Histopathological Methods Research Articles

Multiple Bowen's disease due to long-term narrow-band ultraviolet B phototherapy: A case report and literature review.

By presenting a case study on multiple instances of Bowen's disease and the consistent use of narrow-band ultraviolet B (NB-UVB) phototherapy over a three-year period, our aim is to enhance the comprehension of domestic clinicians regarding the disease. Additionally, we seek to review existing literature, encouraging dermatologists to consider clinical secondary primary lesion diagnoses. Our approach involves analyzing a diagnosed case of multiple Bowen's disease, examining clinical manifestations, histopathology, imaging results, and treatment methods related to NB-UVB phototherapy. We aim to facilitate discussion and understanding through a comprehensive literature analysis. An elderly male with a 30-year history of psoriasis vulgaris initiated continuous NB-UVB therapy three years ago. A year later, he developed red patches and plaques with distinct borders and scaly surfaces on his face, trunk, lower extremities, and scrotum. Histopathological examination confirmed Bowen's disease. Treatment involved liquid nitrogen cryotherapy, with no recurrence observed during the one-year follow-up. This case highlights that Bowen's disease, typically solitary, can manifest as multiple instances, especially in individuals with a history of psoriasis vulgaris. While NB-UVB stands as the primary treatment for psoriasis vulgaris, caution is warranted due to the potential risk of skin tumor induction with prolonged high-dose usage. Clinicians should be vigilant in monitoring and assessing the long-term implications of such therapies.

Effect of pycnogenol on ischemia/reperfusion-induced oxidative and inflammatory damage in the ovaries of rats

Purpose: To investigate the protective effect of pycnogenol (PYC) against ischemia/reperfusion (I/R) ovarian damage induced by experimental ovarian torsion in rats using biochemical and histopathological methods.
 Methods: Four groups of rats (n = 6) were randomly assigned and designated as follows – SG: sham group, PCG: pycnogenol (40 mg/kg) group, IRG: ovarian ischemia-reperfusion group, and PIR: pycnogenol (40 mg/kg) + ovarian ischemia-reperfusion group. In IRG and PYC treatment groups, ischemia was induced in the right ovary for two hours with vascular clips. The ovary was reperfused for two hours after ischemia was induced. Then, the levels of malondialdehyde (MDA), total glutathione (tGSH), nuclear factor-kappa B (NF-κB), tumor necrosis factor-alpha (TNF-α) and interleukin-1β (IL-1β) in the ovarian tissues of rats were examined. Ovarian tissues were also examined histopathologically and follicle counts were performed.
 Results: Histopathological examination revealed that the developing follicles in ovary of I/R-induced group showed both morphological and cellular degeneration and vascular pathology. The PYC treatment group showed less ovarian damage, edema and vascular pathology than I/R-induced group (p < 0.05). The MDA, NF-κB, TNF-α and IL-1β levels in I/R-induced group were significantly higher than in SG and tGSH levels were significantly reduced by I/R damage (p < 0.05). Pycnogenol treatment reversed these biochemical indices as well as I/R-induced histopathological changes.
 Conclusion: Pycnogenol has a protective effect in rat ovaries by alleviating I/R-induced changes in biochemical parameters and histological disruptions. Further studies, including those in humans, to determine the effect of PYC on ovarian injuries would be required.

Unsupervised domain adaptation with local structure preservation for colon histopathological image classification

The histopathological image classification method, based on deep learning, can be used to assist pathologists in cancer recognition in colon histopathology. The popularization of automatic and accurate histopathological image classification methods in this way is of great significance. However, smaller medical institutions with limited medical resources may lack colon histopathology image training sets with reliable labeled information; thus they may be unable to meet the needs of deep learning for many labeled training samples. Therefore, in this paper, the colon histopathological image set with rich label information from a certain medical institution is taken as the source domain; the colon histopathological image set from a smaller medical institution with limited medical resources is taken as the target domain. Considering the potential differences between histopathological images obtained by different institutions, this paper proposes a classification learning framework, namely unsupervised domain adaptation with local structure preservation for colon histopathological image classification, which can learn an adaptive classifier by performing distribution alignment and preserving intra-domain local structure to predict the labels of the colon histopathological images from institutions with lower medical resources. Extensive experiments demonstrate that the proposed framework shows significant improvement in accuracy and specificity of colon histopathological images without reliable labeled information compared to models without unsupervised domain adaptation. Specifically, in an affiliated hospital in Fuyang City, Anhui Province, the classification accuracy of benign and malignant colon histopathological images reaches 96.21%. The results of comparative experiments also show promising classification performance of our method in comparison with other unsupervised domain adaptation methods.

