A functional proteomic method for the enrichment of peripheral membrane proteins reveals the collagen binding protein Hsp47 is exposed on the surface of activated human platelets

Platelets are small blood cells vital for hemostasis. Following vascular damage, platelets adhere to collagens and activate, forming a thrombus that plugs the wound and prevents blood loss. Stimulation of the platelet collagen receptor glycoprotein VI (GPVI) allows recruitment of proteins to receptor-proximal signaling complexes on the inner-leaflet of the plasma membrane. These proteins are often present at low concentrations; therefore, signaling-complex characterization using mass spectrometry is limited due to high sample complexity. We describe a method that facilitates detection of signaling proteins concentrated on membranes. Peripheral membrane proteins (reversibly associated with membranes) were eluted from human platelets with alkaline sodium carbonate. Liquid-phase isoelectric focusing and gel electrophoresis were used to identify proteins that changed in levels on membranes from GPVI-stimulated platelets. Immunoblot analysis verified protein recruitment to platelet membranes and subsequent protein phosphorylation was preserved. Hsp47, a collagen binding protein, was among the proteins identified and found to be exposed on the surface of GPVI-activated platelets. Inhibition of Hsp47 abolished platelet aggregation in response to collagen, while only partially reducing aggregation in response to other platelet agonists. We propose that Hsp47 may therefore play a role in hemostasis and thrombosis.

The anions [(ZrOH(CO3)3)2]6− and [(ZrOH(C2O4)3)2]6−: single crystal X-ray diffraction studies of the complexes guanidinium zirconium carbonate [(C(NH2)3)3ZrOH(CO3)3·H2O]2 and sodium zirconium oxalate Na6(ZrOH(C2O4)3)2·7H2O

The complex [(C(NH2)3)3ZrOH(CO3)3·H2O]2 (A) has been shown by means of a single crystal X-ray diffraction study to contain [C(NH2)3]+ cations and dimeric anions of formulation [(ZrOH(CO3)3)2]6−. The anion is centrosymmetric with each metal being bonded to two bridging OH groups and three chelating CO2−3 ions. The Zr atoms are thus eight coordinate with a dodecahedral environments. The ZrO distances formed by the bridgng OH groups are shorter than those formed through zirconiu carbonate interactions. The non-bonded Zr…Zr distance is 3.47(2) Å. An infrared spectroscopic investigation of A provides data which support the findings of the crystallographic study. Likewise the complex Na6(ZrOH(CO2O4)3)2·7H2O (B) contains the anion [(ZrOH(C2O4)3)2]6−. This anion is structurally related to the anion in A as each Zr atom has an eight-coordinate dodecahedral environment being bonded to two bridging OH groups and three chelating oxalate ligands, but has no imposed crysallographic symmetry. The Zr…Zr non-bonded distance is 3.50(1) Å. The OZrO bridge angles are 69.7(4)° and A and 67.4(3)° in B.

Calcium carbonate solubilization through H-proton release from some legumes grown in calcareous saline-sodic soils

Increasing levels of CO2 and H+ proton in the rhizosphere from some legumes may play an important role in calcite dissolution of calcareous salt affected soils. Soils planted with white and brown varieties of cowpea (Vigna unguiculata L.) and hyacinth bean (Dolichos lablab L.) relying on either fertilizer N (KNO3) or N-fixation were compared against soils to which gypsum was applied and a control without plants and gypsum application to study the possibility of Ca2+ release from calcite and Na+ leaching. As compared to plants relying on inorganic N, leachates from all pore volumes (0·5, 1·0, 1·5, 2·0 pore volume) in lysimeters planted with N-fixing hyacinth bean contained significantly higher concentrations of HCO with lower concentrations from lysimeters planted with white cowpea relying on N-fixation. However, the lowest concentrations of HCO were recorded in the gypsum and control treatments. In initial leaching, lysimeters planted with N-fixing plants maintained similar leachate Ca2+ and Na+ concentrations compared to gypsum amended soils. However, gypsum amended soils were found to have a prolonged positive effect on Na+ removal. It might be concluded that some legumes that are known to fix N in calcareous salt affected soils may be an alternative ameliorant to the extremely expensive gypsum through calcite solubilization and a consequent release of Ca2+.

