Direct Evidence of Active SARS-CoV-2 Replication in the Intestine

Abstract Background Currently, there is no direct evidence to prove the active replication of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in the intestinal tract and relevant pathological changes in the colon and rectum. We investigated the presence of virions and pathological changes in surgical rectal tissues of a patient with clinically confirmed coronavirus disease 2019 (COVID-19) with rectal adenocarcinoma. Methods The clinical data were collected during hospitalization and follow-up of this patient. Quantitative reverse transcriptase–polymerasechain reaction (RT-PCR) was performed on the rectal tissue specimens obtained from surgical resection, succus entericus and intestinal mucosa of ileostomy, and rectal mucosa during follow-up after recovery. Ultrathin sections of surgical samples were observed for SARS-CoV-2 virions using electron microscopy. Histopathological examination was performed using hematoxylin-eosin stain. Immunohistochemical analysis and immunofluorescence were carried out on rectal tissues to evaluate the distribution of SARS-CoV-2 antigen and immune cell infiltrations. Results The patient had fever and cough on day 3 postoperatively, was diagnosed with COVID-19 on day 7, and was discharged from the hospital on day 41. RNA of SARS-CoV-2 was detected in surgically resected rectal specimens but not in samples collected 37 days after discharge. Notably, coincident with rectal tissues of surgical specimens testing nucleic acid positive for SARS-CoV-2, typical coronavirus virions in rectal tissue were observed under electron microscopy. Moreover, abundant lymphocytes and macrophages (some were SARS-CoV-2 positive) infiltrating the lamina propria were found with no significant mucosal damage. Conclusions We first report the direct evidence of active SARS-CoV-2 replication in a patient’s rectum during the incubation period, which might explain SARS-CoV-2 fecal–oral transmission.

Download Full-text

Metachronous Bilateral Isolated Adrenal Metastasis from Rectal Adenocarcinoma: A Case Report

Case Reports in Gastrointestinal Medicine ◽

10.1155/2014/516403 ◽

2014 ◽

Vol 2014 ◽

pp. 1-4 ◽

Cited By ~ 2

Author(s):

H. Jabir ◽

N. Tawfiq ◽

M. Moukhlissi ◽

M. Akssim ◽

A. Guensi ◽

...

Keyword(s):

Colorectal Cancer ◽

Computed Tomography ◽

Rectal Cancer ◽

Histopathological Examination ◽

Rectal Adenocarcinoma ◽

Adrenal Metastasis ◽

Computed Tomography Scan ◽

Serum Carcinoembryonic Antigen ◽

Tomography Scan

We report a case of adrenal metastasis from colorectal cancer in a 54-year-old woman. Nine months after resection for advanced rectal carcinoma, a computed tomography scan revealed bilateral adrenal metastasis. The level of serum carcinoembryonic antigen was normal. A bilateral adrenalectomy was performed after chemotherapy. Histopathological examination showed adenocarcinoma, compatible with metastasis from the rectal cancer. Adrenal metastasis should be considered in the patients’ follow-up for colorectal cancer.

Download Full-text

Diagnostic Utility of AMCAR and p63 Cocktail Antibody in the Benign and Malignant Lesions of Prostate

Annals of Pathology and Laboratory Medicine ◽

10.21276/apalm.2800 ◽

2020 ◽

Vol 7 (11) ◽

pp. A531-537

Author(s):

Sujitha Chougani ◽

Sunandalakshmi G V ◽

Durga Kharidehal ◽

Ravi Sankar V ◽

Santhi Vissa

Keyword(s):

