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Graefe's Archive For Clinical and... Mar 2024To examine histological characteristics and differences between drusen beneath the retinal pigment epithelium (small hard drusen) located in the macula and located in...
PURPOSE
To examine histological characteristics and differences between drusen beneath the retinal pigment epithelium (small hard drusen) located in the macula and located in the parapapillary region.
METHODS
We histomorphometrically examined human eyes enucleated due to uveal melanomas or secondary angle-closure glaucoma.
RESULTS
The study included 106 eyes (age, 62.6 ± 15.2 years) with macular drusen (n = 7 globes) or parapapillary drusen (n = 29 eyes) and 70 eyes without drusen. In all drusen, periodic-acid-Schiff-positive material was located between the RPE basal membrane and the inner collagenous layer of Bruch's membrane (BM). Macular drusen as compared with parapapillary drusen had lower height (15.2 ± 10.1 µm versus 34.3 ± 19.8 µm; P = 0.003), while both groups did not differ significantly in basal drusen width (74.0 ± 36.3 µm versus 108.7 ± 101.0 µm; P = 0.95). Eyes with macular drusen and eyes without drusen did not differ significantly in BM thickness (2.74 ± 0.44 µm versus 2.55 ± 0.88 µm; P = 0.57) or in RPE cell density (35.4 ± 10.4 cells/480 µm versus 32.8 ± 7.5 cells/480 µm; P = 0.53), neither in the drusen region nor in the drusen vicinity, while BM thickness (4.60 ± 1.490 µm; P < 0.001) and RPE cell density (56.9 ± 26.8 cells/480 µm; P = 0.005) were higher at the parapapillary drusen. Eyes with macular drusen, eyes with parapapillary drusen, and eyes without drusen did not differ significantly in choriocapillaris density (all P > 0.10) and thickness (all P > 0.35). Limitations of the study, among others, were a small number and size of drusen examined, diseases leading to enucleation, lack of serial sections, limited resolution of light microscopy, and enucleation-related and histological preparation-associated artefacts.
CONCLUSIONS
The findings of this study, also taking into account its methodological limitations, suggest that macular drusen and parapapillary drusen shared the morphological feature of periodic-acid-Schiff-positive material between the RPE basal membrane and BM and that they did not vary significantly in choriocapillaris thickness and density. RPE cell density and BM thickness were higher in parapapillary drusen than in macular drusen.
PubMed: 38472430
DOI: 10.1007/s00417-024-06438-5 -
The American Journal of Pathology Jan 2023Convolutional neural network (CNN)-based image analysis applications in digital pathology (eg, tissue segmentation) require a large amount of annotated data and are...
Convolutional neural network (CNN)-based image analysis applications in digital pathology (eg, tissue segmentation) require a large amount of annotated data and are mostly trained and applicable on a single stain. Here, a novel concept based on stain augmentation is proposed to develop stain-independent CNNs requiring only one annotated stain. In this benchmark study on stain independence in digital pathology, this approach is comprehensively compared with state-of-the-art techniques including image registration and stain translation, and several modifications thereof. A previously developed CNN for segmentation of periodic acid-Schiff-stained kidney histology was used and applied to various immunohistochemical stainings. Stain augmentation showed very high performance in all evaluated stains and outperformed all other techniques in all structures and stains. Without the need for additional annotations, it enabled segmentation on immunohistochemical stainings with performance nearly comparable to that of the annotated periodic acid-Schiff stain and could further uphold performance on several held-out stains not seen during training. Herein, examples of how this framework can be applied for compartment-specific quantification of immunohistochemical stains for inflammation and fibrosis in animal models and patient biopsy specimens are presented. The results show that stain augmentation is a highly effective approach to enable stain-independent applications of deep-learning segmentation algorithms. This opens new possibilities for broad implementation in digital pathology.
Topics: Deep Learning; Coloring Agents; Periodic Acid; Neural Networks, Computer; Image Processing, Computer-Assisted; Kidney
PubMed: 36309103
DOI: 10.1016/j.ajpath.2022.09.011 -
Genes Mar 2023Lafora disease (LD) is a progressive neurologic disorder caused by biallelic pathogenic variants in or , leading to tissue accumulation of polyglucosan aggregates...
