Purification and characterization of a subtilisin-like serine protease induced during the senescence of wheat leaves

A senescence-specific protease accounting for almost 70% of the total peptide hydrolytic activity of protein extracts, was isolated from detached wheat leaves induced to senescence by incubation in the dark for 72 h. Purification to apparent homogeneity was performed by ammonium sulphate precipitation, ion exchange chromatography and gel filtration chromatography. The enzymatic activity was followed by its ability to hydrolyse the synthetic peptide Suc-AAPF-pNA. SDS/PAGE and gel filtration analysis indicated that the enzyme was a dimer composed of two identical subunits of 59 kDa. The apparent K _m and V _max for the peptide were 1.18 m m and 2.27 mmol pNA mg⁻¹ h⁻¹, respectively. The enzyme was active at pH values above 8.0 and remained active after heat treatment at 60°C for 10 min. It was inhibited by chymostatin, indicating that the enzyme possesses a chymotrypsin-like activity. Rubisco was readily hydrolysed by the purified protease. A sequenced internal fragment of 17 amino acids showed a high level of similarity (65–75% identity) with a highly conserved region of several plant subtilisin-like serine proteases. The absence of this enzymatic activity in fractionated extracts from non-senescent tissues suggests that it might play a role in the senescing process.     

Archaeosomes made of Halorubrum tebenquichense total polar lipids: a new source of adjuvancy

Background  

Archaeosomes (ARC), vesicles prepared from total polar lipids (TPL) extracted from selected genera and species from the Archaea domain, elicit both antibody and cell-mediated immunity to the entrapped antigen, as well as efficient cross priming of exogenous antigens, evoking a profound memory response. Screening for unexplored Archaea genus as new sources of adjuvancy, here we report the presence of two new Halorubrum tebenquichense strains isolated from grey crystals (GC) and black mood (BM) strata from a littoral Argentinean Patagonia salt flat. Cytotoxicity, intracellular transit and immune response induced by two subcutaneous (sc) administrations (days 0 and 21) with BSA entrapped in ARC made of TPL either form BM (ARC-BM) and from GC (ARC-GC) at 2% w/w (BSA/lipids), to C3H/HeN mice (25 μg BSA, 1.3 mg of archaeal lipids per mouse) and boosted on day 180 with 25 μg of bare BSA, were determined.     

Lipid peroxidation and molecular damage to polyunsaturated fatty acids in rat liver. Recognition of two classes of hydroperoxides formed under conditions in vivo

The diene conjugates formed during the autoxidation of microsomal lipid extracts, and in endoplasmic reticulum in vivo after exposing rats to CCl4 have been examined by second derivative absorption spectrophotometry. Within a few minutes after administering CCl4 to a rat there is a pronounced signal in microsomal lipid extracts that is ascribed to the cis-trans diene conjugates of microsomal polyunsaturated fatty acids. Somewhat later a second signal becomes evident that is ascribed to trans-trans isomers. The appearance of the trans-trans isomer is very strongly suppressed by prior administration of vitamin E to the rat. It is concluded that the relative contents of cis-trans and trans-trans dienes in lipid extracts of tissue reflect the tissue contents of hydrogen donors as already established for model experiments with polyunsaturated fatty acids in vitro.     

The substrate specificity of the snail Lymnaea stagnalis UDP-GlcNAc:GlcNAc{beta}-R {beta}4-N-acetylglucosaminyltransferase reveals a novel variant pathway of complex-type oligosaccharide synthesis

UDP-GlcNAc:GlcNAcß-R ß1     

Floristic diversity of Mexican seasonally dry tropical forests

Studies of the variation in tropical plant species diversity and itsrelationship with environmental factors are largely based on research intropical moist/wet forests. Seasonally dry tropical forests (SDTFs), incontrast, have been poorly investigated. In this paper we present data from 20Mexican SDTF sites sampled to describe the magnitude of floristic diversity inthese forests and to address the following questions: (i) to what extent isspecies diversity related to rainfall? (ii) Are there other climatic variablesthat explain variation in species diversity in SDTFs? (iii) How does speciesidentity vary spatially (species turnover) within the country? We found thatspecies diversity was consistently greater (a ca. twofold difference) than wouldbe expected according to the sites' precipitation. Rainfall did notsignificantly explain the variation in species diversity. Likewise, the numberof dry and wet months per year was unrelated to species diversity. In contrast,a simple measure of potential evapotranspiration (Thornthwaite's index)significantly explained the variation in species diversity. In addition to thegreat diversity of species per site (local diversity), species turnover wasconsiderable: of a total of 917 sampled species, 72% were present only in asingle site and the average similarity (Sorensen's index) among sites wasonly 9%. These aspects of floristic diversity and the high deforestation ratesof these forests in Mexico indicate that conservation efforts should be directedto tropical forests growing in locations of low and seasonal rainfall.