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Plants have been an important source for discovery of anticancer compounds. With the current decline in the number of new molecular entities from the pharmaceutical industry, novel anticancer agents are being sought from traditional medicines; therefore the anticancer efficacy of many plants that are used in traditional medicine is yet to be verified. The objective of the study was to evaluate the cytotoxic potential of ethanolic leaf gall extract of Terminalia chebula are evaluated against buffalo rat liver 3A, MCF-7 (Human mammary gland adenocarcinoma) and A-549 (Human lung cancer) cell lines. The cytotoxic effect of the ethanolic extract was evaluated by MTT assay. The extract was potent and effective in inducing cytotoxic effects in all the cell lines with an IC50 value of 305.18 ± 1.7 μg/mL, 643.13 ± 4.2 μg/mL, and 208.16 ± 3.7 μg/mL, respectively. The extract was more effective against A549 cell lines when compared to others. The presences of phenolics, triterpenoids, and flavonoids were identified in the extract. The extract showed total phenolic and flavonoid content of 478 ± 2.2 mg of gallic acid equivalent/g d.w and 538 ± 1.4 mg of quercetin equivalent/g d.w, respectively. This higher content of total phenolics and flavonoids found in the ethanolic extract was directly associated to higher cytotoxicity activity. Conclusion: The ethanolic leaf gall extract of T. chebula showed effective cytotoxic activities; which might be attributed to the phenolics/flavonoids present in higher concentration. Future work will be interesting to know the chemical composition of the extract and also better understand the mechanism of action of the constituents present in the extract to develop it as drug for therapeutic application. [ABSTRACT FROM AUTHOR]

Objective: Terminalia chebula gall extracts are widely used Asian folk and traditional medicine. The present study was carried out to evaluate the anthelmintic potential of different extracts of leaf galls. Materials and Methods: The anthelmintic activity was assessed by applying five different concentrations of the plant extracts on Indian adult earthworms, and the time of paralysis and death was recorded. Results: The ethanolic extract processed potent anthelmintic activity, when compared to the other extracts. The lowest time for paralysis and death of worms, for test sample at highest concentration (250 mg/ml), were found to be 7.30 ± 2.66 and 14 ± 0.58 min, respectively. Albendazole, which was used as standard, caused paralysis and death of worms at 07.00 ± 3.55 min and 12.60 ± 2.01 min, respectively; whereas no mortality of the worms was observed, when distilled water was used as control. The presences of phenolics, flavonoids, triterpens, saponins, glycosides, phytosterols, reducing sugars were identified in the extracts and the significant anthelmintic property of T. chebula might be due to the presence of alkaloids, phenolic compounds, and flavanoids. Conclusion: The results of this study establishes the antihelmentic activities of T. chebula leaf gall extracts and justify the ethnobotanical approach in the search for novel bioactive compounds. The anthelmintic potential of T. chebula extracts may be due to the presence of phyto-constituents like alkaloids, phenolic compounds, and flavanoids. Future work will be interesting to know the chemical composition and better understand the mechanism of action of the antioxidants present in the extract for development as drug for therapeutic application. [ABSTRACT FROM AUTHOR]

Embryo sac development conforms to Polygonum type. Starch grains are noticed in the embryo sac right from the megaspore mother cell stage. Twin tetrads one with a chalazal functional megaspore and another with a micropylar functional megaspore have been noticed in some ovules. While 75% of the embryo sacs have normal polarity, in 20% of the ovules reverse polarity has been noticed. In the remaining 5% of the ovules, 'bipolarity' has been observed. Reverse polarity in relation to double archegoniate theory has been discussed. [ABSTRACT FROM AUTHOR]

Introduction: Free radicals are implicated in several metabolic diseases and the medicinal properties of plants have been explored for their potent antioxidant activities to counteract metabolic disorders. This research highlights the chemical composition and antioxidant potential of leaf gall extracts (aqueous and methanol) of Syzygium cumini S. cumini), which have been extensively used in traditional medications to treat various metabolic diseases. Methods: The antioxidant activities of leaf gall extracts were examined using diphenylpicrylhydrazyl (DPPH), nitric oxide scavenging, hydroxyl scavenging and ferric reducing power (FRAP) methods. Results: In all the methods, the methanolic extract showed higher antioxidant potential than the standard ascorbic acid. The presence of phenolics, flavonoids, phytosterols, terpenoids, and reducing sugars was identified in both the extracts. When compared, the methanol extract had the highest total phenolic and flavonoid contents at 474±2.2 mg of GAE/g d.w and 668±1.4 mg of QUE/g d.w, respectively. The significant high antioxidant activity can be positively correlated to the high content of total polyphenols/flavonoids of the methanol extract. Conclusion: The present study confirms the folklore use of S. cumini leaves gall extracts as a natural antioxidant and justifies its ethnobotanical use. Further, the result of antioxidant properties encourages the use of S. cumini leaf gall extracts for medicinal health, functional food and nutraceuticals applications.

