Regulatory activation is accompanied by movement in the C terminus of the Na-K-Cl cotransporter (NKCC1)

The Na-K-Cl cotransporter (NKCC1) is expressed in most vertebrate cells and is crucial in the regulation of cell volume and intracellular chloride concentration. To study the structure and function of NKCC1, we tagged the transporter with cyan (CFP) and yellow (YFP) fluorescent proteins at two sites within the C terminus and measured fluorescence resonance energy transfer (FRET) in stably expressing human embryonic kidney cell lines. Both singly and doubly tagged NKCC1s were appropriately produced, trafficked to the plasma membrane, and exhibited (86)Rb transport activity. When both fluorescent probes were placed within the same C terminus of an NKCC1 transporter, we recorded an 11% FRET decrease upon activation of the transporter. This result clearly demonstrates movement of the C terminus during the regulatory response to phosphorylation of the N terminus. When we introduced CFP and YFP separately in different NKCC1 constructs and cotransfected these in HEK cells, we observed FRET between dimer pairs, and the fractional FRET decrease upon transporter activation was 46%. Quantitatively, this indicates that the largest FRET-signaled movement is between dimer pairs, an observation supported by further experiments in which the doubly tagged construct was cotransfectionally diluted with untagged NKCC1. Our results demonstrate that regulation of NKCC1 is accompanied by a large movement between two positions in the C termini of a dimeric cotransporter. We suggest that the NKCC1 C terminus is involved in transport regulation and that dimerization may play a key structural role in the regulatory process. It is anticipated that when combined with structural information, our findings will provide a model for understanding the conformational changes that bring about NKCC1 regulation.     

Bio-informatics based analysis of genes implicated in alcohol mediated liver injury

Alcohol induced liver injury has been studied extensively. Using literature search and bioinformatics tools, the present study characterizes the genes involved in alcohol induced liver injury. The cellular and metabolic processes in which genes involved in alcohol induced liver injury are implicated are also discussed. The genes related to alcohol induced liver injury are also involved in affecting certain molecular functions and metabolism of drugs, besides being associated with diseases. In conclusion, the changes in regulation of genes implicated in alcohol induced liver injury apart from causing alcohol mediated hepatic dysfunction may affect other vital processes in the body.     

The dynamic viscoelasticity and plasticizer leachability of tissue conditioners

doi: 10.1111/j.1741‐2358.2012.00639.x
The dynamic viscoelasticity and plasticizer leachability of tissue conditioners Objectives: Dynamic viscoelasticity is one of the most important characteristics of tissue conditioners. Leaching of plasticizer from tissue conditioners may cause changes in the dynamic viscoelasticity of these materials. The purpose of this study was to determine the dynamic viscoelasticity and plasticizer leachability of commercial tissue conditioners. Materials and methods: Five commercial tissue conditioners were used in this study: COE Comfort (CC), Fit Softer (FS), Hydro‐Cast (HC), Soft Conditioner (SC) and Visco‐Gel (VG). Five specimens of each material were stored in 37°C distilled water. The dynamic viscoelasticity and plasticizer leaching of each specimen were measured at 0, 1, 3, 7 and 14 days after immersion using a dynamic mechanical analyzer and high performance liquid chromatography. Data were analyzed using one‐way anova and Student–Newman–Keuls test (α = 0.05). Results: Significant differences in dynamic viscoelasticity and plasticizer leaching were found among the materials. The dynamic viscoelasticity of all materials increased or decreased significantly with time. Rapid changes in dynamic viscoelasticity were evident on day 1 after water immersion. The material CC recorded the highest level of plasticizer leaching. Conclusion: These results suggest that the dynamic viscoelasticity of commercial tissue conditioners is affected by plasticizer leaching. However, these effects are limited.     

Cationic polypeptide-induced fusion of acidic liposomes

Fusion of acidic liposomes was induced by Mg²⁺, Ca²⁺, polylysine and polymyxin B. The extent of fusion and the concomitant change in liposome permeability induced by divalent cations depended on the concentration of liposomes in the suspension as well as on the cation concentration. In contradistinction, the extent of fusion and the change in permeability induced by the polypeptides depended only on the polycation concentration. The difference in the pattern of interaction, between the liposomes and the various cations, is a result of different binding affinities. The binding of the polypeptides to the liposomes, in contrast to divalent cations, is practically irreversible. The potential of polylysine to induce fusion of acidic phosphatidylethanolamine-devoid liposomes was used to demonstrate that in order to obtain fusion, both membranes involved must be susceptible, at least to a certain degree, to fusion by the proper inducer. When lysophosphatidylcholine substituted for phosphatidylcholine in phosphatidylethanolamine-rich acidic liposomes, extensive polylysine-induced fusion was obtained without concomitant spillage of the liposome contents.     

