Inhibition of the mitochondrial calcium uniporter (MCU) rescues dopaminergic neurons in pink1-/- zebrafish

Mutations in PTEN-induced putative kinase 1 (PINK1) are a cause of early onset Parkinson's disease (PD). Loss of PINK1 function causes dysregulation of mitochondrial calcium homeostasis, resulting in mitochondrial dysfunction and neuronal cell death. We report that both genetic and pharmacological inactivation of the mitochondrial calcium uniporter (MCU), located in the inner mitochondrial membrane, prevents dopaminergic neuronal cell loss in pink1Y431* mutant zebrafish (Danio rerio) via rescue of mitochondrial respiratory chain function. In contrast, genetic inactivation of the voltage dependent anion channel 1 (VDAC1), located in the outer mitochondrial membrane, did not rescue dopaminergic neurons in PINK1 deficient Danio rerio. Subsequent gene expression studies revealed specific upregulation of the mcu regulator micu1 in pink1Y431* mutant zebrafish larvae and inactivation of micu1 also results in rescue of dopaminergic neurons. The functional consequences of PINK1 deficiency and modified MCU activity were confirmed using a dynamic in silico model of Ca2+ triggered mitochondrial activity. Our data suggest modulation of MCU-mediated mitochondrial calcium homeostasis as a possible neuroprotective strategy in PINK1 mutant PD

Crystallographic Study Of The Phosphoethanolamine Transferase EptC required For Polymyxin Resistance And Motility In Campylobacter jejuni

The foodborne enteric pathogen Campylobacter jejuni decorates a variety of its cell-surface structures with phosphoethanolamine (pEtN). Modifying lipid A with pEtN promotes cationic antimicrobial peptide resistance, whereas post-translationally modifying the flagellar rod protein FlgG with pEtN promotes flagellar assembly and motility, which are processes that are important for intestinal colonization. EptC, the pEtN transferase required for all known pEtN cell-surface modifications in C. jejuni, is a predicted inner-membrane metalloenzyme with a five-helix N-terminal transmembrane domain followed by a soluble sulfatase-like catalytic domain in the periplasm. The atomic structure of the catalytic domain of EptC (cEptC) was crystallized and solved to a resolution of 2.40 angstrom. cEptC adopts the alpha/beta/alpha fold of the sulfatase protein family and harbors a zinc-binding site. A phosphorylated Thr266 residue was observed that was hypothesized to mimic a covalent pEtN-enzyme intermediate. The requirement for Thr266 as well as the nearby residues Asn308, Ser309, His358 and His440 was ascertained via in vivo activity assays on mutant strains. The results establish a basis for the design of pEtN transferase inhibitors.National Institutes of Health (grants AI064184, AI076322, GM106112Army Research Office (grantW911NF-12-1-0390)College of Natural SciencesOffice of the Executive Vice President and ProvostInstitute for Cellular and Molecular Biology at the University of Texas at AustinUS DOE DE-AC02-06CH11357National Institute of General Medical SciencesHoward Hughes Medical InstituteOffice of Science, Office of Basic Energy Sciences of the US Department of Energy DE-AC02-05CH11231Maria Person and the Proteomics Facility at the University of Texas at Austin ES007784 (CRED) and RP110782 (CPRIT)Molecular Bioscience

Restriction of mitochondrial calcium overload by mcu inactivation renders a neuroprotective effect in zebrafish models of Parkinson's disease

The loss of dopaminergic neurons (DA) is a pathological hallmark of sporadic and familial forms of Parkinson's disease (PD). We have previously shown that inhibiting mitochondrial calcium uniporter (mcu) using morpholinos can rescue DA neurons in the PTEN-induced putative kinase 1 (pink1)−/− zebrafish model of PD. In this article, we show results from our studies in mcu knockout zebrafish, which was generated using the CRISPR/Cas9 system. Functional assays confirmed impaired mitochondrial calcium influx in mcu−/− zebrafish. We also used in vivo calcium imaging and fluorescent assays in purified mitochondria to investigate mitochondrial calcium dynamics in a pink1−/− zebrafish model of PD. Mitochondrial morphology was evaluated in DA neurons and muscle fibers using immunolabeling and transgenic lines, respectively. We observed diminished mitochondrial area in DA neurons of pink1−/− zebrafish, while deletion of mcu restored mitochondrial area. In contrast, the mitochondrial volume in muscle fibers was not restored after inactivation of mcu in pink1−/− zebrafish. Mitochondrial calcium overload coupled with depolarization of mitochondrial membrane potential leads to mitochondrial dysfunction in the pink1−/− zebrafish model of PD. We used in situ hybridization and immunohistochemical labeling of DA neurons to evaluate the effect of mcu deletion on DA neuronal clusters in the ventral telencephalon of zebrafish brain. We show that DA neurons are rescued after deletion of mcu in pink1−/− and the 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) zebrafish model of PD. Thus, inactivation of mcu is protective in both genetic and chemical models of PD. Our data reveal that regulating mcu function could be an effective therapeutic target in PD pathology