Histopathological Examination ◽

Immunohistochemical Analysis ◽

Cost Effective ◽

Basal Cells ◽

Variable Intensity ◽

Atypical Small Acinar Proliferation ◽

Positive Immunostaining ◽

Malignant Lesions ◽

Made In

Background: Histopathological examination of prostatic specimen is gold standard for the diagnosis of prostate cancer. Current study evaluates the expression of AMACR and p63 in the prostate lesions using AMACR and p63 cocktail. Materials and Method: Total of 180 cases were collected and Haematoxylin and Eosin staining performed followed by immunohistochemical analysis using AMACR and p63 antibody. Result: Out of 180 cases, Benign Prostatic Hyperplasia is the most common lesion noted in about 120 cases. In this study, the predominant population was in the 6th to 7th decade of age. Most of the patients presented with difficulty in micturition. Immunohistochemistry revealed that p63 expression is positive in all normal basal cells, 118 cases (98.33%) were negative for AMACR and only 2 cases were showing focal and weak AMACR immunoreactivity. AMACR was positive in all the 6 HGPIN cases with variable intensity. Out of 5 LGPIN cases AMACR was positive in 3 cases with low intensity, remaining 2 cases shows AMACR negative. p63 is positive in all 11 PIN cases (LGPIN & HGPIN) showing discontinuous staining pattern. All 22 cases of Prostatic adenocarcinomas were negative for p63 and all cases expressed positive immunostaining with AMACR. A diagnosis of adenocarcinoma was made in 48% of atypical cases. Cases which were negative for both AMACR and p63 were diagnosed as Atypical Small Acinar Proliferation, for which further follow-up is required. Conclusion: AMACR/p63 Cocktail antibody is very much useful as it saves time, tissue and is cost-effective.

Download Full-text

A Peripheral Ameloblastic Fibro-Odontoma in a 3-Year-Old Girl: Case Report, Immunohistochemical Analysis, and Literature Review

Case Reports in Dentistry ◽

10.1155/2014/321671 ◽

2014 ◽

Vol 2014 ◽

pp. 1-6 ◽

Cited By ~ 1

Author(s):

Yi-Chun Lin ◽

Hsiu-Ming Hsu ◽

Chiang-Shin Liu ◽

Kuo Yuan

Keyword(s):

Histopathological Examination ◽

Correct Diagnosis ◽

Immunohistochemical Analysis ◽

Clinical Appearance ◽

Jaw Bones ◽

Tissue Sections ◽

Odontogenic Epithelium ◽

Histological Characteristics ◽

Children And Young Adults

Ameloblastic fibro-odontoma (AFO) predominantly occurs in the jaw bones of children and young adults. Extraosseous AFO is extremely rare. We describe a peripheral ameloblastic fibro-odontoma in the maxillary gingiva of a 3-year-old girl. The clinical appearance resembled fiery red reactive gingival lesions. The histopathological examination of the excised lesion showed small islands and cords of odontogenic epithelium with cellular myxoid stroma in the subepithelial tissue. The mass contained calcified material and an enamel-like deposit. Many small blood vessels appeared in the connective tissue surrounding the odontogenic epithelium. The immunohistochemical assays showed strong reactivity for amelogenin,β-catenin, CD44, and CD31 in the tissue sections. There was no recurrence after the 1-year follow-up. Because this lesion clinically resembles other nonneoplastic lesions and is very rare in gingiva, establishing a correct diagnosis is achieved only based on specific histological characteristics. Conservative excision of the tumor is the treatment of choice.

Download Full-text

Responses of Cattle to Gastrointestinal Colonization by Escherichia coli O157:H7

Infection and Immunity ◽

10.1128/iai.01223-07 ◽

2008 ◽

Vol 76 (11) ◽

pp. 5366-5372 ◽

Cited By ~ 32

Author(s):

Pablo Nart ◽

Stuart W. Naylor ◽

John F. Huntley ◽

Iain J. McKendrick ◽

David L. Gally ◽

...

Keyword(s):

