MCP-1 is overexpressed in triple-negative breast cancers and drives cancer invasiveness and metastasis.

BACKGROUND:Triple-negative breast cancer (TNBC) is the most aggressive type of breast cancer that lacks ER/PR and HER2 receptors. Hence, there is urgency in developing new or novel therapeutic strategies for treatment of TNBC. Our study shows that the Monocyte Chemoattractant Protein-1 (MCP-1) is a marker associated with TNBC and may play a key role in TNBC disease progression. EXPERIMENTAL DESIGN:ELISA method was used to measure secreted MCP-1, and mRNA levels were determined by Real-time PCR in numerous cancer cell lines, representing various breast cancer subtypes. Cellular invasiveness was determined by Boyden chamber assay. RESULTS:Our data show that MCP-1 is upregulated in TNBC cell lines both transcriptionally as well as in secreted protein levels compared to ER-positive luminal cell line, MCF-7. Breast cancer patients, with Basal or Claudin-low subtypes, also showed high expression of MCP-1. MCP-1 treatment induced cell invasion in various breast cancer cell types, without affecting cell proliferation. Small molecule antagonists against Chemokine Receptor 2 (CCR2), cognate receptor for MCP-1 as well as the MAP kinase pathway inhibitor U0126 negatively affected MCP-1 induced MCF-7 cell invasion. This suggests that MCP-1-CCR2 axis may regulate invasiveness via the MAP Kinase pathway. Knocking down MCP-1 decreased cell invasion in TNBC cell line BT-549, along with downregulation of key epithelial to mesenchymal transition markers, N-cadherin and Vimentin. CONCLUSION:Our study suggests that MCP-1 mediated pathways could be potential therapeutic targets for the treatment of TNBC, and could reduce cancer health disparities

An Essential Protein that Interacts with Endosomes and Promotes Movement of the SHORT-ROOT Transcription Factor

SummaryPlant cells can communicate through the direct transport of transcription factors [1–7]. One of the best-studied examples of this phenomenon is SHORT-ROOT (SHR), which moves from the stele cells into the endodermis and root tip of Arabidopsis, where it specifies endodermal cell identity and stem cell function, respectively [8–10]. In the endodermis, SHR upregulates the transcription factors SCARECROW (SCR) [2] and JACKDAW (JKD), which in turn inhibit movement of SHR from the endodermis [11]. Although much is known about the regulatory pathways that mediate expression and activity of SHR [1, 8–14], little is known about the factors that promote its movement or the movement of other transcription factors. We have identified a novel protein, SHORT-ROOT INTERACTING EMBRYONIC LETHAL (SIEL), that interacts with SHR, CAPRICE (CPC), TARGET OF MONOPTEROUS 7 (TMO7), and AGAMOUS-LIKE 21 (AGL21). Null alleles of SIEL are embryonic lethal. Hypomorphic alleles produce defects in root patterning and reduce SHR movement. Surprisingly, both SHR and SCR regulate expression of SIEL, so that siel/scr and siel/shr double mutants have extremely disorganized roots. SIEL localizes to the nucleus and cytoplasm of root cells where it is associated with endosomes. We propose that SIEL is an endosome-associated protein that promotes intercellular movement

Assessing the Utility of Photoswitchable Fluorescent Proteins for Tracking Intercellular Protein Movement in the Arabidopsis Root

One way in which cells communicate is through the direct transfer of proteins. In plants, many of these proteins are transcription factors, which are made by one cell type and traffic into another. In order to understand how this movement occurs and its role in development, we would like to track this movement in live, intact plants in real-time. Here we examine the utility of the photoconvertible proteins, Dendra2 and (to a lesser extent) EosFP as tags for studying intracellular and intercellular protein movement in the Arabidopsis root. To this end, we made fusions between Dendra2 and six mobile transcription factors. Our results show that Dendra2 is an effective tool for studying protein movement between plant cells. Interestingly, we found that Dendra2 could not simply be swapped into existing constructs that had originally contained GFP. Most of the fusions made in this way failed to produce a fluorescent fusion. In addition we found that the optimal settings for photoconversion of Dendra2 in stably transformed roots were different from what has been published for photoconversion in transient assays in plants or in animal cells. By modifying the confocal setting, we were able to photoconvert Dendra2 in all cell layers in the root. However the efficiency of photoconversion was affected by the position of the cell layer within the root, with more internal tissues requiring more energy. By examining the Dendra2 fusions, we confirmed the mobility of the SHORT-ROOT (SHR) and CAPRICE (CPC) transcription factors between cells and we further discovered that SHR movement in stele and CPC movement in the epidermis are non-directional

Data Sharing by Scientists: Practices and Perceptions.

