression of OPCML and AXL was determined by immunohistochemistry in 90 CCA tissue samples. The study of CCA cell line sensitivity to R428 was performed by cell viability assay. The expression of OPCML was significantly lower while AXL expression was substantially higher in CCA than in adjacent normal tissue (p<0.001). Furthermore, high AXL expression was significantly associated with lymph node metastasis (p=0.035). Interestingly, patients with combined low OPCML/high AXL expression had significantly shorter overall survival (p=0.007). OPCML enhanced the effect of AXL inhibitor R428 in AXL-expressing CCA cell lines. Combined expression of OPCML and AXL shows potential value as a prognostic marker and OPCML as an agent enhancing the effect of R428 may contribute to better prognosis for patients with CCA.