The Multifaceted Proprotein Convertases: Their Unique,Redundant, Complementary,and Opposite Functions

The secretory proprotein convertase (PC) family comprises nine members: PC1/3, PC2, furin, PC4, PC5/6, PACE4, PC7, SKI-1/S1P, and PCSK9. The first seven PCs cleave their substrates at single or paired basic residues, and SKI-1/S1P cleaves its substrates at non-basic residues in the Golgi. PCSK9 cleaves itself once, and the secreted inactive protease escorts specific receptors for lysosomal degradation. It regulates the levels of circulating LDL cholesterol and is considered a major therapeutic target in phase III clinical trials. In vivo, PCs exhibit unique and often essential functions during development and/or in adulthood, but certain convertases also exhibit complementary, redundant, or opposite functions.     

Properties of polylactic acid composites reinforced with oil palm biomass microcrystalline cellulose

In this work, polylactic acid (PLA) composites filled with microcrystalline cellulose (MCC) from oil palm biomass were successfully prepared through solution casting. Fourier transform infrared (FT-IR) spectroscopy indicates that there are no significant changes in the peak positions, suggesting that incorporation of MCC in PLA did not result in any significant change in chemical structure of PLA. Thermogravimetric analysis was conducted on the samples. The T₅₀ decomposition temperature improved with addition of MCC, showing increase in thermal stability of the composites. The synthesized composites were characterized in terms of tensile properties. The Young's modulus increased by about 30%, while the tensile strength and elongation at break for composites decreased with addition of MCC. Scanning electron microscopy (SEM) of the composites fractured surface shows that the MCC remained as aggregates of crystalline cellulose. Atomic force microscopy (AFM) topographic image of the composite surfaces show clustering of MCC with uneven distribution.     

The genotypic variation of the antioxidant potential of different tomato varieties

There is increasing interest in the ability of diets rich in polyphenols to modulate age-related diseases and promote healthy ageing. We have conducted a pilot experiment with eight tomato varieties to correlate the total antioxidant capacity of the tomato variants with the specific constituent flavonoids present. A strong correlation was observed with the flavonol rhamnoglucoside rutin but not with other flavonoids, such as naringenin chalcone, or hydroxycinnamates, such as chlorogenic, which are also present in the tomato. To test the rigor of this correlation a second study was undertaken with a further 37 tomato varieties selected for low, medium and high rutin levels. We show that the flavonol rutin contributes to the greatest extent to the antioxidant capacity of tomatoes and suggest that this flavonoid may be a useful target for up-regulation in tomatoes in order to improve their antioxidant status.     

Cold-Adapted RTX Lipase from Antarctic Pseudomonas sp. Strain AMS8: Isolation, Molecular Modeling and Heterologous Expression

A new strain of psychrophilic bacteria (designated strain AMS8) from Antarctic soil was screened for extracellular lipolytic activity and further analyzed using molecular approach. Analysis of 16S rDNA showed that strain AMS8 was similar to Pseudomonas sp. A lipase gene named lipAMS8 was successfully isolated from strain AMS8, cloned, sequenced and overexpressed in Escherichia coli. Sequence analysis revealed that lipAMS8 consist of 1,431 bp nucleotides that encoded a polypeptide consisting of 476 amino acids. It lacked an N-terminal signal peptide and contained a glycine- and aspartate-rich nonapeptide sequence at the C-terminus, which are known to be the characteristics of repeats-in-toxin bacterial lipases. Furthermore, the substrate binding site of lipAMS8 was identified as S²⁰⁷, D²⁵⁵ and H³¹³, based on homology modeling and multiple sequence alignment. Crude lipase exhibited maximum activity at 20 °C and retained almost 50 % of its activity at 10 °C. The molecular weight of lipAMS8 was estimated to be 50 kDa via sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The optimal expression level was attained using the recombinant plasmid pET32b/BL21(DE3) expressed at 15 °C for 8 h, induced by 0.1 mM isopropyl β-D thiogalactoside (IPTG) at E. coli growth optimal density of 0.5.     