Objective: To study the detailed pharmacognostic profile of galls of Quercus infectoria Olivier (Q. infectoria olivier) (Fagaceae), an important medicinal plant used in the Indian system of medicine. Methods: Samples of galls of Q. infectoria were studied by macroscopical, microscopical, physiochemical, phytochemical, fluorescence analysis and othjer methods for standardization as recommended by WHO. Results: Macroscopically, the crude drug is globose with horny appearances on external surface (1.4–2.3 cm in length and 1–1.5 cm in diameter), with greyish-brown to brownish-black in colour externally and dark brown buff colored. Surface is smooth with numerous horny protuberances giving rough touch, and with unpleasant odour. Microscopically, a wide zone of radially elongated parenchyma cells between upper and lower epidermis were found. The vascular strands were present at all places and radially elongated sclerides touched the lower epidermis. In physico-chemical studies, the moisture, total ash, acid insoluble ash, alcohol soluble, water soluble, petroleum ether, chloroform extractive value and tannin content were found to be 2.790, 5.020, 0.110, 38.780, 41.210, 0.402, 1.590 and 49.200 percentage respectively. Preliminary phytochemical screening showed the presence of phenols, flavonoids, steroids, triterpenes, tannins, saponins and alkaloids. Conclusions: The results of the present study serve as a valuable source of information and provide suitable standards for identification of this medicinally important plant drug material for future investigations and applications.

Introduction: The worldwide increased bacterial resistance to antibiotics and the undesirable side effects associated with constant use of synthetic drugs has prompted the search for novel antimicrobial agents, particularly those manufactured from plants. This study is designed to ascertain the antibacterial potential of Rhus succedanea leaf gall extracts on the growth of gram-positive and gram–negative bacteria. Methods: The methanolic and hexane extract of different concentrations (100, 250, and 500 μg/ml) were prepared and their antibacterial efficacy was tested against clinical isolates of Escherichia coli, Salmonella typhi, Micrococcus luteus, and Staphylococcus aureus using agar well diffusion method and the size of inhibition zone was measured in millimeters. Results: The methanol and hexane extracts differed significantly in their antimicrobial activity with methanol extract showing a potent inhibitory activity in the range of 16±2 to 23±1, which was almost equal to the values of ciprofloxacin (25±3), used as a standard. Further, the methanol extract was mostly potent and effective in inhibiting the growth of gram-negative bacteria, namely, E. coli, when compared to gram –positive bacteria stains, which are responsible for antimicrobial activities. The phytochemical screening showed positive results for the presence of steroids, triterpenes, alkaloids, and carbohydrates. Conclusion: The potent antibacterial activity of Rhus succedanea leaf gall extracts indicates its useful therapeutic application against bacterial infection. Furthermore, this study indicates that the extract might be exploited as natural drug for the treatment of infectious diseases and could be useful in understanding the relations between traditional cures and current medications.

Disruption of DNA damage repair via impaired homologous recombination is characteristic of Ewing sarcoma (EWS) cells. We hypothesize that this disruption results in increased reliance on nonhomologous end joining to repair DNA damage. In this study, we investigated if pharmacologic inhibition of the enzyme responsible for nonhomologous end joining, the DNA-PK holoenzyme, alters the response of EWS cells to genotoxic standard of care chemotherapy. We used analyses of cell viability and proliferation to investigate the effects of clinical DNA-PK inhibitors (DNA-PKi) in combination with six therapeutic or experimental agents for EWS. We performed calculations of synergy using the Loewe additivity model. Immunoblotting evaluated treatment effects on DNA-PK, DNA damage, and apoptosis. Flow cytometric analyses evaluated effects on cell cycle and fate. We used orthotopic xenograft models to interrogate tolerability, drug mechanism, and efficacy in vivo. DNA-PKi demonstrated on-target activity, reducing phosphorylated DNA-PK levels in EWS cells. DNA-PKi sensitized EWS cell lines to agents that function as topoisomerase 2 (TOP2) poisons and enhanced the DNA damage induced by TOP2 poisons. Nanomolar concentrations of single-agent TOP2 poisons induced G2M arrest and little apoptotic response while adding DNA-PKi-mediated apoptosis. In vivo, the combination of AZD7648 and etoposide had limited tolerability but resulted in enhanced DNA damage, apoptosis, and EWS tumor shrinkage. The combination of DNA-PKi with standard of care TOP2 poisons in EWS models is synergistic, enhances DNA damage and cell death, and may form the basis of a promising future therapeutic strategy for EWS., (©2024 American Association for Cancer Research.)