The effects of particulate and ozone pollution on mortality in Moscow, Russia

The objectives of this study were (1) to evaluate how acute mortality responds to changes in particulate and ozone (O3) pollution levels, (2) to identify vulnerable population groups by age and cause of death, and (3) to address the problem of interaction between the effects of O3 and particulate pollution. Time-series of daily mortality counts, air pollution, and air temperature were obtained for the city of Moscow during a 3-year period (2003–2005). To estimate the pollution-mortality relationships, we used a log-linear model that controlled for potential confounding by daily air temperature and longer term trends. The effects of 10 μg/m3 increases in daily average measures of particulate matter ≤10 μm in aerodynamic diameter (PM10) and O3 were, respectively, (1) a 0.33% [95% confidence interval (CI) 0.09–0.57] and 1.09% (95% CI 0.71–1.47) increase in all-cause non-accidental mortality in Moscow; (2) a 0.66% (0.30–1.02) and 1.61% (1.01–2.21) increase in mortality from ischemic heart disease; (3) a 0.48% (0.02–0.94) and 1.28% (0.54–2.02) increase in mortality from cerebrovascular diseases. In the age group >75 years, mortality increments were consistently higher, typically by factor of 1.2 – 1.5, depending upon the cause of death. PM10-mortality relationships were significantly modified by O3 levels. On the days with O3 concentrations above the 90th percentile, PM10 risk for all-cause mortality was threefold greater and PM10 risk for cerebrovascular disease mortality was fourfold greater than the unadjusted risk estimate

Research Quality at ICRISAT: Separating the grain from the chaff

The International Crops Research Institute for the Semi-Arid Tropics (ICRISAT) is one of the agriculture research centers of the Consultative Group on International Agricultural Research (CGIAR), which is an informal association of over 50 members that supports a network of 15 international research centers in agriculture, forestry and fisheries. The CGIAR aims, through its support to the Centers, to contribute to promoting sustainable agriculture for food security in developing countries. Because the Centers constitute the core of the CGIAR, the effectiveness of ICRISAT and other Centers is crucial to the continued success of the CGIAR as a system. The CGIAR has established a tradition of external reviews to provide a mechanism of transparency and accountability to the members and other stakeholders of the CGIAR. The monitoring of quality of science is important to ensure that the Centers are producing transparent and quality research outputs that align with the agenda of the CGIAR. External Program Reviews (EPR) provide a measure of central oversight and serve as an essential component of the CGIAR’s accountability system. It complements the Center Commissioned External Reviews (CCERs) by providing a CGIARcommissioned comprehensive external assessment of the Center’s program and management, especially its relevance and quality of research and future directions. These reviews assess the mission, priorities and strategies, relevance and quality of science, effectiveness and efficiency of research leadership and program management and accomplishments and impact

Exploitation and Survival of Reared Salmon Released into the Burrishoole River System

Hatchery reared salmon smolts have been microtagged using binary coded nose tags and released into Irish rivers since 1980. These tagged fish are intercepted and identified in high seas coastal fisheries and in Irish rivers as adult salmon. The tag recovery programme provides valuable information on the marine survival and exploitation rates of these tagged fish and the return rates back to the river of origin. This leaflet summarises the results for release groups of reared indigenous salmon from the Burrishoole system in Co. Mayo. Marine survival prior to homewater exploitation has been very variable in the period examined. Although homewater marine exploitation rates have varied considerably depending on the fishing areas, they remained high for all areas combined between the years 1982 to 1989 with exploitation rates by coastal fisheries up to 87%. The greater part of the catch is taken in the Mayo area. Exploitation rates have decreased since 1989 and approximately 60% of the returning stock is estimated to have been caught in coastal fisheries in 1993. Survival to the river has also varied with an average of 2.5% of the total number of smolts released returning as adults to the river

The state of stocks of cod, whiting, sole and plaice on the west and southwest coasts of Ireland

Stocks of cod, whiting, plaice and sole in ICES Divisions VIIb,c and VIIj,k are an important part of the Irish fishing resource yet, until 1993, were not subject to any assessment. Landings of these stocks in 1996 were valued at £8.6 million, representing 17% of the overall value of all Irish landings of demersal species. In 1993, the Fisheries Research Centre initiated a stock monitoring programme with the aim of providing adequate data to enable an assessment of these stocks. This paper presents the results of growth, catch curve and yield per recruit analyses from the monitoring programme conducted between 1993 and 1996. Preliminary results show that all of these stocks but sole in VIIb,c are over-exploited. TAC levels (based on reported catches in previous years) may be inappropriate for the current stock sizes. It is also important to compare biological characteristics between ICES Divisions to determine the appropriate assessment areas. Comparisons of biological parameters between areas have so far been inconclusive.Funder: Marine Institut

