HER-2 Expression in Immunohistochemistry Has No Prognostic Significance in Gastric Cancer Patients

The role of HER-2 expression as a prognostic factor in gastric cancer (GC) is still controversial. The aim of the study was to asses HER-2 status, its correlations with clinicopathological parameters, and prognostic impact in GC patients. Tumor samples were collected from 78 patients who had undergone curative surgery. In order to evaluate the intensity of immunohistochemical (IHC) reactions two scales were applied: the immunoreactive score according to Remmele modified by the authors and standardised Hercep test score modified for GC by Hofmann et al. The HER-2 overexpression was detected by IHC in 23 (29.5%) tumors in Hercep test (score 2+/3+) and in 24 (30.7%) in IRS scale (IRS 4–12). The overexpression of HER-2 was associated with poorly differentiated tumors, but this correlation was not significant (P = 0.064). No relationship was found between HER-2 expression and primary tumor size and degree of spread to regional lymph nodes. Both univariate and multivariate analyses revealed that TNM stage and patient's age were the crucial negative prognostic factors. No correlation was observed between patient survival and expression of HER-2 estimated using both scales. This research did not confirm HER-2 expression (evaluated with immunohistochemistry) value as a prognostic tool in GC

Advanced ovarian cancer imitating deep infiltrating endometriosis. Radical resection and reconstructive surgery of the anterior abdominal wall

Endometriosis is a disease affecting approximately 10–15% of the female population of reproductive age [1]. A rare location is endometriosis in the scar after caesarean section — CSE (caesarean scar endometriosis) accounting for 0.5–1.0 % of all cases. Although endometriosis is usually a benign condition, its malignant transformation affects 0.7–1% of cases. In women diagnosed with ovarian cancer, foci of endometriosis are present in up to 30% of patients. This paper presents the case of a 36-year-old patient initially diagnosed with extensive endometriosis involving the anterior abdominal wall and the pelvis minor. After biopsy, a diagnosis of advanced low-grade serous ovarian cancer was established. The diagnostic methods used and the extent of surgery with reconstruction of the anterior abdominal wall were described

Landscape of oncoplastic breast surgery across Poland

Techniki onkoplastyczne i rekonstrukcyjne stanowią podstawowe narzędzia pracy współczesnych chirurgów piersi. Celem badania było ustalenie rodzajów rekonstrukcji onkoplastycznych przeprowadzanych w ośrodkach leczenia raka piersi w Polsce. Drogą e-mailową rozesłano kwestionariusz zawierający 18 pytań do członków Polskiego Towarzystwa Chirurgii Onkologicznej oraz Polskiego Towarzystwa Chirurgii Plastycznej, Rekonstrukcyjnej i Estetycznej poprzez ich portale internetowe. Liczba pacjentek z rakiem piersi poddawana operacji sięgała od 120 do 904 rocznie w każdym z ośrodków. Wykonywano głównie operacje oszczędzające pierś (breast conserving surgery - BCS) z wyjątkiem jednego ośrodka (zakres 50 – 70%). Jednoczasową rekonstrukcję piersi (immediate breast reconstruction - IBR) wykonywano w 6-42% zabiegów. Najczęstszym rodzajem IBR była dwuetapowa rekonstrukcja z użyciem ekspandera i wszczepieniem implantu lub jednoetapowa rekonstrukcja przy użyciu implantu z lub bez wszczepienia siatki syntetycznej. Najczęściej wykonywanym zabiegiem odroczonej rekonstrukcji piersi (delayed breast reconstruction - DBR) była dwuetapowa rekonstrukcja z użyciem ekspandera i następnie z wszczepieniem implantu. W żadnym z badanych ośrodków nie wykonywano rekonstrukcji z użyciem wolnego płata. W ośrodku chirurgii plastycznej wykonywano rekonstrukcję z wykorzystaniem płata perforatorów głębokich naczyń nabrzusznych dolnych (DIEP). W ośrodkach onkologicznych wykonywano rekonstrukcje z użyciem płatów uszypułowanych. W wybranych ośrodkach stosowano bezkomórkowe macierze skórne (ADM) oraz przeszczep tkanki tłuszczowej. Oceniono wyniki na podstawie opinii pacjentów (patient-reported outcome measures - PROM) oraz powikłania po zabiegach. Nasze wyniki mogą stanowić podstawę do dalszego doskonalenia umiejętności, akredytacji, zbierania danych i audytu, w tym oceny na podstawie opinii pacjentów. Istnieje również pilna potrzeba rozwiązania problemu nierówności w refundacji procedur w różnych państwach Europy.Oncoplastic and reconstructive techniques are essential tools in the armamentarium of contemporary breast surgeons. The aim of the study was to identify oncoplastic reconstructive patterns in breast cancer centers across Poland. A questionnaire of 18 questions was sent by email to the members of the Polish Society of Surgical Oncology and the Polish Society of Plastic, Reconstructive and Esthetic Surgery via their dedicated websites. The numbers of breast cancer patients operated on in each center ranged from 120 to 904 per year. Breast-conserving surgery (BCS) predominated in all but one center (range 50-70%). Immediate breast reconstructions (IBR) accounted for 6-42% of procedures, The most frequent type of IBR was either a two-stage expander followed by a permanent implant or one-stage implant- based with or without synthetic mesh. The most frequent type of delayed breast reconstruction (DBR) was a two-stage expander followed by implant-based reconstruction. None of the surveyed cancer centers performed free flap reconstruction. Deep inferior epigastric perforator (DIEP) flaps were performed in the plastic surgery department. Reconstructions based on pedicled flaps were performed in cancer centers. Acellular dermal matrices (ADM) and fat transfer were used in selected centers. In the clinical scenario of adjuvant radiotherapy, delayed breast reconstruction was favored. The full range of oncoplastic BCS was performed. Patient-reported outcome measures (PROM) and complications were assessed. Our findings can act as a platform for further improvement in skills, certification, data collection and audit, including patient reported expectation measures. There is also an urgent need to address pan-European inconsistencies in procedural reimbursement