The mechanistic basis for the metastasis of Ewing sarcomas remains poorly understood, as these tumors harbor few mutations beyond the chromosomal translocation that initiates the disease. Instead, the epigenome of Ewing sarcoma cells reflects the regulatory state of genes associated with the DNA-binding activity of the fusion oncoproteins EWSR1::FLI1 or EWSR1::ERG. In this study, we examined the EWSR1::FLI1/ERG's repression of transcription factor genes, concentrating on those that exhibit a broader range of expression in tumors than in Ewing sarcoma cell lines. Focusing on one of these target genes, ETS1, we detected EWSR1::FLI1 binding and an H3K27me3-repressive mark at this locus. Depletion of EWSR1::FLI1 results in ETS1's binding of promoter regions, substantially altering the transcriptome of Ewing sarcoma cells, including the upregulation of the gene encoding TENSIN3 (TNS3), a focal adhesion protein. Ewing sarcoma cell lines expressing ETS1 (CRISPRa) exhibited increased TNS3 expression and enhanced movement compared with control cells. Visualization of control Ewing sarcoma cells showed a distributed vinculin signal and a network-like organization of F-actin; in contrast, ETS1-activated Ewing sarcoma cells showed an accumulation of vinculin and F-actin toward the plasma membrane. Interestingly, the phenotype of ETS1-activated Ewing sarcoma cell lines depleted of TNS3 resembled the phenotype of the control cells. Critically, these findings have clinical relevance as TNS3 expression in Ewing sarcoma tumors positively correlates with that of ETS1. Implications: ETS1's transcriptional regulation of the gene encoding the focal adhesion protein TENSIN3 in Ewing sarcoma cells promotes cell movement, a critical step in the evolution of metastasis., (©2024 American Association for Cancer Research.)

EWSR1 is a member of the FET family of nucleic acid binding proteins that includes FUS and TAF15. Here, we report the systematic analysis of endogenous EWSR1's cellular organization in human cells. We demonstrate that EWSR1, which contains low complexity and nucleic acid binding domains, is present in cells in faster and slower-recovering fractions, indicative of a protein undergoing both rapid exchange and longer-term interactions. The employment of complementary high-resolution imaging approaches shows EWSR1 exists in two visual modalities, a distributed state which is present throughout the nucleoplasm, and a concentrated state consistent with the formation of foci. Both EWSR1 visual modalities localize with nascent RNA. EWSR1 foci concentrate in regions of euchromatin, adjacent to protein markers of transcriptional activation, and significantly colocalize with phosphorylated RNA polymerase II. Our results contribute to bridging the gap between our understanding of the biophysical and biochemical properties of FET proteins, including EWSR1, their functions as transcriptional regulators, and the participation of these proteins in tumorigenesis and neurodegenerative disease.

Background: Continual improvement in adjuvant therapies has resulted in a better prognosis for women diagnosed with breast cancer. A surrogate marker used to detect the spread of disease after treatment of breast cancer is local and regional recurrence. The risk of local and regional recurrence after mastectomy increases with the number of axillary lymph nodes affected by cancer. There is a consensus to use radiotherapy as an adjuvant treatment after mastectomy (postmastectomy radiotherapy (PMRT)) in women diagnosed with breast cancer and found to have disease in four or more positive axillary lymph nodes. Despite data showing almost double the risk of local and regional recurrence in women treated with mastectomy and found to have one to three positive lymph nodes, there is a lack of international consensus on the use of PMRT in this group., Objectives: To assess the effects of PMRT in women diagnosed with early breast cancer and found to have one to three positive axillary lymph nodes., Search Methods: We searched the Cochrane Breast Cancer Group's Specialised Register, CENTRAL, MEDLINE, Embase, the World Health Organization (WHO) International Clinical Trials Registry Platform (ICTRP) and ClinicalTrials.gov up to 24 September 2021., Selection Criteria: We included randomised controlled trials (RCTs). The inclusion criteria included women diagnosed with breast cancer treated with simple or modified radical mastectomy and axillary surgery (sentinel lymph node biopsy (SLNB) alone or those undergoing axillary lymph node clearance with or without prior SLNB). We included only women receiving PMRT using X-rays (electron and photon radiation), and we defined the radiotherapy dose to reflect what is currently being recommended (i.e. 40 Gray (Gy) to 50 Gy in 15 to 25/28 fractions in 3 to 5 weeks. The included studies did not administer any boost to the tumour bed. In this review, we excluded studies using neoadjuvant chemotherapy as a supportive treatment before surgery., Data Collection and Analysis: We used Covidence to screen records. We collected data on tumour characteristics, adjuvant treatments and the outcomes of local and regional recurrence, overall survival, disease-free survival, time to progression, short- and long-term adverse events and quality of life. We reported on time-to-event outcome measures using the hazard ratio (HR) and subdistribution HR. We used Cochrane's risk of bias tool (RoB 1), and we presented overall certainty of the evidence using the GRADE approach., Main Results: The RCTs included in this review were subgroup analyses of original RCTs conducted in the 1980s to assess the effectiveness of PMRT. Hence, the type and duration of adjuvant systemic treatments used in the studies included in this review were suboptimal compared to the current standard of care. The review involved three RCTs with a total of 829 women diagnosed with breast cancer and low-volume axillary disease. Amongst the included studies, only a single study pertained to the modern-day radiotherapy practice. The results from this one study showed a reduction of local and regional recurrence (HR 0.20, 95% confidence interval (CI) 0.13 to 0.33, 1 study, 522 women; low-certainty evidence) and improvement in overall survival with PMRT (HR 0.76, 95% CI 0.60 to 0.97, 1 study, 522 women; moderate-certainty evidence). One of the other studies using radiotherapy techniques that do not reflect modern-day practice reported on disease-free survival in women with low-volume axillary disease (subdistribution HR 0.63, 95% CI 0.41 to 0.96, 1 study, 173 women). None of the included studies reported on PMRT side effects or quality-of-life outcome measures., Authors' Conclusions: Based on one study, the use of PMRT in women diagnosed with breast cancer and low-volume axillary disease indicated a reduction in locoregional recurrence and an improvement in survival. There is a need for more research to be conducted using modern-day radiotherapy equipment and methods to support and supplement the review findings., (Copyright © 2023 The Authors. Cochrane Database of Systematic Reviews published by John Wiley & Sons, Ltd. on behalf of The Cochrane Collaboration.)

