Removing human immunodeficiency virus (HIV) from human blood using immobilized heparin

Heparin covalently attached to a water-insoluble resin suspended in HIV-infected aqueous buffer or whole blood captures the virus; subsequent physical separation of the immobilized heparin reduced the viral titers by over 80 and 50%, respectively. The detoxification concept has been validated by both circulating an HIV-1 solution through a column packed with the heparin–sepharose beads and successively mixing an HIV-1 solution with fresh beads.     

Fluorometric determination of DNA in Chlamydomonas

The diaminobenzoic acid dihydrochloride fluorescence method has been examined to determine optimal conditions for the assay of DNA in perchloric acid extracts and in pretreated Chlamydomonas cells on glass-fiber filters. Maximal fluorescence yield was obtained by allowing: (a) the perchloric acid extract to react with 4–6% diaminobenzoic acid dihydrochloride for 20–40 min at 60–80°C at about pH 3.0; (b) the pretreated cells on glass-fiber filters to react with a 20% solution of diaminobenzoic acid dihydrochloride for 40 min at 60°C.     

Tryptophan fluorescence studies of plasma fibronectin: effects of environmental factors

Steady state fluorescence measurements have been used to study tryptophan fluorescence of plasma fibronectin. The native protein has an emission maximum at 337 nm with a quantum yield of 0.03. A red shift of emission maximum was observed in 3–5M urea and a further red shift in 7–8M urea. The emission maximum shifted from 337 to 345 nm when the temperature was changed from 30 to 80°C, with a midpoint of thermal denaturation at 58°C. Similarly, the emission maximum shifted from 337 to 345 nm when the solution pH was increased from 9 to 12, with a midpoint of pH transition at 10.6. The results obtained from difference absorption spectroscopy studies suggest that the unfolding of fibronectin at alkaline pH is related at least in part to ionization of tyrosine residues. Since most of the tryptophan residues are in invariant positions in homology sequences, it is suggested here that tryptophan residues are useful intrinsic probes for elucidating fibronectin structure in solution.     

A sustainable process for adsorptive removal of methylene blue onto a food grade mucilage: kinetics,thermodynamics, and equilibrium evaluation

Abstract

Adsorption of dyes onto natural materials like polysaccharides is considered a green chemistry approach for remediation of wastewater. In this work, the polysaccharide isolated from the corm of Colocasia esculenta (L.) Schott or taro tuber (CEM) was utilized for removing methylene blue (MB) from aqueous solution by batch adsorption method. The CEM adsorbent was characterized by FTIR spectroscopy, Brunauer–Emmett–Teller (BET), and scanning electron microscopy (SEM). The solution pH and adsorbent dose have been found to have a significant positive correlation with the adsorptive removal efficiency of CEM for MB dye. The removal efficiency of CEM was found to be 72.35% under the optimum conditions; 20?mg/L initial concentration of dye, 120?mg of adsorbent dose, solution pH 8.5, 311.2?K temperature and 80?min contact time. The adsorption of MB onto CEM followed best the Freundlich isotherm and pseudo-second-order kinetics. The adsorption was thermodynamically favorable and was endothermic in nature. The desorption/adsorption data justifiably indicated the reuse capability of CEM adsorbent for MB adsorption. Hence, CEM may be regarded as an eco-friendly and cost-effective natural adsorbent for MB dye removal from aqueous solution.     

Flow injection–chemiluminescence determination of acyclovir

The chemiluminescence (CL) reaction of acyclovir (ACV)–potassium permanganate, with formaldehyde as an enhancer, was investigated by the flow‐injection system, and a new method is reported for the determination of ACV on the basis of the reaction. The method is rapid, effective and simple for the determination of acyclovir in the range 0.2–80 mg/L, with a limit of detection of 0.06 mg/L (3 S:N), a relative standard deviation (RSD) of 3.7% for the determination of 1.0 mg/L acyclovir solution in 11 repeated measurements. The method has been applied to the determination of acyclovir in pharmaceuticals, with satisfactory results. The possible reaction mechanism is also discussed briefly. Copyright © 2011 John Wiley & Sons, Ltd.     

