Immunohistochemical expression of the cation channel TRPC6 in the submandibular and lacrimal gland and in salivary gland tumors.

BACKGROUND: Canonical transient receptor potential channels play a crucial role in cancer cell proliferation. While TRPC6 subtype detection in submandibular glands and the relevance of some TRPC channels in this gland have been shown in animal models, its histological detection in human lacrimal and submandibular glands, as well as related tumors, lacks systematic study. Studying TRPC6 in humans could lead to new therapeutic options. This research aimed to immunohistochemically detect TRPC6 in human samples of physiological lacrimal and submandibular glands and of adenoid cystic carcinoma and mucoepidermoid carcinoma. METHODS: Seven fixed body donors and samples of six cancer patients were examined. The ten tissue samples collected from the submandibular and lacrimal glands were then processed into histological slides and stained with hematoxylin-eosin. Tumor samples were provided as sections. TRPC6 presence was determined by immunohistochemistry, which was performed by indirect detection with a primary TRPC6 antibody, a secondary HRP-conjugated antibody and the chromogen diaminobenzidine. RESULTS: Results confirm TRPC6 expression in all ten physiological gland samples: all samples showed a immunohistochemical signal with varying intensity. No significant gender-specific differences could be observed. TRPC6 was detected in four of six submandibular adenoid cystic carcinoma and the mucoepidermoid carcinoma samples, especially in tumor cells' cytoplasma and nuclei. Excretory ducts consistently showed TRPC6. Mucous tubules, their nuclei and the nuclei of adipocytes generally showed no signal while serous acini and their nuclei showed a weak TRPC6 signal. CONCLUSION: The discovery of TRPC6 in glandular tissue indicates a role in salivary gland function and calcium homeostasis is a basis for further research into its significance for tumor development in adenoid cystic carcinoma and mucoepidermoid carcinoma of salivary glands. TRPC6 could be used as a target for treatment of these tumors. However, the correlation between TRPC6 and submandibular and lacrimal gland diseases requires further exploration.

Differential miRNA and Protein Expression Reveals miR-1285, Its Targets TGM2 and CDH-1, as Well as CD166 and S100A13 as Potential New Biomarkers in Patients with Diabetes Mellitus and Pancreatic Adenocarcinoma.

Early detection of PDAC remains challenging due to the lack of early symptoms and the absence of reliable biomarkers. The aim of the present project was to identify miRNA and proteomics signatures discriminating PDAC patients with DM from nondiabetic PDAC patients. Proteomics analysis and miRNA array were used for protein and miRNA screening. We used Western blotting and Real-Time Quantitative Reverse Transcription polymerase chain reaction (qRT-PCR) for protein and miRNA validation. Comparisons between experimental groups with normal distributions were performed using one-way ANOVA followed by Tukey's post hoc test, and pairwise tests were performed using t-tests. p ≤ 0.05 was considered statistically significant. Protein clusters of differentiation 166 (CD166), glycoprotein CD63 (CD63), S100 calcium-binding protein A13 (S100A13), and tumor necrosis factor-ß (TNF-ß) were detected in the proteomics screening. The miRNA assay revealed a differential miRNA 1285 regulation. Previously described target proteins of miR-1285 cadherin-1 (CDH-1), cellular Jun (c-Jun), p53, mothers against decapentaplegic homolog 4 (Smad4), human transglutaminase 2 (TGM2) and yes-associated protein (YAP), were validated via Western blotting. miR-1285-3p was successfully validated as differentially regulated in PDAC + DM via qRT-PCR. Overall, our data suggest miRNA1285-3p, TGM2, CDH-1, CD166, and S100A13 as potential meaningful biomarker candidates to characterize patients with PDAC + DM. Data are available via ProteomeXchange with the identifier PXD053169.

Impact of the COVID-19 Pandemic on Tumor Stage and Pathohistological Parameters of Vulvar Cancer.

