Identification of a conserved cluster in the RH domain of GRK critical for activation by GPCRs

One of the most critical aspects of G Protein Coupled Receptors (GPCRs) regulation is their rapid and acute desensitization following agonist stimulation. Phosphorylation of these receptors by GPCR kinases (GRK) is a major mechanism of desensitization. Considerable evidence from studies of rhodopsin kinase and GRK2 suggests there is an allosteric docking site for the receptor distinct from the GRK catalytic site. While the agonist-activated GPCR appears crucial for GRK activation, the molecular details of this interaction remain unclear. Recent studies suggested an important role for the N- and C-termini and domains in the small lobe of the kinase domain in allosteric activation; however, neither the mechanism of action of that site nor the RH domain contributions have been elucidated. To search for the allosteric site, we first indentified evolutionarily conserved sites within the RH and kinase domains presumably deterministic of protein function employing evolutionary trace (ET) methodology and crystal structures of GRK6. Focusing on a conserved cluster centered on helices 3, 9, and 10 in the RH domain, key residues of GRK5 and 6 were targeted for mutagenesis and functional assays. We found that a number of double mutations within helices 3, 9, and 10 and the N-terminus markedly reduced (50–90%) the constitutive phosphorylation of the β-2 Adrenergic Receptor (β2AR) in intact cells and phosphorylation of light-activated rhodopsin (Rho*) in vitro as compared to wild type (WT) GRK5 or 6. Based on these results, we designed peptide mimetics of GRK5 helix 9 both computationally and through chemical modifications with the goal of both confirming the importance of helix 9 and developing a useful inhibitor to disrupt the GPCR-GRK interaction. Several peptides were found to block Rho* phosphorylation by GRK5 including the native helix 9 sequence, Peptide Builder designed-peptide preserving only the key ET residues, and chemically locked helices. Most peptidomimetics showed inhibition of GRK5 activity greater than 80 % with an IC50 of ∼ 30 µM. Alanine scanning of helix 9 has further revealed both essential and non-essential residues for inhibition. Importantly, substitution of Arg 169 by an alanine in the native helix 9-based peptide gave an almost complete inhibition at 30 µM with an IC50 of ∼ 10 µM. In summary we report a previously unrecognized crucial role for the RH domain of GRK5 and 6, and the subsequent identification of a lead peptide inhibitor of protein-protein interaction with potential for specific blockade of GPCR desensitization. ^

AN EMPIRICAL ESTIMATION OF MEDICAL AND NON-MEDICAL INPUTS IN THE PRODUCTION OF CARDIO-RESPIRATORY PHYSICAL HEALTH STATUS

The purpose of this study was to examine, in the context of an economic model of health production, the relationship between inputs (health influencing activities) and fitness.^ Primary data were collected from 204 employees of a large insurance company at the time of their enrollment in an industrially-based health promotion program. The inputs of production included medical care use, exercise, smoking, drinking, eating, coronary disease history, and obesity. The variables of age, gender and education known to affect the production process were also examined. Two estimates of fitness were used; self-report and a physiologic estimate based on exercise treadmill performance. Ordinary least squares and two-stage least squares regression analyses were used to estimate the fitness production functions.^ In the production of self-reported fitness status the coefficients for the exercise, smoking, eating, and drinking production inputs, and the control variable of gender were statistically significant and possessed theoretically correct signs. In the production of physiologic fitness exercise, smoking and gender were statistically significant. Exercise and gender were theoretically consistent while smoking was not. Results are compared with previous analyses of health production. ^