Comparison of isolation and histopathology methods in diagnosis of pigeon tuberculosis

Comparison of isolation and histopathology methods in diagnosis of pigeon tuberculosis

An automatic classification method of testicular histopathology based on SC-YOLO framework.

The pathological diagnosis and treatment of azoospermia depend on precise identification of spermatogenic cells. Traditional methods are time-consuming and highly subjective due to complexity of Johnsen score, posing challenges for accurately diagnosing azoospermia. Here, we introduce a novel SC-YOLO framework for automating the classification of spermatogenic cells that integrates S3Ghost module, CoordAtt module and DCNv2 module, effectively capturing texture and shape features of spermatogenic cells while reducing model parameters. Furthermore, we propose a simplified Johnsen score criteria to expedite the diagnostic process. Our SC-YOLO framework presents the higher efficiency and accuracy of deep learning technology in spermatogenic cell recognition. Future research endeavors will focus on optimizing the model's performance and exploring its potential for clinical applications.

Comparative study of parasite load in the spleen, lymph node, and skin of dogs with visceral leishmaniasis

Canine visceral leishmaniasis (VL) is a zoonosis caused by the protozoan Leishmania infantum. The lymph nodes, spleen, and skin are essential organs in the immunopathogenesis of the disease. This study aimed to investigate the histomorphological alterations and parasite load in the popliteal lymph node, spleen, and skin of eleven VL-positive dogs in the fine needle aspiration (FNA), Dual-path Platform chromatographic immunoassay (DPP® CVL rapid test) and Enzyme-linked immunosorbent assay (ELISA). Histopathological and immunohistochemical methods were used to evaluate the samples, and the results showed variable histopathological changes and parasite load. The popliteal lymph nodes and spleen exhibited granulomatous reaction, lymphoid atrophy, presence of plasma cells, and disorganization of the architecture was marked. The skin showed multifocal to diffuse inflammation in the superficial dermis, composed of lymphoplasmacytic infiltrate and granulomatous reaction. Immunodetection of the parasite Leishmania sp. was observed in all organs. The intensity of histological changes was not associated with the higher number of parasitized macrophages. The popliteal lymph node had the highest median parasite load (11.2) compared to the skin and spleen. Statistically, the Pearson correlation test revealed a highly significant correlation in the parasite load between the popliteal lymph node and spleen (r=0.89081, p=0.0002) and between the popliteal lymph node and skin (r=0.71185, p=0.0140). The study concludes that VL-positive dogs’ lymph nodes, spleen, and skin suffer histomorphological alterations that could be one of the aspects that favor the maintenance of the infection.

A Novel Colorectal Histopathological Image Classification Method Based on Progressive Multi-Granularity Feature Fusion of Patch

A Novel Colorectal Histopathological Image Classification Method Based on Progressive Multi-Granularity Feature Fusion of Patch

Effects of dose-dependent chronic caffeine consumption in a rat burn wound model: Histopathological and immunohistochemical evaluation.

Using histopathological and immunohistochemical methods, we aimed to examine the dose-dependent effects of chronic caffeine consumption on the recovery of burn wounds in an in vivo rat model. Forty-five rats were randomly assigned to a high-dose group (20 mg/kg per day for eight weeks; n=15), a low-dose group (10 mg/kg per day for eight weeks; n=15), or a control group (n=15). The burn model was created in rats. The groups were separated into three subgroups (n=5) based on the day after injury (7th, 14th, or 21st day). The wound area, wound closure percentage, and histopathological and immunohistochemical reactivity were evaluated. Successful wound healing was noted in rats treated with low doses of caffeine, similar to the control group. Pathology revealed low re-epithelization, low inflammation, and high granulation in the high-dose group. In addition, there was a significant difference between the control and high-dose groups regarding the immunohistochemical reactivity of αVβ3 integrin, vascular endothelial growth factor (VEGF), and matrix metalloproteinase 9 (MMP-9) (P<0.05). We demonstrated that chronic caffeine consumption in rats adversely affects the recovery process of wounds in a dose-dependent manner. This effect may occur through delayed wound healing via the molecules MMP-9, αVβ3 integrin, and VEGF. Treatment that modulates these molecules can lead to enhanced and quicker recovery of damaged skin in coffee lovers.

