Immunoreactive Score Research Articles

Vesicular monoamine transporters expression in pheochromocytomas and paragangliomas according to scintigraphy and positron emission tomography behavior.

The expression of vesicular catecholamine transporters (VMAT1 and 2) in pheochromocytomas (PHEOs) and paragangliomas (PGLs) and the possible relationships with [18F]FDOPA PET/CT and [123I]MIBG scintigraphy uptake are unknown. Our purpose was to investigate possible correlations of either VMAT1 and VMAT2 expression with the functional imaging in patients with PHEOs and PGLs. An observational 3-year time study was performed by enrolling 31 consecutive patients with PHEO (N.=17) or PGL (N.=14). They underwent the same diagnostic work-up; moreover, [123I]MIBG SPECT/CT (N.=20) and [18F]FDOPA PET/CT (N.=14) were performed in a subset of patients. After surgery, routine histology and semiquantitative analysis of VMAT1/VMAT2 immunoreactivity were carried out in all cases. VMAT1 immunoreactivity was found in all tumors, but two PHEOs. VMAT1 immunoreactivity was higher in PGLs than in PHEOs, though at not significant extent. Elevated VMAT2 immunoreactivity score was present in all but two negative tumors. Normal [123I]-MIBG uptake was independent from VMAT1/2 immunoreactivity. Patients undergoing [18F]FDOPA PET/CT showed a high score level of both VMATs and were detected by the technique in all cases. VMAT1 and VMAT2 are highly expressed in most tumors, though VMAT1 immunoreactivity is apparently prevalent in PGLs as compared to PHEOs. Presence and expression of VMAT1 and VMAT2 are not limiting factors for MIBG uptake. The status of VMAT expression might help to understand why the more frequently used radiotracers do not always have the expected diagnostic performance. Finally, the present study points out the importance of developing new radiotracers with higher sensitivity, specificity and accuracy consequently reducing healthcare costs.

BCL3 expression is strongly associated with the occurrence of breast cancer relapse under tamoxifen treatment in a retrospective cohort study

Patients with estrogen receptor positive breast cancer are usually receiving an anti-estrogen therapy by either aromatase inhibitors or selective estrogen receptor mediators such as tamoxifen. Nevertheless, acquired resistance to tamoxifen under treatment frequently hampers therapy. One proposed explanation for this phenomenon is the interaction of the tumor cells with cells of the tumor microenvironment via the Insulin-like growth factor RNA binding protein 5/B-cell lymphoma 3 (IGFBP5/BCL3) axis. Here we investigated whether a high expression of BCL3 either cytoplasmic or nuclear is associated with the occurrence of a relapse under anti-estrogen therapy in patients. Formaldehyde-fixed, paraffin-embedded samples of 180 breast cancer patients were analyzed for BCL3 expression by immunohistochemistry. An immunoreactive score (IRS) was calculated from staining intensity in cytoplasm and nucleus as well as the percentage of positive tumor cells. These scores were correlated with clinico-pathological parameters using cross-tabulation analysis and patients’ relapse free and overall survival by Kaplan–Meier analysis and Cox regression. A tamoxifen-adapted MCF-7 derived cell line was investigated for BCL3 localization by immunofluorescence. The cytosolic BCL3-IRS significantly correlated with the proliferation marker Ki-67, and with the occurrence of a relapse under tamoxifen treatment. Nuclear score correlated only with tamoxifen-relapse. In survival analysis, both scores were highly significant prognostic factors for relapse free, but not for overall survival. This was especially obvious for estrogen receptor positive and HER2/NEU negative cases as well as lobular breast cancer. Tamoxifen-treated, but not aromatase-treated patients had a poor survival when BCL3 scores were high. A tamoxifen adapted cell line exhibited a reduced expression and mainly nuclear localization of BCL3, compared to the parental estrogen receptor positive cell-line MCF-7. Altogether, these data strongly support a function of BCL3 in tamoxifen resistance and its potential use as a predictive biomarker for tamoxifen resistance.

Frequent FGFR1 hotspot alterations in driver-unknown low-grade glioma and mixed neuronal-glial tumors

