FEATURES OF THE PATHOMORPHOLOGICAL DIAGNOSIS OF MICROCRYSTALLINE ARTHROPATHIES IN THE PRACTICE OF SURGICAL MATERIAL EXAMINATION

2020 ◽  
Vol 58 (3) ◽  
pp. 286-289
Author(s):  
N. S. Migalkin ◽  
T. A. Stupina ◽  
A. V. Kaminsky ◽  
D. S. Mokhovikov ◽  
D. A. Shabalin ◽  
...  
Keyword(s):  
Staining Technique ◽  
Calcium Pyrophosphate ◽  
Alcohol Solution ◽  
Formalin Fixation ◽  
Histological Techniques ◽  
Tissue Sections ◽  
Stereo Microscope ◽  
Hematoxylin And Eosin ◽  
Tissue Specimens ◽  
Msu Crystals

The development of microcrystalline arthritides is most frequently associated with the formation of monosodium urate (MSU) and calcium pyrophosphate (CPP) crystals. Their identification is of crucial importance in recognizing these diseases. Objective: to determine the possibilities of histological techniques in identifying MSU and CPP crystals and to evaluate the effectiveness of the techniques. Subjects and methods. Twenty-four tissue blocks (fragments of the affected areas of the elbow joint, the interphalangeal joint of the index finger, and hip joint) from 7 patients were examined. Paraffin sections were stained with a 0.5% alcohol solution of eosin, as well as with hematoxylin and eosin. Tissue specimens were examined and digitized using an AxioScope.A1 stereo microscope with Zenblue software (Carl Zeiss MicroImaging GmbH, Germany). Results and discussion. When staining the tissue sections with hematoxylin and eosin, microcrystals were not visualized; the major portions of MSU crystals was dissolved during fixation and staining, whereas CPP crystals were masked with hematoxylin as focal basophilic aggregates. The staining technique with an alcohol solution of eosin and short formalin fixation (within 12 hours) made it possible to avoid dissolution of MSU crystals and to visualize both MSU and CPP crystals, and to determine their shape and color. Conclusion. Light microscopy of the tissue sections stained with an alcohol solution of eosin along with short formalin fixation is a reliable method to differentiate MSU and CPP crystals. In patients undergoing endoprosthetic replacement, the significance of this technique for the pathomorphological study of surgical material consists in assessing inflammatory activity and in eliminating a disease, such as microcrystalline arthropathy.

Identification of Fasciola Species Using Tegumental Spines in Tissue Sections

2020 ◽  
Author(s):  
Arezoo FADAVI ◽  
Keyhan ASHRAFI ◽  
Hamid HASSANPOUR ◽  
Mohamad Bagher ROKNI ◽  
Seyyed Mostafa HOSSEINI ◽  
...  
Keyword(s):  
Staining Technique ◽  
Morphological Characters ◽  
Image Analysis System ◽  
Scanning Electron Micrographs ◽  
Tissue Sections ◽  
Fasciola Species ◽  
Accuracy Of Measurement ◽  
Hematoxylin And Eosin ◽  
Analysis System ◽  
Liver Flukes

Background: Efforts to find a reliable non-molecular means of identification has been the main purpose of the current work that always is persuaded by researchers interested in the field of parasitology. Methods: Adult fasciolids were obtained from the slaughterhouses in different parts of Iran in 2017, and investigated using the classical old fashion morphological appearances of the worms implementing a camera lucida equipped microscope. Histological procedure was subsequently performed for almost the entire collected adult worms followed by Hematoxylin and Eosin (H&E) staining technique. DNA extraction and RFLP-PCR technique were carried out for the entire fasciolid liver flukes. To attain more comparable morphological conclusions, Scanning Electron Micrographs were also implemented for two molecularly identified fasciolids. Results: Based on spine morphology observed in worm’s tissue sections two types of tegumental spines, “pointed” and “molar” shapes have been identified addressing to distinguish F. hepatica and F. gigantica species respectively. The present identification has been also supported by Molecular analysis using RFLP-PCR technique. Conclusion: There are some hidden morphological characters implemented in species identification for certain helminths. Meanwhile, the emergence of computer image analysis system (CIAS) on the scene of taxonomy, has revolutionized the accuracy of measurement in morphology by employing detailed parameters that have not been regarded before. The current study has illustrated the tegumental spines of two Fasciola species in tissue sections which has not been enough considered in helminthological publications so far.

