Biomedical subjects

C Pinzino

Publications and source records attributed to C Pinzino.

8 recordsLinked to original sources

PubMed2006

Enzymatic and non-enzymatic protective mechanisms in recalcitrant seeds of Araucaria bidwillii subjected to desiccation.

The changes in several antioxidants as well as in the level of C-centered free radicals and thiobarbituric acid reactive substances (TBARS) were studied in seeds of Araucaria bidwillii Hook desiccated to 37%, 28% and 21% moisture content. The lowest-safe moisture content for the seedling establishment was 37%. The embryo, besides double amounts of free radicals, showed higher levels of both enzymatic and non-enzymatic antioxidants than endosperm. Lutein decreased in both organs whereas alpha-tocopherol values were not affected by desiccation. In the embryo at 37% seed moisture content the antioxidant defense system increased giving rise to a decrease in free radicals. Beyond this point, free radicals and TBARS increased in agreement with the umpiring of the ascorbate/glutathione cycle by the decrease in reduced glutathione and glutathione reductase activity (GR, EC 1.6.4.2). At 21% moisture GR decreased. In the endosperm during desiccation, the consumption of ascorbate, total glutathione and lutein prevented the rise in free radicals and TBARS till 28% moisture, at which an increase in oxidized glutathione was also observed.

Antioxidants↗

PubMed2001

Structure and dynamics of high molecular weight glutenin subunits of durum wheat (Triticum durum) in water and alcohol solutions studied by electron paramagnetic resonance and circular dichroism spectroscopies.

Complementary information on the structure and dynamics of high molecular weight glutenin subunits (HMW-GS) of durum wheat (Triticum durum) was obtained by means of two spectroscopic techniques. Electron paramagnetic resonance was used to investigate the dynamics of the HMW-GS hydrated with two 2-propanol/water mixtures at temperatures between 268 and 308 K by specific spin labeling of their cysteine residues. Spectra were of a composite type, resulting from two populations of spin labels differing in molecular mobility, both undergoing isotropic rotational diffusion. Diffusional coefficients and populations of the fast- and slow-moving spin labels, determined by an accurate spectral line shape analysis, are discussed as a function of temperature and water content in the solvent systems. Far-UV circular dichroism was employed to provide information on the secondary structure of the HMW-GS in three different solvents [aqueous 50% (v/v) 2-propanol, aqueous 0.1% (v/v) trifluoroacetic acid, and trifluoroethanol]. For the first one, the influence of temperature on HMW-GS structure was also investigated.

2-Propanol↗

PubMed2000

Degradation of gluten by proteases from dry and germinating wheat (Triticum durum) seeds: An in vitro approach to storage protein mobilization.

Vital gluten was used as an ideal substrate to investigate the role of some proteases in storage protein degradation. Aspartic proteinase and carboxypeptidase were identified as endogenous enzymes adsorbed on gluten and their optimum pH values determined. SDS-PAGE of soluble products released by gluten digestion revealed that the activity of these proteases plays a minor role in protein mobilization, whereas cysteine proteinase, purified from wheat seeds at the fourth day of germination, is extremely effective, producing a remarkable protein degradation in short times. Synergistic effects of aspartic and cysteine proteinase were not observed. Spin labeling of the sulfhydryl groups of gluten proteins enabled a comparative EPR investigation of the consequences of proteolytic degradation on gluten elasticity. It was found that storage protein mobilization brings a loss of elasticity to the polymeric network of gluten, which is particularly marked when the hydrolysis is performed by cysteine proteinase.

Endopeptidases↗

PubMed2000

Protein dynamics in thylakoids of the desiccation-tolerant plant Boea hygroscopica during dehydration and rehydration.

Plants of Boea hygroscopica F. Muell were dehydrated to 9% relative water content (RWC) by withholding water for 26 d, and afterward the plants were rehydrated. Leaves were taken from control plants after 7, 12, and 26 d from the beginning of dehydration, and after 6 and 48 h from rehydration. The RWC decreased by 80% during dehydration, but the leaves regained RWC with rehydration. Dehydrated plants showed lesser amounts of proteins, lipids, and chlorophyll, all of which increased following rewatering. The lipid-to-protein ratio, which decreased during dehydration, returned to control level after 48 h of rehydration. Thylakoid lipids were more unsaturated when RWC reached the value of 9%. EPR measurements of spin-labeled proteins showed the presence of three different groups of proteins with different mobility in thylakoid membranes. The rotational correlation time of groups 1 and 2 increased with dehydration and decreased upon rehydration, whereas group 3 showed little changes. Desiccation did not cause thylakoid swelling or breakage, but the membrane system assemblage showed changes in thylakoid stacking. After 48 h of rehydration the membrane system recovered completely the organization of the fully hydrated state, showing several well-defined and regularly distributed grana.

