Oral Squamous Cells and Age Estimation in Exfoliative Cytology with Hematoxylin and Eosin Stain– A Quantitative Study

Introduction: Exfoliative cytology in age estimation is a simple, painless, less invasive collection of exfoliative cells from epithelial layers, used as a diagnostic aid for age estimation. The oral cavity is an ideal site for exfoliative epithelial cells with a physiological turnover of cells, turnover decreases as the age increases show age variation with cellular morphological changes. Age estimation is one of the important factors to identify an individual and also helps to know the chronological age of a person. Aim: To analyze and estimate the age from buccal smear and comparing the average cellular size under Image morphometric analysis. Materials and Methods: Buccal mucosal smears are taken using a wooden spatula in gentle motion of scraping and smeared on a clean glass slide and fixed in 95% ethanol immediately after smearing a minimum of around 15 minutes and stained with Haematoxylin and eosin stain. After staining, the cells were observed by microscope and measured by a paint tool. Pearson correlation analysis was done using SPSS software. Results: The cell and nuclear size difference values observed using a Pearson correlation coefficient were statistically significant with p value<0.05 revealing that there is shrinkage in cells with increase in age. Conclusion: Exfoliative cytology is a successful and vastly growing technology that is used for the detection of premalignant lesions.

Histomorphological spectrum of intracranial space occupying lesions: Experience at tertiary care centre

An “Intra-cranial space occupying lesion” (ICSOL) is defined as a mass in the cranial cavity with a diverse etiology like benign or malignant neoplasm, inflammatory or parasitic lesion, haematoma or arterio-venous malformation. The aim was to study the histomorphological spectrum of intracranial space occupying lesions at a tertiary care centre hospital in order to give accurate pretherapeutic diagnosis to facilitate better therapeutic results. This prospective and descriptive type of study of two years duration was carried out from November 2017 to October 2019 in Govt. Medical College and Superspeciality Hospital, Nagpur in the Department of Pathology. A total of 300 samples were received from Department of Neurosurgery. Detailed collection of clinical data was done in all patients regarding age, sex, clinical symptoms and radiological findings.The samples were received in 10% formalin and subjected to routine histopathological processing. Slides prepared were stained with hematoxylin and eosin stain. Special stains were done wherever required. Total 300 cases were studied of which 261 cases (87%) were neoplastic in nature and 39 cases (13%) were non neoplastic. Amongst primary tumours, most common were gliomas (48%) followed by meningiomas (20%) and schwannomas (9%). ICSOLs can present with nonspecific clinical features. Radio diagnostic investigations are helpful adjunct tools but cannot give exact diagnosis. Histopathological examination remains a gold standard for diagnosing and grading of tumours on which basis clinician can decide further line of management.

Robust and generalizable segmentation of human functional tissue units

The Human BioMolecular Atlas Program aims to compile a reference atlas for the healthy human adult body at the cellular level. Functional tissue units (FTU, e.g., renal glomeruli and colonic crypts) are of pathobiological significance and relevant for modeling and understanding disease progression. Yet, annotation of FTUs is time consuming and expensive when done manually and existing algorithms achieve low accuracy and do not generalize well. This paper compares the five winning algorithms from the "Hacking the Kidney" Kaggle competition to which more than a thousand teams from sixty countries contributed. We compare the accuracy and performance of the algorithms on a large-scale renal glomerulus Periodic acid-Schiff stain dataset and their generalizability to a colonic crypts hematoxylin and eosin stain dataset. Results help to characterize how the number of FTUs per unit area differs in relationship to their position in kidney and colon with respect to age, sex, body mass index (BMI), and other clinical data and are relevant for advancing pathology, anatomy, and surgery.

