Salivary gland pathologies: evolution in classification and association with unique genetic alterations.

PURPOSE: The correct classification of salivary gland pathologies is crucial for choosing a treatment method and determining the prognosis. Better outcomes are now achievable thanks to the introduction of new therapy approaches, such as targeted therapies for malignant salivary gland tumors. To apply these in clinical routine, a clear classification of the lesions is required. METHODS: The following review examines all changes from the first World Health Organization (WHO) Classification of salivary gland pathologies from 1972 to fifth edition from 2022. Possible developments in the diagnosis and classification of salivary gland pathology are also presented. RESULTS: The current WHO classification is the fifth edition. With the development of new diagnostic methods, based on genetic alterations, it provides insight into the molecular basis of lesions. This has resulted in the evolution of classification, introduction of new entities and reclassification of existing ones. CONCLUSIONS: Genetic alterations will become increasingly more significant in the identification of salivary gland pathologies in the future. These alterations will be helpful as prognostic and predictive biomarkers, and may also serve as targets for anti-cancer therapies.

Solitary breast cancer metastasis to the facial nerve.

Facial nerve (FN) palsy, as a solitary symptom, resulting from metastatic tumor is not frequent. In this article, we report an unusual case of the breast cancer metastasis to the labyrinthine segment of a facial nerve.

Non-neoplastic adrenal pseudocysts: A clinicopathologic study of 44 cases with potential insights into pathogenesis.

BACKGROUND: Literature on non-neoplastic adrenal pseudocysts (NNAPC) remains limited and to date no large series have been reported. The pathogenesis of these lesions remains poorly defined, however a vascular origin is most often suggested in the literature. We aimed to evaluate the clinicopathological features and the spectrum of vascular changes within NNAPC, in order to better understand the mechanisms and circumstances of their pathogenesis. METHODS AND RESULTS: We reviewed 44 cases of surgically resected NNAPC. There were 30 females and 14 males ranging from 23 to 82 years (median, 53 years). On the basis of histopathologic and immunohistochemical analysis of the vascular changes the following types were defined: pseudocysts with lymphatic-related changes (type 1, n = 16), pseudocysts with large vein-related changes (type 2, n = 15) and pseudocysts with blood vessel microvasculature-related changes (type 3, n = 13). The median patient age of the latter group was higher than that of type 1 and 2 (64 years versus 51 and 50 years, respectively; p = 0.0002). Type 3 pseudocysts were more frequently associated with a history of systemic vascular and vascular-related disorders than type 1 and type 2 pseudocysts (92% versus 33% and 64%, respectively; p = 0.008). Type 1 pseudocysts were more frequently connected with a history of previous intra-abdominal surgical procedures than type 2 and 3 pseudocysts (60% versus 7% and 25%, respectively; p = 0.0079). CONCLUSIONS: NNAPC are clinically heterogenous and can arise on a background of various vascular changes. They may represent end-stage processes related to lymphangiomatous lesions, changes in adrenal venous structures or microvasculature.

High cytoplasmic HuR expression is associated with advanced pT stage, high grade and increased microvessel density in urothelial bladder carcinoma.

PURPOSE: HuR (human antigen R) protein is a RNA binding protein that stabilizes the mRNA and controls the translation of genes involved in cell proliferation, differentiation, and carcinogenesis. Overexpression of HuR was reported in a variety of cancers, however its clinical significance in urothelial bladder cancer (UBC) is still unknown. Our aim is to investigate the association between HuR expression and selected histopathological factors, such as tumor grade, pT stage, regional lymph nodes status and microvessel density (MVD). METHODS: We studied expression of HuR protein in 119 patients with UBC in stages pTis and pTa-pT4 using immunohistochemistry (IHC). Tumor MVD was evaluated immunohistochemically using anti-CD31 antibody. RESULTS: We observed no association between nuclear HuR immunoreactivity and tumor grade, stage or MVD. We found a significant association between cytoplasmic HuR positivity and high tumor grade, pT stage and MVD (p<0,001). We also observed significantly higher MVD values in cases with positive cytoplasmic HuR expression (p<0,001). No association between HuR immunoreactivity and lymph nodes status was found. CONCLUSIONS: Our results may suggest that HuR is involved in the process of acquiring malignant histopathological features and ability to invade the muscularis propria by UBC cells. Considering frequent difficulties in diagnosing UBC in specimens obtained from transurethral tumor resection and the risk of understaging, cytoplasmic HuR expression would suggest an advanced disease and necessitate serial sectioning of the specimen in search of muscle invasion. Association between HuR expression and MVD could suggest HuR involvement in the process of angiogenesis in UBC.

