Immunohistochemistry In Relation Research Articles

Focal adhesion proteins in hepatocellular carcinoma: RSU1 a novel tumour suppressor with prognostic significance

AimHepatocellular carcinoma (HCC) is a common cause a cancer-related death. Focal adhesions (FAs) represent multiprotein complexes at integrin-mediated cell-extracellular matrix adhesion sites that orchestrate vital cellular functions. The heterotrimeric ILK-PINCH-PARVB (IPP) complex, RSU1, a PINCH binding protein and CTEN, a member of the tensin family of proteins exert a critical role in FAs, where they regulate important cancer related functions such as cell adhesion, migration, proliferation and survival. Previous studies implicate these FA proteins in liver pathophysiology but their detailed role in human HCC is not fully understood. Here in we investigated expression and function of IPP, RSU1 and CTEN in human HCC. MethodsThe expression of focal adhesion proteins was studied in human HCC by immunohistochemistry in relation to clinicopathological parameters, previous studied genomic instability markers and patient’s survival. Effects on cell proliferation and FA proteins expression upon ILK inhibition and RSU1 silencing were also investigated in HCC in vitro. ResultsIPP complex and CTEN proteins are overexpressed while RSU1 expression is decreased in human HCC. CTEN expression correlates with reduced patients’ survival while RSU1 represents an independent favorable prognostic indicator in human HCC. Nuclear ILK expression correlates with markers of genomic instability. Pharmacological targeting of ILK suppresses, while RSU1 silencing promotes cell growth of HCC cells in vitro, while in both experimental conditions expression and/or localization of focal adhesion proteins is deregulated. ConclusionOur results suggest that FA signaling is implicated in hepatocellular carcinogenesis with prognostic significance. RSU1 seems to exert tumor suppressive functions in HCC and represents a novel favorable prognostic indicator.

Expression of adhesion molecules CD44 and epithelial cadherin in epithelial ovarian cancer and relation to clinicopathological parameters

Background Ovarian cancer is the most lethal gynecological malignancy. Most cases are diagnosed in advanced metastatic stage. Alteration of cellular-adhesion molecules is involved in invasion and metastasis of ovarian cancer. Patients and methods In 56 patients with epithelial ovarian cancer, CD44 and E-cadherin expression was determined by immunohistochemistry in relation to clinicopathological characteristics. Results CD44 expression was significantly related to high tumor grade (P=0.003), advanced tumor stage (P=0.011), and the presence of omental metastasis (P=0.004). Epithelial cadherin expression was inversely related to tumor grade (P=0.044) and omental metastasis (P=0.020). Among CD44/E-cadherin expression groups, the CD44high/E-cadherinlow group was significantly related to high tumor grade, advanced stage, omental metastasis, capsular invasion, and vascular invasion. Conclusion Since the CD44high/E-cadherinlow group was related to the features of aggressive tumor behavior, we speculate that assessment of the adhesion molecules CD44 and epithelial cadherin in epithelial ovarian cancer may be of considerable prognostic value and such markers can be promising therapeutic targets.

Study on the correlation between the expression of basic transcription factor 3 (BTF3) and the efficacy and prognosis of patients with malignant melanoma (MM).