Survival of canine kidneys after treatment with dimethyl-sulfoxide, freezing at --80 degrees C, and thawing by microwave illumination.

Forty canine kidneys were the subject of this pilot study where control groups perfused with Perfudex plus DMSO (1.4 m), modified Collins' solution with DMSO (1.4 m) and modified Sacks' solution with DMSO (1.4 m) showed little toxicity and life-sustaining conservation. In the experimental group, 16 kidneys were frozen for 15 min to ?80 °C, thawed by microwave illumination, and reimplanted. Of the 16 dogs, eight survived 2–14 months on their single kidney. The technique of inducing freezing by using intra-arterial cold helium and thawing with high-power microwave illumination gave an overall success rate of 50% long-term life-sustaining survival.     

Modification of Oligo (Poly) Nucleotide Phosphomonoester Groups in Aqueous Solutions

Abstract

Selective modification of oligo (poly) nucleotide phosphomonoester groups in an aqueous medium by N-(3-dimethylaminopropyl)-N′-ethylcarbodiimide in the presence of various nucleophilic agents has been investigated. Optimal conditions of the modification by amino- and hydroxycompounds have been found. Based on these studies a general efficient method for preparation of oligo (poly) nucleotide phosphoamidates and phosphodiesters in an aqueous solution has been developed. The method allows to prepare both oligodeoxyribonucleotide derivatives at 3′- and 5′-terminal phosphate groups and oligoribonucleotide derivatives at 5′-terminal phosphate groups with 80–100% yields.     

Action of combined magnetic fields on aqueous solution of glutamic acid: the further development of investigations

In the present work the results of the known investigation of the influence of combined static (40 μT) and alternating (amplitude of 40 nT) parallel magnetic fields on the current through the aqueous solution of glutamic acid, were successfully replicated. Fourteen experiments were carried out by the application of the combined magnetic fields to the solution placed into a Plexiglas reaction vessel at application of static voltage to golden electrodes placed into the solution. Six experiments were carried out by the application of the combined magnetic fields to the solution placed in a Plexiglas reaction vessel, without electrodes, within an electric field, generated by means of a capacitor at the voltage of 27 mV. The frequency of the alternating field was scanned within the bounds of 1.0 Hz including the cyclotron frequency corresponding to a glutamic acid ion and to the applied static magnetic field. In this study the prominent peaks with half-width of ~0.5 Hz and with different heights (till 80 nA) were registered at the alternating magnetic field frequency equal to the cyclotron frequency (4.2 Hz). The general reproducibility of the investigated effects was 70% among the all solutions studied by us and they arose usually after 40–60 min. after preparation of the solution. In some made-up solutions the appearance of instability in the registered current was noted in 30–45 min after the solution preparation. This instability endured for 20–40 min. At the end of such instability period the effects of combined fields action appeared practically every time. The possible mechanisms of revealed effects were discussed on the basis of modern quantum electrodynamics.     

An immuno-marking technique for thrips

Rabbit immunoglobulin G (R‐IgG) was used successfully as an external mark for thrips. Females of both Thrips tabaci Lindeman and Frankliniella occidentalis (Pergande) (Thysanoptera: Thripidae) were marked with 1 mg ml^?1 R‐IgG solution with 1%‘Tween 20’ by the contact exposure method. Determining the retention of the mark was by running the rinsing solution of individual thrips in an enzyme‐linked immunosorbent assay (ELISA). The sandwich ELISA method was used with an additional biotin–avidin step. The threshold for a positive marking score was defined as three times the mean optical density readings of the negative control thrips. Under laboratory conditions, on bean pods, all marked thrips scored positive up to 6 days after marking (DAM). When marked thrips were kept in the laboratory on marigold flowers for 2 days, they all scored positive. When marked and unmarked thrips were placed together on these flowers, the mark was transferred to 10–20% of the unmarked thrips and they became positive. Under field conditions, on sticky traps covered with water‐base glue, 100, 80, and 20% of the marked T. tabaci scored positive by the 3rd, 6th, and 9th DAM, respectively. Under the same conditions 100, 90, and 10% of the marked F. occidentalis scored positive by the 3rd, 6th, and 9th DAM, respectively. The retention of the R‐IgG decreased significantly under conditions of wetness and high humidity. After 6 days on chive plants kept at 80–100% r.h., all marked thrips scored negative while on plants kept at 40–60% r.h., 85% of the marked thrips scored positive. Rabbit IgG can be used as an external marker for thrips. The suitability of this marking method for dispersal studies of these important pests needs to be evaluated.     