Seasonal production patterns of planktonic foraminifera in the NE Atlantic Ocean

Sediment trap samples from OMEX 2 (49°N, 13°W) provide a continuous record of the seasonal succession of planktonic foraminifera in the midlatitude North Atlantic and reveal a complex relationship between periods of production and specific hydrographic conditions. Neogloboquadrina pachyderma dextral coiling (d.), Globigerina bulloides, and Globorotalia inflata are found in great numbers during both the spring and summer seasons, whereas Globigerina quinqueloba, Globorotalia hirsuta, Globorotalia scitula, and Globigerinita glutinata are associated predominantly with the increase in productivity during the spring bloom. Globigerinella aequilateralis, Orbulina universa, and Globigerinoides sacculifer are restricted to late summer conditions following the establishment of a warm, well-stratified surface ocean. An annually integrated fauna from the sediment trap, comprising ~13,000 individuals, is used to evaluate the accuracy of five faunal-based statistical methods of paleotemperature estimation. All of the temperature reconstruction techniques produce estimates of ~16°C and ~11°C for summer and winter surface temperature, respectively, which are in excellent agreement with regional hydrographic data and suggest that the sediment trap assemblage is well represented in the core top faunas. Analysis of the key species that dominate the OMEX 2 sediment trap fauna, G. bulloides, G. inflata, and N. pachyderma d., based on d18O derived temperatures from North Atlantic core top samples, suggests that seasonal variations in planktonic foraminiferal production are nonuniform across the midlatitudes and that this is likely to complicate reconstructing past seasonal hydrographic dynamics using these taxa.

Calcium isotopic composition of planktonic foraminifera from ODP Site 154-925

An 18 million year record of the Ca isotopic composition (d44/42Ca) of planktonic foraminiferans from ODP site 925, in the Atlantic, on the Ceara Rise, provides the opportunity for critical analysis of Ca isotope-based reconstructions of the Ca cycle. ?44/42Ca in this record averages +0.37+/-0.05 (1 sigma SD) and ranges from +0.21? to +0.52?. The record is a good match to previously published Neogene Ca isotope records based on foraminiferans, but is not similar to the record based on bulk carbonates, which has values that are as much as 0.25? lower. Bulk carbonate and planktonic foraminiferans from core tops differ slightly in their d44/42Ca (i.e., by 0.06+/-0.06? (n = 5)), while the difference between bulk carbonate and foraminiferan values further back in time is markedly larger, leaving open the question of the cause of the difference. Modeling the global Ca cycle from downcore variations in d44/42Ca by assuming fixed values for the isotopic composition of weathering inputs (d44/42Ca_w) and for isotope fractionation associated with the production of carbonate sediments (D_sed) results in unrealistically large variations in the total mass of Ca2+ in the oceans over the Neogene. Alternatively, variations of +/-0.05? in the Ca isotope composition of weathering inputs or in the extent of fractionation of Ca isotopes during calcareous sediment formation could entirely account for variations in the Ca isotopic composition of marine carbonates. Ca isotope fractionation during continental weathering, such as has been recently observed, could easily result in variations in d44/42Ca_w of a few tenths of permil. Likewise a difference in the fractionation factors associated with aragonite versus calcite formation could drive shifts in D_sed of tenths of permil with shifts in the relative output of calcite and aragonite from the ocean. Until better constraints on variations in d44/42Ca_w and D_sed have been established, modeling the Ca2+ content of seawater from Ca isotope curves should be approached cautiously.

(Table 2) Production parameters of phytoplankton and some associated parameters in the area of the Titanic Polygon

