ER Markers Research Articles

A study of the prevalence of PGR gene expression with ER and PR Receptor markers positivity in female breast cancer cases in Al-Najaf province, Iraq

A study of the prevalence of PGR gene expression with ER and PR Receptor markers positivity in female breast cancer cases in Al-Najaf province, Iraq

PLASMA CALPROTECTIN VERSUS CONVENTIONAL INFECTION BIOMARKERS IN HIP FRACTURE PATIENTS: A PROSPECTIVE COHORT STUDY

After a hip fracture, infections are common, but signs of infection resemble those of systemic inflammatory response to trauma and surgery, and conventional infection markers lack specificity. Plasma-calprotectin, a novel marker of neutrophil activation, has shown potential as an infection marker in ER and ICU settings.To investigate if plasma-calprotectin is superior compared to conventional infection biomarkers after hip fracture.Prospective cohort study of hip fracture patients admitted to our department. Calprotectin, procalcitonin (PCT), C-reactive protein (CRP), and white blood cell (WBC) count were measured in blood plasma upon admission and on day 3 post-surgery. Patients with infection (pneumonia, UTI, sepsis, SSI, other soft tissue infections) pre- or post-surgery were compared to a control group without infection within 30 days.Statistics: Wilcoxon rank-sum test, medians with interquartile range, and area under the curve (AUC) with 95% confidence intervals.Pilot study comprises calprotectin obtained at least once for 60 patients at admission and 48 on day 3. Mean age 84 years (SD 8.4), 65% women.9/60 patients (23%) were admitted with infections. They had higher levels of CRP (median 111 [73-149]) and PCT (0.35 [0.18–0.86]) compared to the control group (29 [16-64], p=0.037; 0.10 [0.07–0.17], p=0.007). Calprotectin (2.67 vs 2.51) and WBC (12.2 vs 9.3) did not differ significantly. AUC was highest for PCT (0.79 [CI 0.60–0.97]), followed by CRP (0.71 [0.46–0.96]), WBC (0.60 [0.35–0.84]), and calprotectin (0.58, [0.33–0.83]).Day 3, 6/48 (13%) had infections, without significant differences between groups in any marker. The median levels were: calprotectin 3.5 vs 3.1, CRP 172 vs 104, WBC 12 vs 9, PCT 0.16 vs 0.17. Calprotectin had highest AUC 0.68 (0.41–0.93, n.s.). AUC for WBC was 0.67 (0.31–1.00), CRP 0.66 (0.38–0.94), PCT 0.56 (0.29–0.82).Preliminary data show no significant associations with postoperative infection for any of the studied biomarkers. However, plasma-calprotectin might perform slightly better compared to conventional markers.

ATF4 as a Prognostic Marker and Modulator of Glutamine Metabolism in Oestrogen Receptor-Positive Breast Cancer.

ATF4, a stress-responsive transcription factor that upregulates adaptive genes, is a potential prognostic marker and modulator of glutamine metabolism in breast cancer. However, its exact role remains to be elucidated. ATF4 expression was evaluated at genomic and transcriptomic levels using METABRIC (n = 1,980), GeneMiner (n = 4,712), and KM-Plotter datasets. Proteomic expression was assessed via immunohistochemistry (n = 2,225) in the Nottingham Primary Breast Cancer Series. ATF4 genomic copy number (CN) variation and mRNA/protein in association with clinicopathological parameters, amino acid transporters (AATs), and patient outcome were investigated. Genomic, transcriptomic, and proteomic overexpression of ATF4 was associated with more aggressive ER-negative tumours. ATF4 mRNA and protein expression were significantly associated with increased expression of glutamine related AATs including SLC1A5 (p < 0.01) and SLC7A11 (p < 0.02). High ATF4 and SLC1A5 protein expression was significantly associated with shorter breast cancer-specific survival (p < 0.01), especially in ER+ tumours (p < 0.01), while high ATF4 and SLC7A11 protein expression was associated with shorter survival (p < 0.01). These findings suggest a complex interplay between ATF4 and AATs in breast cancer biology and underscore the potential role for ATF4 as a prognostic marker in ER+ breast cancer, offering a unique opportunity for risk stratification and personalized treatment strategies.

Autophagy Promotes Enrichment of Raft Components within Extracellular Vesicles Secreted by Human 2FTGH Cells.