The path to fertility: Current approaches to mare endometritis and endometrosis.

The path to fertility in the mare requires an understanding of the hormonal influences, the immune response, genetics, and epigenetic mechanisms involved not only in physiological reproductive processes, but also such pathologies as endometritis and endometrosis. Endometritis may lead to endometrosis establishment. In the presence of endometritis, neutrophils arrive at the mare endometrium, and form neutrophil extracellular traps. While NETosis plays pivotal roles, prolonged inflammation can lead to chronic endometritis, endometrosis, and fertility issues. Matrix metalloproteinases and epigenetic changes influence the course of endometrosis. Inhibitors of specific enzymes involved in NETosis and epigenetic inhibitors have shown potential in reducing pro-fibrotic effects. Collagen type III (COL3) has emerged as a putative biomarker, correlating with endometrosis and useful in fertility assessment. Thus, COL3 may offer a non-invasive diagnostic tool, as a complement to histopathological methods. Epigenetic modifications and miRNA expressions offer new avenues for therapeutic strategies, emphasizing the importance of understanding the cellular mechanisms at play in mare endometrial fibrosis.

Enfeksiyon hastalıkları polikliniğinde ekstrapulmoner tüberküloz hastalarının epidemiyolojik ve klinik tutulumlarının yıllara göre dağılımı

Aim: It was aimed to examine the patients diagnosed with extrapulmonary tuberculosis (EPTB) in terms of diagnostic methods and demographic characteristics. Materials and Methods: The files of patients with EPTB who were followed up in the infectious disease’s outpatient clinic between 2012 and 2022 in our study were retrospectively reviewed. Results: Of the patients diagnosed with EPTB, 70.8% (102) were female and 29.2% (42) were male. The ages of the patients ranged from 20 to 88, the mean age of women was 54.2, and the mean age of men was 55. The most common site of involvement in these patients was lymph node involvement. This was followed by bone-joint, peritoneal, central nervous system and genitourinary system involvement, respectively. Histopathological methods were used most frequently in 81 (56.5%) of the patients. Conclusion: The signs and symptoms of EPTB differ according to the organs and tissues involved in the body. We believe that EPTB should be considered in the differential diagnosis in endemic regions.

Investigation of the tumor microenvironment, hypoxia and angiogenesis by immunohistochemical and histopathological methods in canine mammary tumors

The tumor microenvironment (TME) is an important component for studying tumor behavior in several cancers in human beings. However, little information regarding the role of the TME in canine mammary tumors (CMTs) is available compared to humans. In this study, the aim was to investigate the relationship between the TME, hypoxia and angiogenesis through CD31, VEGF, HIF-1a, CD68 and CD163 expression by using immunohistochemical (IHC) methods in formalin-fixed paraffin-embedded canine mammary tumor samples [(n=34: malignant (n=28) and benign (n=6)], to compare them with the clinicopathological features of tumors, and to analyze the relationship between them. There was no significant relationship between CD31, VEGF, HIF-1a, CD68 and CD163 expression in malignant tumors compared to benign tumors (P&gt;0.05). There was an association between microvessel density and clinicopathological variables (the tumor size P=0.013, the presence of necrosis P=0.022) and individual histological grade (G2 vs. G3 P=0.028) in malignant tumors. While there was a positive correlation between CD68 and CD163 in malignant tumors in the dogs (P&lt;0.01), no correlation was determined between other antibodies. Immunohistochemical determination of the level of angiogenesis in the TME may give further useful information about the angiogenic potential and grading of the clinical aggressiveness of some CMTs.