Lipoprotein inflammatory properties and serum amyloid A levels but not cholesterol levels predict lesion area in cholesterol-fed rabbits

Rabbits on a 1% cholesterol diet received injections of vehicle with or without D-4F or L-4F. After 1 month, the percent of aorta with atherosclerotic lesions was 24 +/- 15% (vehicle), 10 +/- 6% (D-4F) (P < 0.01 vs. vehicle), and 13 +/- 9% (L-4F) (P < 0.05 vs. vehicle). Inflammatory indexes for HDL and LDL were determined by measuring monocyte chemotactic activity after adding rabbit lipoproteins to human endothelial cells. HDL-inflammatory index (HII) and LDL-inflammatory index (LII), respectively, were 1.39 +/- 0.24; 1.35 +/- 0.29 (vehicle), 0.67 +/- 0.26; 0.63 +/- 0.38 (D-4F) (P < 0.001 vs. vehicle), and 0.67 +/- 0.2; 0.68 +/- 0.32 (L-4F) (P < 0.01 vs. vehicle). Serum amyloid A (SAA) levels were 95 +/- 39, 8 +/- 22, and 7 +/- 19 mug/ml, respectively, for vehicle, D-4F, and L-4F (P < 0.001 vs. vehicle). There was no correlation between lesion area and total plasma or HDL-cholesterol levels. In contrast, there was a positive correlation with HII, LII, and SAA (P = 0.002, P = 0.0026, P = 0.0079, respectively). HII correlated closely with SAA levels (r = 0.6616; r(2) = 0.4377, P < 0.0001). Thus, HII, LII, and SAA are better predictors of lesion area than are total plasma or HDL-cholesterol levels in cholesterol-fed rabbits.     

Fis targets assembly of the Xis nucleoprotein filament to promote excisive recombination by phage lambda

The phage-encoded Xis protein is the major determinant controlling the direction of recombination in phage lambda. Xis is a winged-helix DNA binding protein that cooperatively binds to the attR recombination site to generate a curved microfilament, which promotes assembly of the excisive intasome but inhibits formation of an integrative intasome. We find that lambda synthesizes surprisingly high levels of Xis immediately upon prophage induction when excision rates are maximal. However, because of its low sequence-specific binding activity, exemplified by a 1.9 A co-crystal structure of a non-specifically bound DNA complex, Xis is relatively ineffective at promoting excision in vivo in the absence of the host Fis protein. Fis binds to a segment in attR that almost entirely overlaps one of the Xis binding sites. Instead of sterically excluding Xis binding from this site, as has been previously believed, we show that Fis enhances binding of all three Xis protomers to generate the microfilament. A specific Fis-Xis interface is supported by the effects of mutations within each protein, and relaxed, but not completely sequence-neutral, binding by the central Xis protomer is supported by the effects of DNA mutations. We present a structural model for the 50 bp curved Fis-Xis cooperative complex that is assembled between the arm and core Int binding sites whose trajectory places constraints on models for the excisive intasome structure.     

How can LCA include prospective elements to assess emerging technologies and system transitions? The 76th LCA Discussion Forum on Life Cycle Assessment, 19 November 2020

This paper summarizes the 76th LCA Discussion Forum end its main findings. Main issues when addressing emerging technologies identified were: the lack of primary data, the need for (shared) future background scenarios and (guidlines for) a common methodology. The following recommendations have been derived by the organizers: 1) Specific foreground inventories are always tailor-made, but consistency can be improved through lists of mandatory considerations. 2) Continue sharing (future) technology data and proxy processes, that can be readily replicated to new studies and assist in developing inventories. 3) Streamline and unify the process of including scenarios for background systems. New approaches may provide first important solutions to efficiently include consistent future scenarios in prospective LCA.