The year 2021 marks the 20th anniversary of the first publications reporting the discovery of the gene silencing mechanism, RNA interference (RNAi) in mammalian cells. Along with the many studies that delineated the proteins and substrates that form the RNAi pathway, this finding changed our understanding of the posttranscriptional regulation of mammalian gene expression. Furthermore, the development of methods that exploited the RNAi pathway began the technological revolution that eventually enabled the interrogation of mammalian gene function-from a single gene to the whole genome-in only a few days. The needs of the cancer research community have driven much of this progress. In this perspective, we highlight milestones in the development and application of RNAi-based methods to study carcinogenesis. We discuss how RNAi-based functional genetic analysis of exemplar tumor suppressors and oncogenes furthered our understanding of cancer initiation and progression and explore how such studies formed the basis of genome-wide scale efforts to identify cancer or cancer-type specific vulnerabilities, including studies conducted in vivo. Furthermore, we examine how RNAi technologies have revealed new cancer-relevant molecular targets and the implications for cancer of the first RNAi-based drugs. Finally, we discuss the future of functional genetic analysis, highlighting the increasing availability of complementary approaches to analyze cancer gene function., (© 2020 The Authors. Molecular Carcinogenesis published by Wiley Periodicals LLC.)

Gene fusions are an important class of mutations in several cancer types and include genomic rearrangements that fuse regulatory or coding elements from two different genes. Analysis of the genetics of cancers harboring fusion oncogenes and the proteins they encode have enhanced cancer diagnosis and in some cases patient treatment. However, the effect of the complex structure of fusion genes on the biogenesis of the resulting chimeric transcripts they express is not well studied. There are two potential RNA-related vulnerabilities inherent to fusion-driven cancers: (a) the processing of the fusion precursor messenger RNA (pre-mRNA) to the mature mRNA and (b) the mature mRNA. In this study, we discuss the effects that the genetic organization of fusion oncogenes has on the generation of translatable mature RNAs and the diversity of fusion transcripts expressed in different cancer subtypes, which can fundamentally influence both tumorigenesis and treatment. We also discuss functional genomic approaches that can be utilized to identify proteins that mediate the processing of fusion pre-mRNAs. Furthermore, we assert that an enhanced understanding of fusion transcript biogenesis and the diversity of the chimeric RNAs present in fusion-driven cancers will increase the likelihood of successful application of RNA-based therapies in this class of tumors. This article is categorized under: RNA Processing > RNA Editing and Modification RNA Processing > Splicing Regulation/Alternative Splicing RNA in Disease and Development > RNA in Disease., (© 2019 Centre National De La Recherche Scientifique (CNRS). WIREs RNA published by Wiley Periodicals, Inc.)

Male breast cancer (MBC) is an uncommon malignancy. We have previously reported that the expression of the Hippo transducers TAZ/YAP and their target CTGF was associated with inferior survival in MBC patients. Preclinical evidence demonstrated that Axl is a transcriptional target of TAZ/YAP. Thus, we herein assessed AXL expression to further investigate the significance of active TAZ/YAP-driven transcription in MBC. For this study, 255 MBC samples represented in tissue microarrays were screened for AXL expression, and 116 patients were included. The association between categorical variables was verified by the Pearson's Chi-squared test of independence (2-tailed) or the Fisher Exact test. The relationship between continuous variables was tested with the Pearson's correlation coefficient. The Kaplan-Meier method was used for estimating survival curves, which were compared by log-rank test. Factors potentially impacting 10-year and overall survival were verified in Cox proportional regression models. AXL was positively associated with the TAZ/CTGF and YAP/CTGF phenotypes (P = 0.001 and P = 0.002, respectively). Patients with TAZ/CTGF/AXL- or YAP/CTGF/AXL-expressing tumors had inferior survival compared with non-triple-positive patients (log rank P = 0.042 and P = 0.048, respectively). The variables TAZ/CTGF/AXL and YAP/CTGF/AXL were adverse factors for 10-year survival in the multivariate Cox models (HR 2.31, 95%CI:1.02-5.22, P = 0.045, and HR 2.27, 95%CI:1.00-5.13, P = 0.050). Nearly comparable results were obtained from multivariate analyses of overall survival. The expression pattern of AXL corroborates the idea of the detrimental role of TAZ/YAP activation in MBC. Overall, Hippo-linked biomarkers deserve increased attention in this rare disease. J. Cell. Physiol. 232: 2246-2252, 2017. © 2016 Wiley Periodicals, Inc., (© 2016 Wiley Periodicals, Inc.)