Scientific research in the 21st century is more data intensive and collaborative than in the past. It is important to study the data practices of researchers - data accessibility, discovery, re-use, preservation and, particularly, data sharing. Data sharing is a valuable part of the scientific method allowing for verification of results and extending research from prior results. A total of 1329 scientists participated in this survey exploring current data sharing practices and perceptions of the barriers and enablers of data sharing. Scientists do not make their data electronically available to others for various reasons, including insufficient time and lack of funding. Most respondents are satisfied with their current processes for the initial and short-term parts of the data or research lifecycle (collecting their research data; searching for, describing or cataloging, analyzing, and short-term storage of their data) but are not satisfied with long-term data preservation. Many organizations do not provide support to their researchers for data management both in the short- and long-term. If certain conditions are met (such as formal citation and sharing reprints) respondents agree they are willing to share their data. There are also significant differences and approaches in data management practices based on primary funding agency, subject discipline, age, work focus, and world region. Conclusions/Significance Barriers to effective data sharing and preservation are deeply rooted in the practices and culture of the research process as well as the researchers themselves. New mandates for data management plans from NSF and other federal agencies and world-wide attention to the need to share and preserve data could lead to changes. Large scale programs, such as the NSF-sponsored DataNET (including projects like DataONE) will both bring attention and resources to the issue and make it easier for scientists to apply sound data management principles

A Novel Polymorphism of FcgammaRIIIa (CD16) Alters Receptor Function and Predisposes to Autoimmune Disease

A novel polymorphism in the extracellular domain 2 (EC2) of FcgammaRIIIA affects ligand binding by natural killer (NK) cells and monocytes from genotyped homozygous normal donors independently of receptor expression. The nonconservative T to G substitution at nucleotide 559 predicts a change of phenylalanine (F) to valine (V) at amino acid position 176. Compared with F/F homozygotes, FcgammaRIIIa expressed on NK cells and monocytes in V/V homozygotes bound more IgG1 and IgG3 despite identical levels of receptor expression. In response to a standard aggregated human IgG stimulus, FcgammaRIIIa engagement on NK cells from V/V (high-binding) homozygotes led to a larger rise in [Ca2+]i, a greater level of NK cell activation, and a more rapid induction of activation-induced cell death (by apoptosis). Investigation of an independently phenotyped normal cohort revealed that all donors with a low binding phenotype are F/F homozygotes, while all phenotypic high binding donors have at least one V allele. Initial analysis of 200 patients with SLE indicates a strong association of the low binding phenotype with disease, especially in patients with nephritis who have an underrepresentation of the homozygous high binding phenotype. Thus, the FcgammaRIIIa polymorphism at residue 176 appears to impact directly on human biology, an effect which may extend beyond autoimmune disease characterized by immune complexes to host defense mechanisms

A social work roundtable examining impacts and lessons learned from the COVID-19 pandemic

As have other disciplines, social work has been affected by the COVID-19 pandemic. Societal shifts and service provision gaps have emerged or been amplified over the course of the pandemic. An online roundtable was convened with five Canadian social work leaders to explore impacts of the pandemic on social work as well as to reflect on lingering effects of the pandemic and lessons learned for moving forward. Panelists’ varied substantive areas of social work practice and/or research included youth advocacy, healthcare, social work education and field education, and community development and disaster response. This paper offers a verbatim reproduction of the roundtable including panelists’ reflections on client and community experiences, social worker experiences, workforce impacts, shifts in the way service and practice are conceptualized and delivered, and implications for moving forward. Recommendations are offered in considered disciplinary, interdisciplinary and community advancement.Comme d’autres disciplines, le travail social a été touché par la pandémie de COVID-19. Des changements sociétaux et des lacunes dans la prestation de services sont apparus ou se sont amplifiés au cours de la pandémie. Une table ronde en ligne a été organisée avec cinq leaders canadiens du travail social pour explorer les impacts de la pandémie sur le travail social ainsi que pour réfléchir aux effets persistants de la pandémie et aux leçons apprises pour aller de l'avant. Les divers domaines de fond de la pratique et/ou de la recherche en travail social des panélistes comprenaient la défense des droits des jeunes, les soins de santé, la formation en travail social et la formation sur le terrain, ainsi que le développement communautaire et la réponse aux catastrophes. Cet article propose une reproduction textuelle de la table ronde comprenant les réflexions des panélistes sur les expériences des clients et de la communauté, les expériences des travailleurs sociaux, les impacts sur la main-d'oeuvre, les changements dans la façon dont les services et les pratiques sont conceptualisés et fournis, et les implications pour l'avenir. Des recommandations sont proposées en matière d’avancement disciplinaire, interdisciplinaire et communautaire

Defining genetic intra-tumor heterogeneity: a chronological annotation of mutational pathways

