Insights into the mechanism of human erythrocyte hexose transport : a transferred NOE study of glucose binding to GLUT1

This study examines binding of α- and β-D-glucose in their equilibrium mixture to the glucose transporter (GLUT1) in human erythrocyte membrane preparations by an ^1H NMR method, the transferred NOE (TRNOE). This method is shown theoretically and experimentally to be a sensitive probe of weak ligand-macromolecule interactions. The TRNOEs observed are shown to arise solely from glucose binding to GLUT1. Sites at both membrane faces contribute to the TRNOEs. Binding curves obtained are consistent with a homogeneous class of sugar sites, with an apparent K_D which varies (from ~30 mM to ~70 mM for both anomers) depending on the membrane preparation examined. Preparations with a higher proportion of the cytoplasmic membrane face exposed to bulk solution yield higher apparent KK_Ds. The glucose transport inhibitor cytochalasin B essentially eliminates the TRNOE. Nonlinearity was found in the dependence on sugar concentration of the apparent inhibition constant for cytochalasin B reversal of the TRNOE observed in the α anomer (and probably the β anomer); such nonlinearity implies the existence of ternary complexes of sugar, inhibitor and transporter. The inhibition results furthermore imply the presence of a class of relatively high-affinity (K_D < 2mM) sugar sites specific for the α anomer which do not contribute to NMR-observable binding. The presence of two classes of sugar-sensitive cytochalasin B sites is also indicated. These results are compared with predictions of the alternating conformer model of glucose transport. Variation of apparent K_D in the NMR-observable sites, the formation of ternary complexes and the presence of an anomer-specific site are shown to be inconsistent with this model. An alternate model is developed which reconciles these results with the known transport behavior of GLUT1. In this model, the transporter possesses (at minimum) three classes of sugar sites: (i) transport sites, which are alternately exposed to the cytoplasmic or the extracellular compartment, but never to both simultaneously, (ii) a class of sites (probably relatively low-affinity) which are confined to one compartment, and (iii) the high-affinity α anomer-specific sites, which are confined to the cytoplasmic compartment.

Reaction of glucose catalyzed by framework and extraframework tin sites in zeolite beta

The isomerization of glucose into fructose is a large-scale reaction for the production of high-fructose corn syrup, and is now being considered as an intermediate step in the possible route of biomass conversion into fuels and chemicals. Recently, it has been shown that a hydrophobic, large pore, silica molecular sieve having the zeolite beta structure and containing framework Sn⁴⁺ (Sn-Beta) is able to isomerize glucose into fructose in aqueous media. Here, I have investigated how this catalyst converts glucose to fructose and show that it is analogous to that achieved with metalloenzymes. Specifically, glucose partitions into the molecular sieve in the pyranose form, ring opens to the acyclic form in the presence of the Lewis acid center (framework Sn⁴⁺), isomerizes into the acyclic form of fructose and finally ring closes to yield the furanose product. Akin to the metalloenzyme, the isomerization step proceeds by intramolecular hydride transfer from C2 to C1. Extraframework tin oxides located within hydrophobic channels of the molecular sieve that exclude liquid water can also isomerize glucose to fructose in aqueous media, but do so through a base-catalyzed proton abstraction mechanism. Extraframework tin oxide particles located at the external surface of the molecular sieve crystals or on amorphous silica supports are not active in aqueous media but are able to perform the isomerization in methanol by a base-catalyzed proton abstraction mechanism. Post-synthetic exchange of Na⁺ with Sn-Beta alters the glucose reaction pathway from the 1,2 intramolecular hydrogen shift (isomerization) to produce fructose towards the 1,2 intramolecular carbon shift (epimerization) that forms mannose. Na⁺ remains exchanged onto silanol groups during reaction in methanol solvent, leading to a near complete shift in selectivity towards glucose epimerization to mannose. In contrast, decationation occurs during reaction in aqueous solutions and gradually increases the reaction selectivity to isomerization at the expense of epimerization. Decationation and concomitant changes in selectivity can be eliminated by addition of NaCl to the aqueous reaction solution. Thus, framework tin sites with a proximal silanol group are the active sites for the 1, 2 intramolecular hydride shift in the isomerization of glucose to fructose, while these sites with Na-exchanged silanol group are the active sites for the 1, 2 intramolecular carbon shift in epimerization of glucose to mannose.

