Phosphate transport in mitochondria: Past accomplishments,present problems,and future challenges

The requirement of inorganic phosphate (P_i) for oxidative phosphorylation in eukaryotic cells is fulfilled through specific P_i transport systems. The mitochondrial proton/phosphate symporter (P_ic) is a membrane-embedded protein which translocates P_i from the cytosol into the mitochondrial matrix. P_ic is responsible for the very rapid transport of most of the P_i used in ATP synthesis. During the past five years there have been advances on several fronts. Genomic and cDNA clones for yeast, bovine, rat, and human P_ic have been isolated and sequenced. Functional expression of yeast P_ic in yeast strains deficient in P_i transport and expression inEscherichia coli of a chimera protein involving P_ic and ATP synthase subunit have been accomplished. P_ic, in contrast to other members of the family of transporters involved in energy metabolism, was demonstrated to have a presequence, which optimizes the import of the precursor protein into mitochondria. Six transmembrane segments appear to be a structural feature shared between P_ic and other mitochondrial anion carriers, and recent-site directed mutagenesis studies implicate structure-functional relationships to bacteriorhodopsin. These recent advances on P_ic will be assessed in light of a more global interpretation of transport mechanism across the inner mitochondrial membrane.     

Benthic invertebrates of some saline lakes of the Sud Lipez region,Bolivia

The benthic invertebrates fauna of most of the saline lakes of the Sud Lipez region (Bolivia, Altiplano) has been until now quite unstudied. Samples collected during an extensive survey of 12 lakes and two small inflow rivers allow a first list of the main macroinvertebrates living in these biotopes.The heterogeneous nature of these saline lakes with their freshwater springs and phreatic inflows offers a variety of habitats to macroinvertebrates. The benthic fauna in lakes with salinities > 10 g l^–1 is not so low in density but includes few species and is dominated by Orthocladiinae and Podonominae larvae. In contrast, the freshwater springs and inflows are colonized by a diverse fauna, with a mixture of both freshwater and saline taxa, but dominated by Elmidae and Amphipoda. The lakes are quite isolated and, apart from some cosmopolitan organisms, their fauna can be quite distinctive.     

Seasonal variations of P compounds and their concentrations in two coastal lagoons (Herault,France)

This article concerns seasonal variations in the phosphate concentrations in two coastal lagoons near Montpellier (Mediterranean coast, France). The o-P concentration in the overlying water is highest during summer. The role of the sediment, particularly that of the different P fractions in the sediment, is discussed. Significant variations, especially in the FeOOH ≈ P fraction, occur. For both Tot-P_sed and the Fe00H≈P fraction a gradient from surface to bottom is observed, as well as a distinct decrease in the FeOOH≈P fraction in the surface sediments during summer and autumn. Variations in the FeOOH≈P fraction appear to be compensated by variations in the CaC0₃≈P fraction. These variations appear to be determined by the ferric hydroxide concentration. This compound represents only a small part (maximally 15%) of the total iron in the sediments and is related to the dissolved oxygen content of the immediately overlying water. Besides the fractions o-P, Fe(OOH)≈P, a large part of the CaC0₃≈P fraction is potentially bioavailable. A large proportion of the Tot-P_sed is therefore bioavailable.[/p]     

A population model for the alkali fly at Mono Lake: depth distribution and changing habitat availability

The densities of alkali fly larvae and pupae were measured in relation to depth and substrate type at six locations around Mono Lake. Samples representing a mixture of different bottom features were taken to a depth of 10 m (33 ft) using SCUBA. This is at or near the depth limit of fly larvae and pupae. The biomass of larvae and pupae on hard substrate were maximum and approximately equal at depths of 0.5 m and 1 m, substantially lower at intermediate depths of 3 m and 5 m, and over an order of magnitude further reduced at 10 m. Densities of flies on hard or rocky substrates (mainly calcareous tufa deposits), were significantly greater than those found on soft substrates such as mud or sand, at all but the greatest depth surveyed.Bathymetric maps of the areas of hard and soft substrate occurring at different lake depths were used to estimate the fly population size over the whole lake, based on the density distribution of larvae and pupae with depth on different substrates. The mapped areas of soft and hard substrates were also calculated for different lake levels, and applying the same procedure, a population model comparing the abundance of flies at different lake levels was developed. This habitat-based population model predicts that the abundance of the alkali fly is maximized at 6380 ft (1945 m) lake surface elevation. Most of the tufa substrate submerged at this lake level will become exposed and unavailable as habitat as the lake declines to 6370 ft (1942 m). In late 1991, the lake level was just over 6374 ft (1943 + m).     

Changes in intracellular pH during repeated exercise

The rates of change in intracellular pH during repeated exercise sessions with rest periods was determined by 31 phosphorus-nuclear magnetic resonance spectroscopy (³¹P-MRS). Five long-distance runners and six healthy male subjects as controls performed a 2-min femoral flexion at 20 kg · m · min^–1 in a 2.1 T superconducting magnet with a 67-cm bore and repeated this exercise four times with 2-min rest periods intervening. In all cases during exercise the inorganic phosphate (P_i) peak split into two, the earlier increased rapidly (high-pH P_i) and the later (low-pH P_i) increased more slowly. The P_i peaks were separated by a fitting procedure using the least square mean method. The high-pH P_i area during exercise decreased as the number of repeated exercise periods increased, while the low-pH P_i area gradually increased. Although the total P_i area decreased exponentially during the recovery period, the high-pH P_i area decreased first and then the low-pH P_i area reduced gradually. The pH values were estimated from the chemical shift between the phosphocreatine peak and each split peak in the P_i. The high-pH in pooled data ranged from 6.6 to 7.0 during exercise and recovery, while the low pH decreased to 6.2 during exercise. As the number of exercise periods increased, each pH value gradually became less acidic, although there was a tendency to more acidity in the control subjects than in the long-distance runners. In conclusion, it was possible to obtain by non-invasive, continuous³¹P-MRS, a split pattern of P_i peaks during exercise and there were at least tow different intracellular pH values during exercise, suggesting that each P_i peak might be attributed to the types of muscle fibre recruited.     