Tumor heterogeneity is believed to be important in tumor progression and its response to therapies. However, despite numerous mutations being reported in human tumors, genetic intra-tumor heterogeneity remains poorly defined. We have developed a novel strategy to provide a chronological annotation of mutational events in a tumor. We used an endometrial tumor from a patient and transplanted it into athymic mice to create many tumor xenografts. While the patient tumor xenografts were initially responsive to raloxifene treatment, xenografts created with cancer cell clones isolated from the same patient tumor showed dramatic differences in response to raloxifene, indicating existence of intra-tumor heterogeneity with some subpopulations inherently resistant to the drug. A 250K single nucleotide polymorphism (SNP) array from Affymetrix was used to profile genotype changes on 3 xenografts and 10 single cells from another 10 xenografts. We found 797 SNP sites containing loss of heterozygosity (LOH) common to all these specimens, indicating that genetic mutations in these regions may contain the earliest genetic events in the original patient tumor. Based upon the genotype information from the 10 single cancer cells, we developed a phylogenetic tree using neighbor-joining method. We showed that there are at least 3 distinct subpopulations in the patient tumor. Additionally, the phylogenetic tree was used to determine the order of genetic events, thus providing a chronological annotation to genetic mutations. Our approach represents an important analytic strategy for defining genetic intra-tumor heterogeneity and providing chronological annotations to the genetic landscape revealed by future whole genome sequencing in tumors

Characterisation of production, marketing and consumption patterns of farmed tilapia in the Nile Delta of Egypt

AbstractEgypt has one of the world’s largest aquaculture sectors which makes a significant contribution to income, employment creation and food security. However, there are very limited data available on the farmed tilapia value chain. The aim of this study therefore was to characterise production, marketing and consumption patterns of farmed tilapia in the Nile Delta of Egypt. A cross sectional study was conducted to collect data from tilapia producers (100), transporters (32), retailers (100), fish fry shops (20) and households (300) in three case study communities (fish producing, peri-urban and rural community). We conducted structured questionnaire interviews and participatory assessments for producers and consumers. Focus group discussions with mothers were also held to collect data for the availability, sources and consumption patterns of tilapia.Results showed that, more than half of producers were small scale, having a farm size of 10feddan or less (1feddan=4200m2). The main water supply for almost all farms was agricultural drain water, a potential source of contamination with chemical and biological hazards. The main production constraints were reported to be feed prices, water quality and availability, land rent, fuel and energy sources and environmental conditions. The farmed tilapia value chain was short with some value added in the form of marketing fresh and live fish as well as selling tilapia in fried or grilled form. The majority of produced tilapia was transported to retail sale and sold to consumers as fresh, while only a small proportion was processed by cleaning, grilling or frying. A lack of hygiene during transportation and marketing of farmed tilapia was found that could be potential sources for post-harvesting contamination. The availability and frequency of tilapia consumption were higher in the community in the production areas than in other communities. In non-producing areas, tilapia may be available in the market once a week during the village market day. Potential areas for further research in order to improve safety, quality and production of farmed tilapia were identified

Crystallization and preliminary X-ray diffraction analysis of the complex between a human anti-interferon antibody fragment and human interferon α-2A

Crystals of the complex between the Fab fragment of a human anti-interferon α therapeutic antibody and human interferon α-2A have been obtained and diffracted to 3.0 Å resolution

The 3′ processing of antisense RNAs physically links to chromatin-based transcriptional control

Noncoding RNA plays essential roles in transcriptional control and chromatin silencing. At Arabidopsis thaliana FLC, antisense transcription quantitatively influences transcriptional output, but the mechanism by which this occurs is still unclear. Proximal polyadenylation of the antisense transcripts by FCA, an RNA-binding protein that physically interacts with RNA 3′ processing factors, reduces FLC transcription. This process genetically requires FLD, a homolog of the H3K4 demethylase LSD1. However, the mechanism linking RNA processing to FLD function had not been established. Here, we show that FLD tightly associates with LUMINIDEPENDENS (LD) and SET DOMAIN GROUP 26 (SDG26) in vivo, and, together, they prevent accumulation of monomethylated H3K4 (H3K4me1) over the FLC gene body. SDG26 interacts with the RNA 3′ processing factor FY (WDR33), thus linking activities for proximal polyadenylation of the antisense transcripts to FLD/LD/SDG26-associated H3K4 demethylation. We propose this demethylation antagonizes an active transcription module, thus reducing H3K36me3 accumulation and increasing H3K27me3. Consistent with this view, we show that Polycomb Repressive Complex 2 (PRC2) silencing is genetically required by FCA to repress FLC. Overall, our work provides insights into RNA-mediated chromatin silencing