Storage of environmental samples for guaranteeing nucleic acid yields for molecular microbiological studies

The purpose of this study is to evaluate whether sample preservation can affect the yield of nucleic acid extracts from environmental samples. Storage of microbial samples was studied using three sediment types of varying carbon contents (10–57% carbon of dry weight). Four different storage solutions were tested at three temperatures. Freezing of samples at ?20 °C or ?80 °C, either without preservative or in phenol–chloroform solution, retained nucleic acid quantities very efficiently. Storage of samples in phenol–chloroform solution at +4 °C also gave good yields except for sediment with extremely high-carbon content. Ethanol and RNAlater® preservation decreased nucleic acid yields drastically at all temperatures. To study how sample preservation may affect the result of microbial community analysis, one type of sediment was selected for length heterogeneity-PCR analysis and PCR cloning of the 16S rRNA genes. Ethanol and RNAlater® preservation caused a slight bias towards certain microbial types in the community analyses shown by underrepresentation of Bacteroidetes, Betaproteobacteria and Gammaproteobacteria-affiliated peak sizes and overrepresentation of Actinobacteria, Chloroflexi and Alphaproteobacteria-affiliated peak sizes. Based on the results of this study, preservation in phenol–chloroform solution can be recommended as an alternative storage method when freezing is not possible such as during extended field sampling; however, ethanol and RNAlater® may cause serious problems when used as preservatives for environmental samples containing humic acids.     

Liquid chromatography–fast atom bombardment mass spectrometry for detection and determination of pentazocine in human tissues

A reliable and sensitive method was developed for the detection and determination of pentazocine in human solid tissues using liquid chromatography–dynamic fast atom bombardment (FAB) mass spectrometry, combined with a three-step liquid–liquid extraction procedure. Levallorphan tartrate served as an internal standard. The extract was evaporated to dryness and dissolved in the mobile phase, acetonitrile–10 mM ammonium acetate solution (20:80, pH 4.0) containing 0.5% glycerol as FAB matrix. The eluent was pumped at a flow rate of 25 μl/min and split before introduction to FAB mass spectrometer. Quantitative analysis was carried out by means of monitoring quasi-molecular ions with m/z 286 for pentazocine and m/z 284 for levallorphan. The lower limit of detection of pentazocine in each tissue tested was 1 ng/g with scan mode and 0.1 ng/g with SIM mode. Using this method, the concentrations of pentazocine were determined in the tissues of an autopsied individual to perform toxicological evaluation.     

In vitro assembly complex formation of TRAIP CC and RAP 80 zinc finger motif revealed by our study

