Morphometric and Immunohistochemical Characteristics of the Adult Human Soft Palate Muscles

The soft palate is the only structure that reversibly separates the respiratory and gastrointestinal systems. Most species can eat and breathe at the same time. Humans cannot do this and malfunction of the soft palate may allow food to enter the lungs and cause fatal aspiration pneumonia. Speech is the most defining characteristic of humans and the soft palate, along with the larynx and tongue, plays the key roles. In addition, palatal muscles are involved in snoring and obstructive sleep apnea. Considering the significance of the soft palate, its function is insufficiently understood. The objectives of this study were to document morphometric and immunohistochemical characteristics of adult human soft palate muscles, including fiber size, the fiber type, and myosin heavy chain (MyHC) composition for better understanding muscle functions. In this study, 15 soft palates were obtained from human autopsies. The palatal muscles were separated, cryosectioned, and stained using histological and immunohistochemical techniques. The results showed that there was a fast type II predominance in the musculus uvulae and palatopharyngeus and a slow type I predominance in the levator veli palatine. Approximately equal proportions of type I and type II fibers existed in both the palatoglossus and tensor veli palatine. Soft palate muscles also contained hybrid fibers and some specialized myofibers expressing slow-tonic and embryonic MyHC isoforms. These findings would help better understand muscle functions.

Inflammatory Cell Composition and Immune-Related microRNA Signature of Temporal Artery Biopsies From Patients With Giant Cell Arteritis

ObjectivesThe aim of this study was to quantitatively assess distinct immune cell subsets comprising inflammatory infiltrate in temporal artery biopsies (TABs) from patients with giant cell arteritis (GCA), and to link the obtained histopathological data with expression profiles of immune-related microRNAs (miRNAs).MethodsThe study included 68 formalin-fixed, paraffin-embedded TABs from treatment-naïve patients, including 30 histologically positive GCA and 16 negative GCA TABs, and 22 control non-GCA TABs. Quantitative assessment of histological parameters was performed using histopathological and immunohistochemical techniques. miRNA expression analysis was performed by quantitative real-time PCR.ResultsIntense transmural mononuclear inflammatory infiltrates in TAB-positive GCA arteries were predominantly composed of CD3+, CD4+ and CD8+ T lymphocytes, and CD68+ macrophages, accompanied by a strong nuclear overexpression of the nuclear factor of activated T cells, cytoplasmic 1 (NFATC) in the lymphocyte infiltrate fraction. Furthermore, TAB-positive GCA arteries were characterized by significant overexpression of nine pro-inflammatory miRNAs (miR-132-3p/-142-3p/-142-5p/-155-5p/-210-3p/-212-3p/-326/-342-5p/-511-5p) and a significant under-expression of six regulatory immune-related miRNAs (miR-30a-5p/-30b-5p/-30c-5p/-30d-5p/-30e-5p/-124-3p), whose expression levels significantly associated with most evaluated histopathological parameters. Notably, we revealed miR-132-3p/-142-3p/-142-5p/-155-5p/-212-3p/-511-5p as major promoters of arterial inflammation and miR-30a-5p/-30c-5p/-30d-5p as putative regulators of NFATC signaling in TAB-positive GCA arteries.ConclusionOverall, we demonstrated that an altered arterial tissue-specific pro-inflammatory miRNA signature favors enhanced T cell-driven inflammation and macrophage activity in TAB-positive GCA arteries. Moreover, dysregulation of several immune-related miRNAs seems to contribute crucially to GCA pathogenesis, through impairing their regulatory activity towards T cell-mediated immune responses driven by the calcineurin (CaN)/NFAT signaling pathway, indicating their therapeutic, diagnostic and prognostic potential.