Autophagy plays a key role in removing protein aggregates and damaged organelles. In addition to its conventional degradative functions, autophagy machinery contributes to the release of cytosolic proteins through an unconventional secretion pathway. In this research, we analyzed autophagy-induced extracellular vesicles (EVs) in HT1080-derived human fibrosarcoma 2FTGH cells using transmission electron microscopy and atomic force microscopy (AFM). We preliminary observed that autophagy induces the formation of a subset of large heterogeneous intracellular vesicular structures. Moreover, AFM showed that autophagy triggering led to a more visible smooth cell surface with a reduced amount of plasma membrane protrusions. Next, we characterized EVs secreted by cells following autophagy induction, demonstrating that cells release both plasma membrane-derived microvesicles and exosomes. A self-forming iodixanol gradient was performed for cell subfractionation. Western blot analysis showed that endogenous LC3-II co-fractionated with CD63 and CD81. Then, we analyzed whether raft components are enriched within EV cargoes following autophagy triggering. We observed that the raft marker GD3 and ER marker ERLIN1 co-fractionated with LC3-II; dual staining by immunogold electron microscopy and coimmunoprecipitation revealed GD3-LC3-II association, indicating that autophagy promotes enrichment of raft components within EVs. Introducing a new brick in the crosstalk between autophagy and the endolysosomal system may have important implications for the knowledge of pathogenic mechanisms, suggesting alternative raft target therapies in diseases in which the generation of EV is active.

Overexpression of Mammaglobin-A in Primary Breast Tissue Tumor and High Concentration of mRNA Mammaglobin-A in Peripheral Blood as Risk Factors for Metastatic Breast Cancer

Objective: There is still limited evidence for the use of biomarkers in breast cancer. However, mammaglobin-A in tissue and blood has recently been found as a promising biomarker for detecting metastases. Therefore, this study will examine the overexpression of mammaglobin-A in primary breast tissue tumors and the elevated concentration of mammaglobin-A messenger RNA (mRNA0 in peripheral blood as potential risk factors for breast cancer metastasis. Methods: The study was conducted at Sanglah General Hospital in Bali, Indonesia, from July 2017 to March 2018. It employed a hybrid research design, combining both cross-sectional (n = 70) and case-control (metastasis = 20 and non-metastasis = 20) approaches. Peripheral blood samples were collected using specialized L6 tubes and were analyzed for the expression of mammaglobin-A mRNA using quantitative real-time PCR. The diagnosis of breast cancer was established through open biopsy, considered the gold standard. Biopsy specimens underwent histopathological examination and standard immunohistochemistry (IHC) analysis to assess ER, PR, HER2, and Ki67 markers. The evaluation of mammaglobin-A protein expression was conducted via IHC at the Anatomical Pathology Laboratory of Prima Medika Hospital. Result: The mean mammaglobin-A mRNA level in the metastatic group was 11.59±1.37, while in the non-metastatic group was 8.17±1.27-fold change relative to the housekeeping gene beta microglobulin. The mean mammaglobin-A mRNA levels in the two groups were significantly different (p<0.05). Overexpression of mammaglobin-A in cancer tissue was 7.36 times more likely for metastasis compared to non-metastatic (OR = 7.36; 95% CI = 1.34-40.55; p = 0.013). Additionally, mammaglobin-A mRNA concentration was found to be nine times higher for breast cancer metastasis compared to the non-metastatic group (OR = 9.00; 95% CI = 2.15-37.66; p = 0.002). Conclusion: The mean mammaglobin-A level in the metastatic group significantly differs from that in the non-metastatic group. Overexpression of mammaglobin-A and a high concentration of Mammaglobin-A mRNA are influential risk factors for metastatic breast cancer.

Differences in Histological Subtypes of Invasive Lobular Breast Carcinoma According to Immunohistochemical Molecular Classification.

The aim of this study was to compare the differences between histological and molecular subtypes of invasive lobular carcinoma (ILC) of the breast, in order to be able to predict the behavior and prognosis of the disease, as well as to effectively determine therapy. This study included 263 cases of breast ILC diagnosed over a seven-year period. The diagnosis of invasive lobular carcinoma is based on the characteristic growth pattern and phenotype of cancer cells with the respective subtypes: classic, alveolar, solid, tubulolobular, pleomorphic and mixed lobular type. The examined cases were divided into five groups according to molecular classification based on the expression of ER, PR, HER2 and Ki67 immunohistochemical markers. It was found that the pleomorphic subtype occurred statistically significantly less often as the luminal A subtype compared to others (p = 0.00027), and the HER2-enriched subtype occurred statistically significantly more often in the pT4 stage (p = 0.024). The results of this study significantly singled out the luminal A subtype, and among them classic ILC, as the subtype with the most favorable expression ratio of the investigated predictive/prognostic immunohistochemical markers.