Antioxidants↗

PubMed1999

Aging, free radicals, and antioxidants in wheat seeds.

Free radical oxidative attack is considered a major cause of disruption and deteriorative changes observed in aged seeds. Antioxidant defense mechanisms may remove potentially damaging molecular species, and carotenoids may act as radical scavengers. The content of lutein, the major carotenoid in wheat seeds, was determined in the flours. It showed a rapid decrease during seed aging. In addition, the content of free radicals in glutens made from flours of wheat seeds after long-term storage was studied. The concentration of radicals appeared to be age dependent, because the highest content of radicals was detected between 13 and 15 years of aging over 36 years of storage. Specific spin labeling of the sulfhydryl groups of gluten proteins enabled comparative EPR studies of the rigidity of the protein chains. A progressive stiffening of polymeric gluten with seed storage was found.

Antioxidants↗

PubMed1999

Superoxide and hydroxyl radical generation, and superoxide dismutase in PSII membrane fragments from wheat.

Illumination of photosystem II (PSII) membrane fragments of wheat (Triticum durum Desf. cv. Adamello) gave rise to both O2*- and *OH radicals adducts of the novel spin trap 5-(diethoxyphosphoryl)-5-methyl-1-pyrroline-N-oxide (DEPMPO). With time, *OH became predominant displaying the conversion of O2*- into *OH. An intrinsic activity of superoxide dismutase (SOD) was found in PSII. Photoreduction of nitroblue tetrazolium (NBT) by PSII membrane fragments was induced by the addition of sodium azide and hydrogen peroxide. Western blotting of PSII proteins showed that a 29 kDa protein was recognised by an antibody against chloroplastic Fe-SOD from water lily. An increased formation rate of O2*- was observed in damaged PSII where the SOD activity decreased following a treatment with a free radical-generating system. Damage in PSII consisted also in a decrease in chlorophyll and in carotenoids as well as in a change in the lipid : chlorophyll : protein ratio. Upon treatment a decrease in the unsaturation of PSII membrane fragments was also monitored together with a degradation towards more saturated molecular species of monogalactosyldiacylglycerol.

Electron Spin Resonance Spectroscopy↗

PubMed1999

Activated oxygen generation from thylakoids: a novel spin trap.

Using a novel phosphorylated spin trap, 5-(diethoxyphosphoryl)-5-methyl-1-pyrroline-N-oxide (DEPMPO), an analogue of the commonly used spin trap 5,5'-dimethyl-1-pyrroline N-oxide (DMPO), we have investigated the production of oxygen radical species under illumination of thylakoids from wheat (Triticum durum Desf cv. Ofanto). DEPMPO reacted with superoxide (O2*-) and hydroxyl radical (HO*) forming distinctive spin trap adducts. Spectra of (O2*-) and HO* adducts of DEPMPO were recorded in the presence of xanthine/xanthine oxidase and FeSO4/H2O2, respectively, and computer simulation of spectra was performed. During illumination of thylakoids both O2*- and HO* were detected as well. Transition metals catalysed transformation of O2*- into HO*. The conversion was enhanced by H2O2 and prevented by exogenous superoxide dismutase and catalase. The presence of a thylakoid-bound superoxide dismutase, whose activity was inhibited by H2O2 and diethyldithiocarbamic acid, was responsible for H2O2 production from O2*- and thus for HO* generation.

Cyclic N-Oxides↗

PubMed1991

Activity and conformational changes of alpha-chymotrypsin in reverse micelles studied by spin labeling.

alpha-Chymotrypsin (CT), spin-labeled at the active site by using an acylating label which constitutes a substrate for this protein, has been investigated in reverse micelles formed by AOT in isooctane. The electron spin resonance spectra provided information on conformation, dynamics and deacylation activity. The dynamics of the label bound to CT appears to be more hindered in reverse micelles than in aqueous solution, probably owing to the effect of the micellar environment on protein conformation. The deacylation rate in reverse micelles does not show the characteristic bell-shaped dependence on water content which is generally found for CT enzymatic activity.

Acylation↗