PurposeLow-grade gliomas (LGG) and mixed neuronal-glial tumors (MNGT) show frequent MAPK pathway alterations. Oncogenic fibroblast growth factor receptor 1 (FGFR1) tyrosinase kinase domain has been reported in brain tumors of various histologies. We sought to determine the frequency of FGFR1 hotspot mutations N546 and K656 in driver-unknown LGG/MNGT and examined FGFR1 immunohistochemistry as a potential tool to detect those alterations.MethodsWe analyzed 476 LGG/MNGT tumors for KIAA-1549-BRAF fusion, IDH1/2, TERT promotor, NF1, H3F3A and the remaining cases for FGFR1 mutation frequency and correlated FGFR1 immunohistochemistry in 106 cases.Results368 of 476 LGG/MNGT tumors contained non-FGFR1 alterations. We identified 9 FGFR1 p.N546K and 4 FGFR1 p.K656E mutations among the 108 remaining driver-unknown samples. Five tumors were classified as dysembryoplastic neuroepithelial tumor (DNT), 4 as pilocytic astrocytoma (PA) and 3 as rosette-forming glioneuronal tumor (RGNT). FGFR1 mutations were associated with oligodendroglia-like cells, but not with age or tumor location. FGFR1 immunohistochemical expression was observed in 92 cases. FGFR1 immunoreactivity score was higher in PA and DNT compared to diffuse astrocytoma, but no correlation between FGFR1 mutation in tumors and FGFR1 expression level was observed.ConclusionFGFR1 hotspot mutations are the fifth most prevailing alteration in LGG/MNGT. Performing FGFR1 sequencing analysis in driver-unknown low-grade brain tumors could yield up to 12% FGFR1 N546/K656 mutant cases.

Distribution of transient receptor potential vanilloid-1 channels in gastrointestinal tract of patients with morbid obesity.

BACKGROUNDTransient receptor potential vanilloid-1 (TRPV1), a nonselective cation channel, is activated by capsaicin, a pungent ingredient of hot pepper. Previous studies have suggested a link between obesity and capsaicin-associated pathways, and activation of TRPV1 may provide an alternative approach for obesity treatment. However, data on the TRPV1 distribution in human gastric mucosa are limited, and the degree of TRPV1 distribution in the gastric and duodenal mucosal cells of obese people in comparison with normal-weight individuals is unknown.AIMTo clarify gastric and duodenal mucosal expression of TRPV1 in humans and compare TRPV1 expression in obese and healthy individuals.METHODSForty-six patients with a body mass index (BMI) of > 40 kg/m2 and 20 patients with a BMI between 18-25 kg/m2 were included. Simultaneous biopsies from the fundus, antrum, and duodenum tissues were obtained from subjects between the ages of 18 and 65 who underwent esophagogastroduodenoscopy. Age, sex, history of alcohol and cigarette consumption, and past medical history regarding chronic diseases and medications were accessed from patient charts and were analyzed accordingly. Evaluation with anti-TRPV1 antibody was performed separately according to cell types in the fundus, antrum, and duodenum tissues using an immunoreactivity score. Data were analyzed using SPSS 17.0.RESULTSTRPV1 expression was higher in the stomach than in the duodenum and was predominantly found in parietal and chief cells of the fundus and mucous and foveolar cells of the antrum. Unlike foveolar cells in the antrum, TRPV1 was relatively low in foveolar cells in the fundus (4.92 ± 0.49 vs 0.48 ± 0.16, P < 0.01, Mann-Whitney U test). Additionally, the mucous cells in the duodenum also had low levels of TRPV1 compared to mucous cells in the antrum (1.33 ± 0.31 vs 2.95 ± 0.46, P < 0.01, Mann-Whitney U test). TRPV1 expression levels of different cell types in the fundus, antrum, and duodenum tissues of the morbidly obese group were similar to those of the control group. Staining with TRPV1 in fundus chief cells and antrum and duodenum mucous cells was higher in patients aged ≥ 45 years than in patients < 45 years (3.03 ± 0.42, 4.37 ± 0.76, 2.28 ± 0.55 vs 1.9 ± 0.46, 1.58 ± 0.44, 0.37 ± 0.18, P = 0.03, P < 0.01, P < 0.01, respectively, Mann-Whitney U test). The mean staining levels of TRPV1 in duodenal mucous cells in patients with diabetes and hypertension were higher than those in patients without diabetes and hypertension (diabetes: 2.11 ± 0.67 vs 1.02 ± 0.34, P = 0.04; hypertension: 2.42 ± 0.75 vs 1.02 ± 0.33, P < 0.01 Mann-Whitney U test).CONCLUSIONThe expression of TRPV1 is unchanged in the gastroduodenal mucosa of morbidly obese patients demonstrating that drugs targeting TRPV1 may be effective in these patients.

Nanog expression in patients with squamous cell carcinoma of oropharynx in relation to immunohistochemical score.

