Quantification of microtubule stutters: dynamic instability behaviors that are strongly associated with catastrophe

Microtubules (MTs) are cytoskeletal fibers that undergo dynamic instability (DI), a remarkable process involving phases of growth and shortening separated by stochastic transitions called catastrophe and rescue. Dissecting DI mechanism(s) requires first characterizing and quantifying these dynamics, a subjective process that often ignores complexity in MT behavior. We present a Statistical Tool for Automated Dynamic Instability Analysis (STADIA) that identifies and quantifies not only growth and shortening, but also a category of intermediate behaviors that we term “stutters.” During stutters, the rate of MT length change tends to be smaller in magnitude than during typical growth or shortening phases. Quantifying stutters and other behaviors with STADIA demonstrates that stutters precede most catastrophes in our in vitro experiments and dimer-scale MT simulations, suggesting that stutters are mechanistically involved in catastrophes. Related to this idea, we show that the anticatastrophe factor CLASP2γ works by promoting the return of stuttering MTs to growth. STADIA enables more comprehensive and data-driven analysis of MT dynamics compared with previous methods. The treatment of stutters as distinct and quantifiable DI behaviors provides new opportunities for analyzing mechanisms of MT dynamics and their regulation by binding proteins.     

The Giardia ventrolateral flange is a lamellar membrane protrusion that supports attachment

Attachment to the intestinal epithelium is critical to the lifestyle of the ubiquitous parasite Giardia lamblia. The ventrolateral flange is a sheet-like membrane protrusion at the interface between parasites and attached surfaces. This structure has been implicated in attachment, but its role has been poorly defined. Here, we identified a novel actin associated protein with putative WH2-like actin binding domains we named Flangin. Flangin complexes with Giardia actin (GlActin) and is enriched in the ventrolateral flange making it a valuable marker for studying the flanges’ role in Giardia biology. Live imaging revealed that the flange grows to around 1 μm in width after cytokinesis, then remains uniform in size during interphase, grows in mitosis, and is resorbed during cytokinesis. A flangin truncation mutant stabilizes the flange and blocks cytokinesis, indicating that flange disassembly is necessary for rapid myosin-independent cytokinesis in Giardia. Rho family GTPases are important regulators of membrane protrusions and GlRac, the sole Rho family GTPase in Giardia, was localized to the flange. Knockdown of Flangin, GlActin, and GlRac result in flange formation defects. This indicates a conserved role for GlRac and GlActin in forming membrane protrusions, despite the absence of canonical actin binding proteins that link Rho GTPase signaling to lamellipodia formation. Flangin-depleted parasites had reduced surface contact and when challenged with fluid shear force in flow chambers they had a reduced ability to remain attached, confirming a role for the flange in attachment. This secondary attachment mechanism complements the microtubule based adhesive ventral disc, a feature that may be particularly important during mitosis when the parental ventral disc disassembles in preparation for cytokinesis. This work supports the emerging view that Giardia’s unconventional actin cytoskeleton has an important role in supporting parasite attachment.     

Anthropology in the Public Sphere, 2008: Emerging Trends and Significant Impacts

ABSTRACT   The themes, trends, and significant events of 2008 demonstrate that anthropology has established a new foothold in the public sphere—one that makes the most of novel forms of communication to reach far beyond the ivory tower to disseminate knowledge widely and freely. This review focuses on six topical areas of robust anthropological research in 2008 that also addressed some of the year's most pressing problems and issues, including the following: (1) war and peace; (2) climate change; (3) natural, industrial, and development-induced disaster recovery; (4) human rights; (5) health disparities; and (6) racial understanding, politics, and equity in the United States. It concludes by addressing some emerging issues in 2009 that especially require anthropological attention and insight, if we are to move beyond "business as usual."[Keywords: practicing anthropology, public anthropology, 2009 trends, anthropological impacts]     

Three research priorities for just and sustainable urban systems: Now is the time to refocus

Now is the time to refocus efforts in urban research and design. A changing climate and extreme weather events are presenting unique challenges to urban systems around the world. These challenges illuminate the social barriers that accompany disruptive events such as resource inequities and injustices. In this perspective, we provide three research priorities for just and sustainable urban systems that help to address these matters. The three research priorities are: (1) social equity and justice, (2) circularity, and (3) digital twins. Conceptual context and future research directions are provided for each. For social equity and justice, the future directions are mandatory equity analysis and inclusionary practices, understanding and reconciling historical injustices, and intentional integration with diverse community stakeholders. For circularity applications, they are better metrics for integration, more robust evaluation frameworks, and dynamic modeling at multiple spatial and temporal scales. Future directions for digital twins include developing principles to reduce complexity, integrating model and system components, and reducing barriers to data access. These research priorities are core to meeting several of the United Nations Sustainable Development Goals (i.e., 1—No Poverty, 8—Decent Work and Economic Growth, 10—Reduced Inequalities, and 11—Sustainable Cities and Communities). Useful social and technical matters are discussed throughout, where we highlight the importance of prioritizing localized research efforts, provide guidance for community-engaged research and co-development practices, and explain how these priorities interact to align with the evolving field of industrial ecology.     