Cultivation of circulating tumor cells in esophageal cancer

The presence of circulating tumor cells (CTCs) in patients with metastatic carcinoma is generally associated with poor clinical outcome. There have been many investigations showing a possible use of CTCs as minimally invasive predictive and prognostic biomarker in cancer medicine. In this report a size-based method (MetaCell®) for quick and easy enrichment and cultivation of CTCs is presented to enable possible CTCs use in esophageal cancer (EC) management. In total, 43 patients with diagnosed EC, 20 with adenocarcinoma (AdenoCa) and 23 with squamous cell carcinoma (SCC), were enrolled into the adaptive prospective-like study .All the patients were candidates for surgery. The CTCs were detected in 27 patients (62.8%), with a higher rate in adenocarcinoma (75%) than SCC (52%). Finally, there were 26 patients with resectable tumors exhibiting CTCs-positivity in 69.2% and 17 patients with non-resectable tumors with 41.7% CTCs-positivity. Interestingly, in the patients undergoing neoadjuvant therapy, the CTCs were detected at time of surgery in 55.5% (10/18). The overall size-based filtration approach enabled to isolate viable CTCs and evaluate to their cytomorphological features by means of vital fluorescent staining. The CTCs were cultured in vitro for further downstream applications including immunohistochemical analysis. This is the first report of the successful culturing of esophageal cancer CTCs. The detection of CTCs presence could help in the future to guide timing of surgical treatment in EC patients

Serum-derived extracellular vesicles from breast cancer patients contribute to differential regulation of T-cell-mediated immune-escape mechanisms in breast cancer subtypes