The effect of hippocampal overexpression of dopamine neurotrophic factor (CDNF) on the behavior of mice with genetic predisposition to depressive-like behavior

Cerebral dopamine neurotrophic factor (CDNF) is the promising tool for treatment of Parkinson’s disease. However, its role in the regulation of non-motor behavior, including various psychopathologies, remains unclear. In this regard, the aim of present work was to study the effect of CDNF overexpression in the hippocampus on the behavior of ASC mice (Antidepressant Sensitive Cataleptics) with a genetic predisposition to depression-like behavior. CDNF overexpression in mouse hippocampal neurons was induced using an adeno-associated viral vector. Four weeks after stereotaxic injection of the AAV-CDNF construct into the dorsal hippocampus, home cage behavior, exploratory, anxiety and depressive-like behaviors, as well as spatial and associative learning were assessed. We found significant improvements in the dynamics of spatial learning in the Morris water maze in CDNF-overexpressing animals. At the same time, no effect of CDNF was found on other studied behaviors. The behavior of the experimental animals in the home cage conditions did not differ from that in the control group, except for a decrease in the total amount of food eaten and a slight increase in the number of sleep episodes in the light phase of the day. In the present study we also attempted to determine the molecular basis for the above mentioned changes by genes expression assessment. We did not find significant changes in the mRNA level of key kinases, genes involved in neuroplasticity and neuronal survival as well as genes encoding receptors for the main neurotransmitter systems. However, animals overexpressing CDNF show increased lever of spliced Xbp indicating activation of the Ire1α/Xbp1 pathway traditionally associated with ER stress. Immunohistochemical analysis showed that CDNF was co-localized with the ER marker calreticulin. Thus, the effects of endogenous CDNF on behavior that we have found may be mediated by a specific molecular cascade, which emphasizes its difference from classical neurotrophic factors.

Investigating The Expression of ER, PR, And Ki-67 In Different Grades of Meningioma

Introduction: Tumor grade is one of the significant factors in determining the prognosis of meningioma. Ki-67 is a marker expressed in the active phase of the cell cycle and indicates cell division and can be used as an auxiliary tool in determining the grade of meningioma. Steroid hormones are one of the factors that are effective in the pathogenesis of meningioma. This study evaluates the expression level of ER, PR, and ki-67 markers in different grades of meningioma.
 Materials and Methods: In a descriptive study, slides, and paraffin blocks of 80 meningioma patients who underwent surgery over 10 years at Besat Hospital, Hamedan, were extracted from the patient’s medical records and were evaluated in terms of the expression of ER markers, PR and ki-67 after immunohistochemical staining. Results: In meningioma grades 1, 2, and 3, mean PR expression was 17.33, 11.66, and zero (P=0.033), respectively, and mean Ki-67 expression was 0.67, 5.22, and 10.50, respectively ( P<0.001), and mean ER expression was zero in all three grades. There was an inverse and significant correlation between tumor grade and PR rate (p=0.04, r=-0.21). Conclusion: ki-67 expression can be used as an auxiliary method in meningioma grading, and more PR positivity indicates lower grades, and thus, better tumor prognosis.

The Value of ER∝ in the Prognosis of GH- and PRL-Secreting PitNETs: Clinicopathological Correlations.