Geochemical mapping of carbonate sediments in the Makgadikgadi basin, Botswana using moderate resolution remote sensing data

This paper maps the carbonate geochemistry of the Makgadikgadi Pans region of northern Botswana from moderate resolution (500 m pixels) remotely sensed data, to assess the impact of various geomorphological processes on surficial carbonate distribution. Previous palaeo-environmental studies have demonstrated that the pans have experienced several highstands during the Quaternary, forming calcretes around shoreline embayments. The pans are also a significant regional source of dust, and some workers have suggested that surficial carbonate distributions may be controlled, in part, by wind regime. Field studies of carbonate deposits in the region have also highlighted the importance of fluvial and groundwater processes in calcrete formation. However, due to the large area involved and problems of accessibility, the carbonate distribution across the entire Makgadikgadi basin remains poorly understood. The MODIS instrument permits mapping of carbonate distribution over large areas; comparison with estimates from Landsat Thematic Mapper data show reasonable agreement, and there is good agreement with estimates from laboratory analysis of field samples. The results suggest that palaeo-lake highstands, reconstructed here using the SRTM 3 arc-second digital elevation model, have left behind surficial carbonate deposits, which can be mapped by the MODIS instrument. Copyright (c) 2006 John Wiley & Sons, Ltd.

Influence of sodium chloride on seed germination and seedling root growth of cotton (Gossypium hirsutum L.)

Response of cotton (Gossypium hirsutum L. cv. NIAB-78) to salinity, in terms of seed germination, seedling root growth and root Na+ and K+ content was determined in a laboratory experiment. Cotton seeds were exposed to increasing salinity levels using germination water with Sodium chloride concentrations of 0, 50, 100, 150 and 200 mM, to provide different degrees of salt stress. Germinated seeds were counted and roots were harvested at 24, 48, 72 and 96 h after the start of the experiment. It appeared that seed germination was only slightly affected by an increase in salinity (in most cases the differences between treatment were non-significant), whereas root length, root growth rate, root fresh and dry weights were severely affected, generally highly significant differences in these variables were found for comparisons involving most combinations of salinity levels, in particular with increased incubation period. K+ contents decreased with increasing salinity levels, although differences in K+ content were only significant when comparing the control and the 4 salinity levels. Na+ content of the roots increased with increasing levels of NaCl in the germination water, suggesting an exchange of K+ for Na+. The ratio K+/Na+ strongly decreased with rising levels of salinity from around 4.5 for the control to similar to 1 at 200 mM NaCl.

The application of ichnofabrics towards bridging the dichotomy between siliciclastic and carbonate shelf facies: examples from the Upper Jurassic Fulmar Formation (UK) and the Jubaila Formation (Saudi Arabia)

An initial study of the ichnofabrics of the Upper Jurassic (Kimmeridgian) Jubaila Formation of Saudi Arabia shows that the ichnofabrics are closely matched to the relatively well-described ichnofabrics of the contemporary Fulmar Formation of the UK Continental Shelf (North Sea), in respect of the lower shoreface/offshore transition facies to offshore facies. The ichnology and ichnofabrics of the Lower Jubaila Formation show that deposition took place on an open-marine platform on the Arabian craton subject to periodic storm activity, but under a persisting equilibrium between sediment accumulation and subsidence. This is consistent with the moderately deep-marine foraminiferal assemblages and the presence of calcareous nannofossils. Cyclicity is absent, though storm beds may be grouped, in contrast with the genetic sequences present in the rift and halokinetic scenario of the North Sea. In contrast with the siliciclastic setting hardgrotinds (with Gastrochaenolites), more common firmground omission surfaces, and micritic mudstones with Chondrites and Zoophycos are notable features of the carbonate facies. In siliciclastic successions (parasequences) the latter ichnotaxa are generally regarded as having been deposited in rather deeper water, but in the carbonate Jubaila Formation are interpreted as being associated with local areas of lower turbulence. Likewise, the hardgrounds and firmgrounds, which have not been traced laterally, are tentatively regarded to be of local significance.