BackgroundIntracellular communication within the tumour is complex and extracellular vesicles (EVs) have been identified as major contributing factors for the cell-to-cell communication in the local and distant tumour environments. Here, we examine the differential effects of breast cancer (BC) subtype-specific patient serum and cell-line derived EVs in the regulation of T cell mediated immune responses. MethodsUltracentrifugation was used to isolate EVs from sera of 63 BC patients, 15 healthy volunteers and 4 human breast cancer cell lines. Longitudinal blood draws for EV isolation for patients on neoadjuvant chemotherapy was also performed. Characterization of EVs was performed by Nanoparticle Tracking Analysis (NTA), transmission electron microscopy (TEM) and immunoblotting. CD63 staining was performed on a tissue microarray of 218 BC patients. In-house bioinformatics algorithms were utilized for the computation of EV associated expression scores within The Cancer Genome Atlas (TCGA) and correlated with tumour infiltrating lymphocyte (TIL) scores. In vitro stimulation of PBMCs with EVs from serum and cell-line derived EVs was performed and changes in the immune phenotypes characterized by flow cytometry. Cytokine profiles were assessed using a 105-plex immunoassay or IL10 ELISA. ResultsPatients with triple negative breast cancers (TNBCs) exhibited the lowest number of EVs in the sera; whilst the highest was detected in ER+HER2+ cancers; reflected also in the higher level of CD63+ vesicles found within the ER+HER2+ local tumour microenvironment. Transcriptomic analysis of the TCGA data identified that samples assigned with lower EV scores had significantly higher abundance of CD4+ memory activated T cells, T follicular cells and CD8 T cells, plasma, and memory B cells; whilst samples with high EV scores were more enriched for anti-inflammatory M2 macrophages and mast cells. A negative correlation between EV expression scores and stromal TIL counts was also observed. In vitro experiments confirmed that circulating EVs within breast cancer subtypes have functionally differing immunomodulatory capabilities, with EVs from patients with the most aggressive breast cancer subtype (TNBCs) demonstrating the most immune-suppressive phenotype (decreased CD3+HLA-DR+ but increased CD3+PD-L1 T cells, increased CD4+CD127-CD25hi T regulatory cells with associated increase in IL10 cytokine production). In depth assessment of the cytokine modulation triggered by the serum/cell line derived exosomes confirmed differential inflammatory cytokine profiles across differing breast cancer subtypes. Studies using the MDA-231 TNBC breast cancer cell-line derived EVs provided further support that TNBC EVs induced the most immunosuppressive response within PBMCs.DiscussionOur study supports further investigations into how tumour derived EVs are a mechanism that cancers can exploit to promote immune suppression; and breast cancer subtypes produce EVs with differing immunomodulatory capabilities. Understanding the intracellular/extracellular pathways implicated in alteration from active to suppressed immune may provide a promising way forward for restoring immune competence in specific breast cancer patient populations

Serum-derived extracellular vesicles from breast cancer patients contribute to differential regulation of T-cell-mediated immune-escape mechanisms in breast cancer subtypes

Background: Intracellular communication within the tumour is complex and extracellular vesicles (EVs) have been identified as major contributing factors for the cell-to-cell communication in the local and distant tumour environments. Here, we examine the differential effects of breast cancer (BC) subtype-specific patient serum and cell-line derived EVs in the regulation of T cell mediated immune responses. Methods: Ultracentrifugation was used to isolate EVs from sera of 63 BC patients, 15 healthy volunteers and 4 human breast cancer cell lines. Longitudinal blood draws for EV isolation for patients on neoadjuvant chemotherapy was also performed. Characterization of EVs was performed by Nanoparticle Tracking Analysis (NTA), transmission electron microscopy (TEM) and immunoblotting. CD63 staining was performed on a tissue microarray of 218 BC patients. In-house bioinformatics algorithms were utilized for the computation of EV associated expression scores within The Cancer Genome Atlas (TCGA) and correlated with tumour infiltrating lymphocyte (TIL) scores. In vitro stimulation of PBMCs with EVs from serum and cell-line derived EVs was performed and changes in the immune phenotypes characterized by flow cytometry. Cytokine profiles were assessed using a 105-plex immunoassay or IL10 ELISA. Results: Patients with triple negative breast cancers (TNBCs) exhibited the lowest number of EVs in the sera; whilst the highest was detected in ER+HER2+ cancers; reflected also in the higher level of CD63+ vesicles found within the ER+HER2+ local tumour microenvironment. Transcriptomic analysis of the TCGA data identified that samples assigned with lower EV scores had significantly higher abundance of CD4+ memory activated T cells, T follicular cells and CD8 T cells, plasma, and memory B cells; whilst samples with high EV scores were more enriched for anti-inflammatory M2 macrophages and mast cells. A negative correlation between EV expression scores and stromal TIL counts was also observed. In vitro experiments confirmed that circulating EVs within breast cancer subtypes have functionally differing immunomodulatory capabilities, with EVs from patients with the most aggressive breast cancer subtype (TNBCs) demonstrating the most immune-suppressive phenotype (decreased CD3+HLA-DR+ but increased CD3+PD-L1 T cells, increased CD4+CD127-CD25hi T regulatory cells with associated increase in IL10 cytokine production). In depth assessment of the cytokine modulation triggered by the serum/cell line derived exosomes confirmed differential inflammatory cytokine profiles across differing breast cancer subtypes. Studies using the MDA-231 TNBC breast cancer cell-line derived EVs provided further support that TNBC EVs induced the most immunosuppressive response within PBMCs. Discussion: Our study supports further investigations into how tumour derived EVs are a mechanism that cancers can exploit to promote immune suppression; and breast cancer subtypes produce EVs with differing immunomodulatory capabilities. Understanding the intracellular/extracellular pathways implicated in alteration from active to suppressed immune state may provide a promising way forward for restoring immune competence in specific breast cancer patient populations

