Adaptation of proteomic techniques for the identification and characterization of protein species from murine heart

Disturbed energy metabolism with impaired fatty acid oxidation, ATP synthesis and changing levels of contractile proteins has been observed during the development and manifestation of cardiovascular diseases, with the latter showing sexual differences in terms of onset, manifestation and progress. Estrogenic compounds, such as estrogens and phytoestrogens, are known to exert beneficial effects on several cardiovascular parameters. However, global studies implying the normal, non-failing myocardium are rare. Thus, identifying and characterizing protein patterns involved in the maintenance of normal heart physiology at the protein species level will help understanding disease conditions. In this study, we performed an adapted 2-DE/MS approach in order to identify and characterize post-translational modified and truncated protein species from murine heart. Female and male animals of different age were receiving the phytoestrogen genistein and comparative analyses were performed to identify sex and genistein treatment-related effects. Selected 2-DE spots that exposed varying abundance between animal groups and identified as identical proteins were subject to multi-protease cleavage to generate an elevated sequence coverage enabling characterization of post-translational modifications and truncation loci via high-resolution MS. Several truncated, phosphorylated and acetylated species were identified for mitochondrial ATP synthase, malate dehydrogenase and trifunctional enzyme subunit alpha. However, confirmation of several of these modifications by manual spectra interpretation failed. Thus, our results warrant caution for the blind trust in software output. For the regulatory light chain of myosin, we identified an N-terminal processed species, which so far has been related to ischemic conditions only. We tried to unravel the information content of protein species separated by high-resolution 2-DE as an alternative to high-throughput proteomics, which mainly is interested in lists of protein names, ignoring the protein species identity.     

^14CO2 Fixation and Translocation of Photoassimilates as Selection Criteria of Egyptian Taro Genotypes

The growth characterlstlcs, different physlological parameters, photosynthetic activity （^14CO2 fixation）, and the translocatlon rate of photoassimllates In different taro （Colocasia esculenta L. Schott） genotypes was studled In order to determlne the posslble use of these parameters as selectlon crlterla for dlfferent wldely used genotypes of taro （Delta No. 9, 15, 20, 21, and balady）. The results obtalned suggest that Delta No. 21 shows the most slgnlflcant increase In all parameters tested compared wlth the control （balady）, followed by Delta No. 9, 15, and 20, respectively. The results show a positive correlation between photosynthetlc actlvlty, translocatlon efflclency, and total yield. The selected clone Delta No. 21 Is recommended for cultlvatlon In the delta reglon of Egypt.     

Bioaccumulation of heavy metals in the terrestrial isopod Porcellionides pruinosus in the vicinity of Gabes-Ghannouch industrial complex

Abstract

The current study reports the effects of heavy metals issued from the emissions of phosphate and their bioaccumulation in the soils, leaf litter, and the terrestrial isopod Porcellionides pruinosus in the vicinity of Gabes-Ghannouch industrial complex. Cd, Pb, Zn, and Cu concentrations were measured in soils, leaf litter, and P. pruinosus individuals, collected from four stations located at different distances from the factory. Results showed that heavy metal concentrations in soils varied from one site to another and did not reflect those measured in individuals. In contrast, a positive correlation was found between the isopods’ body length and the distance from the pollution source, revealing the isopod sensitivity to the soil metal concentrations. The bioaccumulation factor was calculated using soil and leaf litter as a solid support with the aim of ordering the metal accumulation in P. pruinosus for each site. Overall, P. pruinosus could be defined as a macroconcentrator of Cd, Zn, and Cu (BAF?>?2) and as a deconcentrator of Pb (BAF?<?1).     

Phagocytosis of autologous lymphocytes by cervical preneoplastic and neoplastic cells

Eighty-nine random Pap smears of the uterine cervix were examined to evaluate the phagocytic abnormal cells (PACs) of atypical, dysplastic and neoplastic tissues against infiltrated blood cells. The results revealed that none of the PACs have been identified in atypical (II) and mild dysplastic cells (IIIa). Low levels (1.2%) of PACs were initially demonstrated in patients with moderate dysplasia (IIIb) that increased 1.7-fold in subsequent severe dysplasia (IIIc) and further increased 2.8-fold in invasive carcinomas of the cervix (V). In addition, the data showed age-related responsiveness toward the development of PACs, where 82% of old patients have developed phagocytic activity in moderate dysplastic cells compared with 27% of young patients. However, the difference became less significant at the subsequent classes of the disease. These data further demonstrated that PACs are class-dependent and it may explain one mechanism by which precancerous cells escape immunosurveillance.     

Constitution of sargassan,a sulphated heteropolysaccharide from sargassum linifolium

The behaviour towards periodate of the brown-algal polysaccharide sargassan before and after partial hydrolysis, alkali treatment, and methanolysis has been studied. Evidence is thereby provided that the sargassan backbone is composed of (1→4)-linked β-D-glucuronic acid and β-D-mannose residues. Heteropolymeric, partially sulphated branches are attached to the backbone, and these branches comprise various proportions of(l→4)-linked, β-D-galactose, β-D-galactose 6-sulphate, and β-D-galactose 3,6-disulphate residues, (1→2)-linked α-L-fucose 4-sulphate residues, and (1→3)-linked β-D-xylose residues.     

Ecto-5′-nucleotidase regeneration after chemical modification of the plasma membrane

Treatment of the C6 glioblastoma cell with trinitrobenzenesulfonic acid (TNBS) resulted in the selective inactivation of ecto-5'-nucleotidase under conditions which maintained cell viability. Cells respond to ecto-enzyme inactivation by replacing 80% of lost activity within 24 hrs. A lag time of 4-6 hrs before ecto-5'-nucleotidase replacement began and its complete blockage by cycloheximide indicated that the source of replaced enzyme was de novo synthesis and not an intracellular pool. Release of 5'-nucleotidase activity into culture medium in the form of membraneous vesicles slowed during the active recovery period and then steadily increased with time as the plasma membrane enzyme level approached normal. TNBS did not exert a direct inhibitory action upon the exfoliative process as release of vesicular GM1 and protein were little affected. Decrease in exfoliated 5'-nucleotidase activity may be due to a selective conservation of the enzyme in the exfoliative process.     

Synthesis and biological activity of some new 3,6-dinitro-1:8-naphthaloyl- and 3,6-diamino-1:8-naphthaloylamino acids and dipeptide derivatives

Synthesis of a series of 3,6-dinitro-1:8-naphthaloylamino acids (II-IX) and some of their corresponding methyl esters (X-XVI) and 3,6-diamino-1:8-naphthaloylamino acid derivatives (XXIX-XXXVI) is described. Coupling of 3,6-dinitro-1:8-naphthaloylamino acids with amino acid methyl ester hydrochlorides in dioxane-DMF-Et3N medium using DCC method furnishes the desired 3,6-dinitro-1:8-naphthaloyldipeptide methyl esters (XVII-XXVIII). Most of the synthesized 3,6-dinitro-1:8-naphthaloylamino acids, esters and dipeptide derivatives (compounds III-VI, XI-XV, XVII, XIX-XXI, XXIII and XXV) and 3,6-diamino-1:8-naphthaloylamino acid derivatives (XXIX-XXXV) were found to be active against a number of microorganisms.