Pathways for the uptake of dissolved organic matter (DOM) by aquatic animals

In this article, pathways from freshwater and marine environments are described. DOM is defined operationally as all the organic compounds which pass through a filter of pore size 0.45 microm., those retained on the surface of the filter being particulate organic matter (POM). DOM can be taken up directly by animals by transfer across the body wall, but more commonly DOM is obtained from ingested food. Once ingested POM from food particles are broken down in the gut, small molecules of DOM are released for transfer across the gut wall. Some ingested particles are attacked by micro-organisms living in the gut, thereby making the DOM available to the host animal. The importance of the microbial loop is discussed, as well as aggregation processes between the fractions of DOM which are more obviously particulate in nature. (DBO)

Different patterns of uptake and depuration of cadmium by periphyton community and a grazer species (Physa sp.): A mesocosm evalution

Widespread pollution by heavy metals generated by various industries has serious adverse effects on human health and the environment. Cadmium is a heavy metal recognised as one of the most hazardous environmental pollutants. It is a non-essential and non-beneficial element to organisms, causing toxicity and other deleterious effects on various components of the aquatic environment. The ability of algal periphyton to concentrate cadmium from fresh water is well known. Moreover, periphyton communities are able to accumulate large amounts of cadmium despite its low concentration in fresh water. Many researchers use algal periphyton as an indicator of water quality in aquatic environments. In the present study, the authors ask two basic questions: Does cadmium accumulate along a food chain consisting of the periphyton community and a grazer species (Physa sp.) under semi-natural conditions provided by artificial streams? If not, which one can better indicate the water quality?

A nonlinear systems model for the control mechanism of free fatty acid-glucose metabolosm in normal humans

A mathematical model is proposed in this thesis for the control mechanism of free fatty acid-glucose metabolism in healthy individuals under resting conditions. The objective is to explain in a consistent manner some clinical laboratory observations such as glucose, insulin and free fatty acid responses to intravenous injection of glucose, insulin, etc. Responses up to only about two hours from the beginning of infusion are considered. The model is an extension of the one for glucose homeostasis proposed by Charette, Kadish and Sridhar (Modeling and Control Aspects of Glucose Homeostasis. Mathematical Biosciences, 1969). It is based upon a systems approach and agrees with the current theories of glucose and free fatty acid metabolism. The description is in terms of ordinary differential equations. Validation of the model is based on clinical laboratory data available at the present time. Finally procedures are suggested for systematically identifying the parameters associated with the free fatty acid portion of the model.

Neutral red uptake (NRU) entseguaren optimizazioa lur-zizareen sistema immuneko zelulen kultibo primarioetan

Azken urteotan eman diren kutsatzaile desberdinen gehiegizko isurpen edo erabilpena dela eta, lurzoruetan metal astunak metatu dira eta horrek , kate trofikoaren oinarrian dauden organismoen gainean k altea k sorrarazi d itu. Eisenia fetida zizarea espezie zent inela gisa erabili da lur entsegu ekotoxikologi koet an . Esperimentu honetan Eisenia fetida - ren jariakin zelomikoan dauden eta babes funtzioa duten zelomozitoak pH desberdinetara edo Kadmio bezalako metalera esposatzeak Neutral Red (NR) edo gorri neutro tindatzaile kationikoaren harreran eta fagoz itosi ahalmenean , eragina duen aztertu nahi izan da. Esperimentua in vitro egin zen 3R filosofia jarraituz. Alde batetik, pH desberdinen eragin ari dagokio nez gorri neutroaren harrera ( NRU ) entsegua n erantzun bimodal bat behatu zen eta beste alde bate tik, f agozitatzeko ahalmenari dagokiola zelomozitoak Eagle medioan kultibatzean ez zen aldaketa esangarria ikusi kadmioa ren, kontr olaren eta kontrol positi boaren artean. Bai ordea L - 1 5 medioarekin, non kontrol posit iboa k beste biak baino absorbantzia handiagoa erakutsi zuen. Emaitza hauek erakusten dute zelomozitoe n mintza gradualki kaltetzen doala pH neutrotik aldentzen doan heinean eta puntu batetik aurrera NRU emendatzen da zelomozito batzuen (amebozitoen) fagozitosia aktibatzen d el a ko .