Pituitary neuroendocrine tumors (PitNETs) are divided into multiple histological subtypes, which determine their clinical and biological variable behavior. Despite their benign evolution, in some cases, prolactin (PRL) and growth hormone (GH)-secreting PitNETs may have aggressive behavior. In this study, we investigated the potential predictive role of ER∝, alongside the clinicopathological classification of PitNETs (tumor diameter, tumor type, and tumor grade). A retrospective study was conducted with 32 consecutive cases of PRL- and mixed GH- and PRL-secreting PitNETs (5 patients with prolactinomas and 27 with acromegaly, among them, 7 patients with GH- and PRL- co-secretion) who underwent transsphenoidal intervention. Tumor specimens were histologically and immunohistochemical examined: anterior pituitary hormones, ki-67 labeling index, CAM 5.2, and ER∝; ER∝ expression was correlated with basal PRL levels at diagnosis (rho = 0.60, p < 0.01) and postoperative PRL levels (rho = 0.58, p < 0.001). In our study, the ER∝ intensity score was lower in female patients. Postoperative maximal tumor diameter correlated with Knosp grade (p = 0.02); CAM 5.2 pattern (densely/sparsely granulated/mixed densely and sparsely granulated) was correlated with postoperative PRL level (p = 0.002), and with ki-67 (p < 0.001). The IGF1 level at diagnosis was correlated with the postoperative GH nadir value in the oral glucose tolerance test (OGTT) (rho = 0.52, p < 0.05). Also, basal PRL level at diagnosis was correlated with postoperative tumor diameter (p = 0.63, p < 0.001). At univariate logistic regression, GH nadir in OGTT test at diagnostic, IGF1, gender, and invasion were independent predictors of remission for mixed GH- and PRL-secreting Pit-NETs; ER∝ can be used as a prognostic marker and loss of ER∝ expression should be considered a sign of lower differentiation and a likely indicator of poor prognosis. A sex-related difference can be considered in the evolution and prognosis of these tumors, but further studies are needed to confirm this hypothesis.

Immunohistochemistry-derived subtypes of breast cancer distribution in four regions of Ethiopia.

Different biological characteristics, therapeutic responses, and disease-specific outcomes are associated with different molecular subtypes of breast cancer (BC). Although there have been different studies on BC in the Ethiopian capital city of Addis Ababa, there have been few studies in other parts of the nation, and none have evaluated biological characteristics in other locations in the context of the extensive ethnic and genetic diversity found in Ethiopia. This study was carried out to evaluate the distribution of immunohistochemistry (IHC) subtypes of BCs throughout four Ethiopian regions. A total of 227 formalin-fixed paraffin-embedded (FFPE) tissue blocks were collected from tertiary hospitals in four Ethiopian regions between 2015 and 2021. The IHC staining was performed for subtyping, ER, PR, HER2, and Ki-67 proliferation markers. The mean age at diagnosis was 43.9 years. The percentage of ER and PR-negative tumors were 48.3% and 53.2%, respectively. The IHC subtypes showed the following distribution: 33.1% triple-negative breast cancer (TNBC), 27.6% luminal B, 25.2% luminal A, and 14.1% HER2 enriched. In multiple logistic regression analysis, grade III and HER2 positivity were associated with larger tumor size, and also originating from Jimma compared to Mekele. Patients with ER-negative, PR-negative, and TNBC were found in 48.3%, 53.2%, and 33.1% of cases, respectively, showing that half the patients could potentially benefit from endocrine treatment. A considerably high prevalence of TNBC was reported in our study, demanding additional research that includes genetic predisposition factors. Additionally, aggressive tumors were found in a high percentage of younger age groups, which must be considered when planning personalized treatment strategies.

AI-based intra-tumor heterogeneity score of Ki67 expression as a prognostic marker for early-stage ER+/HER2- breast cancer.

Early-stage estrogen receptor positive and human epidermal growth factor receptor negative (ER+/HER2-) luminal breast cancer (BC) is quite heterogeneous and accounts for about 70% of all BCs. Ki67 is a proliferation marker that has a significant prognostic value in luminal BC despite the challenges in its assessment. There is increasing evidence that spatial colocalization, which measures the evenness of different types of cells, is clinically important in several types of cancer. However, reproducible quantification of intra-tumor spatial heterogeneity remains largely unexplored. We propose an automated pipeline for prognostication of luminal BC based on the analysis of spatial distribution of Ki67 expression in tumor cells using a large well-characterized cohort (n = 2,081). The proposed Ki67 colocalization (Ki67CL) score can stratify ER+/HER2- BC patients with high significance in terms of BC-specific survival (p < 0.00001) and distant metastasis-free survival (p = 0.0048). Ki67CL score is shown to be highly significant compared with the standard Ki67 index. In addition, we show that the proposed Ki67CL score can help identify luminal BC patients who can potentially benefit from adjuvant chemotherapy.

Effect of Hippocampal Overexpression of Dopamine Neurotrophic Factor (CDNF) on Behavior of Mice with Genetic Predisposition to Depressive-Like Behavior.