Background/Objectives: Vulvar cancer (VC) comprises a small fraction of female neoplasms with notable high-incidence clusters among German regions. Despite a proposed impact of nationwide lockdowns in response to the COVID-19 pandemic on oncological diseases, the effect on VC staging and tumor characteristics remains yet to be resolved; therefore, analyzing pathological data from patients with squamous cell VC pre-, during, and post-COVID in a high-incidence region may offer insights into potential epidemiological and clinical trends. Methods: We identified a total of 90 patients who were diagnosed at the Institute of Pathology, University Hospital Saarland, between 2018 and 2023, and defined three distinct cohorts: a pre-COVID cohort (2018-2019), a COVID cohort (2020-2021), and a post-COVID cohort (2022-2023). Histomorphological data were collected from the individual patient reports and statistically analyzed using Fisher's exact test or the Kruskal-Wallis test. Results: Although we found no statistically significant differences in age, T-stage, perineural infiltration, blood vessel infiltration, resection status, grading, or resection margin between our three cohorts, surprisingly, we determined a greater extent of lymphovascular infiltration (Fisher's exact test; p = 0.041), as well as deeper tumor infiltration depth (Kruskal-Wallis test; p < 0.001) before the COVID-19 pandemic. Furthermore, we did not identify any soft indications of abnormalities in patient care within our center (unchanged status of the resection margins across all three cohorts). Conclusions: Our results clearly do not support a negative affection of clinical or pathobiological characteristics of VC during or after the pandemic. However, final assessments regarding the pandemic's effect on VC require additional study approaches in various regions, preferably with future extended timeframes of a longer follow-up.

Distribution of TRPC3 and TRPC6 in the human exocrine and endocrine pancreas.

BACKGROUND: Expression and function of TRPC3 and TRPC6 in the pancreas is a controversial topic. Investigation in human tissue is seldom. We aimed to provide here a detailed description of the distribution of TRPC3 and TRPC6 in the human exocrine and endocrine pancreas. METHODS: We collected healthy samples from cadavers (n = 4) and visceral surgery (n = 4) to investigate the respective expression profiles using immunohistochemical tracing with knockout-validated antibodies. RESULTS: TRPC3- and TRPC6-proteins were detected in different pancreatic structures including acinar cells, as well as epithelial ductal cells from intercalate, intralobular, and interlobular ducts. Respective connective tissue layers appeared unstained. Endocrine islets of Langerhans were clearly and homogenously immunolabeled by the anti-TRPC3 and anti-TRPC6 antibodies. Insular α, ß, γ, and δ cells were conclusively stained, although no secure differentiation of cell types was performed. CONCLUSIONS: Due to aforementioned antibody specificity verification, protein expression in the immunolabeled localizations can be accepted. Our study in human tissue supports previous investigations especially with respect to acinar and insular α and ß cells, while other localizations are here reported for the first time to express TRPC3 and TRPC6, ultimately warranting further research.

Mind the Pitfall: Solitary Nodular Fasciitis Mimicking Extra-Nodal Manifestation of Hodgkin Lymphoma on [18F]FDG PET/CT.

We report a [18F]fluorodeoxyglucose positron emission tomography/computed tomography ([18F]FDG PET/CT) scan of a 17-year-old male presenting increased focal glucose metabolism of a histologically proven solitary nodular fasciitis mimicking an extranodal manifestation of Hodgkin lymphoma. This interesting image should draw attention to considering nodular fasciitis as a possible pitfall in the staging of malignant diseases.

TRPC3 Is Downregulated in Primary Hyperparathyroidism.

Transient receptor potential canonical sub-family channel 3 (TRPC3) is considered to play a critical role in calcium homeostasis. However, there are no established findings in this respect with regard to TRPC6. Although the parathyroid gland is a crucial organ in calcium household regulation, little is known about the protein distribution of TRPC channels-especially TRPC3 and TRPC6-in this organ. Our aim was therefore to investigate the protein expression profile of TRPC3 and TRPC6 in healthy and diseased human parathyroid glands. Surgery samples from patients with healthy parathyroid glands and from patients suffering from primary hyperparathyroidism (pHPT) were investigated by immunohistochemistry using knockout-validated antibodies against TRPC3 and TRPC6. A software-based analysis similar to an H-score was performed. For the first time, to our knowledge, TRPC3 and TRPC6 protein expression is described here in the parathyroid glands. It is found in both chief and oxyphilic cells. Furthermore, the TRPC3 staining score in diseased tissue (pHPT) was statistically significantly lower than that in healthy tissue. In conclusion, TRPC3 and TRPC6 proteins are expressed in the human parathyroid gland. Furthermore, there is strong evidence indicating that TRPC3 plays a role in pHPT and subsequently in parathyroid hormone secretion regulation. These findings ultimately require further research in order to not only confirm our results but also to further investigate the relevance of these channels and, in particular, that of TRPC3 in the aforementioned physiological functions and pathophysiological conditions.

Endothelial PTP1B Deletion Promotes VWF Exocytosis and Venous Thromboinflammation.