Uptake and factors that influence the use of 'sit less, move more' occupational intervention strategies in Spanish office employees

Background: Little is known about the types of 'sit less, move more' strategies that appeal to office employees, or what factors influence their use. This study assessed the uptake of strategies in Spanish university office employees engaged in an intervention, and those factors that enabled or limited strategy uptake. Methods: The study used a mixed method design. Semi-structured interviews were conducted with academics and administrators (n = 12; 44 +/- 12 mean SD age; 6 women) at three points across the five-month intervention, and data used to identify factors that influenced the uptake of strategies. Employees who finished the intervention then completed a survey rating (n = 88; 42 +/- 8 mean SD age; 51 women) the extent to which strategies were used [never (1) to usually (4)]; additional survey items (generated from interviewee data) rated the impact of factors that enabled or limited strategy uptake [no influence (1) to very strong influence (4)]. Survey score distributions and averages were calculated and findings triangulated with interview data. Results: Relative to baseline, 67% of the sample increased step counts post intervention (n = 59); 60% decreased occupational sitting (n = 53). 'Active work tasks' and 'increases in walking intensity' were the strategies most frequently used by employees (89% and 94% sometimes or usually utilised these strategies); 'walk-talk meetings' and ` lunchtime walking groups' were the least used (80% and 96% hardly ever or never utilised these strategies). 'Sitting time and step count logging' was the most important enabler of behaviour change (mean survey score of 3.1 +/- 0.8); interviewees highlighted the motivational value of being able to view logged data through visual graphics in a dedicated website, and gain feedback on progress against set goals. 'Screen based work' (mean survey score of 3.2 +/- 0.8) was the most significant barrier limiting the uptake of strategies. Inherent time pressures and cultural norms that dictated sedentary work practices limited the adoption of 'walk-talk meetings' and ` lunch time walking groups'. Conclusions: The findings provide practical insights into which strategies and influences practitioners need to target to maximise the impact of 'sit less, move more' occupational intervention strategies.

Uptake and intracytoplasmic storage of pigmented particles by human CD34+stromal cells/telocytes: endocytic property of telocytes

We studied the phagocytic-like capacity of human CD34+ stromal cells/telocytes (TCs). For this, we examined segments of the colon after injection of India ink to help surgeons localize lesions identified at endoscopy. Our results demonstrate that CD34+ TCs have endocytic properties (phagocytic-like TCs: phTCs), with the capacity to uptake and store India ink particles. phTCs conserve the characteristics of TCs (long, thin, bipolar or multipolar, moniliform cytoplasmic processes/telopodes, with linear distribution of the pigment) and maintain their typical distribution. Likewise, they are easily distinguished from pigment-loaded macrophages (CD68+ macrophages, with oval morphology and coarse granules of pigment clustered in their cytoplasm). A few c-kit/CD117+ interstitial cells of Cajal also incorporate pigment and may conserve the phagocytic-like property of their probable TC precursors. CD34+ stromal cells in other locations (skin and periodontal tissues) also have the phagocytic-like capacity to uptake and store pigments (hemosiderin, some components of dental amalgam and melanin). This suggests a function of TCs in general, which may be related to the transfer of macromolecules in these cells. Our ultrastructural observation of melanin-storing stromal cells with characteristics of TCs (telopodes with dichotomous branching pattern) favours this possibility. In conclusion, intestinal TCs have a phagocytic-like property, a function that may be generalized to TCs in other locations. This function (the ability to internalize small particles), together with the capacity of these cells to release extracellular vesicles with macromolecules, could close the cellular bidirectional cooperative circle of informative exchange and intercellular interactions.

Analytical method development for the uptake study of different organic pollutants by crops cultivated in compost-amended soils

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