Vacuolar Na+/H+ antiporter cation selectivity is regulated by calmodulin from within the vacuole in a Ca2+- and pH-dependent manner

The selective movement of ions between intracellular compartments is fundamental for eukaryotes. Arabidopsis thaliana Na(+)/H(+) exchanger 1 (AtNHX1), the most abundant vacuolar Na(+)/H(+) antiporter in A. thaliana, has important roles affecting the maintenance of cellular pH, ion homeostasis, and the regulation of protein trafficking. Previously, we have shown that the AtNHX1 C-terminal hydrophilic region localized in the vacuolar lumen plays an important role in regulating the antiporter's activity. Here, we have identified A. thaliana calmodulin-like protein 15 (AtCaM15), which interacts with the AtNHX1 C terminus. When expressed in yeast, AtCaM15 is localized in the vacuolar lumen. The transient expression of AtCaM15 in Arabidopsis leaf protoplasts showed that AtCaM15 is present in the central vacuole. The binding of AtCaM15 to AtNHX1 was Ca(2+)- and pH-dependent and decreased with increasing pH values. Our results also show that the binding of AtCaM15 to AtNHX1 modified the Na(+)/K(+) selectivity of the antiporter, decreasing its Na(+)/H(+) exchange activity. Taken together, the presence of a vacuolar calmodulin-like protein acting on the vacuolar-localized AtNHX1 C terminus in a Ca(2+)- pH-dependent manner suggests the presence of signaling entities acting within the vacuole.     

Regulatory element copy number differences shape primate expression profiles

Gene expression differences are shaped by selective pressures and contribute to phenotypic differences between species. We identified 964 copy number differences (CNDs) of conserved sequences across three primate species and examined their potential effects on gene expression profiles. Samples with copy number different genes had significantly different expression than samples with neutral copy number. Genes encoding regulatory molecules differed in copy number and were associated with significant expression differences. Additionally, we identified 127 CNDs that were processed pseudogenes and some of which were expressed. Furthermore, there were copy number-different regulatory regions such as ultraconserved elements and long intergenic noncoding RNAs with the potential to affect expression. We postulate that CNDs of these conserved sequences fine-tune developmental pathways by altering the levels of RNA.     

Enhanced cellulose degradation by targeted integration of a cohesin-fused β-glucosidase into the Clostridium thermocellum cellulosome

The conversion of recalcitrant plant-derived cellulosic biomass into biofuels is dependent on highly efficient cellulase systems that produce near-quantitative levels of soluble saccharides. Similar to other fungal and bacterial cellulase systems, the multienzyme cellulosome system of the anaerobic, cellulolytic bacterium Clostridium thermocellum is strongly inhibited by the major end product cellobiose. Cellobiose-induced inhibition can be relieved via its cleavage to noninhibitory glucose by the addition of exogenous noncellulosomal enzyme β-glucosidase; however, because the cellulosome is adsorbed to the insoluble substrate only a fraction of β-glucosidase would be available to the cellulosome. Towards this end, we designed a chimeric cohesin-fused β-glucosidase (BglA-CohII) that binds directly to the cellulosome through an unoccupied dockerin module of its major scaffoldin subunit. The β-glucosidase activity is thus focused at the immediate site of cellobiose production by the cellulosomal enzymes. BglA-CohII was shown to retain cellobiase activity and was readily incorporated into the native cellulosome complex. Surprisingly, it was found that the native C. thermocellum cellulosome exists as a homooligomer and the high-affinity interaction of BglA-CohII with the scaffoldin moiety appears to dissociate the oligomeric state of the cellulosome. Complexation of the cellulosome and BglA-CohII resulted in higher overall degradation of microcrystalline cellulose and pretreated switchgrass compared to the native cellulosome alone or in combination with wild-type BglA in solution. These results demonstrate the effect of enzyme targeting and its potential for enhanced degradation of cellulosic biomass.     

LASIK versus PRK for high astigmatism

Purpose

To compare the refractive outcomes of laser in situ keratomileusis (LASIK) and photorefractive keratectomy (PRK) for myopic astigmatism of 3 diopters (D) or more.

Study design

Retrospective matched comparative study.

Methods

This study include consecutive myopic patients (SE 0 to –10 D) undergoing LASIK or PRK between 2007 and 2016 with astigmatism of 3 to 6 D, and postoperative follow-up of at least 30 days for LASIK and 60 days for PRK, compared outcomes of LASIK and PRK eyes.

Results

The LASIK and PRK groups comprised 175 eyes of 175 patients each, with median follow-up of 39 and 139 days, respectively (P?<?0.001). Mean preoperative manifest astigmatism was –3.35?±?0.46 and –3.42?±?0.51 D (P?=?0.92), postoperative SE was –0.43?±?0.55 and –0.16?±?0.64 D (P?<?0.001), and arithmetic astigmatism was –0.59?±?0.46 and –0.88?±?0.60 D (P?<?0.001), for the LASIK and PRK groups, respectively. Fifty-seven and 64.0% eyes had postoperative SE within?±?0.5 D of emmetropia (P?=?0.19), and 57.7 and 38.8% eyes were within 0.5 D of attempted astigmatic correction (P?<?0.001) for the LASIK and PRK groups, respectively. More PRK eyes were overtreated regarding both SE and astigmatism than LASIK eyes (P?<?0.001). The efficacy and safety indices were close to 1.0 in both groups. The surgically induced astigmatism, magnitude of error, index of success, correction index and flattening index were all better in the LASIK group.

Conclusion

Both LASIK and PRK achieve good outcomes in high astigmatism. LASIK achieved mild superiority over PRK.