HIF-1α expression is inversely associated with tumor stage, grade and microvessel density in urothelial bladder carcinoma.

Urothelial bladder carcinoma (UBC) is the most common urinary tract malignancy. The most important histopathological factors affecting prognosis are cancer stage and grade. Studies show that microvessel density (MVD) reflecting angiogenesis is also associated with clinicopathological features and affects the outcome in UBC. One of the most important regulators of angiogenesis is hypoxia inducible factor 1 (HIF-1). Previous reports describing expression of the HIF-1α subunit in UBC showed unclear and inconsistent results. Our study attempted do evaluate the association between HIF-1α expression and tumor stage, grade, lymph nodes status and MVD in UBC. We performed immunohistochemical staining in 99 UBC cases, including 38 non-muscle invasive (NMIBC) and 61 muscle invasive tumors (MIBC). We observed inverse relationships between HIF-1α immunoreactivity score (IRS) and tumor stage, grade and MVD. Significantly lower HIF-1α IRS values were observed in MIBC and high grade cancers. We found a significant negative correlation between HIF-1α IRS and MVD. These results suggest that HIF-1α pathway is not involved in UBC growth and progression, and that angiogenesis in high grade MIBC is not regulated by HIF-1. Our findings contradict previous reports regarding HIF-1α, MVD and UBC which shows the necessity of additional molecular studies in this field.

Expression of E-cadherin, ß-catenin, and epithelial membrane antigen does not predict survival in patients with high-risk non-muscle-invasive bladder cancer.

THE AIM OF THE STUDY: The aim of the study was to validate the value of E-cadherin and ß-catenin expression and to test an alternative prognostic marker, epithelial membrane antigen (EMA). MATERIAL AND METHODS: Forty-nine consecutive patients with primary stage T1 non-muscle-invasive bladder cancer (NMIBC) were enrolled in this study. Tissue specimens were stained with the following mouse anti-human antibodies: anti-E-cadherin, anti-ß-catenin, and anti-EMA. Reaction intensity within cancer cells was assessed according to the immunoreactive score (IRS). Finally, the association between the expression of selected proteins and patient survival was assessed. RESULTS: The mean follow-up was 34.8 months. Recurrence-free survival, progression-free survival, and overall survival (OS) were 47.5%, 72.5%, and 72.5%, respectively. Differences in the IRS for ß-catenin and EMA were found clinically, but were not statistically significant in prediction of the risk of disease progression (p > 0.05). No difference in protein expression was observed regarding the risk of recurrence, OS, or cancer-specific mortality (p > 0.05). Stratification of patients based on the IRS into three groups (poor, moderate, and intensive reaction) failed to identify a prognostic marker among the tested proteins (p > 0.05). CONCLUSIONS: Expression of E-cadherin, ß-catenin, and EMA cannot reliably predict survival in patients with high-risk NMIBC. Further searches are needed to identify tissue markers of progression and recurrence in NMIBC.

The role of apelin in central cardiovascular regulation in rats with post-infarct heart failure maintained on a normal fat or high fat diet.