e21554 Background: BTF3 is an RNA polymerase II transcription factor, studies have confirmed that BTF3 is abnormally expressed in several types of tumors and closely related to the proliferation and prognosis of gastrointestinal tumors. However, the function of BTF3 in MM remains unknown. The present study aim to detect the expression and proliferation of BTF3，to explore the correlation between the BTF3 expression and the efficacy and prognosis of MM patients. Methods: We explored the effect of BTF3 through silencing BTF3 expression by siRNA transfection with lentivirus in human melanoma cell lines. The cell proliferation, cell cycle and apoptosis were determined by methyl-thaizolyl-tetrazolium assay, flow cytometry and western blot, respectively. Then we investigated the effect of BTF3 in nude mice by tumor formation experiment. The expression of BTF3 was determined by immunohistochemistry (IHC) from 32 patients with different subtypes of MM during October 2014 to October 2020. Among them, 28 cases could be resected (stage I-III), 4 cases could not be resected (stage III-IV). Cutaneous MM 7 cases, Acral MM 14 cases, mucosal MM 9 cases, unknown primary MM 2 cases. The correlation of BTF3 expression and RFS (recurrence-free survival) /OS (overall survival) in 28 patients with resected MM was explored by Student’s t test. Results: The increased viability of negative control virus infection group was 4.706 times than that of BTF3-siRNA group. Compared with the former, BTF3-siRNA group had more cells in G1 and S phase (P< 0.05) and more apoptotic cells (P< 0.05). BTF3-siRNA reduced tumor formation in the nude mice (P<0.01). The relative IHC scores of BTF3 was 5.50±3.19 in tumor tissues (the higher the score, the higher the expression of BTF3), 6.52±4.13 in adjacent noncancerous tissues. The expression of BTF3 in different subtypes of MM tissues are shown in the table. In the median follow-up time of 27 months (6-68 months), 13 cases survived without recurrence or metastasis, 10 cases survived with recurrence or metastasis, 8 cases died. In 28 resectable MM patients, the median RFS/OS of high BTF3 expression (IHC score>5) and low expression (IHC score:0-4) were 18.57 / 24.62 months and 20.42 / 22.78 months. The corresponding RFS/OS P values of patients with high and low expression were 0.323 and 0.607. Conclusions: The study suggests that BTF3 play a role in the proliferation, cell cycle regulation, apoptosis and tumorigenicity of MM. Preliminary small-sample retrospective clinical study did not suggest that BTF3 was related to the efficacy and prognosis of patients, which needs to be further verified by large-sample prospective studies.[Table: see text]

High expression of Ras-specific guanine nucleotide-releasing factor 2 (RasGRF2) in lung adenocarcinoma is associated with tumor invasion and poor prognosis.

The expression of Ras‐specific guanine nucleotide‐releasing factor 2 (RasGRF2) in lung adenocarcinomas was examined using immunohistochemistry in relation to clinicopathological characteristics and prognosis. In comparison to low expression, high expression of RasGRF2 was more closely associated with poor prognosis. Interestingly, expression of phosphorylated epithelial cell transforming 2 (pECT2), which – like RasGRF2 – is also a guanine‐nucleotide exchange factor, was also associated with prognosis, and patients with high expression of both RasGRF2 and pECT2 had a much poorer outcome than those who were negative for both.

The simultaneous analysis of mesenchymal stem cells and early osteocytes accumulation in osteoarthritic femoral head sclerotic bone.

ObjectiveOA subchondral bone is a key target for therapy development. Osteocytes, the most abundant bone cell, critically regulate bone formation and resorption. Their progenitors, mesenchymal stem cells (MSCs), display altered behaviour in osteoarthritic subchondral bone. This study investigated the relationships between native osteocytes and native MSCs in osteoarthritic femoral heads.MethodsTo avoid culture manipulations, a bone treatment procedure was developed to simultaneously obtain pure osteocyte-enriched fragments and matched native CD45-CD271+ MSCs. Gene expression in osteocytes and MSCs was compared between healthy and OA bone and selected molecules were examined by immunohistochemistry in relation to OA tissue pathology. Cell sorting and standard trilineage differentiation assays were employed to test OA MSC functionality.ResultsNative osteocyte enrichment was confirmed histologically and by higher-level osteocyte maturation transcripts expression, compared with purified MSCs. Compared with healthy bone, native OA osteocytes expressed 9- and 4-fold more early/embedding osteocyte molecules E11 and MMP14, and 6-fold more osteoprotegerin (P<0.01). CD271+ MSCs accumulated in the regions of bone sclerosis (9-fold, P<0.0001) in close juxtaposition to trabeculae densely populated with morphologically immature E11-positive osteocytes (medians of 76% vs 15% in non-sclerotic areas, P<0.0001), and osteoblasts. Gene expression of OA MSCs indicated their bone formation bias, with retained multipotentiality following culture-expansion.ConclusionsIn human late-stage OA, osteogenically-committed MSCs and adjacent immature osteocytes exhibit a marked accumulation in sclerotic areas. This hitherto unappreciated MSC-early osteocyte axis could be key to understanding bone abnormalities in OA and represents a potential target for novel therapy development in early disease.