Regional differences in glutaminase activation by phosphate and calcium in rat brain: Impairment in aged rats and implications for regional glutaminase isozymes

Regional regulation of glutaminase by phosphate and calcium was examined in the temporal cortex (TCX), striatum (STR) and hippocampus (HIPP) from adult and aged male F344 rats. Phosphate-dependent glutaminase activity in adult rats was significantly lower (35–43%) in the HIPP (100 and 150 mM) and STR (150 mM) compared to PAG activity in the TCX. Phosphate activation in aged rats was 50–60% lower in the HIPP at concentrations greater than 25 mM compared to the aged TCX or STR. PAG activity in the TCX and STR was unaffected by age, but was significantly reduced (30–50%) in the HIPP from aged rats at phosphate concentrations of 25 mM and greater when compared to adult rats. In adult rats at concentrations of CaCl₂ above 1 mM, PAG activity was significantly lower (60–75%) in the STR and HIPP when compared to the TCX. In aged rats, PAG activity (1 mM CaCl₂) in the HIPP was significantly less (50%) than STR PAG activity in aged rats. Diminished PAG activity was seen only in the TCX (2.5 mM; 32%), and the HIPP (0.5 mM; 25% and 1 mM; 38%) at higher calcium concentrations compared to adult. Phosphate-independent calcium activation of PAG occurred in the HIPP but not in either the TCX or the STR. Addition of phosphate resulted in a synergistic activation of PAG in the STR and TCX, but not in the HIPP. These findings suggest that PAG is regionally regulated by phosphate and calcium, and this regulation is impaired in aged rats. These data also support the hypothesis that isozymes of PAG exist with different regulatory properties.Abbreviation PAG Phosphate-activated glutaminase - L-glutamine amidohydrolase EC 3.5.1.2 - TCX temporal cortex - STR striatum - HIPP hippocampus - F344 Fischer-344 rat     

The role of mineral compounds and chemical conditions in the binding of phosphate in the Ems estuary

In the Ems estuary the gradients in dissolved phosphorus and iron are non-conservative and opposite. The relations between different mineral compounds, environmental conditions and phosphate concentrations were studied, in an attempt to explain this. Laboratory experiments on water samples from the reaches with high concentrations of phosphate and of suspended matter revealed a low concentration of calcite-bound phosphorus in the middle reaches of the estuary (only 0.07%). Therefore, it is concluded that calcite possibly plays only a modest role in phosphate distribution in the Ems estuary. Further experiments revealed that nearly 70% of the particulate inorganic phosphorus is iron-bound. It is demonstrated that organic coatings on minerals inhibit phosphate release at a low redox potential. Moreover, the organic matter itself contained c. 20% of particulate phosphorus. Approximately 10% of the particulate phosphorus may be associated with clay minerals. It is concluded that in addition to biological processes, the phosphorus associated with ironoxyhydroxides may be crucial for the seasonal variation of the phosphate concentrations in the water.     

Nanodiamond-promoted white light emission in Eu,Dy, and Cu-containing phosphate glass: quantitative analysis and colorimetric study

A thorough analysis of glass containing Eu₂O₃ and Dy₂O₃, or Eu₂O₃, Dy₂O₃, and CuO melted together with nanodiamond powder was pursued based on measurements of optical absorption, photoluminescence (PL) emission and excitation spectra, and colorimetry. Nanodiamond facilitated the stabilization of Cu⁺ and Eu²⁺ ions with blue-emitting characteristics that, along with yellow-emitting Dy³⁺ and red-emitting Eu³⁺ led to the white light-emitting glass. Novel intensity notations implemented in intensity-based spectral ratios, and difference intensity correlation analysis were proposed for the assessment of PL properties. The chromaticity and correlated colour temperature of the emission were ultimately investigated as a two-parametric problem based on: (1) the different ionic components; and (2) the various excitation wavelengths employed. The optical analysis approach adds to the characterization methods to further fundamental understanding and provide helpful analytical tools for designing materials for tunable white light-emitting devices.     

Environmentally friendly protocol for the determination of sitagliptin phosphate in pharmaceutical preparations and biological fluids using l-tyrosine as a fluorescence probe

A responsive spectrofluorometric method was developed for the determination of sitagliptin phosphate using l -tyrosine as a fluorescence probe. The fluorescence intensity of l -tyrosine was quenched with sitagliptin phosphate. The fluorescence intensity was recorded at 307 nm using a 272 nm excitation wavelength. The calibration plot between fluorescence intensity and the concentration of drug was linear in the range of 0.1 to 2.0 mM with a good correlation value of 0.997. The limit of detection and quantification were established to be 3.7 × 10⁻⁴ and 1.23 × 10⁻³ mM, respectively. Commonly used excipients did not interfere with sitagliptin phosphate measurement. The proposed method was used to measure the sitagliptin phosphate in its standard type, dosage form, and biological samples. The percent recovery ranged from 97.41–103.36%. The static quenching was shown to be responsible for quenching as indicated by the Stern–Volmer plot. The method was validated using ICH guidelines and profitably applied for the content uniformity test, resulting in a high percent recovery and small relative standard deviation. The proposed approach is effortless, susceptible, selective, economic, and provides a high precision and accuracy, and can be used to determine sitagliptin phosphate in the pharmaceutical industry.