Based on the available literature, it can be assumed that in cases of post-infarct heart failure (HF) and obesity, a significant change in the central regulation of the cardiovascular system takes place with, among others, the involvement of the apelinergic system. The main objective of the present study was to clarify the role of apelin-13 in the central regulation of the cardiovascular system in Sprague Dawley rats with HF or sham operated (SO) and fed on a normal fat (NFD) or a high fat diet (HFD). The study was divided into two parts: Part I, hemodynamic studies; and Part II, biochemical and molecular studies. The animals were subjected to the following research procedures. Part I and II: feeding NFD or HFD; experimental induction of HF or SO; Part I: intracerebroventricular (ICV) infusion of the examined substances, monitoring of mean arterial blood pressure (MABP) and heart rate (HR); Part II: venous blood and tissue samples collected. ICV infusion of apelin-13 caused significantly higher changes in ΔMABP in the SO NFD group. No changes were noted in ΔHR in any of the studied groups. Apelin and apelin receptor (APJ) mRNA expression in the brain and adipose tissues was higher in the HF rats. HFD causes significant increase in expression of apelin and APJ mRNA in the left ventricle. In conclusion, HF and HFD appear to play an important role in modifying the activity of the central apelinergic system and significant changes in mRNA expression of apelin and APJ receptor.

Tunnel technique and subepithelial connective tissue graft, with or without cross-linked hyaluronic acid, in the treatment of multiple gingival recessions: 12-month outcomes of a randomized clinical trial.

BACKGROUND: This study evaluated the influence of the adjunctive application of a cross-linked hyaluronic acid (HA) in the treatment of multiple gingival recessions, using a modified coronally advanced tunnel (MCAT) technique and subepithelial connective tissue graft (SCTG) (MCAT+SCTG±HA). METHODS: A randomized, split-mouth, double-masked comparison of the effects of MCAT+HA+SCTG (test) versus MCAT+SCTG (control) in the treatment of multiple, contralateral gingival recessions with clinical, esthetic, and histological evaluations was carried out. All samples were stained with hematoxylin and eosin, Masson's trichrome, Verhoeff-Van Gieson, and Alcian blue stain for semiquantitative evaluation. The primary outcome variable was 12-month mean root coverage (MRC). RESULTS: Twenty-four patients with 266 gingival recessions received both control and test treatments (133 recessions per group). 12-month MRC of the MCAT+HA+SCTG group was not significantly different from the MCAT+SCTG group with 84.32%±  34.46% and 85.71%±  36.43%, respectively (p = 0.991). Both treatment modes produced favorable esthetic outcomes (root coverage esthetic score [RES] 9.51±  1.01 tests vs. 9.26±  1.10 controls, p = 0.7292). However, the application of HA improved soft tissue texture (p = 0.0091). The remaining end point measures did not differ significantly between groups. Histological evaluation showed a significantly greater number of elastic fibers and a moderate increase in collagen fiber density in biopsy samples taken from the test sides when compared to the control sides (p = 0.0419 and p = 0.300, respectively). CONCLUSIONS: MCAT+SCTG is an effective procedure in the treatment of multiple recession Type 1 (RT1) and RT2 recessions. There were no statistically significant differences in evaluated clinical treatment outcomes in the MCAT+HA+SCTG group compared to the MCAT+SCTG group within a period of 12 months. The application of HA increased collagen and elastic fiber density.

E-Cadherin, Integrin Alpha2 (Cd49b), and Transferrin Receptor-1 (Tfr1) Are Promising Immunohistochemical Markers of Selected Adverse Pathological Features in Patients Treated with Radical Prostatectomy.