B6 Mouse Strain: The Best Fit for LPS-Induced Interstitial Cystitis Model

Interstitial cystitis (IC) is a chronic inflammatory disease characterized by bladder pain and increased urinary frequency. Although the C57BL/6J (B6) and FVB/NJ (FVB) mouse strains are commonly used as animal models for studies involving the urinary system, few reports have compared their lower urinary tract anatomy, despite the importance of such data. Our study aimed to characterize bladder function changes in FVB and B6 mouse strains with lipopolysaccharide (LPS)-induced IC, to understand mouse model-based bladder research. The bladder function parameters were measured by cystometrogram. Histological assay was examined by hematoxylin and eosin stain, Masson’s trichrome stain, and immunofluorescence staining. Results indicated that the two strains in the control group exhibited different bladder structures and functions, with significant anatomical differences, including a larger bladder size in the FVB than in the B6 strain. Furthermore, cystometry tests revealed differences in bladder function pressure. LPS-treated B6 mice presented significant changes in peak pressure, with decreased intercontraction intervals; these results were similar to symptoms of IC in humans. Each strain displayed distinct characteristics, emphasizing the care required in choosing the appropriate strain for bladder-model studies. The results suggested that the B6 mouse strain is more suitable for IC models.

Histopathological Distribution of the Gastrointestinal Tract Lesions

Introduction: Gastrointestinal (GI) diseases are common and can affect any portion of the gastrointestinal tract from the mouth to the anus. Diseases can just shows clinical conditions like stomach pain, constipation, diarrhea which can be self- limiting. But sometimes disease may be life-threatening like malignancy. Biopsy is necessary for confirmatory diagnosis and further treatment of the patient. So histopathologic examination is a must for all surgical procedures for confirmation and categorization of GI disorders. Objectives: This study was done to find out the various patterns of lesions of the gastrointestinal tract with its commonest age group and sex involvement Methodology: This was a prospective study of all the surgically resected GI tissue received in the Department of Pathology Histopathology unit in Birat Medical College and Teaching Hospital (BMCTH) from 1st February 2021 to 30 thApril 2021.The hematoxylin and eosin stain slides of the GI tissue received were studied and the lesions were diagnosed on their histomorphology. According to organ, age and sex, the lesions were categorized. The data were entered in Microsoft excel and the percentage value was calculated. Results: Out of the total 344 cases 146(42.44%) were male and 198(57.56%) were female patients. The most common age range for GI lesions was 41 to 60 years comprising of 126(36.62%) of total cases. Maximum numbers of cases 160 (46.50%) were of cholecystectomies followed by appendectomies 95(27.60%). Inflammatory and benign lesions comprised 332(96.51%), 9(2.61%) were malignant tumor and 3(0.88%) were premalignant lesions. The most common inflammatory lesions and malignant tumors were chronic cholecystitis and gastric adenocarcinoma respectively Conclusions: The study identifies that gastrointestinal lesions comprise of the most common biopsies received in the histopathology department. Early diagnosis of premalignant and malignant lesions can improve the overall survival rate of patients.

Effect of the Fungal Immunomodulatory Protein FIP-fve in the Neutrophilic Asthma Animal Model

<b><i>Background:</i></b> Asthma animal models provide valuable information about the pathogenesis and the treatment of asthma. An ovalbumin (OVA)/complete Freund’s adjuvant (CFA)-sensitized model was developed to induce neutrophil-dominant asthma and to investigate whether fungal immunomodulatory peptide-<i>fve</i> (FIP-<i>fve</i>) could improve asthma features in the OVA/CFA-sensitized model. <b><i>Methods:</i></b> We used female BALB/c mice and sensitized them intraperitoneally with OVA/CFA on days 1, 2, and 3. On days 14, 17, 21, 24, and 27, they were challenged with intranasal OVA. The airway hyper-responsiveness (AHR) was detected by BUXCO, inflammatory cells were stained with Liu’s stain, the cytokines were detected using ELISA, and the airway inflammation was analyzed with hematoxylin and eosin stain. <b><i>Results:</i></b> According to the results, OVA/CFA sensitization could induce AHR, high levels of IgE, and inflammatory cells especially neutrophils infiltration in the lung and airway inflammation. IL-4, IL-5, IL-6, IL-8, IL-10, IL-13, IL-17, IL-25, IL-33, and transforming growth factor-β (TGF-β) increased in the OVA/CFA-sensitized mice. OVA/CFA-sensitized mice treated with FIP-<i>fve</i> not only increased IL-12 and IFN-γ but also decreased IL-4, IL-5, IL-6, IL-8, IL-13, IL-17, IL-25, IL-33, and TGF-β in the bronchoalveolar lavage fluid. Moreover, FIP-<i>fve</i> significantly decreased neutrophil infiltration in the lung. <b><i>Conclusion:</i></b> The OVA/CFA model induced neutrophilic asthma successfully, and FIP-<i>fve</i> improved neutrophil-dominant asthma.