BackgroundTumor necrosis factor interacting protein (TRAIP/TRIP) is an important cell-signaling molecule that prevents the TNF-induced-nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) activation via direct interaction with TRAF 2 protein. TRAIP is a crucial downstream signaling molecule, implicated in several signaling pathways. Due to these multifunctional effects, TRAIP is more related to cellular mitosis, chromosome segregation, and DNA damage response. Tumor necrosis factor interacting protein is a downstream signaling molecule that contains a RING domain with E3 ubiquitin ligase activity at the N terminal side followed by coiled-coil and C terminal leucine zipper domain. Human TRAIP is constituted of 469 amino acids with 76% sequence similarity with the mouse TRAIP protein. Although, the main inhibitory function of TRAIP has been known for decades, however, in vitro interaction of TRAIPCC domain with RAP80 Zinc finger motif has not been reported yet. Besides, RAP80, the binding partner of TRAIPCC protein has been implicated in DNA damage response.ResultsOur in vitro study shows that the TRAIP CC (64–166) associates with the RAP80 zinc finger of corresponding amino acid 490–584. However, TRAIP CCLZ (66–260) and TRAIP RINGCC (1 = 157) failed to interact with the RAP80 zinc finger of corresponding amino acid 490–584. The current study reinforces TRAIP CC (64–166) and RAP80 zinc finger of corresponding amino acid 490–584 associates to form a complex. Moreover, SDS PAGE arbitrated the homogeneity of RAP80 Zinc finger and TRAIP CC of corresponding amino acid 490–584 and 64–166, respectively.ConclusionIn vitro, a specific interaction was observed between the TRAIP CC (64–166) and the RAP80 zinc finger of the corresponding amino acid 490–584 and a specific binding area of the RAP80 zinc finger motif were investigated. The TRAIPCC region is required for the complex to bind to the RAP80-Zn finger motif. This strategy may be necessary for the RAP80 zinc finger activity to the TRAIP CC protein.     

Reducing the microcapsule diameter by micro-emulsion to improve the insecticidal activity of Bacillus thuringiensis encapsulated formulations

The emulsion/internal gelation method is highly effective to produce microcapsules of Bacillus thuringiensis (Bt) in a short time; however, it has the limitation to produce microcapsules within a wide range of diameters (1–1000?µm). The aim of this study was to reduce the range of small microcapsule diameters by using a water/corn-oil (W/CO) micro-emulsion as the dispersing medium and the mixture Tween 80–Span 80 as the surfactant. It involved the development of the W/CO micro-emulsion and the determination of the suitable agitation time to disperse the gelling medium (sodium alginate) through the micro-emulsion. A micro-emulsion formulation that allowed reduction of the microcapsule diameter was composed of 82% corn oil, 12% alginate solution and 6% surfactant mixture Tween80–Span80 (31:69). Evaluation of four dispersing times showed that 45 min was suitable to produce 75% of microcapsules of an average diameter of 3.1?±?1?µm containing the spore–protein complex (SPC) produced by Bt. Bioassays carried out at low concentrations of microencapsulated formulations of cry proteins allowed determination of how its insecticidal effect increased if the range of microcapsule diameters was reduced in the range 1–9?µm. Furthermore, the SPC formulation in alginate microcapsules showed high resistance to extreme irradiation (2.9?±?0.5?×?10⁸ erg) of a long wavelength (365?nm), which made the microencapsulated formulation profitable and of high yield since repeated applications of the biopesticide during the same harvest period may not be necessary.     

Glycophyte salt resistance

By perfusion of entire sunflower stems with NaCl solutions of various concentrations, we studied the phenomenon of sodium decrement, i.e., sodium retaining in the stem and leaf petioles. Such retaining could comprise up to 50–80% of initial sodium concentration. It depended on the rate of perfusion, the length of xylem vessels, and NaCl concentration. When perfusion with 100–500 mM NaCl concentrations (high for glycophytes) lasted for 10–12 days, we did not observe any decrease in the degree of sodium decrement. Simultaneously with sodium decrement, other ions (K⁺ and Ca²⁺) were secreted into the perfusate, thus providing for physiological equilibrating the monosalt solution supplied to the stem base. The high salt concentration in the perfusate induced a decrease in the hydraulic conductance of the vessels. The conclusion is that stressful NaCl solutions attain the shoot meristem and reproductive organs as an “equilibrated” salt solution and at a declined rate of xylem flow. The mechanisms of observed phenomenon of glycophyte salt resistance are discussed, the main of them being related to osmosis-dependent responses of stem living cells and the processes of ion exchange between the cells and xylem vessel content.     