Bioleaching of a Complex Co-Ni-Cu Sulfide Flotation Concentrate by Bacillus megaterium QM B1551 at Neutral pH

Bioleaching of sulfide minerals at neutral pH has been rarely reported. In this study, a bacterium, Bacillus megaterium QM B1551, was isolated from Jinchuan sulfide tailings and used to leach a complex sulfide flotation concentrate for the extraction of Co²⁺, Ni²⁺ and Cu²⁺ at near neutral pH. A total of 38.2% Co, 44.7% Ni and 3.6% Cu were extracted from the sulfide concentrate in 5 days with an initial pH of 6. An enhanced Co²⁺, Ni²⁺ and Cu²⁺ extraction extent was achieved by first bioleaching the concentrate with Bacillus megaterium QM B1551 at 35°C and then followed by chemical leaching with 4 M sulfuric acid at 90°C. As a result, a total of 60.7% Co²⁺, 76.3% Ni²⁺ and 39.8% Cu²⁺ were extracted. On an industrial scale, the profits from the metal recovery by such a combined leaching procedure are optimum if considering the cost-benefit ratio.     

Metal Leaching and Reductive Dissolution of Iron from Contaminated Soil and Sediment Samples by Indigenous Bacteria and Bacillus Isolates

The purpose of this study was to leach Cu, Zn, As, and Fe from contaminated soil and sediment samples with indigenous heterotrophic bacteria isolated from the study sites. The sediment contained Fe in the form of goethite and low concentrations of other metals. The soil contained hematite and high concentrations of other metals. The environmental conditions affected the bacterial activity in the metals dissolution. As and Fe were the major metals leached from the sediment sample while a minor fraction of Cu was solubilized. Cu and Zn were the major metals leached from the soil sample while only a minor fraction of Fe was dissolved. As a control, a disinfectant was used for partial inactivation of indigenous bacteria. This treatment had a negative effect on the leaching of Fe, Zn and As from soil and sediment samples, but it increased Cu dissolution from the sediment. Bacterial different dissolution of Fe during soil and sediment bioleaching was also investigated with ferrihydrite. The iron concentration was much higher during ferrihydrite dissolution when indigenous bacteria from sediment were used compared to indigenous bacteria isolated from soil. The indigenous bacterial inoculum provided more biological and metabolic diversity which may account for the difference in reductive iron reduction from ferrihydrite. The Bacillus cultures isolated from soil and sediment samples showed similar efficiencies in reductive dissolution of ferrihydrite. The synergetic bacterial inhibition effect created by the environmental conditions can influence bioremediation effect.     

Acid-Generating and Leaching Potential of Soils in a Coal Waste Rock Pile in Northeastern China

The present study involved the assessment of potential generation of acid drainage and also metal leaching from an abandoned large (175 m) low sulfur waste rock pile—the dominant mine waste at the site—at the Haizhou coal mine. Laboratory-based static and column leaching tests on waste rock samples were conducted. The static tests were done for 8 composite samples collected from different parts of pile. A column study was performed using mixture of waste rock soil samples to assess metals attenuation process of background soils in vicinity of pile when rainwater reacts with low-sulfide waste rocks. Total concentration of major elements in solid samples was determined using dispersive X-ray fluorescence spectrometry (XRF) and chemical analysis of leachate were measured by inductively coupled plasma optical emission spectrometry (ICP-OES), ion chromatography and titration methods. The inverse geochemical modeling using PHREEQC was applied to explain possible mass transfer processes between column leachates of waste pile and background soils. Static tests (including acid–base accounting (ABA) and net acid generation (NAG)) and mineralogical information suggest that the waste rock is non-acid generating. The large amount of aluminosilicate minerals and probably trace amounts of carbonates with respect to low sulfur content of waste rock pile represents a potentially large source of neutralization potential according to static test results. It was also found that presence of inherent neutralizing materials in waste rock and also in surrounding background soils provide sufficient neutrality and possibly immobilize the trace and heavy metal contents of waste rocks and potentially protects water resources.     

