Recent advances and challenges in the recovery and purification of cellular exosomes

Exosomes are nanovesicles secreted by most cellular types that carry important biochemical compounds throughout the body with different purposes, playing a preponderant role in cellular communication. Because of their structure, physicochemical properties and stability, recent studies are focusing in their use as nanocarriers for different therapeutic compounds for the treatment of different diseases ranging from cancer to Parkinson's disease. However, current bioseparation protocols and methodologies are selected based on the final exosome application or intended use and present both advantages and disadvantages when compared among them. In this context, this review aims to present the most important technologies available for exosome isolation while discussing their advantages and disadvantages and the possibilities of being combined with other strategies. This is critical since the development of novel exosome‐based therapeutic strategies will be constrained to the effectiveness and yield of the selected downstream purification methodologies for which a thorough understanding of the available technological resources is needed.     

Non-isothermal short-range-order kinetics of binary alloys as influenced by solute-vacancy complexes

A model describing the roles of bound and unbound vacancies is proposed in order to predict defect decay and short-range-order kinetics of quenched binary alloys during linear heating experiments. This is an alternative treatment of a previous approach. The model has been applied to the differential scanning calorimetry (DSC) curves of Cu-5 at.% Zn quenched from different temperatures. An expression to calculate the activation energy for migration of solute-vacancy complexes was also developed which make use of DSC trace data. A value of 89.12±0.32 kJ mol^-1 was obtained for the above alloy. The relative contribution of bound and unbound vacancies to partition of effective activation energy corresponding to the ordering process as influenced by quenching temperature was also assessed. This revised version was published online in July 2006 with corrections to the Cover Date.     

Fleet configuration subject to stochastic demand: an application in the distribution of liquefied petroleum gas

We study the problem of configuring a fleet, in which vehicles receive information on-line about the demand that they should fulfil while they are on the road. In each district it must be decided the number of vehicles and their capacity. The objective function is to minimise the operational cost subject to constraints for the minimum delivery capacity, the maximum vehicle size and the average waiting time for customers. The last constraint is modelled as a queuing system that is adjusted according to the simulation of the delivery process of a Chilean company that distributes liquefied petroleum gas in portable cylinders. We provide the analytical form of all the components of the model, so it can be solved using a standard non-linear programming package. We show that the fleet may increase its sales by 3% and reduce the waiting time of customers 10% by allowing a set of vehicles to share the buffer of orders rather than having vehicles to exclusively serve smaller sectors.     

The oscillopolarographic determination of uranium and thorium in supporting electrolytes containing cupferron

Uranium(VI and IV) and thorium(IV) give cathodic indentations in supporting electrolytes prepared from 0.1M perchloric acid, 0.5 M ammonium thiocyanate and 5·10^-3M cupferron (solution A) or from 0.1 M succinic-succinate buffer pH 4, 0.1 M sodium chloride, 10^-3M cupferron and 0.05% gelatine (solution B). The uranium indentation on the dE/dt = f(E) curve (Q=0.75 and 0.73) permits its detection at the 3·10^-7M level. The thorium indentation (Q=0.78) permits its detection at the 4·10^-7M level in solution B. Methods for the elimination of interfering ions for the uranium determination are described. In the determination of thorium, Ga(III), Fe(III), Ti(IV) and U(VI) interfere.     

SOLVENT EFFECTS ON THE TAUTOMERIZATION OF 5′-DEOXYPYRIDOXAL. A PHOTOPHYSICAL STUDY

Abstract— Absorption and fluorescence spectra of 5′-deoxypyridoxal (DPL) in various pure solvents and mixtures were recorded both at room temperature and over the range10–65°C. The areas under the absorption bands were analyzed to obtain the mole fraction (f_N, f_z) of two tautomers (the zwitterionic, Z, and neutral, N, forms) in the ground state. The following spectral parameters were determined from the fluorescence spectra: Stokes shift (Δv), fluorescence quantum yield of the neutral form (Q_N), fluorescence ratio of the neutral to the zwitterionic form (ø_N/ø_Z) and the rate constant of tautomerization (k₁) from Z to N in the excited state. Some of these parameters (f_N, Δv, Q_N, k₁) were found to depend on the proton donor character of the solvent, whereas others (ø_N/ø_Z) depended on its dipole moment. Thus, the absorption and fluorescence spectra of DPL allow one to obtain information on the polarity and the concentration of –OH groups on its environment.     