This study aimed to compare prognostic potential of Nanog expression analysed by three immunohistochemical scores in the group of 63 squamous cell carcinomas of oropharynx. Immunoreactivity of Nanog expression was analyzed by semiquantitative score, immunoreactive score and H-score. For all three scores, the cut-off points for Nanog overexpression and its lack, allowing for optimal separation of overall and disease free survival curves, were search by minimal p-value method. In semiquantitative score, the best separation of overall and disease free survival curves was obtain by cut-off point lack of staining vs. week/moderate/strong staining, although statistical significance was not reach (OS: HR = 1.016, p = 0.081, DFS: HR = 6.876, p = 0.061). The cut-off points for immunoreactive score and H-score were, respectively: 1 (OS: HR = 6.977, p = 0.014, DFS: HR = 6.002, p = 0.019) and 50 (OS: HR = 6.977, p = 0.014, DFS: HR = 6.002, p = 0.019). The cut-off points found for these two scores allow to identify the same subgroups of patients with lack of Nanog expression (11.1%) and its overexpression (88.9%). All patients with tumors characterized by lack of Nanog overexpression identifying by immunoreactive score and H-score survived 5 years without evidence of cancer progression. In multivariate analysis Nanog immunoreactivity analysed by QRS and IRS was independent prognostic factor for OS (HR = 10.195, p = 0.024). Immunohistochemical score using to distinguish Nanog overexpression or its lack has influence on prognostic potential of this biomarker.

Expression of Septin 2 and Her2/neu in Colorectal Cancer.

Colorectal cancer (CRC) is a common and lethal disease. Septin 2 belongs to the same class of GTPases as the RAS oncogenes influence the invasion and metastasis of many types of tumor cells. Furthermore, HER2/neu is involved in the tumor genesis and progression of various types of tumors. The role of both molecules is still questionable in CRC. The aim of the study is to examine the expression of septin 2 and Her2/neu in patients with CRC. The study was conducted on 2 groups; the first group consisted of 70 paraffin blocks for CRC patients and the second group was formed of 24 blocks from patients diagnosed as colorectal adenoma. For each adenoma and carcinoma case, a section was immunohistochemically stained using antihuman SEPT2 polyclonal antibody. For each carcinoma case, another section was immunostained using monoclonal anti-HER2/neu. The results were statistically analyzed and compared with the collected clinicopathologic data of the cases. For the carcinoma patients, there was a significant association between SEPT2 staining intensity and histologic type (P = 0.001) and grade (P < 0.001), tumor T (P = 0.001) and N (P = 0.011) stages and the presence of lymphovascular invasion (P < 0.001) and a significant association between Her2/neu immunoreactivity scores (IRSs) and histologic grade (P = 0.048), tumor T (P < 0.001) and N (P = 0.019) stages and the presence of perineural (P = 0.004) and lymphovascular (P = 0.003) invasion. In colonic adenoma patients, there was a significant relation between septin 2 IRSs and the grade of dysplasia in the adenoma (P < 0.001) and significant relation with its expression in carcinoma group (P < 0.001). A potential prognostic role of septin 2 and Her2/neu for patients with CRC is suggested as expression of both markers was associated with many important prognostic clinicopathologic variables in patients of CRC.

Do immunohistochemical studies have a role in predicting prognosis of laryngeal squamous cell carcinomas? CD44 and Fascin experience.

Background and objectives:The diagnosis of laryngeal squamous cell carcinoma (LSCC) can be made easily based on histopathological findings, but the relationship between morphological findings and prognosis is not clear. In addition to morphological findings, the use of novel markers may contribute to the development of new treatment strategies and improved patient prognosis. CD44, which is a cancer stem cell marker, and Fascin-1, an actin-binding protein has been associated with poor prognosis in many tumors. The aim of this study was to investigate the relationship between CD44 and Fascin-1 expression and clinicopathologic parameters in LSCC and their roles in the determination of clinical behavior and prognosis. The aim of this study is to investigate whether CD44 and Fascin have a relationship with clinicopathological parameters and have a role in determining clinical behavior and prognosis in LSCC.Methods:130 patients who were operated in our hospital for LSCC between 2012 and 2018 were included in this study. Fascin-1 and CD44 stains were applied immunohistochemically to the paraffin blocks of the tumors. Immunostained specimens were scored according to the intensity of staining and the percentage of staining for each marker. Overall scores were summed and was designated as immunoreactivity score (IRS). Finally, IRS was categorized into two groups; Low and High CD44/Fascin IRS.Results:There were no statistically significant differences between low and high CD44 and Fascin IRS groups in terms of clinicopathologic parameters, overall and disease-free survival (p> 0.05).Conclusion:Immunhistochemical studies are not yet sufficient to predict patient prognosis. Morphological findings still remain of priority and importance for pathologists. (www.actabiomedica.it)

Evaluation of CD9 Expression of Tumour Cells and Stromal Immune Cells in Breast Carcinoma by Immunohistochemistry