N-acetyl cysteine and zinc sulphate ameliorated crude oil-induced hepatotoxicity in Wistar rats

Even while crude oil offers many advantages, its toxic chemical composition has been shown to negatively interfere with the body's ability to maintain homeostasis. This investigation examined the effects of crude oil on the liver function of Wistar rats as well as the protective effects of N-acetyl cysteine (NAC) and zinc sulphate (ZnSO4). The experimental group and control group consisted of 42 Wistar rats, which were divided into groups of six each. The experimental group had six groups, while the control group only consumed food and water. In the experimental group, three groups received a diet contaminated with crude oil for two, three, and four weeks, respectively. In addition, another three groups received NAC (100 mg/kg), ZnSO4 (0.5 mg/kg), and a combination of NAC and ZnSo4 for three weeks each. Rats' body weights were recorded both before and after the administration of crude oil. Using commercial kits and histological examinations carried out using conventional histopathological methods, the liver's toxicity was assessed. As a result of eating food contaminated with crude oil, the results showed a significant drop (p &lt; 0.05) in body weight. Additionally, there was liver cell injury. The levels of alkaline phosphatase and alanine transaminase considerably increased. Histological analysis revealed abnormal changes to the liver's typical structure. NAC and ZnSO4 therapy reduced the elevated Alanine transaminase and alkaline phosphatase levels. The study's findings highlight the negative impacts of crude oil and the therapeutic effects of NAC and ZnSO4.

DNase improves the efficacy of antimicrobial photodynamic therapy in the treatment of candidiasis induced with Candida albicans.

The study evaluated the association of DNase I enzyme with antimicrobial photodynamic therapy (aPDT) in the treatment of oral candidiasis in mice infected with fluconazole-susceptible (CaS) and -resistant (CaR) Candida albicans strains. Mice were inoculated with C. albicans, and after the infection had been established, the tongues were exposed to DNase for 5 min, followed by photosensitizer [Photodithazine®(PDZ)] and light (LED), either singly or combined. The treatments were performed for 5 consecutive days. Treatment efficacy was evaluated by assessing the tongues via fungal viable population, clinical evaluation, histopathological and fluorescence microscopy methods immediately after finishing treatments, and 7 days of follow-up. The combination of DNase with PDZ-aPDT reduced the fungal viability in mice tongues immediately after the treatments by around 4.26 and 2.89 log10 for CaS and CaR, respectively (versus animals only inoculated). In the fluorescence microscopy, the polysaccharides produced by C. albicans and fungal cells were less labeled in animals treated with the combination of DNase with PDZ-aPDT, similar to the healthy animals. After 7 days of the treatment, DNase associated with PDZ-aPDT maintained a lower count, but not as pronounced as immediately after the intervention. For both strains, mice treated with the combination of DNase with PDZ-aPDT showed remission of oral lesions and mild inflammatory infiltrate in both periods assessed, while animals treated only with PDZ-aPDT presented partial remission of oral lesions. DNase I enzyme improved the efficacy of photodynamic treatment.

Enhancing oral squamous cell carcinoma prediction: the prognostic power of the worst pattern of invasion and the limited impact of molecular resection margins.

Oral squamous cell carcinoma (OSCC) originates from the mucosal lining of the oral cavity. Almost half of newly diagnosed cases are classified as advanced stage IV disease, which makes resection difficult. In this study, we investigated the pathological features and mutation profiles of tumor margins in OSCC. We performed hierarchical clustering of principal components to identify distinct patterns of tumor growth and their association with patient prognosis. We also used next-generation sequencing to analyze somatic mutations in tumor and marginal tissue samples. Our analyses uncovered that the grade of worst pattern of invasion (WPOI) is strongly associated with depth of invasion and patient survival in multivariable analysis. Mutations were primarily detected in the DNA isolated from tumors, but several mutations were also identified in marginal tissue. In total, we uncovered 29 mutated genes, mainly tumor suppressor genes involved in DNA repair including BRCA genes; however none of these mutations significantly correlated with a higher chance of relapse in our medium-size cohort. Some resection margins that appeared histologically normal harbored tumorigenic mutations in TP53 and CDKN2A genes. Even histologically normal margins may contain molecular alterations that are not detectable by conventional histopathological methods, but NCCN classification system still outperforms other methods in the prediction of the probability of disease relapse.

