Red grape berry-cultured cells reduce blood pressure in rats with metabolic-like syndrome.

Purpose: Cumulative evidence suggests that moderate red wine consumption protects the cardiovascular system. The effect of cultured cells derived from red grape berry (RGC) on blood pressure (BP) has not been investigated. We therefore studied the antihypertensive effects of oral consumption of RGC in experimental rat model of metabolic-like syndrome and assessed its effect on human umbilical vein endothelial cells (HUVECs).

Glutathione peroxidase regulation of reactive oxygen species level is crucial for in vitro plant differentiation.

Phospholipid hydroperoxide glutathione peroxidase (PHGPx) is overexpressed in plants under abiotic and biotic stress conditions that mediate oxidative stress. To study its biological role and its ability to confer stress resistance in plants, we tried to obtain transgenic plants overexpressing citrus (Citrus sinensis) PHGPx (cit-PHGPx). All attempts to obtain regenerated plants expressing this enzyme constitutively failed.

Modification of photosynthetic regulation in tomato overexpressing glutathione peroxidase.

To investigate the function of glutathione peroxidase (GPX) in plants, we produced transgenic tomato plants overexpressing an eukaryotic selenium-independent GPX (GPX5). We show here that total GPX activity was increased by 50% in transgenic plants, when compared to control plants transformed with the binary vector without the insert (PZP111). A preliminary two-dimensional electrophoretic protein analysis of the GPX overexpressing plants showed notably a decrease in the accumulation of proteins identified as rubisco small subunit 1 and fructose-1,6-bisphosphate aldolase, two proteins involved in photosynthesis.

Serum leptin activity in obese and lean patients.

Blood levels of the satiety hormone leptin are directly correlated to fat stores in obese and lean people. Therefore, leptin resistance is the logical explanation for the phenomenon of common obesity. However, the important question of whether or not the intrinsic leptin activity could differ between obese and lean people has not been examined before.

Substituting selenocysteine for catalytic cysteine 41 enhances enzymatic activity of plant phospholipid hydroperoxide glutathione peroxidase expressed in Escherichia coli.

The citrus phospholipid hydroperoxide glutathione peroxidase (cit-PHGPx) was the first plant peroxidase demonstrated to exhibit PHGPx-specific enzymatic activity, although it was 500-fold weaker than that of the pig heart analog. This relatively low activity is accounted for the catalytic residue of cit-PHGPx, which was found to be cysteine and not the rare selenocysteine (Sec) present in animal enzymes. Sec incorporation into proteins is encoded by a UGA codon, usually a STOP codon, which, in prokaryotes, is suppressed by an adjacent downstream mRNA stem-loop structure, the Sec insertion sequence (SECIS).

Molecular cloning and properties of the chicken leptin-receptor (CLEPR) gene.

The mammalian leptin receptor (LEPR) (formerly OB-R) mediates the weight regulatory effects of the circulating hormone leptin. The extreme obese phenotype of recessive mutations in the mouse leptin or LEPR genes (ob/ob and db/db mice, respectively) indicate the high potential of these genes for medical and agricultural research. In this paper, we report on the cloning of the full-length chicken leptin receptor (CLEPR) cDNA, which is the first non-mammalian cloning of a LEPR gene.

Regulation of stress-induced phospholipid hydroperoxide glutathione peroxidase expression in citrus.

Recent findings in our laboratory showed that in citrus cells, salt treatment induced the accumulation of mRNA and a protein corresponding to phospholipid hydroperoxide glutathione peroxidase (PHGPX), an enzyme active in the cellular antioxidant system. The protein and its encoding gene, csa, were isolated and characterized, and the expected enzymatic activity was demonstrated (G. Ben-Hayyim et al.

Purification and identification of two putative autolytic sites in human calpain 3 (p94) expressed in heterologous systems.

Human muscle-specific calpain (CAPN3) was expressed in two heterologous systems: Sf9 insect cells and Escherichia coli cells. Polyclonal antibodies were prepared against peptides whose sequences were taken from the three unique regions of human CAPN3, namely NS, IS1, and IS2, which are not found in other members of the calpain family. Western blot analysis using these antibodies revealed that CAPN3 was well expressed in both systems.

Induction of a gene encoding an oleosin homologue in cultured citrus cells exposed to salt stress.

A cDNA clone (C3) with high homology to plant oleosins was isolated from citrus cultured cells. The 827-bp cDNA insert has an open reading frame of 144 amino-acid residues. The central hydrophobic domain of the protein is nearly identical to oleosins from Brassica napus and maize, and the C-terminal hydrophilic region following the hydrophobic domain is also highly conserved.

A stress-associated citrus protein is a distinct plant phospholipid hydroperoxide glutathione peroxidase.

A protein whose level is markedly increased upon exposure of cultured citrus cells and whole plants to NaCl, was shown to specifically catalyze the reduction of phosphatidylcholine hydroperoxide in the presence of glutathione. This enzymatic activity was shown to be independent of a similar activity exhibited by glutathione S-transferase in plants. This finding corroborates the significant homology (52%) accounted between the deduced amino acid sequence of the gene encoding for this protein and that of mammalian phospholipid hydroperoxide glutathione peroxidases.