Agreement and Intraobserver Variability in Detection of Helicobacter pylori Infection Using Routine Histology, H. pylori Immunohistochemical Stain, and Warthin-Starry Special Stain on Gastric Tissue Biopsy Samples

Introduction/Objective Helicobacter pylori (HP) is a prevalent cause of gastritis in the US and worldwide. Accurate detection of the organism is important for appropriate treatment. Diagnosis is made using immunohistochemistry and special stains including Warthin-Starry stain (WS) which is low cost, technically easy to perform on tissue sections, and can be automated. We aimed to assess interobserver variability, reproducibility and validity of hematoxylin and eosin stain (H&E), WS and anti-Helicobacter pylori immunohistochemical staining (HP-IHC) for the histopathological identification and evaluation of HP organisms on gastric mucosa biopsies. Methods/Case Report We prospectively evaluated gastric biopsies from ten adult patients. These patients underwent upper gastrointestinal endoscopy with subsequent biopsy for various abdominal complaints. Seven of the ten biopsies (7/10) were HP infection positive, and three cases (3/10) were HP negative by H&E stain and HP-IHC (determined by GI Pathologist). The presence or absence and density of HP, were assessed on H&E, HP-IHC and WS in blinded fashion by five general surgical (GS) pathologists. The GS pathologists assessed the cases for presence and density of HP using the semi-quantitative modified Sydney classification (none, mild, moderate, and severe). Percentage agreement and interobserver variablilty using Cohen-Kappas statistics (KS) were calculated. Results (if a Case Study enter NA) The percentage agreement for presence or absence of HP in the biopsies with H&E, HP-IHC and WS stains were 91%, 98% and 99%, respectively. The interobserver agreement for evaluation of presence of HP was excellent with WS staining method (Overall KS = 0.737 95%, Confidence Interval (CI) = 0.501- 0.973) and HP-IHC (Overall KS = 0.783, 95% CI = 0.585-0.980). Agreement for H&E was moderate (Overall KS = 0.532, 95% CI= 0.392-0.672). HP density agreement was excellent using WS and HP-IHC and H&E was in the moderate range. Conclusion This study found excellent interobserver agreement using IHC and WS. While IHC is the most specific stain, WS is more sensitive in identifying the shape and morphology of HP organisms and is an efficient and low-cost alternative with excellent morphology in general surgical practice.

Evaluation of muscle involvement & degeneration in advanced oral submucous fibrosis using three different stains – a retrospective cross-sectional study