Purpose: Breast cancer affects both genders, but is understudied in men. Although still rare, male breast cancer (MBC) is being diagnosed more frequently. Treatments are wholly informed by clinical studies conducted in women, based on assumptions that underlying biology is similar. Experimental Design: A transcriptomic investigation of male and female breast cancer was performed, confirming transcriptomic data in silico Biomarkers were immunohistochemically assessed in 697 MBCs ( n = 477, training; n = 220, validation set) and quantified in pre- and posttreatment samples from an MBC patient receiving everolimus and PI3K/mTOR inhibitor. Results: Gender-specific gene expression patterns were identified. eIF transcripts were upregulated in MBC. eIF4E and eIF5 were negatively prognostic for overall survival alone (log-rank P = 0.013; HR = 1.77, 1.12-2.8 and P = 0.035; HR = 1.68, 1.03-2.74, respectively), or when coexpressed ( P = 0.01; HR = 2.66, 1.26-5.63), confirmed in the validation set. This remained upon multivariate Cox regression analysis [eIF4E P = 0.016; HR = 2.38 (1.18-4.8), eIF5 P = 0.022; HR = 2.55 (1.14-5.7); coexpression P = 0.001; HR = 7.04 (2.22-22.26)]. Marked reduction in eIF4E and eIF5 expression was seen post BEZ235/everolimus, with extended survival. Conclusions: Translational initiation pathway inhibition could be of clinical utility in MBC patients overexpressing eIF4E and eIF5. With mTOR inhibitors that target this pathway now in the clinic, these biomarkers may represent new targets for therapeutic intervention, although further independent validation is required. Clin Cancer Res; 23(10); 2575-83. ©2016 AACR ., (©2016 American Association for Cancer Research.)

Male breast cancer (MBC) is rare. We assembled 446 MBCs on tissue microarrays and assessed clinicopathological information, together with data from 15 published studies, totalling 1984 cases. By immunohistochemistry we investigated 14 biomarkers (ERα, ERβ1, ERβ2, ERβ5, PR, AR, Bcl-2, HER2, p53, E-cadherin, Ki67, survivin, prolactin, FOXA1) for survival impact. The main histological subtype in our cohort and combined analyses was ductal (81%, 83%), grade 2; (40%, 44%), respectively. Cases were predominantly ERα (84%, 82%) and PR positive (74%, 71%), respectively, with HER2 expression being infrequent (2%, 10%), respectively. In our cohort, advanced age (>67) was the strongest predictor of overall (OS) and disease free survival (DFS) (p = 0.00001; p = 0.01, respectively). Node positivity negatively impacted DFS (p = 0.04). FOXA1 p = 0.005) and AR p = 0.009) were both positively prognostic for DFS, remaining upon multivariate analysis. Network analysis showed ERα, AR and FOXA1 significantly correlated. In summary, the principle phenotype of MBC was luminal A, ductal, grade 2. In ERα+ MBC, only AR had prognostic significance, suggesting AR blockade could be employed therapeutically.

Visceral adipose tissue inflammation in obesity is an established risk factor for metabolic syndrome, which can include insulin resistance, type 2 diabetes, hypertension and cardiovascular diseases. With obesity and related metabolic disorders reaching epidemic proportions globally, an understanding of the mechanisms of adipose tissue inflammation is crucial. Within the immune cell cohort, dendritic cells (DC) play a key role in balancing tolerance and immunity. Despite decades of research into the characterization of DC in lymphoid and non-lymphoid organs, their role in adipose tissue function is poorly understood. There is now an increasing interest in identification and characterization of DC in adipose tissue and understanding their function in regulating tissue metabolic homeostasis. This review provides an overview of the study of DC in adipose tissue, focusing on possible mechanisms by which DC may contribute to adipose tissue homeostasis., (© 2016 John Wiley & Sons Ltd.)

Male breast cancer (MBC) is a rare hormone-driven disease often associated with obesity. HMG-CoAR is the central enzyme of the mevalonate pathway, a molecular route deputed to produce cholesterol and steroid-based hormones. HMG-CoAR regulates the oncogenic Hippo transducers TAZ/YAP whose expression was previously associated with shorter survival in MBC. 225 MBC samples were immunostained for HMG-CoAR and 124 were considered eligible for exploring its relationship with hormone receptors (ER, PgR, AR), Hippo transducers and survival outcomes. HMG-CoAR was positively associated with the expression of hormone receptors (ER, PgR, AR) and Hippo transducers. Overall survival was longer in patients with HMG-CoAR-positive tumors compared with their negative counterparts (p = 0.031). Five- and 10-year survival outcomes were better in patients whose tumors expressed HMG-CoAR (p = 0.044 and p = 0.043). Uni- and multivariate analyses for 10-year survival suggested that HMG-CoAR expression is a protective factor (HR 0.50, 95% CI: 0.25-0.99, p = 0.048 and HR 0.53, 95% CI: 0.26-1.07, p = 0.078). Results were confirmed in a sensitivity analysis by excluding uncommon histotypes (multivariate Cox: HR 0.45, 95% CI: 0.21-0.97, p = 0.043). A positive relationship emerged between HMG-CoAR, hormone receptors and TAZ/YAP, suggesting a connection between the mevalonate pathway, the hormonal milieu and Hippo in MBC. Moreover, HMG-CoAR expression may be a favorable prognostic indicator.