Drought, heat and salt stress induce the expression of a citrus homologue of an atypical late-embryogenesis Lea5 gene.

In a search for genes that are induced in citrus cell suspension in response to salt stress, a cDNA clone with high homology to cotton Lea5 gene was isolated. Data base analysis of the protein deduced from the nucleotide sequence indicates that, like in cotton, the protein from citrus contains regions with significant hydropathic character. The gene, designated C-Lea5, is expressed in citrus leaves as well as cell suspension.

Antibodies for the immunochemistry of the human beta 3-adrenergic receptor.

Based on the amino acid sequence deduced from the recently cloned human beta 3-adrenergic receptor (hu beta 3AR) gene, polyclonal antibodies were prepared against synthetic peptides, corresponding to regions of hu beta 3AR presumed to be exposed at the outer or the inner side of the membrane on the basis of the putative three-dimensional structure of the previously characterized beta 1 and beta 2 adrenergic receptors. Affinity-purified antibodies directed against N-terminal, extracellular or intracellular loops and C-terminal peptides reacted specifically with the hu beta 3AR and not with either the human beta 1 or beta 2 adrenergic receptor. Using these antibodies, it was demonstrated that the receptor is present at the surface of Chinese Hamster Ovary (CHO) cells transfected with the hu beta 3AR gene; in addition, the presence of the receptor protein was established in a human tissue (gall bladder).

A novel plant glutathione peroxidase-like protein provides tolerance to oxygen radicals generated by paraquat in Escherichia coli.

Citrus salt-stress associated protein (Cit-SAP) reveals significant sequence homology to mammalian glutathione peroxidase (GP). In an attempt to assign biological function to this protein, transformed E. coli cells expressing Cit-SAP were examined for their ability to tolerate free radicals formed by paraquat, an O2- radical forming agent.

Molecular characterization of salt-stress-associated protein in citrus: protein and cDNA sequence homology to mammalian glutathione peroxidases.

A gene encoding for a citrus salt-stress-associated protein (Cit-SAP) was cloned from Citrus sinensis salt-treated cell suspension. The gene, designated csa, was isolated from a cDNA expression library. The partial amino acid sequence of the protein, as well as that deduced from the nucleotide sequence of csa, revealed a considerable homology to mammalian glutathione peroxidase (GP), and to clone 6P229 from tobacco protoplasts.

Localization and characterization of three different beta-adrenergic receptors expressed in Escherichia coli.

After fusion with the N-proximal portion of the outer membrane protein LamB, three beta-adrenergic receptors, the human beta 1- and beta 2- and turkey beta 1-adrenergic receptor, were expressed in Escherichia coli with retention of their own specific pharmacological properties. Molecular characterization and localization of the three receptors in bacteria and comparison of the behaviour of each hybrid protein are reported. The bacteria were lysed and fractionated on a sucrose gradient.

Chemical characterization of ligand binding site fragments from turkey beta-adrenergic receptor.

Affinity-labeled beta-adrenergic receptor from turkey erythrocyte membranes was specifically cleaved near cysteine residues after S-cyanylation. Analysis of the labeled polypeptide fragments suggests that iodocyanopindolol diazirine reacted with an amino acid residue which is located in the non-glycosylated region containing the sixth and seventh transmembrane domains of the receptor. However, the possibility cannot be excluded that a second residue, located between the third and fifth transmembrane domains, was also labeled.

Human beta 2-adrenergic receptors expressed in Escherichia coli membranes retain their pharmacological properties.

The coding region of the gene for the human beta 2-adrenergic receptor gene was fused to the beta-galactosidase gene of the lambda gt11 expression vector. The Y1089 Escherichia coli strain was lysogenized with this modified vector and transcription of the fusion gene was induced. Expression of this transcription unit was shown by the appearance in the bacteria of proteins of molecular weight higher than that of native beta-galactosidase, which are immunoreactive with anti-beta-galactosidase antibodies.

Recognitory bacterial surface lectins which mediate its mannose-specific adherence to eukaryotic cells.

Cell surface protein were found to play a role in the sugar-specific molecular mechanism by which bacteria adhere to mammalian cells. We have demonstrated that at least three different types of lectin-like proteins mediate the mannose-sensitive adherence of gram negative bacteria to epithelial cells. One group of such lectins was shown in our study to be associated with the bacterial flagellum.

Participation of pili and cell wall adhesion in the yeast agglutination activity of Escherichia coli.

Escherichia coli strain 2699 (O6:K13) which had been isolated from a case of urinary tract infection exhibited pili during the stationary phase (24 to 40 h), but not during the exponential phase (4 h), when grown in static broth culture. The bacteria were also piliated when grown for 24 h on agar. They agglutinated Saccharomyces cerevisiae (baker's yeast) in the piliated as well as in the nonpiliated state.

Dissociation and reassembly of Escherichia coli type 1 pili.

Escherichia coli type 1 pili, which mediate the mannose-sensitive adherence of the bacterium to eucaryotic cells, are comprised of very stable arrays of pilin protein subunits (molecular weight, approximately 17,000). Previous methods for the dissociation of pili caused their irreversible denaturation. We have found that incubation of pili in saturated guanidine hydrochloride at 37 degrees C led to their complete dissociation, as evidenced by nephelometry and electron microscopy.