Introduction: Cluster of Differentiation 9 (CD9), known as Motility Related Protein (MRP-1) regulates cell adhesion, motility, migration and proliferation. Many studies have stated conflicting results on prognostic significance of invasive breast carcinomas with CD9 expression that had performed on tumour tissues. Aim: To assess the inter-relationship of CD9 expression of tumour cells and stromal immune cells in breast carcinoma with clinicopathological parameters which include age, tumour size, grade, histological type, lymph nodes, tumour staging and molecular classification. Materials and Methods: An observational prospective (2020 July to 2021 June) and retrospective (2019 October to 2020 June) study was done in 71 cases of resected primary invasive breast carcinoma over a period of one and half year at Sri Devaraj Urs Medical College, Karnataka, India. Immunohistochemistry (IHC) staining was done using CD9 antibody. Tumour cells (T-CD9) expression was evaluated by Immunoreactivity scoring (IS= Intensity score × Extent of staining). The stromal cells (S-CD9) expression was evaluated by Percentage (%) of stromal area occupied by CD9 stained immune cells. Chi-square test was used as test of significance for qualitative data. The p-value of &lt;0.05 was considered as statistically significant. Results: Out of 71 cases, T-CD9 expression was noticed in 40 cases (56.34%) and IS &lt;4 considered as negative was observed in 31 cases (43.66%). However there was no association with age, tumour size, grade and molecular markers for the expression of both T-CD9 and S-CD9. Human Epidermal growth factor receptor 2 neu (HER2neu) negative was associated with T-CD9 expression (p-value=0.05). Hence CD9 can be used as prognostic marker for Her2neu negative cases. Conclusion: CD9 expression was not significantly associated with tumour cells (T-CD9) and stromal cells (S-CD9) in breast carcinoma cases. However it was significantly associated with Her2neu negative tumour cells. T-CD9 showed more positivity in Luminal A followed by Triple negative whereas S-CD9 showed more positivity in Luminal B. CD9 did not show association with any parameters except Her2neu negative.

The Effect of Fasting during Pregnancy on Brain-derived Neurotrophic Factor Expression in Cerebrum and Cerebellum

Background: Fasting for pregnant women is still a debatable issue among society and clinicians. Fastingdone by pregnant women causes brain neurons to receive more energy thus increases the connectionbetween the neurons. A brain hormone called Brain-derived Neurotrophic Factor will be affectedduring fasting. Brain-derived neurotrophic factor (BDNF) plays an important role in neuronal survivaland growth, serves as a neurotransmitter modulator, and participates in neuronal plasticity, which isessential for learning and memory. Brain-derived Neurotrophic Factor (BDNF) has an important role inbrain development, namely in the formation of new nerve cells and cognitive processes inside the brainwhich helps to optimize cognitive, learning, and memory functions. Fasting during pregnancy willinfluence fetal brain cells due to increase in the body’s metabolic system which is delivered to cerebrumand cerebellum neuron cells. The objective of this research is to analyze the effect of fasting duringthree trimesters pregnancy on Brain-derived Neurotrophic (BDNF) factor expression in cerebrum andcerebellum.Method: The design of this research is true experimental laboratory post-test with control group design.The research samples are rats (Rattus norvegicus) which consists of 3 groups and 1 control group. Firstgroup (X1) was the rats born from mother with 2 days fasting during first trimester pregnancy, secondgroup (X2) was the rats born from mother with 2 days fasting during second trimester pregnancy andthird group (X3) was the rats born from mother with 2 days fasting during third trimester pregnancy.The control group (X0) was the rats born from mother without fasting during the whole pregnancy.Brain dissections of new born rats were taken and the brain-derived Neurotrophic Factor expressionwas studied within munohistochemistry.Conclusion: Brain-derived Neurotrophic Factor expression in cerebrumand cerebellumwas examinedwith immunohistochemical method. Data from each sample was assessed using the Remmele ScaleIndexmethod (Immunoreactive Score). There was significant cerebrum Brain-derived NeurotrophicFactor expression mean found in third trimester (3.90 2.403) with p-value = 0.008 (p&lt;0.05). Therewere no significant cerebellum Brain-derived Neurotrophic Factor expression differences found in alltrimesters. Fasting during pregnancy is safe for both mother and children. Fasting during pregnancywill increase the Brain-derived Neurotrophic Factor expression so that the brain function will improveas well.

Immunohistochemical evaluation of galectin-3 expression in oral squamous cell carcinoma, oral leukoplakia and normal mucosa.

Galectin 3 (Gal-3) has diverse functions critical in cancer biology including cell proliferation, apoptosis, evasion of immune responses and angiogenesis. The expression of Gal-3 is heterogeneous in normal and neoplastic tissues. In oral squamous cell carcinoma (OSCC) and oral leukoplakia (OL), both increased and decreased expressions of Gal-3 were elicited in numerous studies. To evaluate, compare and correlate the immunohistochemical expression of Gal-3 in OSCC, OL and normal oral mucosa. The study was conducted at the Department of Oral Pathology and Microbiology at PMS College of Dental Science and Research, Vattapara, Thiruvananthapuram. This is a retrospective analytical study. Clinically diagnosed and histopathologically confirmed cases of OSCC (n = 21), OL (n = 21), and normal oral mucosa (n = 21) were included in the study. Paraffin-embedded tissues were subjected to immunohistochemical analysis for Gal-3 expression. Gal-3 staining expression, staining distribution and cellular localisation were evaluated. All sampled categories were compared using immunohistochemical scoring analysis such as the H-score, labelling index (LI), immunoreactive score (IRS) and staining intensity (SI). The results were statistically analysed using multivariate analysis of variance (MANOVA) within and among the groups. The statistical inferences obtained found that the H-score could be used as a guideline for better differentiation between the groups and among the groups. The P value obtained was < 0.0125 and was found to be significant. The observation in our study shows that the immunohistochemical expression of Gal-3 gradually decreased from normal oral mucosa to OL to OSCC.