scholarly journals THU0406 IDENTIFICATION OF INTRACELLULAR VACUOLES IN SYNOVIAL FLUID WITH CALCIUM PYROPHOSPHATE AND MONOSODIUM URATE CRYSTALS

2020 ◽  
Vol 79 (Suppl 1) ◽  
pp. 440.1-441
Author(s):  
M. L. Peral ◽  
I. Calabuig ◽  
A. Martín-Carratalá ◽  
M. Andrés ◽  
E. Pascual
Keyword(s):  
Clinical Practice ◽  
Synovial Fluid ◽  
Phase Contrast ◽  
Calcium Pyrophosphate ◽  
Monosodium Urate ◽  
Fluid Analysis ◽  
Cytoplasmic Vacuoles ◽  
Contrast Microscopy ◽  
Ordinary Light ◽  
Msu Crystals

Background:Synovial fluid analysis using polarized microscopy is the gold standard for the diagnosis of crystal-related arthritis. In our experience, we have noted that, when calcium pyrophosphate (CPP) crystals are observed, they sometimes appear within intracellular vacuoles. However, this phenomenon is not seen in those samples containing monosodium urate (MSU) crystals. This finding has been scantly reported in the literature, but may be useful in clinical practice to ensure accurate crystal identification.Objectives:Our study aims to assess whether the presence of vacuoles contributes to identifying the type of crystal, and also to gauge the frequency of their presentation.Methods:We conducted an observational study in a rheumatology unit between February and June of 2019. Synovial fluids containing CPP or MSU crystals, obtained in daily clinical practice, were consecutively included for analysis. Two observers simultaneously analyzed the presence of vacuoles by ordinary light and phase contrast microscopy in less than 24 hours after their extraction, using a microscope equipped with two viewing stations. The primary study variable was to determine whether CPP and MSU crystals are seen inside intracellular vacuoles, and to calculate the frequency of this finding for each type of crystal, estimating their 95% confidence interval (95% CI) and comparing rates using Fisher’s exact test.Results:Twenty-one samples were obtained. Data is given in the Table. MSU crystals were present in 7 (33.3%) and CPP crystals in 14 (66.6%). Interestingly, none of the MSU samples showed crystal-containing vacuoles (95% CI 0-35.4%). On the contrary, cytoplasmic vacuoles containing crystals were present in all of the CPP samples (95% CI 78.5-100%). The findings were confirmed by phase-contrast microscopy. Differences were statistically significant (p<0.001).Table.SAMPLES ACCORDING TO TYPE OF MICROCRYSTAL(n=21)SAMPLES WITH VACUOLS(UNDER ORDINARY LIGHT)SAMPLES WITH VACUOLS(UNDER PHASE CONTRAST)CPP (14; 66.6%)14 (100%)(95%CI 78.5-100%)14 (100%)(95%CI 78.5-100%)MSU (7; 33.3%)0 (0%)(95%CI 0-35.4%)0 (0%)(95%CI 0-35.4%)Conclusion:The presence of vacuoles may be a useful and easy way to differentiate MSU and CPP crystals when performing synovial fluid microscopy in clinical practice, since it appears to be a distinctive feature in CPP crystal fluids.References:[1]Kohn NN, Hughes RE, McCarty DJ Jr, Faires JS. The significance of calcium phosphate crystals in the synovial fluid of arthritic patients: the «pseudogout syndrome». II. Identification of crystals. Ann InternMed. 1962 May;56:738-45.[2]Pascual E, Sivera F, Andrés M. Synovial Fluid Analysis for Crystals. CurrOpRheumatol 2011;23:161-169.[3]McCarty DJ, Koopman WJ. Arthritis and allied conditions: A textbook of rheumatology, volumen 1. Lea &amp;Febiger. 1993.[4]Pascual E, Sivera F. Synovial fluid crystal Analysis. En Gout and other crystal arthropathies. Terkeltaub R ed. Elsevier; 2012: p.20-34.[5]Hwang HS, Yang CM, Park SJ, Kim HA. Monosodium Urate Crystal-Induced Chondrocyte Death via Autophagic Process. Int J Mol Sci. 2015 Dec 8;16(12):29265-77.Image 1. Microscopy with ordinary light. Cells with cytoplasmic vacuoles are observed, as well as abundant intra and extracellular CPP crystals.Image 2. Microscopy with phase contrast technique. Cells with intracellular vacuoles are observed inside which have microcrystals with parallelepiped morphology, compatible with CPP.Disclosure of Interests: :None declared

