inducible-lap-tagged-stable-cell-lines-for-investigating-protein

Inducible LAP-tagged Stable Cell Lines for Investigating Protein Function, Spatiotemporal Localization and Protein Interaction Networks

Multi-protein complexes, rather than single proteins acting in isolation, often govern molecular pathways regulating cellular homeostasis. Based on this ...

University of California, Los Angeles

In Chile, the recommendation for salt iodination was reduced from 100 to 40 ppm, after detecting high urinary iodine excretion in school age children.

[Iodine nutrition in school children of four areas of Chile during the year 2001].

We studied the prevalence of weight excess and short stature among school-age children in Chile over the past decade.

Trends in nutritional status and stature among school-age children in Chile.

This study explores mediating medical risk factors in the association between racial residential segregation (i.e., racial 'black' isolation) and low birthweight in New York City, adjusting for maternal and infant risk factors and neighborhood poverty. This race-specific cross-sectional multilevel study found that as racial isolation increased in neighborhoods, the odds of having a low birthweight infant also increased for African-American and White women living in these areas. Medical conditions that mediated the racial isolation and low birthweight relationship included chronic hypertension and pregnancy-related hypertension for African-American women and chronic hypertension and lung disease for White women. Although this study was limited by the quality of the birth certificate data, it does provide exploratory pathways by which medical risks and their sequelae are linked to neighborhood environments and reproductive vulnerability.

Mediating medical risk factors in the residential segregation and low birthweight relationship by race in New York City.

This new century addresses several environmental challenges among which distribution of drinking water, global warming and availability of novel renewable energy sources to substitute for fossil fuels are of utmost importance. The last two concerns are closely related because the major part of carbon dioxide (CO(2)), considered as the main cause of the greenhouse effect, is widely produced from fossil fuel combustion. Renewable energy sources fully balanced in CO(2) are therefore of special interest, especially the issue of biological production from organic wastes. Among the possibilities of bioenergy production from wastes, two approaches are particularly interesting: The first one is relatively old and related to the production of biomethane by anaerobic digestion while the second one, more recent and innovative, relies on biohydrogen production by microbial ecosystems.

[Bioenergy production from waste: examples of biomethane and biohydrogen].

The anaerobic digestion process comprises a whole network of sequential and parallel reactions, of both biochemical and physicochemical nature. Mathematical models, aiming at understanding and optimization of the anaerobic digestion process, describe these reactions in a structured way, the IWA Anaerobic Digestion Model No. 1 (ADM1) being the most well established example. While these models distinguish between different microorganisms involved in different reactions, to our knowledge they all neglect species diversity between organisms with the same function, i.e. performing the same reaction. Nevertheless, available experimental evidence suggests that the structure and properties of a microbial community may be influenced by process operation and on their turn also determine the reactor functioning. In order to adequately describe these phenomena, mathematical models need to consider the underlying microbial diversity. This is demonstrated in this contribution by extending the ADM1 to describe microbial diversity between organisms of the same functional group. The resulting model has been compared with the traditional ADM1 in describing experimental data of a pilot-scale hybrid Upflow Anaerobic Sludge Filter Bed (UASFB) reactor, as well as in a more detailed simulation study. The presented model is further shown useful in assessing the relationship between reactor performance and microbial community structure in mesophilic CSTRs seeded with slaughterhouse wastewater when facing increasing levels of ammonia.

Modeling microbial diversity in anaerobic digestion through an extended ADM1 model.

Anaerobic digestion disintegration and hydrolysis have been traditionally modeled according to first-order kinetics assuming that their rates do not depend on disintegration/hydrolytic biomass concentrations. However, the typical sigmoid-shape increase in time of the disintegration/hydrolysis rates cannot be described with first-order models. For complex substrates, first-order kinetics should thus be modified to account for slowly degradable material. In this study, a slightly modified IWA ADM1 model is presented to simulate thermophilic anaerobic digestion of thermally pretreated waste activated sludge. Contois model is first included for disintegration and hydrolysis steps instead of first-order kinetics and Hill function is then used to model ammonia inhibition of aceticlastic methanogens instead of a non-competitive function. One batch experimental data set of anaerobic degradation of a raw waste activated sludge is used to calibrate the proposed model and three additional data sets from similar sludge thermally pretreated at three different temperatures are used to validate the parameters values.

Modified ADM1 disintegration/hydrolysis structures for modeling batch thermophilic anaerobic digestion of thermally pretreated waste activated sludge.

Chronic Chagas disease (15 million patients; annual incidence, 40, 000 patients; annual mortality, 12 ,500 patients) is the most serious parasitic disease in Latin America. Between 10 and 30 years after infection, 30% of patients with Chagas disease develop heart injury, which is the main reason for its high mortality. Consequently, frequent cardiac diagnostics are required for patients with Chagas disease.

Cardiac troponin T measured with a highly sensitive assay for diagnosis and monitoring of heart injury in chronic Chagas disease.