Glial Fibrillary Acid Protein Expression and Behavioral Changes in Hippocampus Following Prenatal Co-Administration of Ethyl Acetate Leaf Fraction of Tamarindus Indica and Aluminum Chloride in Wistar Rats

Background: The endowment of various plant parts with important phytochemicals needed in the management of human health breeds lots of hope. The present study investigated glial fibrillary acid protein (GFAP) expression and hippocampal behavioral changes following prenatal co-administration of ethyl acetate leaf fraction of Tamarindus indica (EATI) and aluminum chloride (AlCl3) in male Wistar rat pups. Methodology: Twenty pregnant Wistar rats were divided into five groups (n = 4). Group 1 received distilled water, while the treatment Groups 2 to5 received 200 mg/kg of AlCl3, followed by 400 and 800 mg/kg EATI in Groups 3 and 4, respectively, and 300 mg/kg Vitamin E in Group 5. All administrations lasted for 14 days from prenatal day 7 till parturition. The male pups (n = 6) were curled for Morris water maze (MWM) from postnatal day (PoND) 16–20, then sacrificed humanely on PoND 21. The brain tissues were harvested for oxidative stress studies (OSS) and the demonstration of GFAP antibody. Result: The result of the MWM showed significantly high mean latency to locate the platform in Groups 2 and 5 when compared to the control group (P &lt; 0.05). The OSS revealed significantly higher superoxide dismutase concentration in Groups 4 and 5 compared to that observed in Group 2. The GFAP studies revealed significantly higher immunoreactivity scores in Group 2 when compared to every other group (P &lt; 0.05) in the CA1 hippocampal region. Conclusion: EATI was associated with some protective potential during prenatal aluminum chloride exposure in Wistar rats.

Evaluation of MUC1, MUC2, MUC5AC, and MUC6 Expression Differences in Lung Adenocarcinoma Subtypes by Using a Final Immunoreactivity Score (FIRS).

Lung adenocarcinomas are divided into acinar, lepidic, papillary, micropapillary, and solid predominant subtypes according to the current World Health Organization (WHO) classification. We designed this retrospective study to demonstrate profiles of MUC expression (MUC1, MUC2, MUC5AC, and MUC6) of different histologic patterns within the same tumor among pulmonary adenocarcinomas and investigate correlations of MUC expression with clinicopathologic features. We analyzed the expression of mucins (MUC1, MUC2, MUC5AC, and MUC6) in a series of 99 resected lung adenocarcinomas, which included a total of 193 patterns (71 acinar, 30 lepidic, 25 papillary, 20 micropapillary, 34 solid and 13 mucinous) and calculated a final immune reactivity score (FIRS) per tumor. MUC1 IRS scores were significantly higher in lepidic and solid patterns compared with mucinous patterns (p=0.013). MUC2 expression was seen only in three cases (1 acinar, 2 mucinous). MUC5AC and MUC2 expression was more common in mucinous patterns (p < 0.001 and p=0.028, respectively). MUC6 expression was only detected in seven patterns and the expression was weak. No significant difference was seen among histologic patterns for the staining scores of MUC6. Mucinous adenocarcinoma differed from other histologic subtypes regarding MUC1 and MUC5AC expression. Mucinous adenocarcinoma showed less MUC1 expression with lower IRS scores and higher MUC5AC expression. Tumor size (p=0.006), lymphatic invasion (p=0.018), vascular invasion (p=0.025), perineural invasion (p=0.019), MUC1 IRS scores (p=0.018), and MUC1 IRS scores > 8.5 (p=0.018) were significant predictors for lymph node metastasis. An alternative scoring for MUC1 can be used as a predictor for lymph node metastasis regardless of the histologic subtype.