Lafora disease (LD) is a progressive neurologic disorder caused by biallelic pathogenic variants in or , leading to tissue accumulation of polyglucosan aggregates termed Lafora bodies (LBs). This study aimed to characterize the retinal phenotype in mice by examining knockout (KO; ) and control (WT) littermates at two time points (10 and 14 months, respectively). In vivo exams included electroretinogram (ERG) testing, optical coherence tomography (OCT) and retinal photography. Ex vivo retinal testing included Periodic acid Schiff Diastase (PASD) staining, followed by imaging to assess and quantify LB deposition. There was no significant difference in any dark-adapted or light-adapted ERG parameters between KO and WT mice. The total retinal thickness was comparable between the groups and the retinal appearance was normal in both groups. On PASD staining, LBs were observed in KO mice within the inner and outer plexiform layers and in the inner nuclear layer. The average number of LBs within the inner plexiform layer in KO mice were 1743 ± 533 and 2615 ± 915 per mm, at 10 and 14 months, respectively. This is the first study to characterize the retinal phenotype in an mouse model, demonstrating significant LB deposition in the bipolar cell nuclear layer and its synapses. This finding may be used to monitor the efficacy of experimental treatments in mouse models.
Topics: Mice; Animals; Lafora Disease; Disease Models, Animal; Retina; Myoclonic Epilepsies, Progressive; Electroretinography
PubMed: 37107612
DOI: 10.3390/genes14040854 -
Ophthalmology Science Jun 2021To evaluate PAX8 expression by immunohistochemistry in the normal pediatric and adult crystalline lens and to assess the usefulness of PAX8 immunohistochemical stain in...
PURPOSE
To evaluate PAX8 expression by immunohistochemistry in the normal pediatric and adult crystalline lens and to assess the usefulness of PAX8 immunohistochemical stain in the diagnosis of morphologically challenging lesions of lenticular origin.
DESIGN
Retrospective, observational case series.
PARTICIPANTS
Fourteen congenital and acquired lens-derived lesions and 10 control crystalline lenses.
METHODS
Hematoxylin-eosin and periodic acid-Schiff stains and an immunohistochemical panel of PAX8, vimentin, S100, smooth muscle actin, AE1/AE3, cytokeratin 7, and cytokeratin 5/6 antibodies were performed on all tissues.
MAIN OUTCOME MEASURES
Distribution of PAX8 expression in normal crystalline lens and in lens-derived lesions.
RESULTS
Records search identified 10 normal pediatric and adult crystalline lenses, 1 phakomatous choristoma, 1 Peters anomaly with adherent leukoma, 1 lens capsule with congenital pyramidal cataract formation, 2 lenses with anterior and posterior subcapsular cataract formation, 3 postsurgical cataractous lenses (Soemmerring ring cataract and capsular fibrosis), and 6 retrocorneal membranes that incorporated various components of metaplastic corneal endothelium, metaplastic lens epithelium, corneal stroma, and epithelial downgrowth. Strong nuclear PAX8 expression was observed in the lens epithelium and in the equatorial lens bow of normal pediatric and adult lenses. Nuclear PAX8 expression also was observed in the lesions that retained some of the epithelial morphologic features, such as phakomatous choristoma, adherent leukoma, congenital pyramidal cataract, and components of intraocular membranes with lens epithelial differentiation. PAX8 expression was lost in lens epithelial lesions that had undergone mesenchymal transition, such as anterior subcapsular cataract and capsular fibrosis.
CONCLUSIONS
PAX8 antibody may be a useful adjunct to the immunohistochemical panels in morphologically challenging lens epithelial-derived lesions that retain epithelial morphologic features. PAX8 is not useful in the diagnosis of lens-derived lesions that feature epithelial-mesenchymal transition.
PubMed: 36249298
DOI: 10.1016/j.xops.2021.100024 -
Current Eye Research Oct 2022To explore the effect of doxycycline on vasculogenic mimicry (VM) formation and the potential mechanism in human pterygium fibroblasts in order to find novel targets for...
PURPOSE
To explore the effect of doxycycline on vasculogenic mimicry (VM) formation and the potential mechanism in human pterygium fibroblasts in order to find novel targets for pterygium therapy.
METHODS
First, we demonstrate the existence of VM in 73 pterygium specimens by CD31 and periodic acid Schiff (PAS) dual staining. Then we used cell counting kit-8, clone formation assay and flow cytometry to prove the inhibitory effect of doxycycline on cell proliferation and apoptosis. The VM formation was evaluated through wound healing assay, cell transwell assay and three-dimensional cell culture combined with PAS staining. Finally, we used Western blot to testify the correlation of the VM and the factors in protein level preliminarily.
RESULTS
Our results showed that VM existed in human pterygium specimens exactly. Otherwise, in human pterygium fibroblasts, doxycycline induced a dose-dependent inhibitory effect on cell proliferation and apoptosis induction. Besides, doxycycline significantly suppressed vasculogenic mimicry tube formation, cell migration and invasion. Furthermore, doxycycline impaired the expression of MMP-9, MMP-2 and VEGF which may related to pterygium VM formation.
