ABSTRACT
Toll-like receptor (TLR) 2/1 signalling is linked to autophagy through transcriptional actions of the 1,25-dihydroxyvitamin D3 (1,25(OH)2D3)-vitamin D receptor (VDR) complex. Population-specific effects have been reported for TLR2/1-VDR signalling. We hypothesized that population effects extend to autophagy and are influenced by vitamin D status. Serum 25(OH)D3 of healthy South Africans (Black individuals n = 10, White individuals n = 10) was quantified by LC-MS/MS. Primary monocytes-macrophages were supplemented in vitro with 1,25(OH)2D3 and stimulated with the lipoprotein Pam3CysSerLys4. TLR2, VDR, hCAP18, Beclin1, LC3-IIB, cytokines and CYP24A1 mRNA were quantified by flow cytometry and RT-qPCR, respectively. Black individuals showed significantly lower overall cumulative LC3-IIB (P < 0.010), but higher Beclin1, VDR, IL6 and TNFA (P < 0.050) than White individuals. 1,25(OH)2D3 enhanced autophagic flux in monocytes-macrophages from Black individuals upon TLR2/1 stimulation and strengthened autophagy in 25(OH)D3 deficient individuals (independent cohort, n = 20). These findings support population-directed vitamin D supplementation.
Subject(s)
Autophagy/physiology , Calcitriol/metabolism , Monocytes/metabolism , Adult , Autophagy/drug effects , Calcitriol/blood , Calcitriol/physiology , Chromatography, Liquid/methods , Female , Humans , Male , Microtubule-Associated Proteins/genetics , Middle Aged , Primary Cell Culture , Receptors, Calcitriol/genetics , Receptors, Calcitriol/metabolism , South Africa , Tandem Mass Spectrometry/methods , Toll-Like Receptor 2 , Toll-Like Receptors/genetics , Toll-Like Receptors/metabolism , Vitamin D/metabolism , Vitamin D/physiologyABSTRACT
An iodine-mediated hydration reaction of alkynes serves as a green alternative to metal-catalyzed procedures. Previous work has shown that this method works well with terminal alkynes on keto-functionalized scaffolds, including 1,3-dicarbonyls and their heteroatom analogues. It was hypothesized that the reaction proceeds through a 5-exo-dig neighboring group participation (NGP) cyclization and an α-iodo intermediate. The work described herein probes the existence of the intermediate through NMR investigations and explores the scope of the hydration process with internal alkynes. The NMR experiments confirm the existence of the α-iodo intermediate, and methodology studies demonstrate that alkyl-capped, asymmetric, internal alkynes undergo a regiospecific hydration, also via the 5-exo-dig NGP pathway.
ABSTRACT
The Hawaiian Islands have long been an important laboratory for evolutionary research because their geological histories offer many natural experiments. For example, the Maui Nui complex, 4 islands that have been repeatedly connected and separated by fluctuating sea levels, lie near Hawaii Island, which has never been connected to another island. Here, we examine the genetic consequences of fluctuating island areas and connectivity using microsatellite analysis of 2 widespread, endemic Hawaiian damselflies. We screened 152 Megalagrion xanthomelas individuals from 5 islands at 14 loci and 34 Megalagrion pacificum from 3 islands at 11 loci to explore dispersal patterns and genetic diversity. Our data suggest that Pleistocene fluctuations in sea level alternated between creating land bridges that facilitated gene flow between once and future islands, and ocean channels that inhibited dispersal. Furthermore, interglacial periods of high sea stands likely reduced suitable habitat availability, causing the loss of genetic diversity on Maui Nui due to bottlenecks and founder events. Finally, we propose that gene flow from Molokai to Lanai may be enhanced by assisted dispersal from the trade winds that are channeled between volcanoes on western Maui and eastern Molokai. Our results emphasize the importance of variable microevolutionary processes in Hawaiian biogeography.
Subject(s)
Biological Evolution , Climate Change , Genetic Variation , Genetics, Population , Odonata/genetics , Animals , Cell Nucleus/genetics , Ecosystem , Gene Flow , Hawaii , Islands , Microsatellite Repeats , Models, Genetic , Sequence Analysis, DNAABSTRACT
Microsatellite loci have been isolated from two species of endemic Hawaiian damselflies, Megalagrion xanthomelas and M. eudytum, that are of conservation concern. Sixteen polymorphic loci were characterized in 32 M. xanthomelas from one population on Molokai and one on Hawaii Island. The total number of alleles per locus ranged from two to 16 and observed population heterozygosity ranged from 0.0 to 0.963. Eleven of these loci amplified successfully in M. eudytum as well. These loci will be used to further conservation efforts and infer genetic consequences of Pleistocene glaciations.