ABSTRACT Background: Oral submucous fibrosis (OSMF) is graded according to various histological factors which include the epithelial changes and the connective tissue changes. These features could be identified in routine Hematoxylin and Eosin (H and E) staining in shades of pink. However, the use of special stains provides a contrast to various connective tissue components thereby aiding in better visualization of these connective tissue changes in advanced OSMF cases. Objective: To compare and evaluate muscle involvement and degeneration in advanced oral submucous fibrosis using three different stains namely, H&E, Van Gieson, and Masson’s Trichrome. Material and Methods: 10 Formalin-fixed paraffin-embedded tissue sections of advanced OSMFwere stained using 3 different stains namely Hematoxylin and Eosin (H&E), Van-Gieson, and Masson trichrome. The results obtained were tabulated and statistically analyzed using Kruskal Wallis ANOVA. Results: The hyalinization and fibrosis involving the skeletal muscle were better visualized in Masson’s Trichrome but were not statistically significant. The muscle degeneration in deeper areas was better visualized in Masson’s trichrome when compared to the H&E and Van Gieson. Conclusion: Hematoxylin and eosin stains all the connective tissue components in various shades of pink, use of special stains bestows contrast between different components of connective tissue, thus improvising grading of OSMF. Masson’s trichrome stain can be used as a single adjunct to hematoxylin and eosin stain as changes in the superficial and deeper connective tissue could be ascertained. Keywords Fibrosis; Masson’s trichrome stain; Muscle degeneration; Oral submucous fibrosis grading.

Isolation and molecular detection of avian mycoplasmosis in selected areas of Mymensingh district in Bangladesh

Avian mycoplasmosis caused by several species of Mycoplasma including Mycoplasma gallisepticum, M synoviae, M. meleagridis and M. iowae. Among these Mycoplasma gallisepticum is the most important poultry pathogen in Bangladesh. For effective control of Mycoplasmosis, proper early diagnosis is the corner stone. The research work was designed, a total of 20 samples, lung exudates, swabs from trachea and air sacs were collected from dead birds of different poultry farms in Mymensingh district during October-December, 2007. Samples were collected in 10% buffered formalin for histopathological study. Swabs were collected in mycoplasma broth supplemented with supplement-G. Additionally Kanamycin solution was added to prevent the growth of gram–Ve bacteria and then the organisms were transferred into mycoplasma agar for isolation. Histopathological studies were conducted using routine procedure in Hematoxylin and Eosin stain. Isolated Mycoplasma were subjected to DNA extraction, Nested PCR was done using a commercial PCR kit. The histopathological study revealed the presence of mycoplasmal related tissue changes, such as severe congestion and infiltration of mononuclear cells in different organs. The extracted DNA accumulated at the upper position of DNA ladder as band without any smear formation. The DNA from avian mycoplasmas was amplified and gave amplified product of 975 bp by outer primer and 395 bp by inner primer which was much smaller than the expected size. In this study, preliminary results from field samples suggest that culture using mycoplasma agar and broth supplement with Supplement-G and Kanamycin solution could be useful for the isolation of pathogenic avian mycoplasmas. Asian J. Med. Biol. Res. 2021, 7 (2), 182-190

Malakoplakia in the Urinary Bladder of 4 Puppies

Malakoplakia in humans most often affects the urinary bladder and is characterized by inflammation with von Hansemann–type macrophages, with or without Michaelis-Gutmann bodies, and is frequently associated with Escherichia coli infection. We describe the microscopic features of malakoplakia in the urinary bladder of 4 puppies. In all cases, the lamina propria of the urinary bladder was markedly expanded by sheets of large, round to polygonal macrophages with intracytoplasmic, periodic acid-Schiff-positive granules and granular inclusions, and rare Prussian blue–positive inclusions. Macrophages were positive for CD18 and Iba1. In 2 cases, Michaelis-Gutmann bodies were detected with hematoxylin and eosin stain and were best demonstrated with von Kossa stain. E. coli infection was confirmed in 2 cases with bacterial culture or polymerase chain reaction (PCR) and sequencing of the bacterial 16S ribosomal RNA gene. Transmission electron microscopy of one case demonstrated macrophages with abundant lysosomes, phagolysosomes, and rod-shaped bacteria. Microscopic features were similar to human cases of malakoplakia. In dogs, the light microscopic characteristics of malakoplakia closely resemble granular cell tumors and histiocytic ulcerative colitis.