Production and purification of polyclonal anti-hamster immunoglobulins in rabbits

Polyclonal antibodies are mixtures of monoclonal antibodies that were produced against different epitops. The goal of this project is to know the production, purification and horseradish peroxidase (HRP) conjugation of polyclonal antibodies against hamster immunoglobulins in rabbits. 300 ìg/300 ìl of ten hamster immunoglobulins was mixed with the same volume (300 ìl) of adjuvant and injected into three 6-month-old white New Zealand rabbits. Anti hamster rich rabbits serums were isolated from whole blood and precipitated with ammonium sulfate in the final concentration of 50%. The precipitate was dialysed against phosphate buffered saline (PBS) (pH: 7.4) and applied to ion exchange chromatography (IEC) on diethylaminoethyl (DEAE)-sepharose 6B with tris-phosphate (pH: 8.1), andtris-phosphate contain 50 mM NaCl buffer. The purity of produced antibody was confirmed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) under reduced condition. Then purifiedimmunoglobulin G (IgG) was conjugated with HRP. For exact measurement of conjugated IgG titer and evaluating of cross reaction, enzyme linked immunosorbent assay (ELISA) test was designed. Since IEC is a more simple and inexpensive method for the purification of IgG, we obtained a protein with approximate purity of 95%. Produced IgG showed high titer and high specificity in the designed ELISA. Purified antibody and its conjugation with HRP are used in research and diagnosis of hamster disease.Key words: Production, purification, hamster immunoglobulins

Effect of salicylic acid treatment on cadmium toxicity and leaf lipid composition in sunflower

The ameliorative effect of salicylic acid (SA) on cadmium (Cd) toxicity in sunflower plants was studied by investigating plant growth and fatty acid composition. Sunflower plants in two leaves stage were exposed to CdCl2 treatment (0, 50, 100, 150 and 200 µM) and then were treated with salicylic acid (0, 250 and 500 µM) as foliage spraying. One week after the last salicylic acid treatment,plants were harvested and growth parameters were measured . Oil of leaf was extracted in a Soxhlet system and fatty acid composition were measured by gas chromatography (GC). Statistical analyses showed excess Cd reduced growth parameters (fresh weight and length of stems and roots, fresh weight and number of leaves)and SA increased them compared with the control. Maximum reduction in these parameters was at 200 µmol Cd and 0µmol of SA. Cd caused a shift in fatty acids composition, resulting in a lower degree of their unsaturation and an increase in saturated fatty acids in sunflower leaves,whereas SA improved them. SA, particularly increased the percentage of linolenic acid and lowered that of palmitic acid by the same proportion. These results sugg membrane integrity due to lipids est that SA could be used as a potential growth regulator and a stabilizer ofprotection of cadmium-induced oxidative stress to improve plant resistance to Cd stres

A different role for hydrogen peroxide and the antioxidative system under short and long salt stress in Brassica oleracea roots

Salinity affects normal growth and development of plants depending on their capacity to overcome the induced stress. The present study was focused on the response and regulation of the antioxidant defence system in Brassica oleracea roots under short and long salt treatments. The function and the implications of hydrogen peroxide as a stressor or as a signalling molecule were also studied. Two different zones were analysed—the elongation and differentiation zone and the fully differentiated root zone—in order to broaden the knowledge of the different effects of salt stress in root. In general, an accumulation of hydrogen peroxide was observed in both zones at the highest (80 mM NaCl) concentration. A higher accumulation of hydrogen peroxide was observed in the stele of salt-treated roots. At the subcellular level, mitochondria accumulated hydrogen peroxide in salt-treated roots. The results confirm a drastic decrease in the antioxidant enzymes catalase, ascorbate peroxidase, and peroxidases under short salt treatments. However, catalase and peroxidase activities were recovered under long salt stress treatments. The two antioxidant molecules analysed, ascorbate and glutathione, showed a different trend during salt treatments. Ascorbate was progressively accumulated and its redox state maintained, but glutathione was highly accumulated at 24 h of salt treatment, but then its concentration and redox state progressively decreased. Concomitantly, the antioxidant enzymes involved in ascorbate and glutathione regeneration were modified under salt stress treatments. In conclusion, the increase in ascorbate levels and the maintenance of the redox state seem to be critical for root growth and development under salt stress

Salt restriction in kidney disease—a missed therapeutic opportunity?

The importance of salt restriction in the treatment of patients with renal disease has remained highly controversial. In the following we marshal the current evidence that salt plays a definite role in the genesis of hypertension and target organ damage, point to practical problems of salt restriction, and report on novel pathomechanisms of how salt affects blood pressure and causes target organ damage