Escherichia Coli ◽

Bacterial Colonization ◽

Immunoglobulin A ◽

Rectal Mucosa ◽

Mucosal Damage ◽

Pathological Changes ◽

E Coli ◽

Transmission Electron ◽

Gastrointestinal Colonization ◽

Negative Controls

ABSTRACT Recent research has established that the terminal rectum is the predominant colonization site of enterohemorrhagic Escherichia coli O157:H7 in cattle. The main aim of the present work was to investigate pathological changes and associated immune responses at this site in animals colonized with E. coli O157:H7. Tissue and gastrointestinal samples from a total of 22 weaned Holstein-cross calves challenged with E. coli O157:H7 were analyzed for bacterial colonization and pathology. Five unexposed age-matched calves were used as comparative negative controls. E. coli O157:H7 bacteria induced histopathological alterations of the rectal mucosa with enterocyte remodeling. This was often associated with removal of the colonized epithelial layer. Immunogold labeling and transmission electron microscopy (TEM) showed E. coli O157 bacteria on pedestals, as part of attaching and effacing lesions. These pathological changes induced a local infiltration of neutrophils that was quantified as larger in infected animals. Rectal mucosal immunoglobulin A responses were detected against the E. coli O157:H7 antigen. This work presents evidence that E. coli O157:H7 is not a commensal bacteria in the bovine host and that the mucosal damage produced by E. coli O157:H7 colonization of the terminal rectum induces a quantifiable innate immune response and production of specific mucosal antibodies.

Download Full-text

Electron Microscopy of Human Gallstones

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100065523 ◽

1971 ◽

Vol 29 ◽

pp. 296-297

Author(s):

C. Wolpers ◽

R. Blaschke

Keyword(s):

Electron Microscopy ◽

Room Temperature ◽

Bile Pigment ◽

X Ray Diffraction ◽

Triclinic Crystal System ◽

X Ray ◽

Follow Up Study ◽

Infra Red ◽

Main Components

Scanning microscopy was used to study the surface of human gallstones and the surface of fractures. The specimens were obtained by operation, washed with water, dried at room temperature and shadowcasted with carbon and aluminum. Most of the specimens belong to patients from a series of X-ray follow-up study, examined during the last twenty years. So it was possible to evaluate approximately the age of these gallstones and to get information on the intensity of growing and solving.Cholesterol, a group of bile pigment substances and different salts of calcium, are the main components of human gallstones. By X-ray diffraction technique, infra-red spectroscopy and by chemical analysis it was demonstrated that all three components can be found in any gallstone. In the presence of water cholesterol crystallizes in pane-like plates of the triclinic crystal system.

Download Full-text

Electron Microscopy in the diagnosis of lysosomal storage diseases

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100156316 ◽

1989 ◽

Vol 47 ◽

pp. 866-867

Author(s):

Carole Vogler ◽

Harvey S. Rosenberg

Keyword(s):

Electron Microscopy ◽

Lysosomal Enzyme ◽

Rectal Mucosa ◽

Lysosomal Storage Diseases ◽

Cell Types ◽

Lysosomal Storage ◽

Storage Material ◽

Ultrastructural Examination ◽

Blood Leukocytes ◽

Storage Diseases

Diagnostic procedures for evaluation of patients with lysosomal storage diseases (LSD) seek to identify a deficiency of a responsible lysosomal enzyme or accumulation of a substance that requires the missing enzyme for degradation. Most patients with LSD have progressive neurological degeneration and may have a variety of musculoskeletal and visceral abnormalities. In the LSD, the abnormally diminished lysosomal enzyme results in accumulation of unmetabolized catabolites in distended lysosomes. Because of the subcellular morphology and size of lysosomes, electron microscopy is an ideal tool to study tissue from patients with suspected LSD. In patients with LSD all cells lack the specific lysosomal enzyme but the distribution of storage material is dependent on the extent of catabolism of the substrate in each cell type under normal circumstances. Lysosmal storages diseases affect many cell types and tissues. Storage material though does not accumulate in all tissues and cell types and may be different biochemically and morphologically in different tissues.Conjunctiva, skin, rectal mucosa and peripheral blood leukocytes may show ultrastructural evidence of lysosomal storage even in the absence of clinical findings and thus any of these tissues can be used for ultrastructural examination in the diagnostic evaluation of patients with suspected LSD. Biopsy of skin and conjunctiva are easily obtained and provide multiple cell types including endothelium, epithelium, fibroblasts and nerves for ultrastructural study. Fibroblasts from skin and conjunctiva can also be utilized for the initiation of tissue cultures for chemical assays. Brain biopsy has been largely replaced by biopsy of more readily obtained tissue and by biochemical assays. Such assays though may give equivical or nondiagnostic results and in some lysosomal storage diseases an enzyme defect has not yet been identified and diagnoses can be made only by ultrastructural examination.