The Expression of Autophagy Markers Beclin1 and LC3B in Prostatic Carcinoma: Immunohistochemical Case-Control Study

Background & Objective:Prostatic carcinoma represents the second most common cancer diagnosed in men worldwide after lung cancer and the fourth common male malignancy in Egypt. Autophagy is a natural process that has both oncogenic and tumor-suppressive activities. This study aimed to evaluate the role of Beclin1 and LC3B in prostatic carcinoma. Methods: This retrospective case-control study was conducted on 110 prostate biopsies divided into three groups (55 prostatic carcinomas, 45 pure benign prostatic hyperplasias (BPH), and 10 BPH with adjacent prostatic carcinoma) retrieved from the archive of the Pathology Department, Faculty of Medicine, Menoufia University, in the period between 2017 and 2020. All biopsies were stained for Beclin1 and LC3B antibodies.Results:There was a highly significant association between higher Beclin1 and LC3B immunoreactivity score and Gleason score (score 8 and 9) (P=0.002 and 0.000, respectively). Moreover, there was a highly significant direct association between Beclin1 and LC3B expression (r=0.52, P=0.000). Also, there was a significant stepwise increase in Beclin1 positivity among the three studied groups starting from BPH to prostatic carcinoma passing through cases of BPH with neighboring tumor (P=0.000). Conclusion:From the results obtained in the present study, autophagy markers Beclin1 and LC3B showed upregulation in prostatic carcinoma. Moreover, both were associated with poor prognostic factors. So, it might be necessary to control autophagy flux in prostatic carcinoma. This might be one of the future therapeutic targets for the management of prostatic carcinoma.

Ephrin Receptors (Ephs) Expression in Thymic Epithelial Tumors: Prognostic Implications and Future Therapeutic Approaches.

Ephrin receptors (Ephs) are receptor tyrosine kinases (RTKs) implicated in tissue development and homeostasis, and they are aberrantly expressed in tumors. Here, immunohistochemical Eph type-A and -B expression in thymic epithelial tumors (TETs) was assessed and correlated with clinicopathological parameters. Tissue microarrays from 98 TETs were stained for EphA1, -A2, -A4 -A6, -B1, -B2, -B4 and -B6. The relationship between neoplastic and lymphoid cell immunoreactivity score (H-score), histopathological parameters (Pearson’s test) and survival of 35 patients (Mantel-Cox model) was explored. Epithelial-rich subtypes showed higher EphA6 cytoplasmic H-score (B2/B3, carcinoma) (p < 0.001) and stronger EphA4 H-score (B3, carcinoma) (p = 0.011). The immature T-cells, especially in subtypes AB/B1, had higher EphB6 H-score than carcinoma-associated mature lymphocytes (p < 0.001); carcinomas had higher lymphocytic EphB1 H-score (p = 0.026). Higher lymphocytic and lower epithelial EphB6 H-score correlated with Masaoka stage ≤II (p = 0.043, p = 0.010, respectively). All cases showed variable epithelial and lymphocytic EphA2 expression, but clinicopathological associations were not reached. Our study confirmed that Eph type-A and -B expression in TETs is associated with established prognostic parameters, i.e., tumor subtype and Masaoka stage, although correlation with patient survival was not reached. Such findings suggest involvement of these RTKs in thymic neoplasia, as well as their potential utility as treatment targets.

PD-1/PD-L1, Treg-related proteins, and tumour-infiltrating lymphocytes are associated with the development of oral squamous cell carcinoma

Cancer immunomodulation has been implicated in the development of oral squamous cell carcinoma (OSCC), however the role of specific immunomodulatory proteins is not completely understood, particularly in the early stages of the disease. Oral potentially malignant disorders such as leukoplakia commonly precede OSCC but not all will undergo malignant transformation. The aim of this study was to evaluate the diagnostic utility of specific immunomodulator proteins and their role in the progression of OSCC. Samples from 101 patients were included in the study. Cases were classified based on histopathology into four groups: non-dysplastic epithelial hyperplasia/keratosis, low-grade dysplasia, high-grade dysplasia, and OSCC. The PD-1/PD-L1 pathway, as well as regulatory T cell (Treg)-related proteins including FOXP3, TGF-β, IL-6, and IL-10 were immunohistochemically quantified. The number of tumour-infiltrating lymphocytes (TILs) was also assessed for each case. Multinominal regression models were undertaken to assess the significance of each protein in predicting the histopathological grade of oral epithelial disorders, and three diagnostic models were assessed. Significant positive associations were found between the immunoreactive score of each protein and the histopathological grade, p<0.05 suggesting that the PD-1/PD-L1 pathway, Treg-related proteins, and TILs are associated with the development of OSCC. Diagnostic models using the investigated proteins and TILs predicted the grade of oral epithelial disorder, p<0.05. The associated accuracy of this approach was 84.92%. Our findings support the notion that immunomodulation events may play a role in evading the immune system and contributing to potential malignant transformation of oral epithelial disorders. Our data also provide supporting evidence for the potential application of immune checkpoint inhibitors in the chemoprevention of OSCC. Further longitudinal studies to assess individual T-cell populations within the immune microenvironment of various oral potentially malignant disorders are warranted.

Association of endocan expression with clinicopathological prognostic parameters in breast carcinoma.