In patients treated for prostate cancer (PCa) with radical prostatectomy (RP), determining the risk of extraprostatic extension (EPE) and nodal involvement (NI) remains crucial for planning nerve-sparing and extended lymphadenectomy. The study aimed to determine proteins that could serve as immunohistochemical markers of locally advanced PCa. To select candidate proteins associated with adverse pathologic features (APF) reverse-phase protein array data of 498 patients was retrieved from The Cancer Genome Atlas. The analysis yielded 6 proteins which were then validated as predictors of APF utilizing immunohistochemistry in a randomly selected retrospective cohort of 53 patients. For univariate and multivariate analysis, logistic regression was used. Positive expression of TfR1 (OR 13.74; p = 0.015), reduced expression of CD49b (OR 10.15; p = 0.013), and PSA (OR 1.29; p = 0.013) constituted independent predictors of EPE, whereas reduced expression of e-cadherin (OR 10.22; p = 0.005), reduced expression of CD49b (OR 24.44; p = 0.017), and PSA (OR 1.18; p = 0.002) were independently associated with NI. Both models achieved high discrimination (AUROC 0.879 and 0.888, respectively). Immunohistochemistry constitutes a straightforward tool that might be easily utilized before RP. Expression of TfR1 and CD49b is associated with EPE, whereas expression of e-cadherin and CD49b is associated with NI. Since following immunohistochemical markers predicts respective APFs independently from PSA, in the future they might supplement existing preoperative nomograms or be implemented in novel tools.

MCM5 urine expression (ADXBLADDER) is a reliable biomarker of high-risk non- muscle-invasive bladder cancer recurrence: A prospective matched case-control study.

BACKGROUND: Mini Chromosome Maintenance 5 (MCM5) is considered as a urinary biomarker of bladder cancer. ADXBLADDER is a commercially available test to detect MCM5 antibodies. OBJECTIVE: External validation of ADXBLADDER test as a urinary biomarker of histopathologically confirmed non-muscle invasive bladder cancer (NMIBC) recurrence. METHODS: The study enrolled 119 consecutive patients with a history of NMIBC and 37 healthy volunteers matched as controls. Single, full-void urine samples were collected from patients before cystoscopy ± TUR. To measure MCM5 expression, Arquer Diagnostics ADXBLADDER test was used. The study protocol was registered within the clinical trials database (NCT03796299). RESULTS: Among patients with NMIBC history, recurrence was diagnosed in 83 patients (69.7%). ADXBLADDER demonstrated sensitivity of 73.5% (95% confidence interval (CI) 62.7%-82.6%), specificity of 33.3% (95% CI 18.6% to 51%), overall negative predictive value (NPV) of 35.3% (95% CI 23.3% to 49.5%) and overall positive predictive value of 71.8% (95% CI 66.1% to 76.8%) for detecting recurrence. In a control group, false positive ADXBLADDER results were noticed in 18 patients (48.6%). The sensitivity and NPV were the highest in invasive tumors (100% and 100%, respectively) and in high-grade recurrences (81.8% and 94.1%, respectively). CONCLUSIONS: ADXBLADDER has a moderate sensitivity and poor specificity in detecting NMIBC recurrence. However, it properly diagnoses patients with T1+ stage recurrence or high-grade tumors.

Expression of Toll-Like Receptors in the Animal Model of Bladder Outlet Obstruction.