Download Full-text

Data Driven Mathematical Model of FOLFIRI Treatment for Colon Cancer

Cancers ◽

10.3390/cancers13112632 ◽

2021 ◽

Vol 13 (11) ◽

pp. 2632

Author(s):

Aparajita Budithi ◽

Sumeyye Su ◽

Arkadz Kirshtein ◽

Leili Shahriyari

Keyword(s):

Mathematical Model ◽

Colon Cancer ◽

Cancer Patients ◽

Immune Cell ◽

Treatment Options ◽

Data Driven ◽

T Helper ◽

Primary Tumors ◽

Cell Fractions

Many colon cancer patients show resistance to their treatments. Therefore, it is important to consider unique characteristic of each tumor to find the best treatment options for each patient. In this study, we develop a data driven mathematical model for interaction between the tumor microenvironment and FOLFIRI drug agents in colon cancer. Patients are divided into five distinct clusters based on their estimated immune cell fractions obtained from their primary tumors’ gene expression data. We then analyze the effects of drugs on cancer cells and immune cells in each group, and we observe different responses to the FOLFIRI drugs between patients in different immune groups. For instance, patients in cluster 3 with the highest T-reg/T-helper ratio respond better to the FOLFIRI treatment, while patients in cluster 2 with the lowest T-reg/T-helper ratio resist the treatment. Moreover, we use ROC curve to validate the model using the tumor status of the patients at their follow up, and the model predicts well for the earlier follow up days.

Download Full-text

Perigastric Hyaline-Vascular Variant Castleman’s Disease

Case Reports in Gastroenterology ◽

10.1159/000513175 ◽

2021 ◽

pp. 632-638

Author(s):

Yu Ming Jin ◽

Gui Ying Jing

Keyword(s):

Histopathological Examination ◽

Immunohistochemical Analysis ◽

Lymphoproliferative Disease ◽

Castleman Disease ◽

Unknown Etiology ◽

Mesenchymal Tumors ◽

Imaging Characteristics ◽

Contrast Enhanced ◽

Hyaline Vascular Variant ◽

Vascular Variant

Castleman disease (CD) is a rare chronic lymphoproliferative disease with unknown etiology and pathogenesis disease. When the lesion is located in the mediastinum, the diagnosis of CD is easy. However, if the lesion presents as a perigastric mass mimicking other subserosal gastric mesenchymal tumors, the diagnosis can be challenging. As few sonographic manifestations of hyaline-vascular variant CD, especially contrast-enhanced ultrasound (CEUS) imaging, as well as computed tomography (CT) and histopathological imaging, have been reported in literature, this case may provide a vivid example of a comprehensive CEUS and CT usage in the diagnosis and surgery with regard to CD. This report presents a case of a 50-year-old female diagnosed with hyaline-vascular variant CD in a random physical examination, the ultrasound examination first revealed a 24.3 mm × 15.4 mm hypoechogenic lesion abutting the stomach, esophagus, and liver, which was under the suspicion of gastrointestinal stromal tumor. Following a series of medical examinations, including CEUS, CT, postoperative histopathological examination, and immunohistochemical analysis, the patient was diagnosed with hyaline-vascular variant unicentric CD. After the mass was completely excised through laparoscopic surgery, the woman recovered very well without recurrence during a follow-up period of 15 months. Thus, mastering ultrasound and CT-imaging characteristics of CD and applying ultrasound and CT examination together would do help to preoperative diagnosis.

Download Full-text

Simultaneous Expression of Th1- and Treg-Associated Chemokine Genes and CD4+, CD8+, and Foxp3+ Cells in the Premalignant Lesions of 4NQO-Induced Mouse Tongue Tumorigenesis

Cancers ◽

10.3390/cancers13081835 ◽

2021 ◽

Vol 13 (8) ◽

pp. 1835

Author(s):

Hana Yamaguchi ◽

Miki Hiroi ◽

Kazumasa Mori ◽

Ryosuke Ushio ◽

Ari Matsumoto ◽

...