Studies were carried out mostly in the area of RMS Titanic wreck site (41°44'N, 49°57'W) located above the continental slope and the south of the Grand Banks of Newfoundland. In a period from 18.06 to 24.09.2001 five surveys of production characteristics of surface phytoplankton were conducted over 5-9 days. Mean values of these characteristics obtained during the surveys were 9.2-11.7 mg C/m**3 per day for primary production (C_phs), 0.102-0.188 mg/m**3 for chlorophyll a (C_chls), and 4.44-7.42 mg C/mg chl. a per hour for assimilation number (AN). The main reason for low C_phs variability was a significant inverse relationship (R=-0.66) between AN and C_chls found over the research area. When cold shelf waters dominated in the area (27.07 to 19.08.2001), C_chls values for the slope region (0.125+/-0.031 µg/l) and for the outer shelf (0.130+/-0.040 µg/l) were similar. During strengthening of influence of warmer slope waters within area (from 29.08 to 13.09.2001), C_chls concentration within surface waters of the outer shelf was 0.152+/-0.039 µg/l and exceeded one for the slope region (0.094+/-0.004 µg/l) by factor 1.6. Against the background of low Cchls values, the High values of integral primary production in the water column (510-1010 mg C/m**2 per day) at low C_chls values measured within the area were determined both by high assimilation activity of phytoplankton and by the deep (30-40 m) maximum of primary production. Main reasons for formation of such a maximum were high chlorophyll concentration within the layer of the deep chlorophyll maximum (up to 0.5-2.5 µg/l) and in the relatively high solar irradiance within this layer varying from 1.4 to 8.6% of subsurface PAR.

CO2 and vitamin B12 interactions determine bioactive trace metal requirements of a subarctic Pacific diatom

Phytoplankton growth can be limited by numerous inorganic nutrients and organic growth factors. Using the subarctic diatom Attheya sp. in culture studies, we examined how the availability of vitamin B(12) and carbon dioxide partial pressure (pCO(2)) influences growth rate, primary productivity, cellular iron (Fe), cobalt (Co), zinc (Zn) and cadmium (Cd) quotas, and the net use efficiencies (NUEs) of these bioactive trace metals (mol C fixed per mol cellular trace metal per day). Under B(12)-replete conditions, cells grown at high pCO(2) had lower Fe, Zn and Cd quotas, and used those trace metals more efficiently in comparison with cells grown at low pCO(2). At high pCO(2), B(12)-limited cells had ~50% lower specific growth and carbon fixation rates, and used Fe ~15-fold less efficiently, and Zn and Cd ~3-fold less efficiently, in comparison with B(12)-replete cells. The observed higher Fe, Zn and Cd NUE under high pCO(2)/B(12)-replete conditions are consistent with predicted downregulation of carbon-concentrating mechanisms. Co quotas of B(12)-replete cells were 5- to 14-fold higher in comparison with B(12)-limited cells, suggesting that >80% of cellular Co of B(12)-limited cells was likely from B(12). Our results demonstrate that CO(2) and vitamin B(12) interactively influence growth, carbon fixation, trace metal requirements and trace metal NUE of this diatom. This suggests the need to consider complex feedback interactions between multiple environmental factors for this biogeochemically critical group of phytoplankton in the last glacial maximum as well as the current and future changing ocean.

Expansion of Asparagus Production and Exports in Peru

Peru is the one of the most important exporters of asparagus in the world. Its export volume of fresh asparagus is ranked number one, and its export volume of preserved asparagus number two, globally. The objective of this paper is to provide an overview of the recent trends in asparagus production and exports around the world and to analyze factors in the development of the Peruvian asparagus industry. The production of asparagus has spread geographically. The center of its production used to be in the principal consuming countries, such as France, Germany and the United States. Afterward, it spread to neighboring countries such as Spain and Mexico where production factors such as climate and labor costs are favorable. After the rise and fall of Taiwan as a major preserved white asparagus exporter, China and Peru replaced its position. Finally, in recent years, Peru expanded its fresh green asparagus exports to the U.S. market by taking advantage of the increasing demand for fresh vegetables and supplying produce in seasons when neither U.S. nor Mexican producers can harvest. In addition to the changing factors in the international market, there are several factors in the development of the industry: high yields of produce due to favorable climatic and soil conditions; the introduction of the drip irrigation system, which enabled desert cultivation; the integration of production and exports, which is indispensable for fresh produce exports; and the collective efforts of the industry with help from the public sector.