scholarly journals PathoFusion: An Open-Source AI Framework for Recognition of Pathomorphological Features and Mapping of Immunohistochemical Data

Cancers ◽  
2021 ◽  
Vol 13 (4) ◽  
pp. 617
Author(s):  
Guoqing Bao ◽  
Xiuying Wang ◽  
Ran Xu ◽  
Christina Loh ◽  
Oreoluwa Daniel Adeyinka ◽  
...  
Keyword(s):  
Neural Network ◽  
Tumor Vasculature ◽  
Integrated System ◽  
Pathological Features ◽  
Adjacent Tissue ◽  
Tissue Sections ◽  
Contextual Feature ◽  
Hematoxylin And Eosin ◽  
Feature Information ◽  
Software Code

We have developed a platform, termed PathoFusion, which is an integrated system for marking, training, and recognition of pathological features in whole-slide tissue sections. The platform uses a bifocal convolutional neural network (BCNN) which is designed to simultaneously capture both index and contextual feature information from shorter and longer image tiles, respectively. This is analogous to how a microscopist in pathology works, identifying a cancerous morphological feature in the tissue context using first a narrow and then a wider focus, hence bifocal. Adjacent tissue sections obtained from glioblastoma cases were processed for hematoxylin and eosin (H&E) and immunohistochemical (CD276) staining. Image tiles cropped from the digitized images based on markings made by a consultant neuropathologist were used to train the BCNN. PathoFusion demonstrated its ability to recognize malignant neuropathological features autonomously and map immunohistochemical data simultaneously. Our experiments show that PathoFusion achieved areas under the curve (AUCs) of 0.985 ± 0.011 and 0.988 ± 0.001 in patch-level recognition of six typical pathomorphological features and detection of associated immunoreactivity, respectively. On this basis, the system further correlated CD276 immunoreactivity to abnormal tumor vasculature. Corresponding feature distributions and overlaps were visualized by heatmaps, permitting high-resolution qualitative as well as quantitative morphological analyses for entire histological slides. Recognition of more user-defined pathomorphological features can be added to the system and included in future tissue analyses. Integration of PathoFusion with the day-to-day service workflow of a (neuro)pathology department is a goal. The software code for PathoFusion is made publicly available.

scholarly journals Deep learning-based transformation of H&E stained tissues into special stains

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Kevin de Haan ◽  
Yijie Zhang ◽  
Jonathan E. Zuckerman ◽  
Tairan Liu ◽  
Anthony E. Sisk ◽  
...  
Keyword(s):  
Visual Inspection ◽  
Kidney Diseases ◽  
Periodic Acid ◽  
Needle Core Biopsy ◽  
Silver Stain ◽  
Periodic Acid Schiff ◽  
Tissue Sections ◽  
Biopsy Tissue ◽  
Hematoxylin And Eosin

AbstractPathology is practiced by visual inspection of histochemically stained tissue slides. While the hematoxylin and eosin (H&E) stain is most commonly used, special stains can provide additional contrast to different tissue components. Here, we demonstrate the utility of supervised learning-based computational stain transformation from H&E to special stains (Masson’s Trichrome, periodic acid-Schiff and Jones silver stain) using kidney needle core biopsy tissue sections. Based on the evaluation by three renal pathologists, followed by adjudication by a fourth pathologist, we show that the generation of virtual special stains from existing H&E images improves the diagnosis of several non-neoplastic kidney diseases, sampled from 58 unique subjects (P = 0.0095). A second study found that the quality of the computationally generated special stains was statistically equivalent to those which were histochemically stained. This stain-to-stain transformation framework can improve preliminary diagnoses when additional special stains are needed, also providing significant savings in time and cost.