The extracellular levels of the neurotransmitter glycine in the brain are tightly regulated by the glycine transporter 1 (GlyT1) and the clearance rate for glycine depends on its rate of transport and the levels of cell surface GlyT1. Over the years, it has been shown that PKC tightly regulates the activity of several neurotransmitter transporters. In the present work, by stably expressing three N-terminus GlyT1 isoforms in porcine aortic endothelial cells and assaying for [(32)P]-orthophosphate metabolic labeling, we demonstrated that the isoforms GlyT1a, GlyT1b, and GlyT1c were constitutively phosphorylated, and that phosphorylation was dramatically enhanced, in a time dependent fashion, after PKC activation by phorbol ester. The phosphorylation was PKC-dependent, since pre-incubation of the cells with bisindolylmaleimide I, a selective PKC inhibitor, abolished the phorbol ester-induced phosphorylation. Blotting with specific anti-phospho-tyrosine antibodies did not yield any signal that could correspond to GlyT1 tyrosine phosphorylation, suggesting that the phosphorylation occurs at serine and/or threonine residues. In addition, a 23-40%-inhibition on V(max) was obtained by incubation with phorbol ester without a significant change on the apparent Km value. Furthermore, pre-incubation of the cells with the selective PKCα/β inhibitor Gö6976 abolished the downregulation effect of phorbol ester on uptake and phosphorylation, whereas the selective PKCβ inhibitors (PKCβ inhibitor or LY333531) prevented the phosphorylation without affecting glycine uptake, defining a specific role of classical PKC on GlyT1 uptake and phosphorylation. Taken together, these data suggest that conventional PKCα/β regulates the uptake of glycine, whereas PKCβ is responsible for GlyT1 phosphorylation.

PKCβ-dependent phosphorylation of the glycine transporter 1.

Chronic Chagas disease afflicts millions of patients in Latin America of which 70% remain asymptomatic but 30% develop fatal heart injury. To evaluate the impact of laboratory medicine for diagnosis and guiding of patients with Chagas' heart disease, we measured N-terminal B-type natriuretic peptide (NT-proBNP) and cardiac troponin T (cTnT).

Combined measurement of N-terminal pro-B-type natriuretic peptide and highly sensitive cardiac troponin T for diagnosis and monitoring of heart injury in chronic Chagas' disease.

In Peru, it was hypothesized that epidemic cholera in 1991 was linked to El Niño, the warm phase of El Niño-Southern Oscillation. While previous studies demonstrated an association in 1997-1998, using cross-sectional data, they did not assess the consistency of this relationship across the decade. Thus, how strong or variable an El Niño-cholera relationship was in Peru or whether El Niño triggered epidemic cholera early in the decade remains unknown. In this study, wavelet and mediation analyses were used to characterize temporal patterns among El Niño, local climate variables (rainfall, river discharge, and air temperature), and cholera incidence in Piura, Peru from 1991 to 2001 and to estimate the mediating effects of local climate on El Niño-cholera relationships. The study hypothesis is that El Niño-related connections with cholera in Piura were transient and interconnected via local climate pathways. Overall, our findings provide evidence that a strong El Niño-cholera link, mediated by local hydrology, existed in the latter part of the 1990s but found no evidence of an El Niño association in the earlier part of the decade, suggesting that El Niño may not have precipitated cholera emergence in Piura. Further examinations of cholera epicenters in Peru are recommended to support these results in Piura. For public health planning, the results may improve existing efforts that utilize El Niño monitoring for preparedness during future climate-related extremes in the region.

El Niño, Climate, and Cholera Associations in Piura, Peru, 1991-2001: A Wavelet Analysis.

The conservation status and taxonomy of the three gadfly petrels that breed in Macaronesia is still discussed partly due to the scarce information on their spatial ecology. Using geolocator and capture-mark-recapture data, we examined phenology, natal philopatry and breeding-site fidelity, year-round distribution, habitat usage and at-sea activity of the three closely-related gadfly petrels that breed in Macaronesia: Zino's petrel Pterodroma madeira, Desertas petrel P. deserta and Cape Verde petrel P. feae. All P. feae remained around the breeding area during their non-breeding season, whereas P. madeira and P. deserta dispersed far from their colony, migrating either to the Cape Verde region, further south to equatorial waters in the central Atlantic, or to the Brazil Current. The three taxa displayed a clear allochrony in timing of breeding. Habitat modelling and at-sea activity patterns highlighted similar environmental preferences and foraging behaviours of the three taxa. Finally, no chick or adult was recaptured away from its natal site and survival estimates were relatively high at all study sites, indicating strong philopatry and breeding-site fidelity for the three taxa. The combination of high philopatry, marked breeding asynchrony and substantial spatio-temporal segregation of their year-round distribution suggest very limited gene flow among the three taxa.

Ivan Ramirez

Department of Chemistry and Biochemistry

University of California, Los Angeles

Inducible LAP-tagged Stable Cell Lines for Investigating Protein Function, Spatiotemporal Localization and Protein Interaction Networks

Ivan Ramirez

.css-o250i3{font-size:18px;font-family:Open Sans,sans-serif;line-height:21.6px;}Department of Chemistry and Biochemistry

University of California, Los Angeles

Inducible LAP-tagged Stable Cell Lines for Investigating Protein Function, Spatiotemporal Localization and Protein Interaction Networks

Department of Chemistry and Biochemistry