Thermal behavior of short-range-order in quenched Cu-12 at% Mn assessed by DSC

Summary The ordering behavior of quenched Cu-12 at% Mn was investigated by differential scanning calorimetry (DSC) under increasing temperature conditions. The ordering process can be better explained in terms of a homogeneous short-range-order (SRO) rather than a disperse-short-range-order (DSRO) model as for concentrated Cu-γMn solid solutions. At the employed high quench rates ordering is established in one stage here termed stage 1, assisted by excess vacancies. This stage obeys a first order kinetics law, being the effective activation energy consistent with a weighed average sum of the activation energy for migration of free and bound vacancies. An estimated solute-vacancy binding energy of 15 kJ mol^-1 seems quite reasonable for this alloy together with the assessed activation energy for complex migration of 92.6 kJ mol^-1. By adopting a first SRO order parameter based in quasi-chemical theory in pair approximation and using boundary values calculated from displayed features of DSC traces, the retained degree of quenched-in order at room temperature was calculated. This procedure also enabled to estimate an ordering energy of -2.7 kJ mol^-1. The effect of quenching temperature demonstrate that for smaller vacancy sink densities, the retained degree of order at room temperature goes through a minimum.     

Degradation of Cephaloridine on Alkaline Hydrolysis

A kinetic study on the alkaline hydrolysis of cephaloridine ( 1 ) at pH 10.5 and 37° was carried out using ion-pair reversed-phase HPLC. The main resulting degradation products, the 7-epimer 2 of 1 , the Δ²-isomer 3 of 1 , and the 3-methylidene compound 4 were identified. The presence of a pyridinio group at C(3¹) results in a slightly increased formation constant for the 3-methylidene compound 4 and the 7-epimer 2, and introduces a new reaction: the isomerization of the double bond at C(3) in the dihydrothiazine ring to C(2).     

Novel Cleavage of Propargylamines by Reaction with Organolithium Compounds

Treatment of secondary aliphatic 2-bromoallylamines with an excess of an organolithium compound led to saturated amines in which the organic group of the organolithium compound is incorporated at the alpha carbon. On the other hand, the successive reaction of the former amines with BuLi and t-BuLi between -80 degrees C and rt gave 1,3-diamines or hexahydropyrimidines depending on the reaction time. The formation of these unexpected products involves initial generation of lithium propargylamides, which subsequently undergo cleavage of the C propargylic-C acetylenic bond induced by the organolithium present in each case in the reaction medium. A mechanism which takes into account all the different reaction products has been proposed and additionally supported by successful trapping of dilithium acetylide.     

Flow-through microdispenser for interfacing micro-HPLC to Raman and mid-IR spectroscopic detection

A flow-through microdispenser has been coupled to a micro HPLC separation system and used as a solvent elimination interface for Fourier transform infrared (FTIR) and Raman spectroscopic detection of the separated compounds. Using the microdispenser picoliter sized droplets can be generated and deposited on an appropriate target placed on a computerized x, y-stage. Evaporation of volatile solvent and buffer is rapid and allows analysis of the obtained dry deposits by various techniques. Due to the destruction free character of Raman and FTIR spectroscopy they can be applied sequentially to interrogate the same deposit. In the reported application five phenolic acids typically present in wine have been separated on a C-18 column technique using a mixture of water, methanol and acetic acid as mobile phase. For spectrum acquisition infrared and Raman microscopes have been used. The spectra recorded from the dried deposits of the separated compounds agreed well with the reference spectra of corresponding components.