BACKGROUND: Endothelial activation promotes the release of procoagulant extracellular vesicles and inflammatory mediators from specialized storage granules. Endothelial membrane exocytosis is controlled by phosphorylation. We hypothesized that the absence of PTP1B (protein tyrosine phosphatase 1B) in endothelial cells promotes venous thromboinflammation by triggering endothelial membrane fusion and exocytosis. METHODS: Mice with inducible endothelial deletion of PTP1B (End.PTP1B-KO) underwent inferior vena cava ligation to induce stenosis and venous thrombosis. Primary endothelial cells from transgenic mice and human umbilical vein endothelial cells were used for mechanistic studies. RESULTS: Vascular ultrasound and histology showed significantly larger venous thrombi containing higher numbers of Ly6G (lymphocyte antigen 6 family member G)-positive neutrophils in mice with endothelial PTP1B deletion, and intravital microscopy confirmed the more pronounced neutrophil recruitment following inferior vena cava ligation. RT2 PCR profiler array and immunocytochemistry analysis revealed increased endothelial activation and adhesion molecule expression in primary End.PTP1B-KO endothelial cells, including CD62P (P-selectin) and VWF (von Willebrand factor). Pretreatment with the NF-κB (nuclear factor kappa B) kinase inhibitor BAY11-7082, antibodies neutralizing CD162 (P-selectin glycoprotein ligand-1) or VWF, or arginylglycylaspartic acid integrin-blocking peptides abolished the neutrophil adhesion to End.PTP1B-KO endothelial cells in vitro. Circulating levels of annexin V+ procoagulant endothelial CD62E+ (E-selectin) and neutrophil (Ly6G+) extracellular vesicles were also elevated in End.PTP1B-KO mice after inferior vena cava ligation. Higher plasma MPO (myeloperoxidase) and Cit-H3 (citrullinated histone-3) levels and neutrophil elastase activity indicated neutrophil activation and extracellular trap formation. Infusion of End.PTP1B-KO extracellular vesicles into C57BL/6J wild-type mice most prominently enhanced the recruitment of endogenous neutrophils, and this response was blunted in VWF-deficient mice or by VWF-blocking antibodies. Reduced PTP1B binding and tyrosine dephosphorylation of SNAP23 (synaptosome-associated protein 23) resulting in increased VWF exocytosis and neutrophil adhesion were identified as mechanisms, all of which could be restored by NF-κB kinase inhibition using BAY11-7082. CONCLUSIONS: Our findings show that endothelial PTP1B deletion promotes venous thromboinflammation by enhancing SNAP23 phosphorylation, endothelial VWF exocytosis, and neutrophil recruitment.

Prediction of lymph node status in patients with surgically treated head and neck squamous cell carcinoma via neck lavage cytology: A pilot study.

BACKGROUND: Neck dissection is a standardized surgical procedure for patients with head and neck squamous cell carcinoma (HNSCC) and plays a critical role in the choice of adjuvant treatment based on histopathological findings. Saline irrigation is routinely performed at the end of surgery. However, this irrigant is not used for diagnostic purposes. METHODS: Intraoperative irrigation of the neck dissection wound was performed in 56 patients with HNSCC (N = 93 neck dissections), and the cytological suspension obtained was processed via the liquid-based cytology (LBC) technique, Papanicolaou staining, and immunocytochemical staining. Microscopic preparations were screened for the presence of tumor cells and classified as positive, borderline, or negative. These results were correlated with the histopathological and clinical data. RESULTS: Neck lavage LBC demonstrated high diagnostic value in detecting lymph node metastases (N+) with extracapsular spread (ECS), with a specificity, sensitivity, negative predictive value, and positive predictive value of 93.1%, 100%, 100%, and 80%, respectively. Tumor cells were detected in 4.8% of N- cases, 20% of N+ cases without ECS, and 100% of N+ cases with ECS. Receiver operating characteristic curve analysis showed an area under the curve of 0.8429 for the prediction of N+ (p < .0001) and 0.9658 for the prediction of N+ with ECS (p < .0001). CONCLUSIONS: Differential lavage cytology can provide valid and rapid information on the lymph node status in patients with HNSCC and showed an excellent correlation with histopathology. Thus, neck lavage LBC may facilitate faster and more reasonable planning of adjuvant treatment and help improve the therapeutic management of patients with HNSCC.

Dual Sec62/Ki67 immunocytochemistry of liquid-based cytological preparations represents a highly valid biomarker for non-invasive detection of head and neck squamous cell carcinomas.