Breast cancer (BC) is a common malignancy in women. Some molecules, including endocan, are still under investigation as potential prognostic factors in BC. In the present study, we aimed to determine the relationship between endocan expression and clinicopathological prognostic parameters in BC. Two hundred and fifty-five patients diagnosed with BC were included in the present study. The immuno-reactivity scoring (IRS) system was used to reveal the tissue endocan expression levels. We found that endocan expression is associated with tumor necrosis, tumor size, and the presence of lobular carcinoma in situ (LCIS) in BC. There was no relationship between endocan expression and survival as well as other clinicopathological prognostic parameters. Endocan overexpression in BC is associated with some prognostic parameters, including tumor necrosis, tumor size, and the presence of LCIS. Further studies with larger series are needed to reveal the clinical and therapeutic implications of endocan in BC.

MTA1 Expression Can Stratify the Risk of Patients with Multifocal Non-Small Cell Lung Cancers ≤3 cm.

PurposeCurrently, there is no uniform standard to guide postoperative adjuvant chemotherapy for patients with multifocal non-small cell lung cancers (NSCLCs) ≤3 cm. Therefore, there is an urgent need to explore prognostic molecular markers to identify high-risk patients with multifocal NSCLCs ≤3 cm. We aimed to explore the potential value of metastasis-associated protein 1(MTA1) expression in risk stratification of patients with multifocal NSCLCs ≤3 cm.MethodsWe retrospectively analyzed the clinical data and postoperative survival data of patients with multifocal NSCLCs ≤3 cm. Paraffin-embedded tissue sections were used for immunohistochemistry. Semiquantitative immunoreactivity scoring (IRS) system was used to evaluate the nuclear expression of MTA1. SPSS software (version 23.0) was used to analyze the data.ResultsThe IRS of MTA1 nuclear expression in 259 lesions of 119 patients ranged from 2.2 to 11.7 (median: 5.6). Our results showed that MTA1 expression was highest in high-risk pathological subtypes of lung adenocarcinoma. MTA1 expression in multiple primary lung cancers (MPLCs) was lower than that in intrapulmonary metastases (IPMs). The median follow-up duration was 25.97 months. The disease-free survival (DFS) of patients with MPLCs was significantly better than that of patients with IPMs, and the DFS of patients with high MTA1 expression was significantly worse than that of patients with low MTA1 expression. Multivariate Cox analysis showed that high MTA1 expression (hazard ratio: 7.937, 95% confidence interval: 2.433–25.64, p =0.001) was a statistically significant predictor of worse DFS in patients with multifocal NSCLCs ≤3 cm.ConclusionMTA1 expression can stratify the risk in patients with multifocal NSCLCs ≤3 cm. Patients with MTA1 immunohistochemical score >5.6 are at a high risk of postoperative recurrence, and these patients may benefit from postoperative adjuvant chemotherapy.

The role and relationship between programmed death ligand 1 and cytotoxic T lymphocyte-associated antigen-4 immunohistochemical expression in colorectal carcinoma patients: an impact on outcome.

BackgroundGlobally, colorectal carcinoma (CRC) is the third most common cancer diagnosed in both men and women. Programmed death ligand 1 (PD-L1) and cytotoxic T lymphocyte-associated antigen-4 (CTLA-4) are immune checkpoints that induce tumour immune escape.AimThis study aimed to evaluate the immunohistochemical expression of PD-L1 and CTLA-4 in CRC and their relationship with clinicopathological parameters and survival data.ResultThis study included 103 CRC, 22 adenoma and 21 non-neoplastic specimens. High PD-L1 epithelial expression was in favour of CRC and high-grade dysplastic adenoma compared to normal specimens. High PD-L1 epithelial expression was associated with larger sized tumours, perforation, advanced T stage, infiltrative tumour border configuration (TBC), high tumour budding (TB) score, low tumour-stroma ratio (TSR) and absence of peritumoural lymphocytes. High PD-L1+ tumour infiltrating lymphocytes (TILs) showed an association with absence of perforation, early T stage, pushing TBC, lower TB score, high TSR and presence of peritumoural lymphocytes. High epithelial CTLA-4 expression was in favour of adenocarcinoma, high-grade dysplastic adenoma and low-grade dysplastic adenoma compared to normal specimens. High CTLA-4 epithelial score showed an association with positive lymph nodes (LNs), presence of an infiltrative TBC and absence of peritumoural lymphocytes. Low CTLA-4+ TILs showed a significant association with advanced tumour stage and increased number of positive LNs. Prolonged survival was associated with low epithelial PD-L1 and CTLA-4, high PD-L1+ TILs and high CTLA-4+ TILs. By multivariate Cox regression analysis, PD-L1+ TILs immunoreactivity score (p = 0.020) and CTLA-4+ TILs H. score (p = 0.036) were independent prognostic factors affecting overall survival among the other prognostic factors.ConclusionPD-L1 and CTLA-4 expression by tumour cells could cooperate with each other in enhancing progression of CRC leading to poor patient outcome, while their expression by TILs could stand against tumour progression.