scholarly journals Discrimination of Aspergillosis, Mucormycosis, Fusariosis, and Scedosporiosis in Formalin-Fixed Paraffin-Embedded Tissue Specimens by Use of Multiple Real-Time Quantitative PCR Assays

2016 ◽  
Vol 54 (11) ◽  
pp. 2798-2803 ◽  
Author(s):  
Elham Salehi ◽  
Mohammad T. Hedayati ◽  
Jan Zoll ◽  
Haleh Rafati ◽  
Maryam Ghasemi ◽  
...  
Keyword(s):  
Fusarium Oxysporum ◽  
Aspergillus Flavus ◽  
Ffpe Tissue ◽  
Content Type ◽  
Real Time Quantitative Pcr ◽  
Tissue Sections ◽  
Formalin Fixed Paraffin ◽  
Formalin Fixed Paraffin Embedded ◽  
Tissue Specimens ◽  
Formalin Fixed

In a retrospective multicenter study, 102 formalin-fixed paraffin-embedded (FFPE) tissue specimens with histopathology results were tested. Two 4- to 5-μm FFPE tissue sections from each specimen were digested with proteinase K, followed by automated nucleic acid extraction. Multiple real-time quantitative PCR (qPCR) assays targeting the internal transcribed spacer 2 (ITS2) region of ribosomal DNA, using fluorescently labeled primers, was performed to identify clinically important genera and species of Aspergillus , Fusarium , Scedosporium , and the Mucormycetes . The molecular identification was correlated with results from histological examination. One of the main findings of our study was the high sensitivity of the automated DNA extraction method, which was estimated to be 94%. The qPCR procedure that was evaluated identified a range of fungal genera/species, including Aspergillus fumigatus , Aspergillus flavus , Aspergillus terreus , Aspergillus niger , Fusarium oxysporum , Fusarium solani , Scedosporium apiospermum , Rhizopus oryzae , Rhizopus microsporus , Mucor spp., and Syncephalastrum . Fusarium oxysporum and F. solani DNA was amplified from five specimens from patients initially diagnosed by histopathology as having aspergillosis. Aspergillus flavus , S. apiospermum , and Syncephalastrum were detected from histopathological mucormycosis samples. In addition, examination of four samples from patients suspected of having concomitant aspergillosis and mucormycosis infections resulted in the identification of two A. flavus isolates, one Mucor isolate, and only one sample having both R. oryzae and A. flavus . Our results indicate that histopathological features of molds may be easily confused in tissue sections. The qPCR assay used in this study is a reliable tool for the rapid and accurate identification of fungal pathogens to the genus and species levels directly from FFPE tissues.

scholarly journals Role of S-100 Immunostain as An Auxiliary Diagnostic Aid in Leprosy

2017 ◽  
Vol 56 (205) ◽  
pp. 141-144 ◽  
Author(s):  
Ramesh Dhakhwa ◽  
Sneh Acharya ◽  
Sailesh Pradhan ◽  
Sanju Babu Shrestha ◽  
Tomoo Itoh
Keyword(s):  
Erythema Nodosum ◽  
Lupus Vulgaris ◽  
Common Pattern ◽  
Tissue Sections ◽  
Histopathologic Diagnosis ◽  
S 100 ◽  
Skin Biopsies ◽  
Hematoxylin And Eosin ◽  
Hematoxylin And Eosin Stain