BACKGROUND: Head and neck squamous cell carcinomas (HNSCC) are frequently diagnosed in advanced stages, which limits therapeutic options and results in persistently poor patient outcomes. The aim of this study was to use liquid-based swab cytology (LBC) in combination with dual immunocytochemical detection of migration and proliferation markers Sec62 and Ki67 in order to allow non-invasive early detection of HNSCC as well as to analyse the diagnostic validity of this method for predicting the malignancy of suspicious oral lesions. METHODS: 104 HNSCC patients and 28 control patients, including healthy patients (n = 17), papilloma (n = 1) and leukoplakia patients (n = 10), were included in this study. For all patients, an LBC swab followed by simultaneous immunocytochemical detection of Sec62 and Ki67 was performed. Immunocytochemical as well as cytopathological results were correlated with histological diagnoses and clinical findings. RESULTS: All HNSCC patients (100%) showed dual Sec62/Ki67 positivity, and all control patients except for the papilloma patient were negative for Sec62/Ki67 (96.4%), resulting in a 100% sensitivity and 96.4% specificity of Sec62/Ki67 dual stain for non-invasive detection of HNSCC. The positive predictive value was 99% and the negative predictive value was 100%. Sec62 expression levels showed a positive correlation with tumour de-differentiation (p = 0.0489). CONCLUSION: Simultaneous immunocytochemical detection of Sec62/Ki67 using LBC represents a promising non-invasive and easy-to-apply tool for the early detection of HNSCC in routine clinical practice. This novel technique can help to avoid incisional biopsies and reduce the frequency with which general anaesthesia is used in diagnostic procedures in patients with suspicious oral lesions.

Expression of 3q Oncogene SEC62 Predicts Survival in Head and Neck Squamous Cell Carcinoma Patients Treated with Primary Chemoradiation.

Primary chemoradiotherapy (CRT) is an established treatment option for locally advanced head and neck squamous cell carcinomas (HNSCC) usually combining intensity modified radiotherapy with concurrent platinum-based chemotherapy. Though the majority of patients can be cured with this regimen, treatment response is highly heterogeneous and can hardly be predicted. SEC62 represents a metastasis stimulating oncogene that is frequently overexpressed in various cancer entities and is associated with poor outcome. Its role in HNSCC patients undergoing CRT has not been investigated so far. A total of 127 HNSCC patients treated with primary CRT were included in this study. The median follow-up was 5.4 years. Pretherapeutic tissue samples of the primary tumors were used for immunohistochemistry targeting SEC62. SEC62 expression, clinical and histopathological parameters, as well as patient outcome, were correlated in univariate and multivariate survival analyses. High SEC62 expression correlated with a significantly shorter overall survival (p = 0.015) and advanced lymph node metastases (p = 0.024). Further significant predictors of poor overall and progression-free survival included response to therapy (RECIST1.1), nodal status, distant metastases, tobacco consumption, recurrence of disease, and UICC stage. In a multivariate Cox hazard proportional regression analysis, only SEC62 expression (p = 0.046) and response to therapy (p < 0.0001) maintained statistical significance as independent predictors of the patients' overall survival. This study identified SEC62 as an independent prognostic biomarker in HNSCC patients treated with primary CRT. The role of SEC62 as a potential therapeutic target and its interaction with radiation-induced molecular alterations in head and neck cancer cells should further be investigated.

Orbital prosthese anchored by dental implants / Prótesis orbitarias fijas mediante implantes dentales

Todo indica que los implantes dentales osteointegrados pueden usarse para fijar prótesis orbitarias en casos de cirugía previa de extirpación amplia de la órbita incluyendo la excenteración de un ojo. Estos implantes se aplican en el primer tiempo y se conectan con refuerzos en el segundo en los procedimientos quirúrgicos de dos etapas. Se estudiaron nueve pacientes caucásicos con un total de 25 implantes periorbitarios osteointegrados. El estudio fue prospectivo, longitudinal para evaluar las variantes integradas (grupo A:14 implantes) y separada (grupo B: 11 implantes) de la cirugía del primer tiempo quirúrgico. Todos los implantes estuvieron estables y las prótesis orbitarias se usaron durante un promedio de tres años (cuatro pacientes del grupo A: 11 implantes; tres pacientes del grupo B: 9 implantes). Sólo se perdió un implante debido a gran carga de estrés sin signos de inflamación (grupo B; después de tres años). Durante el tiempo promedio de observación de 36 meses se observaron únicamente leves reacciones cutáneas alrededor de los refuerzos transdérmicos. No hubo diferencias respecto a los implantes entre el grupo A y el B. El diseño de los implantes regulares demostraron ser apropiados para retener las prótesis craneofaciales con los propósitos antes mencionados. Los resultados de este estudio preliminar sugieren que los implantes dentales son funcionales en la órbita independiente de que sean colocados en el primer tiempo quirúrgico ya sea integrado o separado. Estudios futuros incluirán mayor número de pacientes de implantes