State-of-Art in the Age Determination of Venous Thromboembolism: A Systematic Review

Venous thromboembolism (VTE), consisting of deep vein thrombosis (DVT) and pulmonary embolism (PE), requires a forensic age determination to ascertain their causal relationship with recent events, such as trauma or medical treatment. The main objective of this systematic review is to identify the current state-of-the-art immunohistochemical methods for age determination of fatal VTE. A literature search was performed through different databases, according to Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines. Within the study, we have selected only cases represented by deceased patients for DVT and/or PTE in which thromboembolic material was collected during an autoptic examination and then subjected to a histological and an immunohistochemical investigation. Studies based on animal models were not included. We assessed bias risk. A database-based search produced a total of 19 articles. After excluding duplicate items from the selection, 14 articles were reviewed. Ten articles were excluded because they did not meet the inclusion criteria. The results have pointed out 4 studies that were included in the present analysis for a total of 157 samples of DVT and 171 PTE samples. These were analyzed using traditional histological and immunohistochemical techniques. The results must be interpreted with a critical eye because of their heterogeneity in terms of time, geography, and study design. The present review highlights the importance of associating specific immunohistochemical markers with a histological analysis for the timing of DVT/PTE fatal events. Further future experiences will hopefully endorse actual knowledge on the subject to increase the accuracy in the assessment of thrombus-embolus age.

Homeostatic regulation of axonal Kv1.1 channels accounts for both synaptic and intrinsic modifications in the hippocampal CA3 circuit

Homeostatic plasticity of intrinsic excitability goes hand in hand with homeostatic plasticity of synaptic transmission. However, the mechanisms linking the two forms of homeostatic regulation have not been identified so far. Using electrophysiological, imaging, and immunohistochemical techniques, we show here that blockade of excitatory synaptic receptors for 2 to 3 d induces an up-regulation of both synaptic transmission at CA3–CA3 connections and intrinsic excitability of CA3 pyramidal neurons. Intrinsic plasticity was found to be mediated by a reduction of Kv1.1 channel density at the axon initial segment. In activity-deprived circuits, CA3–CA3 synapses were found to express a high release probability, an insensitivity to dendrotoxin, and a lack of depolarization-induced presynaptic facilitation, indicating a reduction in presynaptic Kv1.1 function. Further support for the down-regulation of axonal Kv1.1 channels in activity-deprived neurons was the broadening of action potentials measured in the axon. We conclude that regulation of the axonal Kv1.1 channel constitutes a major mechanism linking intrinsic excitability and synaptic strength that accounts for the functional synergy existing between homeostatic regulation of intrinsic excitability and synaptic transmission.

Lower Extremity Varicose Veins: An Unusual Presentation of Small Bowel Leiomyosarcoma

Leiomyosarcomas are extremely rare and comprise only 1.2% of small bowel malignancies. Advancements in immunohistochemical techniques have allowed for the differentiation between leiomyosarcomas and gastrointestinal stromal tumors (GISTs). Leiomyosarcomas remain difficult to detect via endoscopy and require a more intricate diagnostic approach. The staging and sizing of these tumors are important prognostic indicators. We report a case of a 67-year-old male who presented with bulging lower extremity veins, abdominal bloating, and weight loss. A CT of the abdomen and pelvis revealed a pelvic mass arising from the small bowel and a metastatic hepatic lesion, which was found to be compressing the inferior vena cava. A biopsy of the hepatic lesion confirmed the diagnosis of metastatic leiomyosarcoma.

Diagnostic Accuracy of Different Histological Stains for Helicobacter Pylori Detection, taking Immunohistochemistry as Gold Standard

Aim: Helicobacter pylori infection has been ascertained to play pivotal role in the pathogenesis of chronic gastritis and gastric neoplasia.1The present study was performed to evaluate the diagnostic accuracy of H&E stain and Giemsa stain for the histological diagnosis of helicobacter pylori by taking immunohistochemical staining as a gold standard. Methods: A total of 155 cases were included in our study. The received biopsies were fixed in 10% buffered formalin, grossed and stained with H&E and giemsa stain. A board of histopathologists analyzed the morphological details to ascertain the diagnosis. The biopsies were stained by using immunohistochemical techniques against H. pylori antigens, and the procedure was performed according to the guidelines provided by the manufacturer considering the appropriate positive and negative controls for staining. IHC staining was evaluated autonomously and recorded on the proforma as positive and negative cases. Results: In our study, mean age was calculated as 38.4±11.57 years, 74(47.74%) were male and 81(52.26%) were females, frequency of H.Pylori on gold standard was recorded as 109(70.32%), the diagnostic accuracy of hematoxylin-eosin stain for helicobacter pylori detection by taking immunohistochemical staining as a gold standard measure was calculated as 63.30%, 65.22%, 81.18%, 42.86% and 63.87% for sensitivity, specificity, positive predictive value, negative predictive value and accuracy rate respectively, while these findings were recorded as 74.31%, 80.43%, 90%, 56.92% and 76.12% for Giemsa stain. Conclusion: We concluded that the diagnostic accuracy of H&E and Giemsa stains for detection of HP is promising and cost-effective method in our population. MeSH words: Helicobacter pylori, Immunohistochemistry, Hematoxylin, Pathology, Diagnosis

