Solubilization of EGF receptor with Triton X-100 alters stimulation of tyrosine residue phosphorylation by EGF and dimethyl sulfoxide.

In isolated hepatic membranes, epidermal growth factor (EGF) and the polar solvent dimethyl sulfoxide (Me2SO) selectively stimulated the phosphorylation of the 170,000-dalton EGF receptor (p170) by 13.6 +/- 2.0- and 10.9 +/- 1.1-fold, respectively. The stimulation by maximally effective concentrations of the two substances was similar in rapidity of onset (less than 30 s at 0 degree C), time course of phosphorylation, and tyrosine residue specificity. These maximal effects were not additive when the substances were combined, indicating that the same kinase/substrate combination is activated by each. The lectin concanavalin A, which inhibits EGF receptor binding, blocked the effect of EGF but not Me2SO. In membranes solubilized with Triton X-100, EGF stimulated p170 phosphorylation by 40- to 55-fold. Me2SO also stimulated phosphorylation, indicating that it acts directly on the protein. However, the effect of the solvent was reduced by half. Additionally, Me2SO blocks the effect of EGF in the solubilized preparation. A room temperature preincubation after addition of either substance was necessary for maximal stimulation of p170 phosphorylation in solubilized membranes. With EGF, 30-40 min was necessary; with Me2SO, only 10 min was required. Thus, a secondary process appears to be involved in EGF receptor/kinase activation

Temporal and spatial drivers of the structure of macroinvertebrate assemblages associated with <em>Laminaria </em><em>hyperborea </em>detritus in the northeast Atlantic

\ua9 2024Kelp forests occur on more than a quarter of the world\u27s coastlines, serving as foundation species supporting high levels of biodiversity. They are also a major source of organic matter in coastal ecosystems, with the majority of primary production released and exported as detritus. Kelp detritus also provides food and shelter for macroinvertebrates, which comprise important components of inshore food-webs. Hitherto, research on kelp detritus-associated macroinvertebrate assemblages remains relatively limited. We quantified spatiotemporal variability in the structure of detritus-associated macroinvertebrate assemblages within Laminaria hyperborea forests and evaluated the influence of putative drivers of the observed variability in assemblages across eight study sites within four regions of the United Kingdom in May and September 2015. We documented 5167 individuals from 106 taxa with Malacostraca, Gastropoda, Isopoda and Bivalvia the most abundant groups sampled. Assemblage structure varied across months, sites, and regions, with highest richness in September compared to May. Many taxa were unique to individual regions, with few documented in all regions. Finally, key drivers of assemblage structure included detritus tissue nitrogen content, depth, sea surface temperature, light intensity, as well as L. hyperborea canopy density and canopy biomass. Despite their dynamic composition and transient existence, accumulations of L. hyperborea detritus represent valuable repositories of biodiversity and represent an additional kelp forest component which influences secondary productivity, and potentially kelp forest food-web dynamics

A quantitative synthesis of approaches, biases, successes, and failures in marine forest restoration, with considerations for future work

1. Marine forests is a term commonly used for coastal marine habitats formed by dense stands of brown macroalgae, typically consisting of kelp and fucoids. These habitats are highly productive, offer habitat to numerous marine organisms, and support a range of invaluable ecosystem services. Despite their importance, marine forests are declining in many regions around the world as a result of interacting global, regional, and local-scale stressors. Consequently, interest in restoration as a tool to mitigate these declines and reinstate marine forests is growing. 2. Recent reviews have provided insights into marine forest restoration; however, for the most part, a synthesis of restoration success is lacking. A meta-analysis and quantitative review of published marine forest restoration efforts was conducted to examine: (i) how restoration affects the abundance and morphology of marine forest species; and (ii) trends in marine forest restoration success. 3. The meta-analysis of 25 studies revealed that restoration positively influences the abundance and morphology of marine forest species. The quantitative review of 63 studies demonstrated that taxa and restoration technique were important factors influencing restoration success, and revealed a bias towards the monitoring and reporting of abundance and morphological response variables. The review also highlighted a lack of monitoring and/or reporting of environmental variables at restoration sites, and limited comparative research across environmental contexts and restored species. 4. It is shown that successful marine forest restoration is possible at experimental scales, but that better monitoring and reporting of restoration efforts, alongside increased project durations, could improve our understanding of restoration success at the ecosystem level. Considerations for future marine forest restoration efforts are also provided. It is hoped that the review will advance marine forest restoration efforts, allowing the preservation of these valuable ecosystems and their associated services

The need, opportunities, and challenges for creating a standardized framework for marine restoration monitoring and reporting