Theory of a Flowchart Approach to Industrial Cluster Policy

A flowchart approach to industrial cluster policy emphasizes the importance ofthe ordering of policy measures. The flow of policy implementation is to establish an industrial zone, to invite an anchor company, and to promote its related companies to invest in the industrial zone. This article delineated "a flowchart approach to industrial cluster policy" by proposing sufficient conditions for forming industrial clusters typical in the manufacturing industry in Asia to enhance regional economic growth. The typical industrial cluster policy was theorized by defining an industrial zone as "quasi-public goods", and it was shown that the policy enhances economic growth under a production function of "increasing returns to scale" of an anchor company. Critical amounts of the production of "scale economies" that are used by the related companies to decide whether or not to invest in clusters were also shown.

Has Low Productivity Constrained Competitiveness of African Firms? : Comparison of the Firm Performances with Asian Firms

It has been argued that poor productive performance is one of critical sources of stagnation of the African manufacturing sector, but firm-level empirical supports are limited. Using the inter-regional firm data of the garment industry, technical efficiency and its contribution to competitiveness measured as unit costs were compared between Kenyan and Bangladeshi firms. Our estimates indicated that there is no significant gap in the average technical efficiency of the two industries despite conservative estimation, although unit costs greatly differ between the two industries. Higher unit cost in Kenyan firms mainly stems from high labour cost, while impact of inefficiency is quite small. Productivity accounts little for the stagnation of garment industry in several African countries.

A New Measurement for International Fragmentation of the Production Process: An International Input-Output Approach

The paper investigates the possibility of constructing a new measurement for analysing international fragmentation of the production process. It asserts that the current usage of relevant data, whether the trade shares of parts and components or the index of Vertical Specialisation, is quite unsatisfactory for measuring the phenomenon, since they critically lack the overall perspective of the entire structure of production chains. 　The new measurement is formulated such that it captures every aspect of the vertical sequence of production linkages. It is based on the input-output model of Average Propagation Lengths, recently developed by Eric Dietzenbacher and others, which show the average number of production stages that are passed through for an exogenous change in one industry to affect another. By applying this model to the data of the Asian International Input-Output Tables, the index is able to measure the international dimension of production sharing and division of labour in East Asia.

Production Networks and Spatial Economic Interdependence: An International Input-Output Analysis of the Asia-Pacific Region

The Asia-Pacific Region has enjoyed remarkable economic growth in the last three decades. This rapid economic growth can be partially attributed to the global spread of production networks, which has brought about major changes in spatial interdependence among economies within the region. By applying an Input-Output based spatial decomposition technique to the Asian International Input-Output Tables for 1985 and 2000, this paper not only analyzes the intrinsic mechanism of spatial economic interdependence, but also shows how value added, employment and CO2 emissions induced are distributed within the international production networks.

Dynamic analysis using finite elements to calculate the critical wear section of the contact wire in suburban railway overhead conductor rails

The purpose of this study is to determine the critical wear levels of the contact wire of the catenary on metropolitan lines. The study has focussed on the zones of contact wire where localised wear is produced, normally associated with the appearance of electric arcs. To this end, a finite element model has been developed to study the dynamics of pantograph-catenary interaction. The model includes a zone of localised wear and a singularity in the contact wire in order to simulate the worst case scenario from the point of view of stresses. In order to consider the different stages in the wire wear process, different depths and widths of the localised wear zone were defined. The results of the dynamic simulations performed for each stage of wear let the area of the minimum resistant section of the contact wire be determined for which stresses are greater than the allowable stress. The maximum tensile stress reached in the contact wire shows a clear sensitivity to the size of the local wear zone, defined by its width and depth. In this way, if the wear measurements taken with an overhead line recording vehicle are analysed, it will be possible to calculate the potential breakage risk of the wire. A strong dependence of the tensile forces of the contact wire has also been observed. These results will allow priorities to be set for replacing the most critical sections of wire, thereby making maintenance much more efficient. The results obtained show that the wire replacement criteria currently borne in mind have turned out to be appropriate, although in some wear scenarios these criteria could be adjusted even more, and so prolong the life cycle of the contact wire.