Blending hard and soft science: the Follow the Technology approach to analyzing and evaluating technology change

Published online: 20 Dec 2001The types of technology change catalyzed by research interventions in integrated natural resource management (INRM) are likely to require much more social negotiation and adaptation than are changes related to plant breeding, the dominant discipline within the system of the Consultative Group on International Agricultural Research (CGIAR). Conceptual models for developing and delivering high-yielding varieties have proven inadequate for delivering natural resource management (NRM) technologies that are adopted in farmers' fields. Successful INRM requires tools and approaches that can blend the technical with the social, so that people from different disciplines and social backgrounds can effectively work and communicate with each other. This paper develops the "follow-the-technology" (FTT) approach to catalyzing, managing, and evaluating rural technology change as a framework that both "hard" and "soft" scientists can work with. To deal with complexity, INRM needs ways of working that are adaptive and flexible. The FTT approach uses technology as the entry point into a complex situation to determine what is important. In this way, it narrows the research arena to achievable boundaries. The methodology can also be used to catalyze technology change, both within and outside agriculture. The FTT approach can make it possible to channel the innovative potential of local people that is necessary in INRM to "scale up" from the pilot site to the landscape. The FTT approach is built on an analogy between technology change and Darwinian evolution, specifically between "learning selection" and natural selection. In learning selection, stakeholders experiment with a new technology and carry out the evolutionary roles of novelty generation, selection, and promulgation. The motivation to participate is a "plausible promise" made by the R&D team to solve a real farming problem. Case studies are presented from a spectrum of technologies to show that repeated learning selection cycles can result in an improvement in the performance of the plausible promise through adaptation and a sense of ownership by the stakeholders

Does crop diversity contribute to dietary diversity? Evidence from integration of vegetables into maize-based farming systems

Background: Maize is the most important staple crop for food security and livelihood of smallholder farmers in many parts of sub-Saharan Africa, but it alone cannot ensure food security. Cropping patterns must be diversified to ensure an adequate supply and economic access to greater variety of foods for smallholder farm households. This study measured the effect of crop diversification on household dietary diversity in a selected study locale using a survey of 300 randomly stratified farm households in 10 villages located in the Babati, Kongwa and Kiteto districts of Tanzania. Results: Based on multiple regression analysis, the study found that simply increasing Simpson’s Index does not influence dietary diversity of farm households due to the presence of interaction effect between Simpson’s Index and crop income. It is much more critical and significant to increase the revenue generated from diversified crops along with other socioeconomic endowment and behavioral characteristics of farm households. This is particularly applicable to poorer smallholder farmers who receive crop income less than US$85 per sales transaction and per season. Particularly, marginal and smallholders might be exposed to the effects of crop diversification and crop income toward increasing in their household dietary diversity score. Conclusion: Under average crop income scenarios, households that diversify their crop production tend to increase their dietary diversity from their existing dietary diversity score at a decreasing rate. However, under below average crop income threshold scenarios, farmers tend to increase their dietary diversity score from their existing score at an increasing rate when they diversify into high-value crops that attract relatively high farm gate values and accrue higher net revenues from the market. Monthly food expenditure also tends to positively influence household dietary diversity, indicating that farm households that spend more on market-purchased food have consistent increases in their dietary diversity scores at the household level. This study concludes that improving economic access to variety of foods at the smallholder household level by diversifying diets through increased crop diversification should be encouraged within maize-based farming systems of the study locale, through integration of micronutrient-rich foods such as vegetables

SBSPKS: structure based sequence analysis of polyketide synthases

Polyketide synthases (PKSs) catalyze biosynthesis of a diverse family of pharmaceutically important secondary metabolites. Bioinformatics analysis of sequence and structural features of PKS proteins plays a crucial role in discovery of new natural products by genome mining, as well as in design of novel secondary metabolites by biosynthetic engineering. The availability of the crystal structures of various PKS catalytic and docking domains, and mammalian fatty acid synthase module prompted us to develop SBSPKS software which consists of three major components. Model_3D_PKS can be used for modeling, visualization and analysis of 3D structure of individual PKS catalytic domains, dimeric structures for complete PKS modules and prediction of substrate specificity. Dock_Dom_Anal identifies the key interacting residue pairs in inter-subunit interfaces based on alignment of inter-polypeptide linker sequences to the docking domain structure. In case of modular PKS with multiple open reading frames (ORFs), it can predict the cognate order of substrate channeling based on combinatorial evaluation of all possible interface contacts. NRPS–PKS provides user friendly tools for identifying various catalytic domains in the sequence of a Type I PKS protein and comparing them with experimentally characterized PKS/NRPS clusters cataloged in the backend databases of SBSPKS. SBSPKS is available at http://www.nii.ac.in/sbspks.html