Marine ecosystems have been used, impacted by, and managed by human populations for millennia. As ecosystem degradation has been a common outcome of these activities, marine management increasingly considers ecosystem restoration. Currently, there is no coherent data recording format or framework for marine restoration projects. As a result, data are inconsistently recorded and it is difficult to universally track progress, assess restoration's global effectiveness, reduce reporting bias, collect a holistic suite of metrics, and share information. Barriers to developing a unified system for reporting marine restoration outcomes include: reaching agreement on a framework that meets the needs of all users, funding its development and maintenance, balancing the need for ‘ease of use’ and detail, and demonstrating the value of using the framework. However, there are opportunities to leverage arising from the United Nation Decades of Ecosystem Restoration and Science for Sustainable Development and with existing processes already developed by restoration groups (e.g. Global Mangrove Alliance, Society for Ecological Restoration). Here we provide guidelines and a roadmap for how such a framework could be developed and the potential benefits of such an endeavor. We call on practitioners to collaborate to develop such a framework and on governing bodies to commit to making detailed reporting a requirement for restoration project funding. Using a standardized marine restoration monitoring framework would enable the application of adaptive management when projects are not progressing as expected, advance our understanding of the state of worldwide marine restoration, and generate knowledge to advance restoration methodologies

Conformational Reorganization of the SARS Coronavirus Spike Following Receptor Binding: Implications for Membrane Fusion

The SARS coronavirus (SARS-CoV) spike is the largest known viral spike molecule, and shares a similar function with all class 1 viral fusion proteins. Previous structural studies of membrane fusion proteins have largely used crystallography of static molecular fragments, in isolation of their transmembrane domains. In this study we have produced purified, irradiated SARS-CoV virions that retain their morphology, and are fusogenic in cell culture. We used cryo-electron microscopy and image processing to investigate conformational changes that occur in the entire spike of intact virions when they bind to the viral receptor, angiotensin-converting enzyme 2 (ACE2). We have shown that ACE2 binding results in structural changes that appear to be the initial step in viral membrane fusion, and precisely localized the receptor-binding and fusion core domains within the entire spike. Furthermore, our results show that receptor binding and subsequent membrane fusion are distinct steps, and that each spike can bind up to three ACE2 molecules. The SARS-CoV spike provides an ideal model system to study receptor binding and membrane fusion in the native state, employing cryo-electron microscopy and single-particle image analysis

HER2 expression in cervical cancer as a potential therapeutic target

BACKGROUND: Trastuzumab, a humanized monoclonal antibody against the HER2 receptor is currently being used in breast and other tumor types. Early studies have shown that a variable proportion of cervical carcinoma tumors overexpress the HER2 receptor as evaluated by diverse techniques and antibodies. Currently it is known that a tumor response to trastuzumab strongly correlates with the level of HER2 expression evaluated by the Hercep Test, thus, it seems desirable to evaluate the status of expression of this receptor using the FDA-approved Hercep Test and grading system to gain insight in the feasibility of using trastuzumab in cervical cancer patients. METHODS: We analyzed a series of cervical cancer cell lines, the primary tumors of 35 cases of cervical cancer patients and four recurrent cases, with the Hercep Test in order to establish whether this tumor type overexpress HER2 at level of 2+/3+ as trastuzumab is currently approved for breast cancer having such level of expression. RESULTS: The results indicate that only 1 out of 35 primary tumors cases overexpress the receptor at this level, however, two out of four recurrent tumors that tested negative at diagnosis shifted to Hercep Test 2+ and 3+ respectively. CONCLUSIONS: The low frequency of expression in primary cases suggests that trastuzumab could have a limited value for the primary management of cervical cancer patients, however, the finding of "conversion" to Hercep Test 2+ and 3+ of recurrent tumors indicates the need to further evaluate the expression of HER2 in the metastatic and recurrent cases

Biologic markers of risk in nipple aspirate fluid are associated with residual cancer and tumour size

We previously demonstrated that nipple aspirate fluid (NAF) can be obtained from virtually all non-Asian women between the ages of 30 and 72. The focus of this report is to (1) determine the association of candidate markers of breast cancer risk in NAF obtained from fresh mastectomy specimens with residual breast carcinoma, and (2) evaluate the association of the markers with breast tumour progression. Nipple aspiration was performed on 97 specimens. Cytology, DNA index (including % hypertetraploid cells), cell cycle parameters (S phase fraction, % cells in G2/M), prostate-specific antigen (PSA), epidermal growth factor (EGF), testosterone, carcinoembryonic antigen (CEA) and prostaglandin D synthase (PGDS) were evaluated in NAF for their association with (1) residual ductal carcinoma in situ (DCIS) or invasive cancer, and (2) pathologic tumour size. NAF was obtained from 99% (96/97) of specimens. Atypical and malignant NAF cytology were significantly associated with residual DCIS or invasive cancer (P = 0.001) and with larger tumours (P = 0.004). One hundred per cent and 88% of subjects with malignant and atypical NAF cytology, respectively, had residual carcinoma. The percentage of cells in G2/M and DNA index were associated both with risk of residual carcinoma (P = 0.01 for each) and larger tumour size (DNA index, P = 0.03; G2/M, P = 0.05), although neither biomarker improved the ability of NAF cytology, to predict residual breast cancer. Higher DNA index was associated with atypical cytology (P = 0.0001). In summary, atypical and malignant NAF cytology are associated with larger tumour size, and are highly predictive of residual carcinoma after needle or excisional biopsy of the breast. © 1999 Cancer Research Campaig