Lymphangioinvasion in routine H&E staining is strongly associated with poor clinical outcome in lymph node-negative cutaneous melanoma patients

Introduction. Lymphatic invasion (LYI) and lymphangiogenesis in the primary tumor are important processes related to the dissemination of neoplasms. The aim of the study was to examine the relationship of LYI status in cutaneous melanoma with patient survival and clinicopathological data. Material and methods. LYI status was assessed in 104 hematoxylin-eosin (H&E) stained melanoma primary tumor samples and analyzed in relation to patient survival and other clinicopathological and histopathological characteristics. Results. LYI was found in 30 (28.8%) patients. It was observed more frequently in ulcerated, proliferating, and thicker tumors. It correlated with the presence of sentinel lymph node, regional and distant metastases. The presence of LYI significantly correlated with shorter overall survival, cancer specific overall survival and disease-free survival in Kaplan-Meier analysis (all P < 0.001). Positive LYI status was a factor of unfavorable prognosis also in patients without regional lymph node metastases. Conclusions. Our results support earlier observations that LYI is a powerful prognostic factor. We recommend an assessment of LYI status during routine review of cutaneous melanoma slides stained with H&E as a standard, potentially informative, yet economically beneficial procedure.

Metabolomics of Interstitial Fluid, Plasma and Urine in Patients with Arterial Hypertension: New Insights into the Underlying Mechanisms

There is growing evidence that lymphatic system plays a pivotal role in the pathogenesis of hypertension. Here, for the first time, the metabolome of interstitial fluid is analyzed in patients with arterial hypertension. Due to ethical issues to obtain human interstitial fluid samples, this study included only oncological patients after axillary lymph node dissection (ALND). These patients were matched into hypertensive (n = 29) and normotensive (n = 35) groups with similar oncological status. Simultaneous evaluation of interstitial fluid, plasma, and urine was obtained by combining high-resolution proton nuclear magnetic resonance (1H NMR) spectroscopy with chemometric analysis. Orthogonal partial least squares discriminant analysis (OPLS-DA) provided a clear differentiation between the hypertension and normotensive group, with the discrimination visible in each biofluid. In interstitial fluid nine potential metabolomic biomarkers for hypertension could be identified (creatinine, proline, pyroglutamine, glycine, alanine, 1-methylhistidine, the lysyl group of albumin, threonine, lipids), seven distinct markers in plasma (creatinine, mannose, isobutyrate, glycine, alanine, lactate, acetate, ornithine), and seven respectively in urine (methylmalonate, citrulline, phenylacetylglycine, fumarate, citrate, 1-methylnicotinamide, trans-aconitate). Biomarkers in plasma and urine allowed for the identification of specific biochemical pathways involved in hypertension, as previously suggested. Analysis of the interstitial fluid metabolome provided additional biomarkers compared to plasma or urine. Those biomarkers reflected primarily alterations in the metabolism of lipids and amino acids, and indicated increased levels of oxidative stress/inflammation in patients with hypertension.Special Issue: Primary and Secondary Hypertension: Novelty in Diagnosis and Treatment</p