The role of FGF-4 and FGFR-2 on preimplantation embryo development in experimental maternal diabetes

Objective Diabetes mellitus can cause spontaneous abortion, neonatal diseases, congenital malformations, and death. There are many studies related to the damage of in vitro hyperglycemia on embryogenesis in literature, but not enough studies on in vivo hyperglycemia effects on embryogenesis. Fibroblast growth factor (FGF) molecules play an essential role in pre-implantation embryo development and diabetes pathogenesis. In our study, we researched whether FGF-4 and FGFR-2 were playing a role in maternal diabetes' effects on embryo development. Material and Methods Thirty adult virgin female BALB/c mice were randomly divided into two groups: control and diabetic. The experimental diabetes model was generated by streptozotocin (55 mg/kg, once, intraperitoneally). The control and the diabetic group were mated. Embryos were collected at the morula and blastocyte stages corresponding to the third and fourth days of pregnancy. Embryo's FGF-4 and FGFR-2 molecules were evaluated by their immunofluorescence staining and immunoreactivity score. Result The results clearly showed that the FGF-4 and FGFR-2 immunofluorescence reactivity was higher in the diabetes group. Conclusion We concluded that FGF-4 and FGFR-2 overexpression might impair mouse pre-implantation embryo development in maternal diabetes and suggest investigating whether they have crucial effects on human embryo development and infertility in maternal diabetes.

Prognostic Significance of Estrogen Receptor Alpha in Oral Squamous Cell Carcinoma

Simple SummaryAlthough the survival rate has improved over the past decades, the prognosis of oral squamous cell carcinoma (OSCC) is still poor, and new treatment strategies are required. The aim of this study was to evaluate estrogen receptor alpha (ERα) expression in OSCC in a large patient cohort as a potential prognostic marker and therapeutic target. The findings indicated a rare expression of ERα that, however, was associated with a dramatic decrease of overall survival in male patients. In ERα-positive OSCC patients, an ER-based therapeutic (adjuvant) approach in the future might be conceivable based on the findings of this study.Introduction: Several studies suggest an estrogen receptor alpha (ERα)-mediated influence on the pathogenesis of oral squamous cell carcinoma (OSCC), as described for other malignancies that are not considered to be primarily hormone-dependent. Recently, an association between ERα expression and improved survival in oropharyngeal squamous cell carcinoma (OPSCC) has been found. However, the prognostic relevance of ERα in OSCC has not been proven to date. Therefore, the aim of this study was to evaluate ERα expression in OSCC in a large patient cohort and analyze its influence on survival and recurrence. Material and Methods: A total of 316 patients with primary OSCC who received initial surgical therapy were included in this analysis. The expression of ERα was evaluated on tissue microarrays by immunohistochemistry in the primary tumor and/or primary lymph node metastases. The expression level was quantified by light microscopy using the immunoreactive score (IRS) for estrogen receptor detection. An IRS equal to or greater than 2 was considered positive. The 5-year overall survival (OS) and relapse-free survival (RFS) were examined by the Kaplan–Meier method and log-rank test. Results: A total of 316 patients (111 females; 205 males) with a mean age of 61.3 years (range 27–96 years) were included in this study. In 16 patients (5.1%; 6 females and 10 males), positive ERα expression was found in the primary tumor (n = 11; 11/302) or lymph node metastases (n = 5; 5/52). Patients with positive ERα expression in primary tumors/primary lymph node metastases had a significantly lower OS and RFS (p = 0.012; p = 0.0053) compared to ERα-negative patients. Sub-group analysis in relation to gender revealed a highly significant influence of ERα expression on OS and RFS in males but not in females, both for the ERα-positive primary tumor cohort (males: p = 0.0013; p < 0.0001; females: p = 0.56; p = 0.89) and the ERα-positive primary tumor/primary lymph node metastasis cohort (males: p < 0.0001; p < 0.0001; females: p = 0.95; p = 0.96). In multivariate cox regression analysis, the ERα IRS of primary tumors (dichotomized; ERα+ vs. ERα−) was an independent risk factor for OS (HR = 4.230; 95%CI 1.616–11.076; p = 0.003) and RFS (HR = 12.390; 95%CI 4.073–37.693; p < 0.001) in the male cohort. There was a significant difference (p = 0.006) of ERα positivity with regard to the localization of the primary tumor. ERα positivity in the primary tumor was significantly associated (p = 0.026) with UICC stage, with most of the cases being diagnosed in stage IV. Furthermore, there was a significantly (p = 0.049) higher rate of bone infiltration in ERα-positive patients. Conclusion: Expression of ERα is rare in OSCC; however, it is associated with a dramatic decrease in OS in male patients. Further studies are necessary to confirm our results and to evaluate the exact mechanism underlying this observation. Hence, ERα-positive OSCC patients might benefit from an ER-based therapeutic (adjuvant) approach in the future.