The role of corneal biopsy in the management of microbial keratitis.

Corneal biopsy helps in diagnosing deep-seated or recalcitrant lesions of microbial keratitis (MK). We aim to analyze its role in managing these challenging cases. This is a retrospective review of 22 cases of corneal biopsy at our institute from January 2010 to December 2021. Data were retrospectively collected using the electronic medical record (EMR) system. Those cases of indolent, progressive MK or deep-seated lesions where cornea scraping was not possible were considered for corneal biopsy to establish the microbiological diagnosis. The primary aims of our study were to analyze the indications, success rates, and outcomes for biopsy patients in our series. Additional outcomes that were analyzed included the average time from presentation to biopsy, the type of causative organism isolated from the biopsy by either histopathological or microbiological method, and the frequency and outcome of surgical interventions performed. Descriptive statistics using mean (±standard deviation) and median (±range) were used to interpret the demographic data. Overall, 15 of 22 patients (68%) had a positive corneal biopsy after microbiological or histopathological examinations. The most identified organism was microsporidia (n = 4,30.7%), followed by mycobacteria (n = 2,15.4%), gram-negative bacilli (n = 2,15.4%), acid-fast bacilli (n = 1,7.6%), fungus (n = 2,15.4%), gram-positive cocci (n = 1,7.6%), and mixed bacterial infection (n = 1,7.6%). Corneal biopsy should be considered a diagnostic modality for patients with deep-seated or unresponsive MK. It can improve the treatment for MK, ensuring targeted therapy.

High-Throughput Multiplex Immunohistochemistry of Glioma Organoids.

The invasive capacity and progression of glioblastoma cells and neoplastic cells in other are dependent on interactions with the surrounding tumor microenvironment. In particular, cancer cells form a reciprocal relationship with noncellular dysregulated extracellular matrix in the tumors. Here, we describe a protocol that can be used to model the functional relationship between tumor cells and extracellular matrix. We demonstrate how 3D organoids, including glioma tumor organoids, can be processed, embedded, and sectioned in a high-throughput setup that enables investigation of the organoids by histopathological methods, multiplex immunohistochemistry, and spatial analysis within the same section.

A case of plasmacytoma with widespread metastasis in a dog

In this study, a case of plasmacytoma with widespread metastasis in a nine-year-old male Rottweiler dog was described by histopathological and immunohistochemical methods. Macroscopic examination revealed multifocal white or pink-red colored masses ranging in size from 0,5 to 3 cm in the lung, liver, mediastinal and mesenterial lymph nodes, spleen, heart, pancreas, intestine and kidneys. In microscopic examination, tumor foci consisting mostly of atypical plasma cells were observed in the lung, liver, spleen, pancreas, mediastinal and mesenteric lymph nodes, intestine, and kidneys. Tumor-type giant cells were observed in tumor foci in the lung, mitotic figures in tumor foci in other organs, including the lung, and abundant tumor cells in capillary vessels. In Methyl Green Pyronin staining of lung, liver, spleen, pancreas, mediastinal and mesenterial lymph nodes, intestine and kidney sections, the cytoplasm of plasma cells was observed to be stained pink. Congo Red staining applied to liver and spleen sections showed amyloid deposits stained in brick red color. In addition, in the immunohistochemical staining of lung, liver, spleen, pancreas, mediastinal and mesenterial lymph nodes, intestine and kidney sections with Proliferating Cell Nuclear Antigen (PCNA) antibody, severe immunoreactivity was detected in tumor cells. As a result, due to the presence of rarely reported widespread metastases in the lung, liver, spleen, pancreas, mediastinal and mesenterial lymph nodes, intestine and kidneys of the diagnosed plasmacytoma case, it was deemed appropriate to publish the case in order to contribute to the literature by attracting the attention of veterinarians.

An Evaluation of the Effectiveness of Melatonin and n-Acetylcysteine in Cerebral Ischemia-Reperfusion Injury in Adult Rats.