Male breast cancer (MBC) is a rare disease and its biology is poorly understood. Deregulated Hippo pathway promotes oncogenic functions in female breast cancer. We herein investigated the expression of the Hippo transducers TAZ/YAP and their target CTGF in MBC. Tissue microarrays containing samples from 255 MBC patients were immunostained for TAZ, YAP and CTGF. One hundred and twenty-nine patients were considered eligible. The Pearson's Chi-squared test of independence was used to test the association between categorical variables. The correlation between TAZ, YAP and CTGF was assessed with the Pearson's correlation coefficient. The Kaplan-Meier method and the log-rank test were used for estimating and comparing survival curves. Cox proportional regression models were built to identify variables impacting overall survival. Statistical tests were two-sided. Tumors were considered to harbor active TAZ/YAP-driven gene transcription when they co-expressed TAZ, or YAP, and CTGF. Patients whose tumors had the TAZ/CTGF and YAP/CTGF phenotypes experienced shorter overall survival compared with their negative counterparts (log rank p = 0.036 for both). TAZ/CTGF and YAP/CTGF tumors were associated with decreased survival in patients with invasive ductal carcinomas, G3 tumors, hormone receptor-positive tumors, and tumors with elevated Ki-67. Multivariate analyses confirmed that the TAZ/CTGF and YAP/CTGF phenotypes are independent predictors of survival (HR 2.03, 95% CI: 1.06-3.90, p = 0.033; and HR 2.00, 95% CI: 1.04-3.84, p = 0.037 respectively). Comparable results were obtained when excluding uncommon histotypes (TAZ/CTGF: HR 2.34, 95% CI: 1.16-4.73, p = 0.018. YAP/CTGF. HR 2.36, 95% CI: 1.17-4.77, p = 0.017). Overall, the TAZ/YAP-driven oncogenic program may be active in MBC, conferring poorer survival., Competing Interests: The authors declare no conflicts of interest.

Biobanks provide a window of opportunity to store and add value to material from rare cases allowing their future use in biomedical research. One such example is the opportunityto obtain good quality tissue from patients undergoing gender re-assignment. Following patient agreement to donate tissue samples to our biobank we catalogued the histological appearance, defined the expression of the hormone receptors ERα, PR, AR and the proliferation marker Ki67, and generated and characterised primary cell cultures in a female to male (FTM) transgender patient referred to our unit for surgery. Immunohistochemistry was performed for ERα, PR and AR and the proliferation marker Ki67. Hormone receptor expression was confined to epithelial cells lining the breast ducts. Ki67 immunoreactivity was sparse indicating little proliferation of luminal epithelium, consistent with normal mammary gland. Cultures of epithelial cells and fibroblasts were derived from surplus tissue. The latter lacked expression of epithelial markers and hormone receptors but exhibited expression of vimentin. Culture of the former on Matrigel saw an outgrowth of more rounded "epithelial-like" cells. Immunofluoresence characterisation showed a mixed phenotype with expression of vimentin and both myoepithelial and luminal epithelial markers. Sporadic weak ERα expression and moderate PR expression was seen. In summary, as well as routinely collecting tissue and blood samples, we have characterised and stored tissue and cells from a FTM transgender patient, adding value to this resource which,available from the Breast Cancer Campaign Tissue Bank for those interested in further studying the biology of FTM transgender tissue.

Women with bacterial vaginosis (BV) display reduced vaginal acidity, which make them susceptible to associated infections such as HIV. In the current study, poly(ethylene glycol) (PEG) nanocarrier-based degradable hydrogels were developed for the controlled release of lactic acid in the vagina of BV-infected women. PEG-lactic acid (PEG-LA) nanocarriers were prepared by covalently attaching lactic acid to 8-arm PEG-SH via cleavable thioester bonds. PEG-LA nanocarriers with 4 copies of lactic acid per molecule provided controlled release of lactic acid with a maximum release of 23% and 47% bound lactic acid in phosphate buffered saline (PBS, pH7.4) and acetate buffer (AB, pH4.3), respectively. The PEG nanocarrier-based hydrogels were formed by cross-linking the PEG-LA nanocarriers with 4-arm PEG-NHS via degradable thioester bonds. The nanocarrier-based hydrogels formed within 20 min under ambient conditions and exhibited an elastic modulus that was 100-fold higher than the viscous modulus. The nanocarrier-based degradable hydrogels provided controlled release of lactic acid for several hours; however, a maximum release of only 10%-14% bound lactic acid was observed possibly due to steric hindrance of the polymer chains in the cross-linked hydrogel. In contrast, hydrogels with passively entrapped lactic acid showed burst release with complete release within 30 min. Lactic acid showed antimicrobial activity against the primary BV pathogen Gardnerella vaginalis with a minimum inhibitory concentration (MIC) of 3.6 mg/ml. In addition, the hydrogels with passively entrapped lactic acid showed retained antimicrobial activity with complete inhibition G. vaginalis growth within 48 h. The results of the current study collectively demonstrate the potential of PEG nanocarrier-based hydrogels for vaginal administration of lactic acid for preventing and treating BV., (Copyright © 2014 Elsevier B.V. All rights reserved.)