Cerebral dopamine neurotrophic factor (CDNF) is a promising agent for Parkinson's disease treatment. However, its role in regulation of non-motor behavior including various psychopathologies remains unclear. In this regard, the aim of the present work was to study effect of CDNF overexpression in hippocampus on behavior of the ASC mice (Antidepressant Sensitive Cataleptics) with genetic predisposition to depressive-like behavior. CDNF overexpression in the mouse hippocampal neurons was induced using an adeno-associated viral vector. Four weeks after stereotaxic injection of the AAV-CDNF construct into the dorsal hippocampus home cage activity, exploratory, anxious and depressive-like types of behavior, as well as spatial and associative learning were assessed. We found significant improvements in the dynamics of spatial learning in the Morris water maze in the CDNF-overexpressing animals. At the same time, no effect of CDNF was found on other types of behavior under study. Behavior of the experimental animals under home cage conditions did not differ from that in the control group, except for the decrease in the total amount of food eaten and slight increase in the number of sleep episodes during the light phase of the day. In the present study we also attempted to determine molecular basis for the above-mentioned changes through assessment of the gene expression pattern. We did not find significant changes in the mRNA level of key kinases genes involved in neuroplasticity and neuronal survival, as well as genes encoding receptors for the main neurotransmitter systems. However, the CDNF-overexpressing animals showed increased level of the spliced Xbp indicating activation of the Ire1α/Xbp-1 pathway traditionally associated with ER stress. Immunohistochemical analysis showed that CDNF was co-localized with the ER marker calreticulin. Thus, the effects of endogenous CDNF on behavior that we have found could be mediated by a specific molecular cascade, which emphasizes its difference from the classical neurotrophic factors.

Obesity Associated with Prediabetes Increases the Risk of Breast Cancer Development and Progression-A Study on an Obese Rat Model with Impaired Glucose Tolerance.

Patients with comorbidities of obesity and diabetes are recognized to be at high risk of breast cancer development and face worse breast cancer outcomes. Though several reports showed the reinforced link between obesity, diabetes, and prediabetes with breast cancer, the underlying molecular mechanisms are still unknown. The present study aimed to investigate the underlying molecular link between increased risks of breast cancer due to coincident diabetes or obesity using a spontaneous obese rat model with impaired glucose tolerance (WNIN/GR-Ob rat). A single dose of solubilized DMBA suspension (40 mg/kg body weight) was orally administered to the animals at the age of 60 days to induce breast tumors. The tumor incidence, latency period, tumor frequency, and tumor volume were measured. Histology, immunohistochemistry, and immunoblotting were performed to evaluate the tumor morphology and expression levels of signal molecules. The development of mammary tumors in GR-Ob rats was characterized by early onset and shorter latency periods compared to control lean rats. While 62% of obese rats developed breast tumors, tumor development in lean rats was only 21%. Overexpression of ER, PR, Ki67, and p53 markers was observed in tumor tissues of obese rats in comparison with lean rats. The levels of the hallmarks of cell proliferation and angiogenesis involved in IGF-1/PI3K/Akt/GSK3β/β-catenin signaling pathway molecules were upregulated in obese rat breast tumors compared to lean rats. Furthermore, obesity with prediabetes is associated with changes in IGF-1 signaling and acts on PI3K/Akt/GSK3β/β-catenin signaling, which results in rapid cell proliferation and development of breast tumors in obese rats than the lean rats. These results indicate that tumor onset and development were faster in spontaneous obese rat models with impaired glucose tolerance than in their lean counterparts.

An Immunohistochemical Study of Breast Cancer Brain Metastases: The Role of CD44 and AKT in the Prognosis.

Breast cancer is a major health burden, and up to one-third of patients with breast cancer develop brain metastases, which are linked to a very poor prognosis. Few biomarkers are available to predict the prognosis of patients with metastases. Assessment by immunohistochemistry may be used as a tool to predict the behavior of these tumors. A retrospective transversal study including 114 patients (diagnosed between 2000 and 2016) with breast cancer brain metastases was carried out using archival biological material from 114 patients with breast cancer brain metastases. Expression of CD44, HER2, ER, PR, CA9, PDL-1, CD133, ALDH1, PTEN, AKT, PI3K, and AR markers was assessed by immunohistochemistry. The overexpression of CD44 and AKT was associated with worse overall survival ( P =0.047 and P =0,034, respectively), on univariate analysis, in the cohort of parenchymal and bone metastases; the impact of AKT expression was also evident in the parenchymal cohort on uni ( P =0.021) and multivariate analysis ( P =0.027). The remaining markers did not exhibit a statistical correlation. Immunohistochemistry markers such as CD44 and AKT may have a prognostic impact on survival in patients with breast cancer brain metastases. The conjugation with other markers may help with the stratification of patients and therapy.