Prolactin-induced and neuronal activation in the brain of mother mice.

Nursing has important consequences on mothers. To separate the prolactin-mediated and the neuronally-mediated actions of nursing, neurons directly affected by prolactin were visualized using pSTAT5 immunohistochemistry in relation to Fos-expressing neurons in suckled mother mice. In response to pup exposure following 22-h pup deprivation, we found a markedly elevated number of pSTAT5-containing neurons in several brain regions, including the lateral septum, medial amygdaloid nucleus, subparafascicular area, caudal periaqueductal gray, dorsal raphe, lateral parabrachial nucleus, nucleus of the solitary tract, and the periventricular, medial preoptic, paraventricular, arcuate and ventromedial nuclei of the hypothalamus. Pup exposure also induced Fos expression in all of these brain regions except the arcuate and ventromedial hypothalamic nuclei. Bromocriptine treatment known to reduce prolactin levels eliminated pSTAT5 from most brain regions while it did not affect Fos activation following suckling. The degree of colocalization for pSTAT5 and Fos ranged from 8 to 80% in the different brain regions suggesting that most neurons responding to pup exposure in mother mice are driven either by prolactin or direct neuronal input from the pups, while the number of neurons affected by both types of inputs depends on the examined brain area. In addition, both pSTAT5 and Fos were also double-labeled with estrogen receptor alpha (ERα) in mother mice, which revealed a very high degree of colocalization between pSTAT5 and ERα with much less potential interaction between Fos- and ERα-containing neurons suggesting that estrogen-sensitive neurons are more likely to be affected by prolactin than by direct neuronal activation.

Asparaginase-like protein 1 is an independent prognostic marker in primary endometrial cancer, and is frequently lost in metastatic lesions

ObjectiveLoss of Asparaginase-like protein 1 (ASRGL1) has been suggested as a prognostic biomarker in endometrial carcinoma. Our objective was to validate this in a prospectively collected, independent patient cohort, and evaluate ASRGL1 expression in endometrial carcinoma precursor lesion and metastases. Methods782 primary endometrial carcinomas, 90 precursor lesions (complex atypical hyperplasia), and 179 metastases (from 87 patients) were evaluated for ASRGL1 expression by immunohistochemistry in relation to clinical and histopathological data. ASRGL1 mRNA level was investigated in 237 primary tumors and related to survival and ASRGL1 protein expression. ResultsLow expression of ASRGL1 protein and ASRGL1 mRNA predicted poor disease specific survival (P<0.001). In multivariate survival analyses ASRGL1 had independent prognostic value both in the whole patient cohort (Hazard ratio (HR): 1.53, 95% confidence interval (CI): 1.04–2.26, P=0.031) and within the endometrioid subgroup (HR: 2.64, CI: 1.47–4.74, P=0.001). Low ASRGL1 expression was less frequent in patients with low grade endometrioid primary tumors compared to high grade endometrioid and non-endometrioid primary tumors, and ASRGL1 was lost in the majority of metastatic lesions. ConclusionsIn a prospective setting ASRGL1 validates as a strong prognostic biomarker in endometrial carcinoma. Loss of ASRGL1 is associated with aggressive disease and poor survival, and is demonstrated for the first time to have independent prognostic value in the entire endometrial carcinoma patient population.

STAT6 Reliably Distinguishes Solitary Fibrous Tumors from Myofibromas.