Immunohistochemical Analysis of the Beclin-1 Expression Predicts the Progression of Oral Squamous Cell Carcinoma

Background: Autophagy is a cellular process responsible for maintaining homeostasis; a dysregulation of this process is involved in the development and progression of neoplasms. Beclin-1 is one of the major proteins linked to autophagy. However, the data regarding the association between the role of Beclin-1 and the progression of Oral Squamous Cell Carcinoma (OSCC) are rather low. For this reason, the objective of this study is to evaluate, through immunohistochemical techniques, the prognostic role of the expression of Beclin-1 autophagy marker in patients with OSCC. Methods: This is a single-centre retrospective study that includes patients with OSCC admitted to the Maxillofacial Unit of “Magna Graecia” University between January 2019 and September 2020. All the samples obtained from surgery were treated with anti Beclin-1 antibodies and subjected to immunohistochemical methods. Results: A total of 26 samples were analysed and the following variables were evaluated for each: percentage of positive Beclin-1 expression by tumour cells, signal strength of tumour cells, and total score. The variables considered were first normalised according to the D’Agostino and Pearson test, then analysed using the Pearson linear correlation coefficient: a statistically significant correlation was found between the parameters infiltration-intensity (p = 0.0088), infiltration-percent (p = 0.0123), intensity-total score (p = 0.0060). Conclusions: The immunohistochemical evaluation of Beclin-1 revealed a statistically significant correlation between the intensity of the molecule’s expression and a greater degree of infiltration of the neoplasm. Beclin-1 can, therefore, be considered a valid prognostic index of disease.

Presence of the Human Cytomegalovirus in Glioblastomas—A Systematic Review

Glioblastoma is a malignant brain tumor with a dismal prognosis. The standard treatment has not changed in the past 15 years as clinical trials of new treatment protocols have failed. A high prevalence of the human cytomegalovirus (HCMV) in glioblastomas was first reported in 2002. The virus was found only in the tumor and not in the surrounding healthy brain tissue. Many groups have confirmed the presence of the HCMV in glioblastomas, but others could not. To resolve this discrepancy, we systematically reviewed 645 articles identified in different databases. Of these, 81 studies included results from 247 analyses of 9444 clinical samples (7024 tumor samples and 2420 blood samples) by different techniques, and 81 articles included 191 studies that identified the HCMV in 2529 tumor samples (36% of all tumor samples). HCMV proteins were often detected, whereas HCMV nucleic acids were not reliably detected by PCR methods. Optimized immunohistochemical techniques identified the virus in 1391 (84,2%) of 1653 samples. These data suggest that the HCMV is highly prevalent in glioblastomas and that optimized immunohistochemistry techniques are required to detect it.

Exploring the role of epidermal growth factor receptor variant III in meningeal tumors

Meningioma is the second most common type of intracranial brain tumor. Immunohistochemical techniques have shown prodigious results in the role of epidermal growth factor receptor variant III (EGFR vIII) in glioma and other cancers. However, the role of EGFR vIII in meningioma is still in question. This study attempt the confer searches for the position attained by EGFR vIII in progression and expression of meningioma. Immunohistochemistry technique showed that EGFR vIII is highly expressed in benign tumors as compared to the atypical meningioma with a highly significant p-value (p<0.05). Further analysis by flow cytometry results supported these findings thus presented high intensity of EGFR vIII in low grades of meningioma. The study revealed that the significant Ki 67 values, to predictor marker for survival and prognosis of the patients. Higher expression of EGFR vIII in low grades meningiomas as compared to high-grade tumors indicate towards its oncogenic properties. To our knowledge, limited studies reported in literature expressing the EGFR vIII in meningioma tumors. Hence, Opinions regarding the role that EGFR vIII in tumorigenesis and tumor progression are clearly conflicting and, therefore, it is crucial not only to find out its mechanism of action, but also to definitely identify its role in meningioma.