Homotypic Entosis as a Potential Novel Diagnostic Marker in Breast Cancer

Homotypic entotic figures, which are a form of "cell-in-cell" structures, are considered a potential novel independent prognostic marker in various cancers. Nevertheless, the knowledge concerning the biological role of this phenomenon is still unclear. Since breast cancer cells are remarkably entosis-competent, we aimed to investigate and compare the frequency of entoses in a primary breast tumor and in its lymph node metastasis. Moreover, as there are limited data on defined molecular markers of entosis, we investigated entosis in correlation with classical breast cancer biomarkers used in routine pathomorphological diagnostics (HER2, ER, PR, and Ki67). In the study, a cohort of entosis-positive breast cancer samples paired into primary lesions and lymph node metastases was used. The inclusion criteria were a diagnosis of NOS cancer, lymph node metastases, the presence of entotic figures in the primary lesion, and/or lymph node metastases. In a selected, double-negative, HER2-positive NOS breast cancer case, entoses were characterized by a correlation between an epithelial-mesenchymal transition and proliferation markers. We observed that in the investigated cohort entotic figures were positively correlated with Ki67 and HER2, but not with ER or PR markers. Moreover, for the first time, we identified Ki67-positive mitotic inner entotic cells in clinical carcinoma samples. Our study performed on primary and secondary breast cancer specimens indicated that entotic figures, when examined by routine HE histological staining, present potential diagnostic value, since they correlate with two classical prognostic factors of breast cancer.

Front Cover

FEBS LettersVolume 597, Issue 5 p. 599-600 Issue InformationFree Access Front Cover First published: 13 March 2023 https://doi.org/10.1002/1873-3468.14383AboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onFacebookTwitterLinkedInRedditWechat Graphical Abstract Cover illustration D-SIM imaging of ER networks labeled by a novel ER marker, RR-mNeonGreen. Image taken from Han et al. ‘A novel fl uorescent endoplasmic reticulum marker for super-resolution imaging in live cells’. Volume597, Issue5March 2023Pages 599-600 RelatedInformation

Breast Cancer Classification According to Immunohistochemical Markers: Clinicopathologic Features in Women Treated at Pietersburg Hospital, Limpopo, South Africa

Background: Breast cancer molecular subtypes can provide critical prognostic information that is useful to individualize care in addition to avoiding unwarranted adjuvant chemotherapy in some patients. Micro-array-based genetic profiling used to type breast tumors is expensive and unavailable for most low- and middle-income countries. Molecular subtyping using immunohistochemical (IHC) markers has proved to be a viable alternative. We therefore used this immunohistochemical technique to classify the breast cancer subtypes in our patient population and evaluated its association with clinicopathologic characteristics in these patients. Patients and Methods: We retrospectively evaluated immunohistochemical marker results (oestrogen, progesterone and Human Epidermal Growth Factor Receptor-2 (HER2) receptors) of 254 breast cancer patients treated at a tertiary referral hospital from 01 January 2010 to 31 December 2011 against their clinicopathological variables. Results: Out of 329 patients with invasive breast cancer treated within the study period 254 who had complete IHC marker test results were evaluated. Of these 136(54%) were luminal A, 69(27%) triple negative breast cancer (TNBC), 29(11%) HER2- enriched, and 20(8%) luminal B. The mean age at diagnosis was 55.6, 54.4, 54.2, and 52.3 years for luminal A, TNBC, HER2, and luminal B, respectively. The rate of lymph node involvement was 80%, 66.7%, 66.2%, and 58.6% for luminal B, TNBC, luminal A, and HER2, respectively. There was no association between molecular subtypes and clinical stage (p=0.578) or distant metastasis (p = 0.940). Grade 3 tumors were most prevalent in TNBC (59.4%) followed by luminal B (40%), HER2 (37.9%), and luminal A (37.5%). Five-year survival was 27.2%, 25%, 24.1% and 23.2% for luminal A, luminal B, HER2 and TNBC respectively (p=0.104). Conclusion: A large proportion or our patients (22.8%, n=75) with missing IHC test results could not be classified into breast cancer molecular subtypes and may have missed an opportunity for an individualized care. There is need for IHC tests in every breast neoplasm biopsy specimen to include ER, PR, HER2, and Ki67 markers for classification of the lesion according to molecular subtypes.