INTRODUCTION: Bladder outlet obstruction (BOO) occurs in more than 20 percent of the adult population and may lead to changes in the structure and function of the bladder. The main objective of the study was to evaluate the expression of Toll-like receptor 4 (TLR 4) and Toll-like receptor 9 (TLR 9) in the animal model of BOO as potential triggers of the inflammation phase in the bladder. In addition, the modulating effect of alpha-1 adrenergic antagonist (tamsulosin) on TLR 4 and TLR 9 expression and inflammatory markers was assessed. Material and Methods. Thirty-two male, 9-week-old Sprague Dawley rats were randomly divided into 4 groups: SOP-sham-operated rats with a placebo (water); SOB-sham-operated rats with an alpha-1 adrenergic antagonist; BOOP-rats with BOO and a placebo; and BOOB-rats with BOO and an alpha-1 adrenergic antagonist. The rats were given a placebo or alpha-1 adrenergic antagonist for 15 days. Next, urine and the bladder were collected from the rats for histopathological and biochemical study. RESULTS: Histopathological analysis showed chronic inflammation without acute inflammation in the bladder. TLR 4 showed positive cytoplasmic reactivity in the urothelium and the smooth muscles of the bladder. TLR 9 showed positive cytoplasmic reactivity only in the urothelium. BOO caused an increase in TLR 4 and TLR 9 expression. Furthermore, treatment with an alpha-1 adrenergic antagonist had no significant effect on TLR 4 and TLR 9 expression in rats with BOO. BOO caused a significant increase in urine concentration of interleukin 6 (IL-6), while alpha-1 antagonist reduced the urine concentration of IL-6 and the concentration of interleukin 18 (IL-18). CONCLUSIONS: The results suggest the participation of TLR 4 and TLR 9 receptors in the induction of inflammation in the bladder, which is the first phase in the development of pathophysiological changes in BOO.

Adrenal cyst with both Müllerian and mesothelial differentiation- a clinicopathological and immunohistochemical study with implications for histogenesis.

True epithelial-lined cysts are rare forms of adrenal cystic lesions, the pathogenesis of which is still not fully understood. In this report we present a case of an adrenal cyst diagnosed incidentally on imaging in a 31-year-old, previously healthy, obese woman. Due to non-specific hormonal disorders and enlargement of the lesion, a right-sided laparoscopic adrenalectomy was performed. The cyst was lined predominantly by ciliated cuboidal-to-columnar, Müllerian-type epithelium, and focally by flat-to-cuboidal, mesothelium-like lining. Immunohistochemistry demonstrated a strong positive reaction in the cells of both types of lining for CKAE1+E3, CK19, CK7 and WT1, and both had a negative reaction for CK20, CD34, Melan-A, SF1, TTF1, SMA and CDX2. The cells of the ciliated cuboidal-to-columnar epithelium were strongly positive for PAX8, ER, Ep-CAM and EMA, focally positive for PR, and were negative for calretinin, whereas the cells of the flat-to-cuboidal lining were positive for calretinin and podoplanin and showed only a weak positive response in individual cells for PAX8, EMA and Ep-CAM, but were negative for ER and PR. This is the first reported case of an adrenal ciliated epithelial cyst with Müllerian differentiation (confirmed immunohistochemically) in the English literature. The differences in morphology and immunophenotype of the two types of lining (epithelial Müllerian phenotype versus mesothelial phenotype), suggest that some adrenal epithelial cysts probably form due to metaplasia of mesothelium-derived lining. A similar mechanism may also be involved in the pathogenesis of at least some of the so-called Müllerian cysts (or inclusions) in other locations.

Clinicopathological and immunohistochemical analysis of epithelial-lined (true) cysts of the adrenal gland with proposal of a new histogenetic categorization.

Epithelial- lined (true) cysts are rare lesions and until now the only information we had about their histogenesis was based on the analysis of a few cases. We retrospectively reviewed 8 cases of cysts with a true epithelial lining (confirmed immunohistochemically). The pathological findings and immunohistochemical analysis of the epithelial linings allowed for categorization of the cysts into 3 groups. Five cysts had pure mesothelial lining, which was flattened to cuboidal, and demonstrated a positive reaction for mesothelial markers (eg. calretinin, WT1), and a negative reaction for EpCAM, EMA, PAX8 and ER. Two cysts had cuboidal to flattened lining, the cells of which were diffusely or focally positive for mesothelial markers, for some epithelial markers (eg. EpCAM and EMA) and despite a lack of müllerian-type epithelium demonstrated a positive reaction for PAX8 and focally for ER. A cyst derived from adreno-hepatic fusion (AHF)-related intra-adrenal bile ductules was diagnosed in a right adrenal gland which was directly adherent to the liver, microscopically features of AHF were visible with intermingling of adrenal and liver parenchymal cells. The immunoreactivity pattern was similar among the preserved cells of the cyst-lining, the intra-adrenal bile ductules and the normal bile ductules in the adjoining liver parenchyma. On the basis of this case series from a single institution (8 presented now and 1 reported before) we propose a new histogenetic categorization of adrenal epithelial cysts into: 1. pure mesothelial cysts (the most common type), 2. mesothelial cysts with incomplete or complete müllerian metaplasia 3. AHF-related cysts.