Solitary fibrous tumors (SFT) and myofibromas (MF) historically have belonged to the same morphologic spectrum and have been lumped together under the nonspecific umbrella term, "hemangiopericytoma" along with other pericytic/myoid tumors. While current evidence shows clear distinction between the two entities, they frequently remain in the same histopathologic differential diagnosis. This diagnostic dilemma especially is common for smaller incisional biopsies from the oral cavity. STAT6 immunohistochemistry (IHC) recently was established as a reliable method to detect solitary fibrous tumor; however, the literature is sparse regarding STAT6 reactivity in MFs. The authors report ten new cases of oral solitary fibrous tumor, discuss histopathologic similarities and differences between the two tumors, and list respective STAT6 IHC expressivity. After IRB approval, 10 cases diagnosed as SFT and 24 cases of MF were collected from the University of Florida Oral and Maxillofacial Pathology Biopsy Service between the years 1994 and 2016. The original hematoxylin and eosin slides and related IHC were reviewed. IHC with STAT6 antibody was performed on all 34 samples, and the findings were analyzed. All cases were from the oral cavity or perioral regions. 10/10 SFTs expressed STAT6 nuclear reactivity, while no cases of MF showed nuclear expression of STAT6. STAT6 is a dependable marker to differentiate SFTs from MFs.

Abstract 4679: Mismatch repair defects in lethal prostate cancer

Abstract INTRODUCTION: A subset of prostate cancers (PCs) has a hypermutated phenotype. We hypothesized that lethal PCs with a high mutational load (ML) have DNA repair defects including mismatch-repair (MMR) aberrations and that such aberrations may impact outcome. METHODS: Diagnostic and metastatic castration resistant prostate cancer (mCRPC) tissues were tested by A) immunohistochemistry (IHC) for MSH2, MSH6, MLH1, and PMS2; B) microsatellite instability (MSI) (Promega v1.2 assay); C) targeted next generation sequencing (NGS) of the coding regions of 113 genes, a panel enriched for DNA-repair genes that includes MMR genes. Utilizing NGS data we also evaluated ML and MSI by NGS (MSINGS; Pritchard et al, 2014). IHC was used as reference for sensitivity (SE) and specificity (SP) of methods B) and C).Youden’s index was used to determine the optimal cut off for the ROC for IHC in relation to MSINGS and ML. Demographics and clinical data were retrospectively collected from electronic records. Relationship between ML in diagnostic and mCRPC samples was evaluated by paired t-test. The relationship between ML and MMR deficient tumors on IHC (MMRd) and/or MSI tumors by PCR (MMRd/MSI) was analyzed by negative binomial regression model. The relationship between MMRd/MSI tumors and overall survival (OS) was analyzed using univariate and multivariate Cox regression, adjusting for radical treatment (prostatectomy or radiotherapy), Gleason score, age, PSA and stage and presence of metastatic disease at diagnosis. RESULTS: We analyzed 306 PC biopsies (180 hormone-sensitive [HS] and 126 castration resistant [CR]) from 208 patients. Overall 16 patients (7.7%) had either MMRd or MSI. Matched, same-patient, HSPC and mCRPC tumor biopsies were available for 82 patients. Only one case with MMRd status in HS was not confirmed in matched mCRPC. This patient was considered MMR proficient for the survival analysis. MSI had a SE of 80% and a SP of 97%. An MSI by NGS cut-off of 0.096 had SE of 60% and SP of 99%, with an area under ROC curve of 0.83. A ML cut-off of 11 mutations per panel had SE of 60% and SP of 99%, with an area under ROC curve of 0.79. Although ML in diagnostic samples correlates with ML in mCRPC, ML was significantly higher in mCRPC MMRd/MSI cases (p &amp;lt;0.001; predicted ML of 14 versus 4 mutations per panel). MMRd/MSI and MMR proficient patients were balanced for type of radical treatments, Gleason score, presence of metastatic disease, PSA, age and stage at diagnosis. Despite this, median OS from start of LHRH for the MMRd/MSI group was significantly shorter than in the MMR proficient group in univariable and multivariable analysis (3.84 vs 6.81 years; aHR: 2.44; 95% CI 1.27 -4.70; p = 0.008). CONCLUSION: We provide evidence that MMRd/MSI status represents a negative prognostic factor for survival for metastatic CRPC. MMRd/MSI status in diagnostic biopsies accurately identifies MMRd/MSI in mCRPC. MSINGS and ML can detect MMRd/MSI tumors. Citation Format: Pasquale Rescigno, Daniel Nava Rodrigues, Wei Yuan, Suzanne Carreira, Maryou Lambros, George Seed, Ruth Riisnaes, Susana Miranda, David Dolling, Matthew Clarke, Mateus Crespo, Claudia Bertan, Gunther Boysen, Joaquin Mateo, Ana Ferreira, Adam Sharp, Ines Figueiredo, Semini Sumanasuriya, Mariane Sousa Fontes, Diletta Bianchini, Zafeiris Zafeiriou, Johann Sebastian de Bono. Mismatch repair defects in lethal prostate cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 4679. doi:10.1158/1538-7445.AM2017-4679