Aims: The handling and examination of sentinel lymph nodes (SLNs) to detect metastasis is critical in the assessment of early breast cancer patients. This survey investigates the variation in practise followed by pathology units across the United Kingdom in the staging evaluation of axillary lymph nodes (ALNs)., Methods and Results: A structured questionnaire, approved by the National Health Service Breast Screening Programme pathology Big 18 committee, was circulated among all pathologists. There were 160 respondents; 92% performed SLN biopsy for staging, 97% had a protocol for processing SLNs and most laboratories examined the ALNs using formalin-fixed, paraffin-embedded (FFPE) samples (85.6%). A few used PCR (7.5%), frozen section (3.8%) or touch imprint cytology (3.1%), with or without subsequent FFPE section examination. Currently, 33% perform serial sectioning, with the majority of the rest (75%) staining three levels using H&E. Most units (85%) undertook immunohistochemistry evaluation only when suspicious cells were detected on H&E-stained sections., Conclusions: The range of practise in UK histopathology departments is described with regard to the dissection and evaluation of ALNs/SLN biopsy. The variation in practise was not very marked and most departments adhered to national guidelines. Any UK study seeking to relate ALN status and outcome would need to be mindful of the variability in nodal processing and examination., (© 2014 John Wiley & Sons Ltd.)

Current treatment options for bacterial vaginosis (BV) have been shown to be inadequate at preventing recurrence and do not provide protection against associated infections, such as that with HIV. This study examines the feasibility of incorporating the antimicrobial peptide subtilosin within covalently cross-linked polyethylene glycol (PEG)-based hydrogels for vaginal administration. The PEG-based hydrogels (4% and 6% [wt/vol]) provided a two-phase release of subtilosin, with an initial rapid release rate of 4.0 μg/h (0 to 12 h) followed by a slow, sustained release rate of 0.26 μg/h (12 to 120 h). The subtilosin-containing hydrogels inhibited the growth of the major BV-associated pathogen Gardnerella vaginalis with a reduction of 8 log10 CFU/ml with hydrogels containing ≥15 μg entrapped subtilosin. In addition, the growth of four common species of vaginal lactobacilli was not significantly inhibited in the presence of the subtilosin-containing hydrogels. The above findings demonstrate the potential application of vaginal subtilosin-containing hydrogels for prophylaxis of BV.

The aim of this study was to validate ImmunoRatio, a web-based automated image analysis application, by comparing the manual and automated analysis scores for oestrogen receptor α (ERα) in breast carcinomas. Tissue microarrays comprising 200 breast cancer cases prestained for ERα were scanned and scored manually using ImageScope viewing software. Corresponding images were then uploaded and assessed according to the web-based ImmunoRatio programme. There was excellent correlation between manual and ImmunoRatio ERα scores (Spearman correlation=0.872; p≥0.001). The manual and ImmunoRatio ERα scores showed only a moderate agreement (κ=0.421; Weighted kappa=0.874 (CI 0.839 to 0.902)), most probably due to lack of specificity of the algorithm to differentiate between cancer and non-cancer nuclei. Further development to enable differentiation of cancer and non-cancer elements should improve the specificity of the application. Our results support the use of ImmunoRatio software for analysing ERα immunohistochemistry in breast cancer tissues for the purposes of research.

Compared with other markers of cell proliferation, geminin is unique being expressed selectively during the proliferative phase of the cell cycle, specifically S, G2 and early mitosis, disappearing completely at the metaphase-anaphase transition. We aimed to compare the prognostic significance of geminin to that of Ki67, a proliferation marker which has been investigated in many breast cancer studies. Breast cancer tissue microarrays containing 368 tumours were stained using anti-geminin and Ki67 antibodies. Labelling index (LI) was calculated for geminin, and the percentage of positive cancer nuclei was determined for Ki67. A receiver operation characteristics analysis was used to determine the optimum cut-off value for geminin (LI ≥ 2), and for Ki67, a score of ≥14 % was considered as positive for survival analysis. Geminin expression correlated positively with Ki67 expression (r = 0.686, p = 0.001). Survival analysis showed only geminin, and not Ki67-positive patients to have poor (breast cancer-specific survival) BCSS [HR 2.85 (1.53-5.32)] and (disease-free survival) DFS [HR 2.63 (1.47-4.71)]. On univariate analysis, along with known clinicopathological variables, both Ki67 and geminin LI were found to be significant predictors of BCSS and DFS. On multivariate analysis, only tumour size, nodal status and adjuvant hormonal therapy were found to be independent predictors for both BCSS and DFS, while geminin positivity (LI ≥ 2 %) was found to be an independent predictor for BCSS [HR 2.27 (1.01-5.06); p = 0.04]. In comparison with Ki67, a more established proliferation marker, geminin expression was a better predictor of adverse outcome in this cohort of breast cancers. Selective expression of geminin during the proliferative phase of the cell cycle and its nuclear specificity increase its potential to be used as an alternative marker of proliferation in breast cancer patients.