Comparison of ammonium lactate, sodium bicarbonate and double calcium lactate methods for extraction of phosphorus from wetland peat soils

Procedures for routine analysis of soil phosphorus (P) have been used for assessment of P status, distribution and P losses from cultivated mineral soils. No similar studies have been carried out on wetland peat soils. The objective was to compare extraction efficiency of ammonium lactate (PAL), sodium bicarbonate (P-Olsen), and double calcium lactate (P-DCaL) and P distribution in the soil profile of wetland peat soils. For this purpose, 34 samples of the 0-30, 30-60 and 60-90 cm layers were collected from peat soils in Germany, Israel, Poland, Slovenia, Sweden and the United Kingdom and analysed for P. Mean soil pH (CaCl2, 0.01 M) was 5.84, 5.51 and 5.47 in the 0-30, 30-60 and 60-90 cm layers, respectively. The P-DCaL was consistently about half the magnitude of either P-AL or P-Olsen. The efficiency of P extraction increased in the order P-DCaL < P-AL &LE; P-Olsen, with corresponding means (mg kg(-1)) for all soils (34 samples) of 15.32, 33.49 and 34.27 in 0-30 cm; 8.87, 17.30 and 21.46 in 30-60 cm; and 5.69, 14.00 and 21.40 in 60-90 cm. The means decreased with depth. When examining soils for each country separately, P-Olsen was relatively evenly distributed in the German, UK and Slovenian soils. P-Olsen was linearly correlated (r = 0.594, P = 0.0002) with pH, whereas the three P tests (except P-Olsen vs P-DCaL) significantly correlated with each other (P = 0.017850.0001). The strongest correlation (r = 0.617, P = 0.0001) was recorded for P-AL vs P-DCaL) and the two methods were inter-convertible using a regression equation: P-AL = -22.593 + 5.353 pH + 1.423 P-DCaL, R-2 = 0.550.

Geochemical mapping of carbonate sediments in the Makgadikgadi basin, Botswana using moderate resolution remote sensing data

This paper maps the carbonate geochemistry of the Makgadikgadi Pans region of northern Botswana from moderate resolution (500 m pixels) remotely sensed data, to assess the impact of various geomorphological processes on surficial carbonate distribution. Previous palaeo-environmental studies have demonstrated that the pans have experienced several highstands during the Quaternary, forming calcretes around shoreline embayments. The pans are also a significant regional source of dust, and some workers have suggested that surficial carbonate distributions may be controlled, in part, by wind regime. Field studies of carbonate deposits in the region have also highlighted the importance of fluvial and groundwater processes in calcrete formation. However, due to the large area involved and problems of accessibility, the carbonate distribution across the entire Makgadikgadi basin remains poorly understood. The MODIS instrument permits mapping of carbonate distribution over large areas; comparison with estimates from Landsat Thematic Mapper data show reasonable agreement, and there is good agreement with estimates from laboratory analysis of field samples. The results suggest that palaeo-lake highstands, reconstructed here using the SRTM 3 arc-second digital elevation model, have left behind surficial carbonate deposits, which can be mapped by the MODIS instrument. Copyright (c) 2006 John Wiley & Sons, Ltd.

Effect of dietary supplementation with selenium-enriched yeast or sodium selenite on selenium tissue distribution and meat quality in beef cattle