Immunohistochemical expression of interferon-γ in different types of granulomatous lesions associated with bovine paratuberculosis.

Animals infected with Mycobacterium avium subsp. paratuberculosis (Map) show a variety of lesions, from focal forms, seen in subclinical stages to diffuse lesions in clinical cases. The purpose of this study was to evaluate the local expression of IFN-γ by immunohistochemistry in relation with the type of lesion in naturally Map-infected cows. The number of immunolabelled cells, -the majority morphologically consistent with lymphocytes-, was higher in focal and diffuse paucibacillary forms than in diffuse multibacillary lesions, where they appeared closely related to epithelioid cells. Diffuse multibacillary lesions had the lowest numbers, but higher than controls, and positive cells were intermingled among the macrophages. The peripheral IFN-γ production was higher in all Map infected cows and a positive correlation was found with the number of immunolabelled cells in the intestine. The findings of this study show that IFN-γ would play a role in the development of the different types of lesions in paratuberculosis, and also points out the importance of adequate sampling of lymphoid tissue containing samples when studying the local immune response in which IFN-γ expression may be involved, especially in cases where focal lesions are present.

Analysis of epithelial-cadherin and human epidermal growth factor receptor 2/ expression in gastric carcinoma using immunohistochemistry.

Gastric adenocarcinoma (GAC) is a common malignancy with high mortality-rate. Analysis of molecular markers could form a foundation for the future use of targeted therapies to reduce morbidity and mortality. To find the prevalence and relation of epithelial cadherin (E-cadherin) and human epidermal growth factor receptor 2 (HER-2/neu) protein expression with histological type and grade of GAC using immunohistochemistry (IHC). A total of 100 cases of GAC diagnosed over a 2 year period were studied. Expression of E-cadherin and HER-2/neu was analyzed by IHC in relation to the histological type and grade. Of the 100 cases of GAC studied, 11 revealed a loss of E-cadherin and over-expression of HER-2/neu was seen in 17 cases. Loss of E-cadherin was seen in 50% of signet ring-cell carcinomas but only in 8% of tubular and none of papillary and mucin-secreting GAC (P = 0.003). Of all the cases of tubular GAC with loss of E-cadherin expression, majority (71.4%) were Grade III (P = 0.04). Of all the tubular GAC cases with an over-expression of HER-2/neu, 20% and 67% were Grade I and II GAC respectively while only 13% were Grade III (P < 0.001). Although poorly-differentiated tumors show loss of E-cadherin, better-differentiated tumors over-express HER-2/neu protein. Signet ring-cell carcinoma is more likely to exhibit a loss of E-cadherin protein. Targeted therapy toward HER-2/neu in GAC should be considered. Novel therapy to block E-cadherin down-regulation is justified.

Abstract P3-05-15: TP53 mutation and nestin protein expression patterns affect the outcome of patients with triple negative breast cancer (TNBC)