CONCLUSIONS
Doxycycline decelerated pterygium progression might be through inhibiting VM formation according to the downregulation of MMP-9, MMP-2 and VEGF, which may provide the basis of further studies involving doxycycline for pterygium treatment.
Topics: Cell Line, Tumor; Conjunctiva; Doxycycline; Fibroblasts; Humans; Matrix Metalloproteinase 2; Matrix Metalloproteinase 9; Neovascularization, Pathologic; Periodic Acid; Pterygium; Vascular Endothelial Growth Factor A
PubMed: 35923150
DOI: 10.1080/02713683.2022.2108455 -
The American Journal of Dermatopathology May 2021Periodic acid-Schiff (PAS) staining of nail clippings is an adjunct diagnostic tool for onychomycosis.
BACKGROUND
Periodic acid-Schiff (PAS) staining of nail clippings is an adjunct diagnostic tool for onychomycosis.
OBJECTIVE
To detect histopathological findings as clues to the presence of PAS-positive (+) fungal elements in nail clippings.
METHODS
Four hundred sixteen consecutive nail clippings suspected of onychomycosis were stained with hematoxylin and eosin, and with PAS stains. All cases were studied histopathologically. The clinical files of the cases with neutrophils were reviewed.
RESULTS
PAS+ staining for fungi were demonstrated in 159 (38%) of the nail clippings. Neutrophils, parakeratosis, plasma globules, and bacteria were observed in 43 (27%), 108 (67%), 80 (50%), and 80 (50%) of the PAS+ cases, respectively, and in 17 (6%), 109 (41%), 84 (32%) and 140 (54%) of the PAS- cases, respectively (P < 0.01). Neutrophils showed by far the highest specificity (93%), although with low sensitivity (27%) for the presence of PAS+ fungi. Among the 43 PAS+ and 17 PAS- specimens with neutrophils, only 1 (2.3%) and 3 (17%) had overt psoriasis, respectively.
CONCLUSION
Neutrophils in nail clippings may serve as a clue for onychomycosis. PAS staining with neutrophils is not necessarily associated with psoriasis.
Topics: Humans; Neutrophils; Onychomycosis; Periodic Acid-Schiff Reaction
PubMed: 32956097
DOI: 10.1097/DAD.0000000000001800 -
Journal of Veterinary Diagnostic... Jul 2022A 10-y-old, castrated male Boxer dog that was born and had lived in Missouri without any travel history to other states, except for a few trips to Kansas, was presented...
A 10-y-old, castrated male Boxer dog that was born and had lived in Missouri without any travel history to other states, except for a few trips to Kansas, was presented with a distended abdomen and declined health. Ultrasonographic examination revealed a large hepatic mass, and the dog was euthanized. A postmortem examination revealed that the left liver lobes were largely replaced by a white-to-tan multilobular mass with a cobblestone surface. The lesion also involved the diaphragm. Histologically, hepatic architecture was effaced by large areas of necrosis with numerous, ≤0.2-cm, cystic structures that stained positively with periodic acid-Schiff stain and contained calcareous corpuscles. Gross and microscopic hepatic lesions were compatible with alveolar echinococcosis (AE) caused by . PCR examination confirmed , and results from genotyping were consistent with the E4 haplotype. To our knowledge, this is only the second canine AE case and the third pet dog that has been confirmed to be infected by in the contiguous United States. is a serious health risk for both pet dogs and humans.
Topics: Animals; Dog Diseases; Dogs; Echinococcosis; Echinococcus multilocularis; Male; Missouri
PubMed: 35678137
DOI: 10.1177/10406387221104754 -
Heliyon Mar 2023The aim of this study was to explore the histopathological and genetic changes in the submandibular glands after duct ligation and provide important clues to functional...
OBJECTIVE
The aim of this study was to explore the histopathological and genetic changes in the submandibular glands after duct ligation and provide important clues to functional regeneration.
DESIGN
We established a rat salivary gland duct ligation model and observed pathological changes in the rat submandibular gland on day 1 and weeks 1, 2, 3, and 4 using hematoxylin and eosin staining, Alcian blue-periodic acid Schiff staining, Masson staining, terminal deoxynucleotidyl transferase dUTP nick-end labeling (TUNEL), and immunohistochemical staining. RNA sequencing was performed on normal salivary glands and those from the ligation model after 1 week. Significantly differentially expressed genes were selected, and Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses were performed.
RESULTS
Apoptosis levels and histological and functional KEGG pathway analyses showed that injury to the salivary gland after ligation gradually increased. The TGF-β pathway was activated and promoted fibrosis. RNA sequencing results and further verification of samples at week 1 showed that the NF-κB pathway plays a vital role in salivary gland atrophy.