Introduction: Histopathologic diagnosis of leprosy is difficult when Bacillary Index (BI) is zero and neural involvement are not easily identifiable on routine Hematoxylin and Eosin stain. This study was undertaken to study the role of S-100 immunostaining in demonstrating different patterns of nerve involvement in various types of leprosy. Methods: Thirty one skin biopsies with clinico-histopathologic diagnoses of leprosy over a period of two years were included in the study. Ten cases of non-lepromatous granulomatous dermatoses (including eight cases of lupus vulgaris and two cases of erythema nodosum) were used as controls. Tissue sections from all cases and controls were stained with Hematoxylin and Eosin (H&E) stain, Fite stain and S-100 immunostain. The H&E stained slides were used to study the histopathological features, Fite stained slides for Bacillary Index and S-100 for nerve changes. Results: Neural changes could be demonstrated in the entire spectrum of leprosy using S-100 immunostaining. The most common pattern of nerve destruction in the tuberculoid spectrum was fragmented and infiltrated whereas lepromatous spectrum showed mostly fragmented nerve twigs. Intact nerves were not detected in any of the leprosy cases. Conclusions:  S-100 immunostain is a useful auxiliary aid to the routine  H&E stain in the diagnosis of leprosy especially tuberculoid spectrum and intermediate leprosy.  Keywords: bacillary index; leprosy; S-100 immunostain.

Persistence of Crystals in Stored Synovial Fluid Samples

2020 ◽  
Vol 47 (9) ◽  
pp. 1416-1423
Author(s):  
Sonia Pastor ◽  
José-Antonio Bernal ◽  
Rocío Caño ◽  
Silvia Gómez-Sabater ◽  
Fernando Borras ◽  
...  
Keyword(s):  
Synovial Fluid ◽  
Light Microscopy ◽  
Room Temperature ◽  
Polarized Light ◽  
Calcium Pyrophosphate ◽  
Polarized Light Microscopy ◽  
Tetraacetic Acid ◽  
Monosodium Urate ◽  
Prospective Longitudinal ◽  
Msu Crystals

Objective.Lack of access to polarized light microscopy is often cited as an argument to justify the clinical diagnosis of crystal-related arthritis. We assessed the influence of time since sampling and preservation methods on crystal identification in synovial fluid (SF) samples under polarized light microscopy.Methods.This was a prospective, longitudinal, observational factorial study, analyzing 30 SF samples: 12 with monosodium urate (MSU) crystals and 18 with calcium pyrophosphate (CPP) crystals. Each SF sample was divided into 4 subsamples (120 subsamples in total). Two were stored in each type of preserving agent, heparin or ethylenediamine tetraacetic acid (EDTA), at room temperature or at 4°C. Samples were analyzed the following day (T1), at 3 days (T2), and at 7 days (T3) by simple polarized light microscopy, and the presence of crystals was recorded.Results.The identification of crystals in the MSU group was similar between groups, with crystals observed in 11/12 (91.7%) room temperature samples and in 12/12 (100%) refrigerated samples at T3. Identification of CPP crystals tended to decrease in all conditions, especially when preserved with EDTA at room temperature [12/18 (66.7%) at T3], while less reduction was seen in refrigerated heparin-containing tubes.Conclusion.Preserving samples with heparin in refrigerated conditions allows delayed microscopic examination for crystals. Avoiding crystal-proven diagnosis because of the immediate unavailability of microscopy no longer appears justified.

scholarly journals Mucormycosis of the hard palate masquerading as carcinoma

2012 ◽  
Vol 2 (1) ◽  
pp. 28 ◽  
Author(s):  
Bhari Sharanesha Manjunatha ◽  
Nagarajappa Das ◽  
Rakesh V. Sutariya ◽  
Tanveer Ahmed
Keyword(s):  
Clinical Presentation ◽  
Fungal Infections ◽  
Survival Rates ◽  
Positive Culture ◽  
Staining Technique ◽  
Necrotic Bone ◽  
Chronic Ulcers ◽  
Hematoxylin And Eosin ◽  
Clinical Differential Diagnosis ◽  
Hematoxylin And Eosin Stain