Abstract Background-aim: Nestin has recently been proposed as an ideal positive biomarker for calling basal-like breast cancer (BL). BL and TNBC overlap in 70-80% of cases and, although very heterogeneous at the genomic and phenotypic levels, most of these carry TP53 mutations. Herein, we investigated the effect of TP53 mutations and Nestin expression on the outcome of early high-risk TNBC patients who had received adjuvant chemotherapy (anthracyclines and/or taxanes). Methods: TP53 mutations were assessed in 190 FFPE TNBC DNA samples with massive parallel sequencing (entire coding region) in Ion Torrent PI chips. Variant calling and annotation (Ion Reporter 1.6) was followed by stringent read quality filtering (p&amp;lt;0.001). Deleterious and hot-spot mutations (Ingenuity &amp; Oncomine databases) in 183 informative tumors were accepted for analysis. Nestin protein expression was examined by immunohistochemistry (IHC) in 283 centrally assessed TNBC on tissue microarrays; positivity was set at 1% of cells regardless of staining intensity. Further BL and TNBC related IHC markers were also examined. Results: Nestin was expressed in 134/283 tumors (47.3%), more frequently in BL-TNBC (CK5+ and/or EGFR+, Fisher's exact p&amp;lt;0.001) and in highly proliferative tumors (p&amp;lt;0.001), while it was positively associated with high tumor grade (p = 0.017) and with P-cadherin IHC (p = 0.001). TP53 mutations were observed in 141/183 tumors (77%), out of which 30 carried truncating TP53 mutations (21.3%) and 59 double damaging mutations (41.8%) usually at similar coverage; among these, truncating mutations occurred in 16 cases. TP53 mutations were more common in BL-TNBC (p = 0.029). Double mutations were positively associated with E- and P-cadherin protein expression (p = 0.016 and p = 0.020, respectively) Although TP53 mutations in general and the presence of double mutations were not associated with p53 IHC, tumors with truncating mutations were almost exclusively p53 IHC negative (p&amp;lt;0.001) in line with the detection capacity of the employed antibody. As single markers, neither Nestin nor TP53 mutations (any pattern) had significant effects on patient outcome. When combined, a DFS-related interaction between Nestin and double TP53 mutations was observed (Wald's p = 0.016). Nestin positive tumors with double TP53 mutations (n = 22) conferred median DFS of 45 mo. This interval was significantly shorter as compared to that of patients with Nestin positive tumors without double TP53 mutations (n = 62; median DFS 56 mo; HR 2.2; 95%CI 1-4.5; p = 0.045) and Nestin negative tumors with double TP53 mutations (n = 25) that were associated with the most favorable DFS (median: 66 mo; HR 4; 95%CI 1.3-12.4; p = 0.016). Nestin negative tumors without double TP53 mutations performed similarly to both positive tumors, although not statistically significant. Conclusions: The presence of double TP53 mutations in TNBC may reflect the genetic instability and heterogeneity characterizing these tumors. This feature in combination with the putative cancer stem cell and BL marker Nestin seems to aggravate the outcome of TNBC patients treated with standard adjuvant chemotherapy regimens. These novel findings might be worthy pursuing for validation in larger patient series. Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr P3-05-15.

Evaluation of the combined expression of chemokine SDF-1α and its receptor CXCR4 as a prognostic marker for gastric cancer

Chemokine stromal cell-derived factor (SDF)-1α and its receptor CXC chemokine receptor 4 (CXCR4) have been shown to impact cancer progression. Accumulating evidence suggests that CXCR4 and SDF-1α expression is useful for evaluating the risk of gastric cancer progression. Thus, combined analysis of SDF-1α and CXCR4 should have high prognostic potential as a molecular marker for gastric cancer. We investigated the expression of SDF-1α and CXCR4 using immunohistochemistry in relation to prognosis, clinicopathological features and clinical outcomes in 221 cases of primary gastric cancer. Patients were categorized into three groups according to CXCR4 and SDF-1α expression: high CXCR4/high SDF-1α, low CXCR4/low SDF-1α, and high CXCR4/low SDF-1α - low CXCR4/high SDF-1α. No significant differences were noted in age, gender, histology, tumor location, lymphovascular invasion or proportion of tumor size >5 cm among the three groups. However, high CXCR4/high SDF-1α expression in tumor cells was significantly associated with depth of invasion of the tumor, lymph node involvement, and higher tumor stage compared to tumors with low CXCR4/low SDF-1α expression or high CXCR4/low SDF-1α - low CXCR4/high SDF-1α expression. Furthermore, patients with high CXCR4/high SDF-1α expression had the worst patient prognosis, whereas patients who had low CXCR4/low SDF-1α expression showed the most favorable prognosis. In conclusion, CXCR4 and SDF-1α are useful prognostic factors in gastric cancer, and the combination of high CXCR4 protein expression with high SDF-1α expression suggests a dismal prognosis.