Background and Objectives: The purpose of this study was to apply histopathological and immunohistochemical methods to compare the protective efficacy of melatonin and N-acetylcysteine (NAC) application in rats with experimental brain ischemia/reperfusion (I/R) injury induced through occlusion of the middle cerebral artery (MCA), and to evaluate the protective effect of their combined use. Materials and Methods: Forty-one young adult male Wistar albino rats were divided into five groups-control (n = 8), I/R group (n = 8), melatonin (n = 8), NAC (n = 8), and melatonin + NAC (n = 9). Results: All scores differed between the groups, apart from vascular congestion (p < 0.05). At two-way comparisons, all histological scores were significantly higher in the I/R group than in the control group (p < 0.05). No change occurred in the vascular congestion scores with the administration of melatonin, although decreases were determined in all other scores. These decreases were statistically significant for cellular eosinophilic pyknotic degeneration, vacuolization, and edema (p < 0.05). All histopathological scores in the group administered NAC together with melatonin were significantly lower than in the I/R group (p < 0.05). Conclusions: The combined use of NAC and melatonin, the neuroprotective efficacy of which on histopathological parameters is shown in this study, now needs to be supported by further research.

Imaging-based characterization of tumoral heterogeneity for personalized cancer treatment.

With personalized tumor therapy, understanding and addressing the heterogeneity of malignant tumors is becoming increasingly important. Heterogeneity can be found within one lesion (intralesional) and between several tumor lesions emerging from one primary tumor (interlesional). The heterogeneous tumor cells may show a different response to treatment due to their biology, which in turn influences the outcome of the affected patients and the choice of therapeutic agents. Therefore, both intra- and interlesional heterogeneity should be addressed at the diagnostic stage. While genetic and biological heterogeneity are important parameters in molecular tumor characterization and in histopathology, they are not yet addressed routinely in medical imaging. This article summarizes the recently established markers for tumor heterogeneity in imaging as well as heterogeneous/mixed response to therapy. Furthermore, a look at emerging markers is given. The ultimate goal of this overview is to provide comprehensive understanding of tumor heterogeneity and its implications for radiology and for communication with interdisciplinary teams in oncology. KEY POINTS:: · Tumor heterogeneity can be described within one lesion (intralesional) or between several lesions (interlesional).. · The heterogeneous biology of tumor cells can lead to a mixed therapeutic response and should be addressed in diagnostics and the therapeutic regime.. · Quantitative image diagnostics can be enhanced using AI, improved histopathological methods, and liquid profiling in the future..

Artificial intelligence-based recurrence prediction outperforms classical histopathological methods in pulmonary adenocarcinoma biopsies

IntroductionBetween 10 and 50% of early-stage lung adenocarcinoma patients experience local or distant recurrence. Histological parameters such as a solid or micropapillary growth pattern are well-described risk factors for recurrence. However, not every patient presenting with such a pattern will develop recurrence. Designing a model which can more accurately predict recurrence on small biopsy samples can aid the stratification of patients for surgery, (neo-)adjuvant therapy, and follow-up. Material and MethodsIn this study, a statistical model on biopsies fed with histological data from early and advanced-stage lung adenocarcinomas was developed to predict recurrence after surgical resection. Additionally, a convolutional neural network (CNN)-based artificial intelligence (AI) classification model, named AI-based Lung Adenocarcinoma Recurrence Predictor (AILARP), was trained to predict recurrence, with an ImageNet pre-trained EfficientNet that was fine-tuned on lung adenocarcinoma biopsies using transfer learning. Both models were validated using the same biopsy dataset to ensure that an accurate comparison was demonstrated. ResultsThe statistical model had an accuracy of 0.49 for all patients when using histology data only. The AI classification model yielded a test accuracy of 0.70 and 0.82 and an area under the curve (AUC) of 0.74 and 0.87 on patch-wise and patient-wise hematoxylin and eosin (H&E) stained whole slide images (WSIs), respectively. ConclusionAI classification outperformed the traditional clinical approach for recurrence prediction on biopsies by a fair margin. The AI classifier may stratify patients according to their recurrence risk, based only on small biopsies. This model warrants validation in a larger lung biopsy cohort.