7T MRI and molecular studies of Dotarem (gadoterate meglumine) retention in macrophages.

Gadolinium-based MRI contrast agent Dotarem is commonly used in the clinic. Free Gd3+ is very toxic. Because the crystallographic radius of Gd3+ and Ca2+ is very similar, Gd3+ can compete with Ca2+ in the regulations of cellular functions. Gadolinium is retained in organs and tissues, but the effects of gadolinium retention are mostly unknown. There is also no information how gadolinium retention impacts immune cells and signaling pathways regulated by Ca2+ such as RhoA/mTORC1 and mTORC2, which control various, including actin cytoskeleton-dependent, cellular functions. We developed the 7T MRI-based method to analyze gadolinium-based MRI contrast agent retention in isolated immune cells (macrophages) and study the effects of gadolinium retention on the expression of RhoA, mTORC1, mTORC2, and mitochondria, Golgi, and ER markers. We showed that macrophages retain gadolinium for at least seven days after exposure. Gadolinium retention downregulated the expression of RhoA, mTORC1 component Raptor and mTORC2 component Rictor proteins, and dysregulated the expression level of organelle markers. The method described here can be used for monitoring gadolinium levels and effects in isolated cells, such as the immune cells in the blood of patients exposed to contrast-enhanced MRI.

A Study of Molecular Subtypes of Carcinoma Breast by Immunohistochemistry at Tertiary Care Center, Jaipur

Introduction: Breast cancer is one of the most common malignant tumors and one of the leading causes of cancer related death amongst females worldwide. The clinical significance of molecular classification of carcinoma breast remains to be established. Molecular subtyping using immunohistochemistry can provide additional prognostic and predictive information. Aim and objectives: To identify morphological variants of carcinoma breast and to determine various proportions of molecular subtypes of carcinoma breast by immunohistochemistry. Materials and Methods: A descriptive type of observational study was done at SMS Medical College, Jaipur. 65 Modified Radical Mastectomy specimens received in the department were included in the study. The surgical specimens were then evaluated histopathologically and immunohistochemically for ER, PR, HER2/neu, and Ki67 markers. Results: Out of 65 breast carcinoma cases the most common histological type encountered in our study was Invasive ductal carcinoma of no special type (NST) (83.09%) followed by medullary carcinoma (4.63%) and invasive lobular carcinoma (3.09%) and a single case each of tubular, cribriform, metaplastic, lymphoepithelioma like carcinoma, carcinoma with apocrine differentiation and oncocytic carcinoma. The cases were further classified into molecular subtypes using protein expression patterns in IHC. The proportion of tumors found were Luminal A (32.31%), Luminal B (18.46%), HER2/neu overexpressed (13.84%), and Triple-negative (35.39%). Conclusion: The most common molecular subtype found in our study was Triple negative. The use of IHC markers in the clinical setting could determine the prognosis and benefit the patients with targeted therapies.&#x0D;

Assessment of estrogen receptor marker and smooth muscle actin marker in oral pyogenic granuloma

Soft tissue enlargement of the oral cavity usually existent a diagnostic challenge because many group of pathologic activity can cause these lesions. An enlargement may display a variation of normal anatomic structure, cyst, inflammation, developmental anomalies and neoplasm. These lesions included a group of reactive hyperplasia which arises in response to recurring; chronic tissue injury that provokes vigorous or extreme tissue repair response .so pyogenic granuloma is one of the many common entities in charge of soft tissue enlargement. Objective: The current study aimed to detect the immunohistochemical expressions of ER and alpha smooth muscle actin (α-SMA) for lobular capillary hemangioma (pyogenic granuloma). Material and Methods: fifty cases of confirmed diagnostic pyogenic granuloma were subjected to immunohistochemically assessment by using ER marker and Smooth Muscle Actin marker. Result: The immunohistochemically finding appeared that all 50 cases were positive for SMA marker while only 14 cases were positive for ER marker. So there is no significant difference between ER marker in relation to clinical parameters (age, gender and location) and intensity.