Fusion and protein-mediated phospholipid exchange studied with single bilayer phosphatidylcholine vesicles of different density

A novel method has been developed for the study of phospholipid exchange and fusion of phospholipid vesicles. Two homogeneous populations of single bilayer phosphatidylcholine vesicles of similar size but markedly different density have been prepared. "Dense" vesicles were made from brominated dioleoyl phosphatidylcholine. "Light" vesicles were prepared from dioleoyl phosphatidylcholine. The two populations were easily separated by density gradient centrifugation. Phosphatidylcholine exchange protein from beef liver was used to promote lecithin exchange between the vesicle populations. Only the lecithin of the external monolayers of the vesicles was available for exchange by exchange protein, implying that flip-flop of vesicle phosphatidylcholine did not take place at a detectable frequency. No spontaneous intervesicle phosphatidylcholine exchange was observed. However, the dense and light vesicles did spontaneously fuse, over several hours, to produce particles of hybrid density.     

Community intervention with the elderly: a social network approach

A technique is described for using the advantages of a social systems approach when working with elderly persons in psychiatric distress. The technique is based on the assumption that the solution to a variety of human predicaments lies within the collective instrumental and affective resources of the client's social network. The vehicle for accomplishing this objective is the "Network Session" during which a mental health professional meets with the elderly person and members of his/her social network to help resolve the difficulty. A case report demonstrating use of the technique is included.     

Molecular cloning, expression, and characterization of a functional single-chain Fv antibody to the mycotoxin zearalenone

The heavy-chain and kappa light-chain variable region genes of an antizearalenone hybridoma cell line (2G3-6E3-2E2) were isolated by PCR and joined by a DNA linker encoding peptide (Gly4Ser)3 as a single-chain Fv (scFv) DNA fragment. The scFv DNA fragment was cloned into a phagemid (pCANTAB5E) and expressed as a fusion protein with E tag and phage M13 p3 in Escherichia coli TG1. In the presence of helper phage M13K07, the scFv fusion protein was displayed on the surfaces of recombinant phages. High-affinity scFv phages were enriched through affinity selection in microtiter wells coated with zearalenone-ovalbumin conjugate. The selected recombinant phages were used to infect E. coli HB2151 for the production of soluble scFv antibodies. One selected clone (pQY1.5) in HB2151 secreted a soluble scFv antibody (QY1.5) with a high zearalenone-binding affinity (concentration required for 50% inhibition of binding, 14 ng/ml), similar to that of parent monoclonal antibody in a competitive indirect enzyme-linked immunosorbent assay. However, scFv QY1.5 exhibited higher cross-reactivity with zearalenone analogs and had greater sensitivity to methanol destabilization than the parent monoclonal antibody did. Nucleotide sequence analyses revealed that the light-chain portion of scFv QY1.5 had a nucleotide sequence identity of 97% to a mouse germ line gene VK23.32 in mouse kappa light-chain variable region subgroup V, whereas the heavy-chain nucleotide sequence was classified as mouse heavy-chain subgroup III (D) but without any closely related members having highly homologous complementarity-determining region sequences. The potential of soluble scFv QY1.5 for routine screening of zearalenone and its analogs was demonstrated with zearalenone-spiked corn extracts.     

Outcome measures of a chronic pain program: a prospective statistical study

Ion beam enhanced deposition (IBED) was adopted to synthesize biocompatible titanium oxide film. Structure characteristics of titanium oxide film were investigated by RBS, AES, and XRD. The blood compatibility of the titanium oxide film was studied by measurements of blood clotting time and platelet adhesion. The results show that the anticoagulation property of titanium oxide film is improved significantly. The mechanism of anticoagulation of the titanium oxide film was discussed.     