Nutrient and salt relations of <Emphasis Type="Italic">Pterocarpus officinalis</Emphasis> L. in coastal wetlands of the Caribbean: assessment through leaf and soil analyses

Pterocarpus officinalis L. is a dominant tree of freshwater coastal wetlands in the Caribbean and the Guiana regions. It is frequently associated with mangroves in areas with high rainfall and/or surface run-off. We hypothesized that P. officinalis is a freshwater swamp species that when occurring in association with mangroves occupies low-salinity soil microsites, or alternatively that it possesses mechanisms preventing accumulation of salt in photosynthetic tissues. To test this we compared the mineral composition of soils and leaves of several species in two Pterocarpus forests in Puerto Rico associated with coastal mangroves, Sabana Seca and Punta Viento. Results indicate that (1) Sabana Seca has low soil salinity values even in the P. officinalis and Laguncularia racemosa mixed zone. In Punta Viento, salinity in the mixed zone was higher than in the Pterocarpus forest, but much lower than in the mangrove area; (2) In both forests, leaves of P. officinalis showed much lower Na concentrations than mangrove species. The K/Na ratios were 16–20 times higher in P. officinalis, indicating preferential absorption of K against Na. The mangrove fern (Acrostichum aureum) growing side by side with P. officinalis and L. racemosa in Punta Viento also revealed high Na exclusion capacity. We found an asymmetric distribution of cations in the blade and rachis of the P. officinalis compound leaves in both sites. The rachis accumulates more Na and Ca, but less Mg than the leaf blade. This sequestration of Na in the rachis prevents salt damage of photosynthetic tissue in the leaf blade.     

A model to predict transformations and losses of nitrogen in UK pastures grazed by beef cattle

The model simulates the cycling of N in grassland systems grazed by beef cattle and predicts the annual amount of N in liveweight gain, and the amounts lost through ammonia volatilization, denitrification and leaching, on the basis of fertilizer application and soil and site characteristics. It aims to provide a better understanding of the way in which these various factors interact in their influence on N transformations. The model has been programmed to run on IBM-compatible personal computers and responds rapidly to changes in input parameters. The model has been constructed from the average annual amounts of N passing through various components of the N cycle in ten field systems grazed by beef cattle. The amounts were either measured directly or were calculated from empirical sub-models, assuming a balance between inputs to, and outputs from the soil inorganic N pool. The model is given wide applicability through the inclusion of a mineralization sub-model which is sensitive to soil texture, sward age, previous cropping history, and climatic zone. Another important sub-model determines the partitioning of soil inorganic N to either plant uptake or the processes of loss: the proportion partitioned to plant uptake decreases as the total amount of soil inorganic N increases. Outputs from the model indicate that fertilizer N has a strong influence on ammonia volatilization, denitrification and leaching at a given site but that, over a range of sites with a given rate of fertilizer N, total loss and the proportions lost by the three processes are greatly influenced by the amount of N mineralized by the soil. The model indicates how fertilizer N should be matched with mineralization to limit gaseous and leaching losses and to achieve optimum efficiency of N use in grazing systems.     

Southeast Asian AE1 associated renal tubular acidosis: Cation leak is a class effect

Anion Exchanger 1 (AE1) is present in the erythrocyte and also in the α-intercalated cell; different mutations can cause either red cell disease or distal renal tubular acidosis (dRTA). Recently, we described a cation leak property in four dRTA-causing AE1 mutants, three autosomal dominant (AD) European mutants, one autosomal recessive (AR) from Southeast Asia, G701D. G701D had a very large leak property and is unusually common in SE Asia. We hypothesized that this property might confer a survival advantage. We characterized three other AR dRTA-associated AE1 mutants found in SE Asia, S773P, Δ850 and A858D via transport experiments in AE1-expressing Xenopus oocytes. These three SE Asian mutants also had cation leaks of similar magnitude to that seen in G701D, a property that distinguishes them as a discrete group. The clustering of these cation-leaky AE1 mutations to malarious areas of SE Asia suggests that they may confer malaria resistance.