Role of angiogenesis in urothelial bladder carcinoma.

INTRODUCTION: Bladder cancer is the most common urinary tract malignancy in western countries. In recent years, extensive research has suggested that angiogenesis plays an important role in bladder cancer biology, contributing to tumor growth and progression. MATERIAL AND METHODS: In this review, we discuss general mechanisms of angiogenesis and highlight the influence of pro- and anti-angiogenic factors, and cancer stem cells on bladder cancer biology, their relation to disease progression, and potential use in novel targeted therapies. RESULTS: Expression of a number of proangiogenic factors, including HIF-1, VEGF, bFGF, IL-8 and MMPs, as well as anti-angiogenic factor TSP-1, was found to be altered in bladder tumors. Involvement of cancer stem cells in bladder cancer development was also proposed. CONCLUSIONS: High expression of most pro-angiogenic factors correlated with disease progression and shorter patient survival, but discrepancies between studies urge us to continue evaluating the significance of angiogenesis in bladder cancer.

Urothelial bladder carcinoma in young patients is characterized by a relatively good prognosis.

INTRODUCTION AND AIM: Urothelial bladder carcinoma (UBC) is a very rare condition in patients aged below 50 years. The aim of the study was to answer the question whether the characteristics of cancer in this group of patients differ from general UBC features. MATERIAL AND METHODS: Altogether 2160 patients treated with primary transurethral resection due to a bladder tumor were included in the study. The mean age of the cohort was 69.1 years (range 11-100). Patients were divided into three subgroups depending on age: age <41 years (group 1), age 41-50 years (group 2), age >50 years (group 3). Sex ratio, tumor grade, and stage of disease were recorded. RESULTS: Women constituted 18.5%, 19.2%, and 25.8% of the patients in groups 1, 2, and 3, respectively (P < 0.05). WHO grade 3 tumors were diagnosed in 0%, 8.5%, and 17.2%, respectively (P < 0.05). Non-invasive papillary carcinoma was found in 100.0%, 76.7%, and 62.7%, respectively (P < 0.05). The incidence of muscle-invasive bladder cancer was 0%, 11.0%, and 15.6%, respectively (P < 0.05). CONCLUSIONS: Pathological characteristics of UBC are dependent on the patients' age. Being a very rare condition, UBC in young patients is characterized by a relatively good prognosis.

Current concepts on pathogenesis and biology of metastatic osteosarcoma tumors.

Osteosarcoma (OS) remains the most common malignancy among orthopaedic neoplasms. Despite advanced surgical techniques and attempts to use second-line chemotherapy, 5 year overall survival in OS patients is still reported to be as low as 60-70%. Progression to metastatic disease is the main cause of treatment failures. Broadening current knowledge on the pathogenesis and biology of metastatic OS tumors is a key element in improving treatment results, i.e. identifying potential therapeutic targets. Recent studies have brought new concepts into this field. This paper outlines the most important issues which may influence treatment methods in the near future. In a few sections, we discuss (1) a model of OS dissemination with special regard to proteins mediating the lysis of the extracellular matrix; (2) the mechanisms protecting circulating OS cells from programmed death; (3) the relationship between angiogenesis, its pathogenesis, and OS metastatic potential; (4) the role of cytokines in OS progression and site-specific metastasis formation; (5) an example of treatment resistance mechanism - the P glycoprotein efflux pump; and, finally, we theorize on (6) whether cancer stem cells may play a role in OS progression.