Effect of Nitrogen Fertilization and Growth Suppression on Pitch Canker Development on Loblolly Pine Seedlings

One-yr-old loblolly pine seedlings of two half-sib families, grown in sand, were fertilized three times per week with nutrient solution containing 20 μg/ml (low) or 80 μg/ml (high) nitrogen. Nitrogen concentration in the nutrient solution was either constant throughout the experiment, or interehanged after the inoculation of stems or shoots with Fusarium subglutinans, 55 days after initiation of fertilization. Growth was suppressed by a weekly excision of shoots branching from the stem apex. Either high nitrogen nutrition or shoot excision generally enhanced canker elongation on stem inoculated plants; the combination of both was extremely conducive for disease development. With intact plants of family 8–68, interchange of pre-inoculation low nitrogen nutrition with high nitrogen after inoculation enhanced canker elongation and rate of wilt. Nitrogen content varied in wood, bark and needles, as well as with time intervals, but was consistently in accordance with nitrogen level in the nutrient solution. In shoot excised plants, nitrogen content was higher than in the respective treatment without shoot excision. The higher nitrogen nutrient accelerated disease development on inoculated shoots, compared to low nitrogen, on both pine families. With respective treatments, stem cankers were larger and rates of shoots exhibiting lesions or wilt were higher on plants of family 8–68 than on 8–61. It is postulated that the disease enhancing effect associated with higher nitrogen content in stem tissues results from an increased nitrogen availability to the pathogen.     

Novel method of niosome generation using supercritical carbon dioxide part I: process mechanics

Abstract

A novel method for the production of non-ionic surfactant vesicles (niosomes) using an rapid expansion of supercritical solution (RESS)-based process coupled with a gas ejector is presented along with an investigation of parameters affecting niosome morphology, size and encapsulation efficiency of a 0.2?M d-glucose solution in Tris buffer at physiological pH. The solubility of the non-ionic surfactant polyoxyethylene(4) sorbitan monostearate in SC-CO₂ was determined at three pressures (10, 15 and 20?MPa) and three temperatures (40, 50 and 60?°C). Mole fraction of Tween61 in the vapor phase increased with pressure at 40?°C, but did not change with pressure at 50 or 60?°C. Solubility data were correlated using the Peng–Robinson equation of state (PREOS) with the Panagiotopoulos and Reid mixing rule. Vesicles were either multilamellar or unilamellar, depending on the degree of precipitation of the lipid formulation at the point of aqueous cargo introduction. Vesicle particle size distributions were bimodal, with the 80–99% of the liposomal volume contributed niosomes ranging in size from 3 to 7?μm and the remaining niosomes ranging from 239 to 969?nm, depending on the system configuration. Encapsulation efficiency as high as 28% using the gas ejector to introduce the glucose cargo solution was achieved. Vesicle particle size and encapsulation efficiency were shown to be dependent on cargo droplet formation.     

Conformation and aggregation of melittin: effect of pH and concentration of sodium dodecyl sulfate

The conformation of melittin, a surface-active polypeptide, in solution was studied by CD spectra between 190 and 240 nm. The molecule was essentially unordered (possibly with a trace of helix) in water without salt at neutral pH. Upon deprotonation of four of the six cationic groups at pH 12 the polypeptide became partially helical (about 35%). The addition of NaDodSO₄ to an aqueous melittin solution first caused the solution to become turbid but it became clear again in excess surfactant solution. The conformational changes depended on the molar NaDodSO₄/melittin ratio, R. With R from 2.34 to 23.4, the melittin solution was turbid and the polypeptide conformation was probably a mixture of α-helix and β-sheets. This was supported by the ir spectrum of the turbid solution, which indicated the presence of both conformations. With R = 46.8 or 468 (1 or 10 mM NaDodSO₄) the polypeptide conformation was characteristic of an α-helix, about 70–80% of the molecule, regardless of whether the surfactant was above or below its critical micelle concentration. This compared well with the x-ray results of 92% helix in crystals. The lower helicity of melittin in NaDodSO₄ solution might be attributed to the end effects that destabilize the first and last turn of an helix at its N- and C-terminus, respectively.     