Population status,habitat dependence,and reproductive ecology of Bahama Orioles: a critically endangered synanthropic species

ABSTRACT Recent elevation of critically endangered Bahama Orioles (Icterus northropi) to species status prompted us to evaluate their population status, habitat use, and breeding ecology. From surveys, we estimated that at least 141 to 254 individuals remain globally, with 90 to 162, 24 to 44, and 27 to 48 individuals remaining on North Andros Island, Mangrove Cay, and South Andros Island, The Bahamas, respectively. Orioles were observed nesting exclusively in anthropogenic habitat (residential and agricultural land), but home ranges also included nearby pine forest and coppice (dry broadleaf forest). Most nests (40 of 46, or 87%) were in nonnative coconut palm (Cocos nucifera), with native Sabal palmetto and Thrinax morrisii, and an introduced Brassaia actinophylla also used. Trees selected by orioles for nesting were significantly taller, less likely to have shrubs underneath, further from cover, and had more palm trees nearby than randomly selected palm trees. Three of eight nests with known contents were parasitized by Shiny Cowbirds (Molothrus bonariensis). Lethal yellowing disease recently devastated coconut palms and reduced the number of orioles on North Andros, but palms on Mangrove Cay and South Andros remain healthy. The juxtaposition of anthropogenic habitat to suitable native habitats may be more important than any single factor for Bahama Orioles, especially for breeding adults and fledged young. Conservation of coppice habitat, at high risk for agricultural and residential development, is crucial for survival of this critically endangered synanthropic species.     

DNA methylation changes in murine breast adenocarcinomas allow the identification of candidate genes for human breast carcinogenesis

Epigenetic inactivation due to aberrant promoter methylation is a key process in breast tumorigenesis. Murine models for human breast cancer have been established for nearly every important human oncogene or tumor suppressor gene. Mouse-to-human comparative gene expression and cytogenetic profiling have been widely investigated for these models; however, little is known about the conservation of epigenetic alterations during tumorigenesis. To determine if this key process in human breast tumorigenesis is also mirrored in a murine breast cancer model, we mapped cytosine methylation changes in primary adenocarcinomas and paired lung metastases derived from the polyomavirus middle T antigen mouse model. Global changes in methylcytosine levels were observed in all tumors when compared to the normal mammary gland. Aberrant methylation and associated gene silencing was observed for Hoxa7, a gene that is differentially methylated in human breast tumors, and Gata2, a novel candidate gene. Analysis of HOXA7 and GATA2 expression in a bank of human primary tumors confirms that the expression of these genes is also reduced in human breast cancer. In addition, HOXA7 hypermethylation is observed in breast cancer tissues when compared to adjacent tumor-free tissue. Based on these studies, we present a model in which comparative epigenetic techniques can be used to identify novel candidate genes important for human breast tumorigenesis, in both primary and metastatic tumors.     

University Multi-User Facility Survey��2010

Multi-user facilities serve as a resource for many universities. In 2010, a survey was conducted investigating possible changes and successful characteristics of multi-user facilities, as well as identifying problems in facilities. Over 300 surveys were e-mailed to persons identified from university websites as being involved with multi-user facilities. Complete responses were received from 36 facilities with an average of 20 years of operation. Facilities were associated with specific departments (22%), colleges (22%), and university research centers (8.3%) or were not affiliated with any department or college within the university (47%). The five most important factors to succeed as a multi-user facility were: 1) maintaining an experienced, professional staff in an open atmosphere; 2) university-level support providing partial funding; 3) broad client base; 4) instrument training programs; and 5) an effective leader and engaged strategic advisory group. The most significant problems were: 1) inadequate university financial support and commitment; 2) problems recovering full service costs from university subsidies and user fees; 3) availability of funds to repair and upgrade equipment; 4) inability to retain highly qualified staff; and 5) unqualified users dirtying/damaging equipment. Further information related to these issues and to fee structure was solicited. Overall, there appeared to be a decline in university support for facilities and more emphasis on securing income by serving clients outside of the institution and by obtaining grants from entities outside of the university.