Supplemental protein plus ruminally protected methionine and lysine for primiparous beef cattle consuming annual rye hay

Primiparous beef cows (n = 35 Bos taurus, average initial BW of 498 kg) were allotted to treatments in a split-plot designed experiment to determine the effects of supplemental ruminally protected amino acids on cow and calf productivity and metabolic changes in the cows. Cows were fed chopped annual rye hay at 1.5% of BW. The following treatments were used: 1) .8 kg soybean hulls, 1.4 kg ground corn, .6 kg soybean meal (CON); 2) 1.4 kg ground corn, 1.4 kg soybean meal (PRO); 3) PRO plus ruminally protected methionine and lysine (supplied 5 and 10 g, respectively; PRO1); and 4) PRO with twice the level of ruminally protected amino acids in PRO1 (PRO2). Cow weight gain was not different (P > .26) among treatments and averaged 1.2 kg/d for the 45 + 6 d before parturition. After parturition, cow weight gain did not differ (P > .47) between CON and PRO treatments, but it decreased quadratically (P < .01) with increasing level of ruminally protected amino acids. Total milk yield, protein, and fat (4 h) were greater (P < .05) for cows consuming PRO supplements than for CON, whereas CON cattle tended (P = .11) to lose less body condition. Total milk protein showed a quadratic increase (P < .05) in response to level of ruminally protected amino acids that was the inverse of the quadratic response noted for cow weight gain. Serum urea N concentration was greater (P = .07) for cattle consuming additional protein. Metabolic hormones were not affected (P > .18) by dietary treatment, but they responded (P < .05) to changes in physiological state. Supplements with additional protein supported greater (P = .0001) milk urea N concentration and output. Milk urea N concentration increased (P < .05) and milk IGF-I decreased (P < .05) as the lactation period progressed. The measurement of CON and PRO diets revealed that supplements with additional soybean meal had greater (P < .05) DM and N degradation; the extent of forage DM and NDF degradation was similar (P > .05) among treatments. Production shifted away from body weight gain to increased milk protein production when daily supplementation of ruminally protected methionine and lysine increased from 5 and 10 to 10 and 20 g, respectively. This shift in production was not reflective of changes in the metabolic regulators measured.     

Antioxidant activity,color chromaticity coordinates,and chemical characterization of monofloral honeys from Morocco

The bioprospecting of several monofloral Moroccan honeys was carried out. The antiradical activity expressed as mmol Trolox equivalents/kg of honey and evaluated by 2,2-diphenyl-1-picrylhydrazyl assay, ranged from 0.15 for euphorb honey to 1.08 for citrus honey. The antioxidant activity expressed as mmol Fe²⁺/kg and evaluated by ferric ion reducing antioxidant power assay, ranged from 0.96 for euphorb honey to 4.74 for orange honey. The total phenol content was evaluated by colorimetric assay, while the color attributes were evaluated as transmittance data. Significant Pearson correlation factors were found between total polyphenol amount and antioxidant activity and between color attributes and antioxidant activity. Furthermore the chemical composition of volatile organic compounds was determined. The volatile organic compounds chemical composition of the studied honeys was mainly represented by terpene and benzene derivatives, Maillard reaction products, isoprenoids, and hydrocarbons. The volatiles fingerprint, as well as a targeted high-performance liquid chromatography analysis of the polar components, was used to tentatively confirm the declared botanical origin of the samples studied.     

Repeated dosing with the peroxisome proliferator clofibrate decreases the toxicity of model hepatotoxic agents in male mice

Pretreatment of mice with clofibrate (CFB) has been shown to protect against acetaminophen (APAP) hepatotoxicity. To determine if pretreatment with CFB prevents the toxicity of other model hepatotoxicants, male C57BL6J or CD-1 mice received 500 mg CFB/kg, i.p., daily for 10 days, and then were challenged with either 250 mg bromobenzene (BrB)/kg, 0.025 ml carbon tetrachloride (CCl4)/kg or 0.5 ml chloroform (CHCl3)/kg. Liver and kidney injury was assessed by plasma sorbitol dehydrogenase activity (SDH) and blood urea nitrogen (BUN), respectively and histopathology. Challenge with BrB significantly elevated plasma SDH activity in C57Bl6J mice. This was prevented in CFB pretreated mice receiving the same dose of BrB. Changes in BUN were not detected in either group of BrB treated mice. Similarly, pretreatment of male CD-1 mice with CFB significantly reduced CCl4-induced elevation in plasma SDH activity, with no BUN elevation detected in either group. CFB pretreatment also diminished elevation in plasma SDH activity produced by CHCl3 in CD-1 mice, while BUN was significantly elevated in both groups, indicating that CFB did not protect against CHCl3-induced nephrotoxicity. Histopathological examination of liver and kidney sections confirmed these results. This study shows that mice pretreated with CFB were protected from toxicity at 24 h after challenge with other model hepatotoxic agents besides APAP.