Considerations on a long-term course of a plexiform amelonlastoma with a recurrence in the soft tissue / Consideraciones de la evolución a largo plazo del ameloblastoma plexiforme con recidiva de tejidos blandos

Los ameloblastomas (ICD-DA.213.XI) son tumores unicéntricos, no funcionales, clínicamente persistentes y benignos, intermitentes en su crecimiento. Desde el punto de vista histológico se pueden distinguir los siguientes patrones: espinocelular, acantomatoso, granular, folicular (simple), basocelular y plexiforme. Hasta ahora no es aún posible determinar los factores pronósticos para el comportamiento de los ameloblastomas. Esto no se puede hacer ni por la clínica, radiología o histología. Sólo los métodos biológicos moleculares, inmunohistoquímicos pueden enfrentar el problema como se muestra durante curso de un ameloblastoma plexiforme (ICD-0-9310.0) muy bien documentado durante 20 años con especial atención a su recidiva en tejidos blandos de mejilla izquierda. Se discuten puntos importantes de la patología y terapéutica. Se requiere de amplia cooperación de las secciones de clínica y patología durante un largo lapso de tiempo en cada caso individual

Chronic fatigue syndrome (CFS): Review of clinical data from 107 cases

Se estudiaron 107 pacientes con SFC de entre 7 y 76 años de edad (promedio 41.8 años). 67 fueron del sexo femenino y 47 del masculino (relación F/M 1.27L1). Los períodos de seguimiento duraron desde 10 meses hasta 7.5 años. Los pacientes fueron reclutados mediante un cuesitonario (anexo) adecuando los criterios de Holmes para SFC, se realizó además examen clínico, biometría hemática, química sanguínea, fenotipificación de linfocitos y control de otros parámetros inmunológicos, así como estudios para detección de infecciones. En un paciente se diagnosticó síndrome miálgico-eosinofílico, 6 pacientes presentaron deficiencia de vitamina D3 (25-OH), cuyos síntomas se revirtieron con tratamiento de reposición. De los 100 pacientes restantes, 4 habían tenido episodios psiquiátricos previos. En 67 por ciento de los pacientes con SFC se comprobó infección persistente o recurrente, otro 16 por ciento sugería infección. La infección activa detectada con más frecuencia fue por virus herpes humano-6 (HHV-6) en 38.6 por ciento de los casos, seguida por infección por virus de Epstein-Barr (VEB) en 19.6 por ciento, menos frecuente fue la detección de virus herpes simple (VHS), chlamydia, campylobacter, coxsackie, citomegalovirus (CMV), yersinia o cándida. En el 46 por ciento de los pacientes se encontraron signos de inmunodeficiencia, mientras que en un 20 por ciento adicional la evidencia fue menos clara, consistiendo por ejemplo en disminución de la estimulación linfocitaria (46 por ciento), baja cantidad de células asesinas naturales (NK) (35 por ciento) y relación de células CD4/CD8 disminuída (21 por ciento). Del 14 al 21 por ciento de los pacientes presentaron datos de laboratorio de infección, alergias o de alteraciones autoinmunes, incluyendo niveles aumentados de factor de necrosis tumoral (sTNFÓ) en 41 por ciento, proteína C reactiva (CRP) positiva en 19.4 por ciento e incremento en los niveles de IgE (14.3 por ciento). Los de pacientes con SFC sin patogénesis identificable. Estos datos confirman el concepto de Demitrak acerca del SFC. De que no se trata de una entidad con una sola etiología sino que existen varios y diversos eventos que coinciden parcialmente y comparten las vías fisiopatológicas que finalmente resultan en este síndrome clínico

Modulation of nuclear factor kappa b (NF-kB) activity in human herpes virus-6 (HHV-6) infection and its sequellae for cell function: Effects of antisense DNA treatment

Varias poblaciones celulares, susceptibles y no susceptibles, fueron infectadas con el virus herpes humano-6 (HHV-6) y tratadas posteriormente con ácido desoxirribonucleico (ADN) -NF-kB antisentido. Algunos cambios celulares secundarios a la integración del genoma viral pudieron ser corregidos con este tratamiento, mientras que aquellos inducidos por efecto de membrana celular del virus no respondieron de manera uniforme. Es así como el tratamiento con NF-kB puede servir para investigaciones posterioes específicas de los efectos virales en la célula. También puede servir para estudiar las actividades competitivas de las proteínas de NF-kB en los sitios genómicos homólogos