Pyelonephritis in slaughter pigs and sows: Morphological characterization and aspects of pathogenesis and aetiology

BackgroundPyelonephritis is a serious disease in pig production that needs to be further studied. The purpose of this study was to describe the morphology, investigate the pathogenesis, and evaluate the aetiological role of Escherichia coli in pyelonephritis in slaughtered pigs by concurrent bacteriological, gross and histopathological examinations.MethodsFrom Danish abattoirs, kidneys and corresponding lymph nodes from 22 slaughtered finishing pigs and 26 slaughtered sows with pyelonephritis were collected and evaluated by bacteriology and pathology. Based on gross lesions, each kidney (lesion) was grouped as acute, chronic, chronic active, or normal and their histological inflammatory stage was determined as normal (0), acute (1), sub-acute (2), chronic active (3), or chronic (4). Immunohistochemical identification of neutrophils, macrophages, T-lymphocytes, B-lymphocytes, plasma cells, E. coli and Tamm-Horsfall protein (THP) in renal sections was performed. The number of E. coli and the proportion of immunohistochemically visualized leukocytes out of the total number of infiltrating leukocytes were scored semi-quantitatively.ResultsLesions in finishing pigs and sows were similar. Macroscopically, multiple unevenly distributed foci of inflammation mostly affecting the renal poles were observed. Histologically, tubulointerstitial infiltration with neutrophils and mononuclear cells and tubular destruction was the main findings. The significant highest scores of L1 antigen+ neutrophils were in inflammatory stage 1 while the significant highest scores of CD79αcy+ B-lymphocytes, IgG+ and IgA+ plasma cells were in stage 3 or 4. Neutrophils were the dominant leukocytes in stage 1 while CD3ε+ T-lymphocytes dominated in stage 2, 3 and 4. Interstitially THP was seen in 82% and 98% of kidneys with pyelonephritis from finishing pigs and sows, respectively. E. coli was demonstrated in monoculture and/or identified by immunohistochemistry in relation to inflammation in four kidneys from finishing pigs and in 34 kidneys from sows.ConclusionsE. coli played a significant role in the aetiology of pyelonephritis. Neutrophils were involved in the first line of defence. CD3ε+ T-lymphocytes were involved in both the acute and chronic inflammatory response while a humoral immune response was most pronounced in later inflammatory stages. The observed renal lesions correspond with an ascending bacterial infection with presence of intra-renal reflux.

Vascular endothelial growth factor A is a putative paracrine regulator in seasonally controlled spermatogenesis: Insights from a ruminant model, the roe deer

Vascular endothelial growth factor A (VEGFA) influences spermatogenesis, but its impact on seasonally regulated sperm production is still not fully understood. Thus, we investigated both expression levels and localisation of VEGFA and its receptors VEGFR1 and 2 in roe buck testis via real-time reverse transcription polymerase chain reaction and immunohistochemistry in relation to seasonal changes in the cellular composition of the testis. VEGFA was expressed by interstitial cells while its receptors were found on endothelial and perivascular cells. Inside the tubules, VEGFA was located in spermatogonia and spermatocytes, VEGFR1 was present on elongating spermatids and VEGFR2 on Sertoli cells. VEGFR1 mRNA was expressed tenfold lower than VEGFR2 and VEGF mRNAs. Relative VEGF and VEGFR2 expression (divided by the number of VEGFA and VEGFR2 expressing cells) showed an increase towards the rut (July/August) and a decrease thereafter. The results suggest involvement of VEGFA in the adjustment of vascular permeability as well as in spermiogenesis and the proliferation of spermatogonia.