The objective was to determine the concentration of total selenium (Se) and the proportion of total Se comprised as selenomethionine (SeMet) and selenocysteine (SeCys) in post mortem tissues of beef cattle offered diets containing graded additions of selenized enriched yeast (SY) [Saccharomyces cerevisae CNCM I-3060]), or sodium selenite (SS). Oxidative stability and tissue glutathione peroxidase (GSH-Px) activity of edible muscle tissue were assessed 10 d post-mortem. Thirty two beef cattle were offered, for a period of 112 d, a total mixed ration which had either been supplemented with SY (0, 0.15 or 0.35 mg Se/kg DM) or SS (0.15 mg Se/kg DM). At enrollment (0 d) and at 28, 56, 84 and 112 d following enrollment, blood samples were taken for Se and Se species determination, as well as whole blood GSH-Px activity. At the end of the study beef cattle were euthanized and samples of heart, liver, kidney, and skeletal muscle (LM and psoas major) were retained for Se and Se species determination. Tissue GSH-Px activity and thiobarbituric acid reactive substances (TBARS) were determined in skeletal muscle tissue (LM only). The incorporation into the diet of ascending concentrations of Se as SY increased whole blood total Se and the proportion of total Se comprised as SeMet, as well as GSH-Px activity. There was also a dose dependant response to the graded addition of SY on total Se and proportion of total Se as SeMet in all tissues and GSH-Px activity in skeletal muscle tissue. Furthermore, total Se concentration of whole blood and tissues was greater in those animals offered SY when compared with those receiving a comparable dose of SS, indicating an improvement in Se availability and tissue Se retention. Likewise, GSH-Px activity in whole blood and LM was greater in those animals offered SY when compared with those receiving a comparable dose of SS. However, these increases in tissue total Se and GSH-Px activity appeared to have little or no effect in meat oxidative stability.

Effects of dietary supplementation with selenium enriched yeast or sodium selenite on selenium tissue distribution and meat quality in lambs

The objective was to determine the concentration of total selenium (Se) and the proportion of total Se comprised as selenomethionine (SeMet) and selenocysteine (SeCys), as well as meat quality in terms of oxidative stability in post mortem tissues of lambs offered diets with an increasing dose rate of selenized enriched yeast (SY), or sodium selenite (SS). Fifty lambs were offered, for a period of 112 d, a total mixed ration which had either been supplemented with SY (0, 0.11, 0.21 or 0.31 mg/kg DM to give total Se contents of 0.19, 0.3, 0.4 and 0.5 mg Se/kg DM for treatments T1, T2, T3 and T4, respectively) or SS (0.11 mg/kg DM to give 0.3 mg Se/kg DM total Se [T5]). At enrolment and at 28, 56, 84 and 112 d following enrolment, blood samples were taken for Se and Se species determination, as well as glutathione peroxidase (GSH-Px) activity. At the end of the study lambs were euthanased and samples of heart, liver, kidney, and skeletal muscle were retained for Se and Se species determination. Tissue GSH-Px activity and thiobarbituric acid reactive substances (TBARS) were determined in Longissimus Thoracis. The incorporation into the diet of ascending concentrations of Se as SY increased whole blood total Se and the proportion of total Se comprised as SeMet, and erythrocyte GSH-Px activity. Comparable doses of SS supplementation did not result in significant differences between these parameters. With the exception of kidney tissue, all other tissues showed a dose dependant response to increasing concentrations of dietary SY, such that total Se and SeMet increased. Selenium content of Psoas Major was higher in animals fed SY when compared to a similar dose of SS, indicating improvements in Se availability and retention. There were no significant treatment effects on meat quality assessments GHS-Px and TBARS, reflecting the lack of difference in the proportion of total Se that was comprised as SeCys. However, oxidative stability improved marginally with ascending tissue Se content, providing an indication of a linear dose response whereby TBARS improved with ascending SY inclusion.

Development of a capillary electrophoresis method for the simultaneous analysis of artificial sweeteners, preservatives and colours in soft drinks

A rapid capillary electrophoresis method was developed simultaneously to determine artificial sweeteners, preservatives and colours used as additives in carbonated soft drinks. Resolution between all additives occurring together in soft drinks was successfully achieved within a 15-min run-time by employing the micellar electrokinetic chromatography mode with a 20 mM carbonate buffer at pH 9.5 as the aqueous phase and 62 mM sodium dodecyl sulfate as the micellar phase. By using a diode-array detector to monitor the UV-visible range (190-600 nm), the identity of sample components, suggested by migration time, could be confirmed by spectral matching relative to standards.