Purification of Bovine α-Lactalbumin by Immobilized Metal Ion Affinity Chromatography

ABSTRACT

The milk protein α-lactalbumin was isolated from bovine whey protein concentrate solution by immobilized metal ion affinity chromatography (MAC) using Cu(II)-Chelating Sepharose Fast Flow. Stepwise pH (5.5–3.8) changes in sodium acetate buffer were used to elute the protein selectively, at which time it was concentrated and reapplied to an uncharged Chelating Sepharose Fast Flow column to remove the contaminating Cu(II) ions. A purity of 90% and recovery of 80% was achieved. The described method appears to be suitable for isolation of a-lactalbumin in a form adequate for milk formula engineering.     

Automatic exposure control at single- and dual-heartbeat CTCA on a 320-MDCT volume scanner: Effect of heart rate,exposure phase window setting,and reconstruction algorithm

PurposeTo investigate whether electrocardiogram (ECG)-gated single- and dual-heartbeat computed tomography coronary angiography (CTCA) with automatic exposure control (AEC) yields images with uniform image noise at reduced radiation doses.Materials and methodsUsing an anthropomorphic chest CT phantom we performed prospectively ECG-gated single- and dual-heartbeat CTCA on a second-generation 320-multidetector CT volume scanner. The exposure phase window was set at 75%, 70–80%, 40–80%, and 0–100% and the heart rate at 60 or 80 or corr80 bpm; images were reconstructed with filtered back projection (FBP) or iterative reconstruction (IR, adaptive iterative dose reduction 3D). We applied AEC and set the image noise level to 20 or 25 HU. For each technique we determined the image noise and the radiation dose to the phantom center.ResultsWith half-scan reconstruction at 60 bpm, a 70–80% phase window- and a 20-HU standard deviation (SD) setting, the imagenoise level and -variation along the z axis manifested similar curves with FBP and IR. With half-scan reconstruction, the radiation dose to the phantom center with 70–80% phase window was 18.89 and 12.34 mGy for FBP and 4.61 and 3.10 mGy for IR at an SD setting SD of 20 and 25 HU, respectively. At 80 bpm with two-segment reconstruction the dose was approximately twice that of 60 bpm at both SD settings. However, increasing radiation dose at corr80 bpm was suppressed to 1.39 times compared to 60 bpm.ConclusionAEC at ECG-gated single- and dual-heartbeat CTCA controls the image noise at different radiation dose.     

The response of soil solution chemistry in European forests to decreasing acid deposition

Acid deposition arising from sulphur (S) and nitrogen (N) emissions from fossil fuel combustion and agriculture has contributed to the acidification of terrestrial ecosystems in many regions globally. However, in Europe and North America, S deposition has greatly decreased in recent decades due to emissions controls. In this study, we assessed the response of soil solution chemistry in mineral horizons of European forests to these changes. Trends in pH, acid neutralizing capacity (ANC), major ions, total aluminium (Al_tot) and dissolved organic carbon were determined for the period 1995–2012. Plots with at least 10 years of observations from the ICP Forests monitoring network were used. Trends were assessed for the upper mineral soil (10–20 cm, 104 plots) and subsoil (40–80 cm, 162 plots). There was a large decrease in the concentration of sulphate () in soil solution; over a 10‐year period (2000–2010), decreased by 52% at 10–20 cm and 40% at 40–80 cm. Nitrate was unchanged at 10–20 cm but decreased at 40–80 cm. The decrease in acid anions was accompanied by a large and significant decrease in the concentration of the nutrient base cations: calcium, magnesium and potassium (Bc = Ca²⁺ + Mg²⁺ + K⁺) and Al_tot over the entire dataset. The response of soil solution acidity was nonuniform. At 10–20 cm, ANC increased in acid‐sensitive soils (base saturation ≤10%) indicating a recovery, but ANC decreased in soils with base saturation >10%. At 40–80 cm, ANC remained unchanged in acid‐sensitive soils (base saturation ≤20%,  ≤ 4.5) and decreased in better‐buffered soils (base saturation >20%,  > 4.5). In addition, the molar ratio of Bc to Al_tot either did not change or decreased. The results suggest a long‐time lag between emission abatement and changes in soil solution acidity and underline the importance of long‐term monitoring in evaluating ecosystem response to decreases in deposition.