Zone‐specific remodeling of tumor blood vessels affects tumor growth

Chaotic organization, abnormal leakiness, and structural instability are characteristics of tumor vessels. However, morphologic events of vascular remodeling in relation to tumor growth are not sufficiently studied yet. By using the rat rhabdomyosarcoma tumor model vascular morphogenesis was studied by light and electron microscopy and immunohistochemistry in relation to tumor regions such as tumor surrounding (TSZ), marginal (TMZ), intermediate (TIZ), and center (TCZ) zones. The analyses revealed that blood vessels of TSZ display a regular ultrastructure, whereas blood vessels of TMZ showed a chaotic organization and unstable structure with a diffuse or even lacking basal lamina, and missing or irregular assembled periendothelial cells. In contrast, blood vessels of TIZ and TCZ exhibited a more or less stabilized vessel structure with increased diameter. Correspondingly, normal assembly of alpha-smooth-muscle-actin (alpha-SMA)-positive cells into the vessel wall was observed in blood vessels of TSZ, TIZ, and TCZ. Also, Ang1 immunostaining was strongest in large vessels of TIZ and TCZ, whereas Ang2 staining was prominent in small vessels of TIZ. Tie2 staining was detectable in small and large vessels of all tumor zones. Immunostaining for alpha(v)beta(3)-integrin was strongest in small vessels of TMZ, whereas large vessels of TIZ and TCZ were almost negative. The results indicate a zone-specific remodeling of tumor blood vessels by stabilization of vessels in TIZ and TCZ, whereas small vessels of these zones obviously undergo regression leading to tumor necrosis. Thus, a better understanding of vascular remodeling and stabilization in tumors would enable new strategies in tumor therapy and imaging.

Clinicopathological characteristics, microsatellite instability, and expression of mucin core proteins and p53 in colorectal mucinous adenocarcinomas in relation to location

It has been suggested that right-sided and left-sided colorectal cancer may arise by different mechanisms. However, there have been few studies of mucinous adenocarcinoma (MA) in relation to location. Therefore, we analyzed clinicopathological characteristics, microsatellite instability (MSI), and expression of MUC1, MUC2, MUC5AC mucin core proteins, and p53 by immunohistochemistry in relation to tumor location. Ninety-six consecutive colorectal MAs and ninety-eight nonmucinous adenocarcinomas (nMAs) were investigated. Right-sided MAs, by comparison with those on the left side, were characterized by older age, larger tumor size, lower stage at presentation, peritumoral lymphocytic response, background of serrated adenoma, MSI-H phenotype, higher MUC2 and MUC5AC expression, and lower p53 protein overexpression. Right-sided nMAs, relative to those on the left side, were associated with MSI-H phenotype, higher MUC2 and MUC5AC expression, and lower p53 protein overexpression. Thus, MSI-H phenotype, expression of MUC2 and MUC5AC, and infrequent p53 protein overexpression are associated with right-sided location as well as mucinous histology. In univariate analysis, right-sided location had a favorable effect on disease specific survival of the patients with MA, although it is not an independent predictor of survival. Our results indicate that MA is a distinctive form of colorectal cancer and has different phenotypes depending on tumor location.

Evaluation of cell cycle and intestinal metaplasia related immunohistochemistry data in esophageal biopsy specimen by optical, virtual microscopy and digital slide based automated counting methods

Evaluation of cell cycle and intestinal metaplasia related immunohistochemistry data in esophageal biopsy specimen by optical, virtual microscopy and digital slide based automated counting methods

Increased cyclin D1 expression in cancer of the ampulla of Vater: relevance to nuclear β catenin accumulation and k-ras gene mutation

Aims: Several studies have reported that dysregulation of β catenin or k-ras mutation promotes cyclin D1 expression. This study investigated the relation between cyclin D1 expression and clinicopathological parameters in...

Calcitonin gene related peptide (CGRP) and calcitonin (CT) immunohistochemistry in relation with the presence of microcalcifications (MC) in invasive and preinvasive breast lesions: Influence in Tc-99m (V)DMSA and Tc-99m MIBI scintimammography (SM)

Calcitonin gene related peptide (CGRP) and calcitonin (CT) immunohistochemistry in relation with the presence of microcalcifications (MC) in invasive and preinvasive breast lesions: Influence in Tc-99m (V)DMSA and Tc-99m MIBI scintimammography (SM)