A growing number of medically compromised patients are encountered by dentists in their practices. Opportunistic fungal infections such as mucormycosis usually occur in immunocompromised patients but can infect healthy individuals as well. Mucormycosis is an acute opportunistic, uncommon, frequently fatal fungal infection, caused by a saprophytic fungus that belongs to the class of phycomycetes. Among the clinical differential diagnosis we can consider squamous cell carcinoma. Such cases present as chronic ulcers with raised margins causing exposure of underlying bone. There is a close histopathological resemblance between mucormycosis and aspergillosis. Microscopically, aspergillosis has septate branching hyphae, which can be distinguished from mucormycotic hyphae by a smaller width and prominent acute angulations of branching hyphae. A definitive diagnosis of mucormycosis can be made by tissue biopsy that identifies the characteristic hyphae, by positive culture or both. The culture of diseased tissue may be negative and histopathologic examination is essential for early diagnosis. Mucormycosis was long regarded as a fatal infection with poor prognosis. However with early medical and surgical management survival rates are now thought to exceed 80%. In the present case, the fungus was identified by hematoxylin and eosin stain and confirmed by Grocott’s silver methenamine special staining technique. Removal of the necrotic bone, which acted as a nidus of infection, was done. Post-operatively patient was advised an obturator to prevent oronasal regurgitation. Since mucormycosis occurs infrequently, it may pose a diagnostic and therapeutic dilemma for those who are not familiar with its clinical presentation.

Enhanced Gross Visualization of Chicken Peyer's Patch: Novel Staining Technique Applied to Fresh Tissue Specimens

2006 ◽  
Vol 50 (2) ◽  
pp. 298-302 ◽  
Author(s):  
L. E. Vaughn ◽  
P. S. Holt ◽  
R. W. Moore ◽  
R. K. Gast
Keyword(s):  
Staining Technique ◽  
Peyer’S Patch ◽  
Peyer's Patch ◽  
Fresh Tissue ◽  
Tissue Specimens

scholarly journals Infiltrated Macrophages Die of Pneumolysin-Mediated Necroptosis following Pneumococcal Myocardial Invasion

2016 ◽  
Vol 84 (5) ◽  
pp. 1457-1469 ◽  
Author(s):  
Ryan P. Gilley ◽  
Norberto González-Juarbe ◽  
Anukul T. Shenoy ◽  
Luis F. Reyes ◽  
Peter H. Dube ◽  
...  
Keyword(s):  
Cardiac Tissue ◽  
Immune Cell ◽  
Kinase Domain ◽  
Cardiac Damage ◽  
Interacting Protein ◽  
Hydropic Degeneration ◽  
Content Type ◽  
Tissue Sections ◽  
Hematoxylin And Eosin

Streptococcus pneumoniae(the pneumococcus) is capable of invading the heart. Herein we observed that pneumococcal invasion of the myocardium occurred soon after development of bacteremia and was continuous thereafter. Using immunofluorescence microscopy (IFM), we observed thatS. pneumoniaereplication within the heart preceded visual signs of tissue damage in cardiac tissue sections stained with hematoxylin and eosin. DifferentS. pneumoniaestrains caused distinct cardiac pathologies: strain TIGR4, a serotype 4 isolate, caused discrete pneumococcus-filled microscopic lesions (microlesions), whereas strain D39, a serotype 2 isolate, was, in most instances, detectable only using IFM and was associated with foci of cardiomyocyte hydropic degeneration and immune cell infiltration. Both strains efficiently invaded the myocardium, but cardiac damage was entirely dependent on the pore-forming toxin pneumolysin only for D39. Early microlesions caused by TIGR4 and microlesions formed by a TIGR4 pneumolysin-deficient mutant were infiltrated with CD11b+and Ly6G-positive neutrophils and CD11b+and F4/80-positive (F4/80+) macrophages. We subsequently demonstrated that macrophages in TIGR4-infected hearts died as a result of pneumolysin-induced necroptosis. The effector of necroptosis, phosphorylated mixed-lineage kinase domain-like protein (MLKL), was detected in CD11b+and F4/80+cells associated with microlesions. Likewise, treatment of infected mice and THP-1 macrophagesin vitrowith the receptor-interacting protein 1 kinase (RIP1) inhibitor necrostatin-5 promoted the formation of purulent microlesions and blocked cell death, respectively. We conclude that pneumococci that have invaded the myocardium are an important cause of cardiac damage, pneumolysin contributes to cardiac damage in a bacterial strain-specific manner, and pneumolysin kills infiltrated macrophages via necroptosis, which alters the immune response.

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