Background: Palpable pure DCIS is a rare entity that presents differently than screen-detected DCIS. The aim of this study was to evaluate the clinical, radiological, and pathological characteristics and management of pDCIS in a retrospective cohort of patients., Patients and Methods: Patients diagnosed with pDCIS from January 1999 to December 2011 were identified from an electronic patient database and were included in this study., Results: During this period, 669 cases of DCIS were diagnosed and 62 (9.3%) were pDCIS (mean age, 56.9 ± 15.1 years). The most common finding on ultrasound was mass in 43 patients (75%) and only 18 (33%) cases had calcification on mammography. The lesion was mammographically occult in 20 patients (37%). Ultrasound was more sensitive and delineated the pDCIS in 45 (80%) cases. Mean size of the pDCIS was 36.9 ± 30.4 mm and most were high grade (n = 42; 68%) and associated with comedo necrosis in 36 (59%). Most were oestrogen receptor (ER)-positive (n = 34; 62%), however 21 patients (38%) were ER-negative. Breast conservation was attempted in 30 patients (48%), however, because of involved margins further therapeutic surgery was needed in 10 patients (33%). Axillary surgery (sentinel lymph node biopsy or axillary nodal sampling) was performed in 34 patients (55%) and no lymph node metastasis was identified. During a medial follow-up of 60 months, 1 patient has developed a mastectomy scar recurrence and the rest remain disease-free., Conclusion: Palpable DCIS is often occult on conventional radiological imaging and is generally associated with aggressive pathological features. Hence, careful individualized surgical planning through a multidisciplinary meeting is necessary for their management., (Copyright © 2013 Elsevier Inc. All rights reserved.)

Objective: To examine the utility of palpation-guided fine-needle aspiration cytology (pgFNAC) in the context of clinically palpable but radiologically occult breast abnormalities in this era of digital mammography and high sensitivity ultrasound., Methods: Women undergoing pgFNAC from January 2005 to December 2007 were identified from the histopathology database and correlated with clinical and radiological findings recorded prospectively in electronic patient records., Results: 142 cases matching our selection criteria were identified with a mean age of 43 (SD ±13.7) years; 83 patients had focal lumps and 59 had non-focal lumpiness. In the latter, pgFNAC showed C1 cytology in 45 (76.3%), C2 in 13 (22%) and C3 in 1 (1.7%) patient. In 83 patients with a focal discrete lump, pgFNAC revealed C1 cytology in 65 (78.3%), C2 in 14 (16.9%), and 2 patients each had C3 and C4 cytology. Core biopsy was undertaken in the latter 4 patients, invasive cancer was found in 1 patient each with C3 and C4 cytology and benign pathology in the rest. To date, none of the patients discharged has developed pre-malignant or malignant lesions in the ipsilateral breast., Conclusion: In patients presenting with clinically palpable but radiologically occult breast abnormality, pgFNAC can identify those who need further investigation or who can be safely discharged., (Copyright © 2012 S. Karger AG, Basel.)

The current study examines the passive pulmonary targeting efficacy and retention of 6μm polystyrene (PS) microparticles (MPs) covalently modified with different surface groups [amine (A-), carboxyl (C-) and sulfate (S-)] or single (PEG(1)-) and double (PEG(2)-) layers of α,ω-diamino poly(ethylene glycol) attached to C-MPs. The ζ-potential of A-MPs (-44.0mV), C-MPs (-54.3mV) and S-MPs (-49.6mV) in deionized water were similar; however PEGylation increased the ζ-potential for both PEG(1)-MPs (-18.3mV) and PEG(2)-MPs (11.5mV). The biodistribution and retention of intravenously administered MPs to male Sprague-Dawley rats was determined in homogenized tissue by fluorescence spectrophotometry. PEG(1)-MPs and PEG(2)-MPs demonstrated enhanced pulmonary retention in rats at 48h after injection when compared to unmodified A-MPs (59.6%, 35.9% and 17.0% of the administered dose, respectively). While unmodified MPs did not significantly differ in lung retention, PEGylation of MPs unexpectedly improved passive lung targeting and retention by modifying surface properties including charge and hydrophobicity but not size., (Copyright © 2010. Published by Elsevier B.V.)

Can quantum dots (QDs) serve as physiologically relevant receptor probes in the interior of cells? We directly visualize endocytosis, redistribution, and shuttling of QD bound-TrkA receptors to PC12 neural processes and far-reaching growth cone tips. Internalized QDs are contained in microtubule-associated vesicles and possess transport properties that reflect TrkA receptor dynamics. This opens up new possibilities for the development of QD platforms as molecular tools to image biochemical signaling and transport cargo in the cell interior.