CONCLUSIONS
Our results detailed the pathological changes in the submandibular gland after ligation and the important functions of the NF-κB pathway.
PubMed: 36950625
DOI: 10.1016/j.heliyon.2023.e14288 -
Biological Trace Element Research Dec 2023Fluoride (F) is widely distributed in the environment and poses serious health risks to humans and animals. Although a good body of literature demonstrates a close...
Fluoride (F) is widely distributed in the environment and poses serious health risks to humans and animals. Although a good body of literature demonstrates a close relationship between F content and renal system performance, there is no satisfactory information on the involved intracellular routes. Hence, this study used histopathology and mitochondrial fission to explore fluorine-induced nephrotoxicity further. For this purpose, mice were exposed to the F ion (0, 25, 50, 100 mg/L) for 90 days. The effects of different F levels on renal pathomorphology and ion metabolism were assessed using hematoxylin and eosin (H&E), periodic acid-Schiff stain (PAS), periodic acid-silver methenamine (PASM), Prussian blue (PB), and alkaline phosphatase (ALP) staining. The results showed that F could lead to glomerular atrophy, tubular degeneration, and vacuolization. Meanwhile, F also could increase glomerular and tubular glycoproteins; made thickening of the renal capsule membrane and thickening of the tubular basement membrane; led to the accumulation of iron ions in the tubules; and increased in glomerular alp and decreased tubular alp. Concomitantly, IHC results showed that F significantly upregulated the expression levels of mitochondrial fission-related proteins, including mitochondrial fission factor (Mff), fission 1 (Fis1), and mitochondrial dynamics proteins of 49 kDa (MiD49) and 51 kDa (MiD51), ultimately caused apoptosis. To sum up, excessive fluorine has a strong nephrotoxicity effect, disrupting the balance of mitochondrial fission and fusion, interfering with the process of mitochondrial fission, and then causing damage to renal tissue structure and apoptosis.
PubMed: 38057486
DOI: 10.1007/s12011-023-03994-5 -
Life Sciences Oct 2022Asthma is a chronic inflammatory airway disease associated with the airway narrowing and obstruction. Sinapic acid (SA), a hydroxycinnamic acid, possesses various...
OBJECTIVE
Asthma is a chronic inflammatory airway disease associated with the airway narrowing and obstruction. Sinapic acid (SA), a hydroxycinnamic acid, possesses various pharmacological properties including antioxidant and anti-inflammatory activity. This research evaluated effects of different doses of SA on murine model of ovalbumin (OVA)-induced allergic asthma.
MATERIALS AND METHODS
Allergic asthma induced by sensitizing mice on days 1 and 14 by intraperitoneal injection of OVA. After initial sensitization, between days 21 and 23, mice were challenged for 30 min with an aerosol of 1 % (wt/vol) OVA. Treatment with dexamethasone (3 mg/kg) or SA (25, 50 or 100 mg/kg) were done by oral gavage on days 15-23. Inflammatory cells infiltration and interferon-γ (IFN-γ), interlukin-4 (IL-4), IL-5 and IL-13 levels were evaluated in the bronchoalveolar lavage fluid (BALF). Serum total and OVA-specific immunoglobulin E (IgE) and lung tissue nitric oxide (NO) levels were measured. Histological changes in lung tissue were examined by staining with hematoxylin and eosin (H&E) for cell infiltration, periodic acid-Schiff (PAS) for mucus production and Masson's trichrome for collagen deposition.
RESULTS
Treatment with SA significantly inhibited inflammatory cell infiltration, enhanced IFN-γ level and decreased IL-4, IL-5 and IL-13 levels in BALF. Serum total and OVA-specific IgE levels and NO level in lung tissue were significantly reduced by SA. Histological examination demonstrated that SA significantly attenuated inflammatory cell infiltration and mucus-producing cells in the lung.
CONCLUSION
These data suggest that SA may be a new therapeutic potential to treat allergic asthma through suppressing T-helper 2 immune responses.
Topics: Animals; Anti-Inflammatory Agents; Antioxidants; Asthma; Coumaric Acids; Cytokines; Dexamethasone; Disease Models, Animal; Eosine Yellowish-(YS); Hematoxylin; Immunoglobulin E; Inflammation; Interferon-gamma; Interleukin-13; Interleukin-4; Interleukin-5; Mice; Mice, Inbred BALB C; Nitric Oxide; Ovalbumin; Periodic Acid; Th2 Cells
PubMed: 35931198
DOI: 10.1016/j.lfs.2022.120858