Keyword(s):

Immune Cell ◽

Tongue Cancer ◽

Immunohistochemical Analysis ◽

Premalignant Lesions ◽

Cytokine Gene Expression ◽

Cell Infiltration ◽

Mrna Levels ◽

Cell Recruitment ◽

T Cell Infiltration ◽

Immunohistochemical Analyses

Chemokines and cytokines in the tumor microenvironment influence immune cell infiltration and activation. To elucidate their role in immune cell recruitment during oral cancer development, we generated a mouse tongue cancer model using the carcinogen 4-nitroquinoline 1-oxide (4NQO) and investigated the carcinogenetic process and chemokine/cytokine gene expression kinetics in the mouse tongue. C57/BL6 mice were administered 4NQO in drinking water, after which tongues were dissected at 16 and 28 weeks and subjected to analysis using the RT2 Profiler PCR Array, qRT-PCR, and pathologic and immunohistochemical analyses. We found that Th1-associated chemokine/cytokine (Cxcl9, Cxcl10, Ccl5, and Ifng) and Treg-associated chemokine/cytokine (Ccl17, Ccl22, and Il10) mRNA levels were simultaneously increased in premalignant lesions of 4NQO-treated mice at 16 weeks. Additionally, although levels of Gata3, a Th2 marker, were not upregulated, those of Cxcr3, Ccr4, and Foxp3 were upregulated in the tongue tissue. Furthermore, immunohistochemical analysis confirmed the infiltration of CD4+, CD8+, and Foxp3+ cells in the tongue tissue of 4NQO-treated mice, as well as significant correlations between Th1- or Treg-associated chemokine/cytokine mRNA expression and T cell infiltration. These results indicate that CD4+, CD8+, and Foxp3+ cells were simultaneously recruited through the expression of Th1- and Treg-associated chemokines in premalignant lesions of 4NQO-induced mouse tongue tissue.

Download Full-text

Autophagic Markers in Chordomas: Immunohistochemical Analysis and Comparison with the Immune Microenvironment of Chordoma Tissues

Cancers ◽

10.3390/cancers13092169 ◽

2021 ◽

Vol 13 (9) ◽

pp. 2169

Author(s):

Georgia Karpathiou ◽

Maroa Dridi ◽

Lila Krebs-Drouot ◽

François Vassal ◽

Emmanuel Jouanneau ◽

...

Keyword(s):

Tumor Cell ◽

Immune Cells ◽

Immune Cell ◽

Positive Association ◽

Immunohistochemical Analysis ◽

Vascular Density ◽

Immune Microenvironment ◽

Significant Positive Association ◽

Sequestosome 1 ◽

Cell Expression

Chordomas are notably resistant to chemotherapy. One of the cytoprotective mechanisms implicated in chemoresistance is autophagy. There are indirect data that autophagy could be implicated in chordomas, but its presence has not been studied in chordoma tissues. Sixty-one (61) chordomas were immunohistochemically studied for autophagic markers and their expression was compared with the expression in notochords, clinicopathological data, as well as the tumor immune microenvironment. All chordomas strongly and diffusely expressed cytoplasmic p62 (sequestosome 1, SQSTM1/p62), whereas 16 (26.2%) tumors also showed nuclear p62 expression. LC3B (Microtubule-associated protein 1A/1B-light chain 3B) tumor cell expression was found in 44 (72.1%) tumors. Autophagy-related 16‑like 1 (ATG16L1) was also expressed by most tumors. All tumors expressed mannose-6-phosphate/insulin-like growth factor 2 receptor (M6PR/IGF2R). LC3B tumor cell expression was negatively associated with tumor size, while no other parameters, such as age, sex, localization, or survival, were associated with the immunohistochemical factors studied. LC3B immune cell expression showed a significant positive association with programmed death-ligand 1 (PD-L1)+ immune cells and with a higher vascular density. ATG16L1 expression was also positively associated with higher vascular density. Notochords (n = 5) showed different immunostaining with a very weak LC3B and M6PR expression, and no p62 expression. In contrast to normal notochords, autophagic factors such as LC3B and ATG16L1 are often present in chordomas, associated with a strong and diffuse expression of p62, suggesting a blocked autophagic flow. Furthermore, PD-L1+ immune cells also express LC3B, suggesting the need for further investigations between autophagy and the immune microenvironment.

Download Full-text