Evolutionary analysis of FAM83H in vertebrates

<div><p>Amelogenesis imperfecta is a group of disorders causing abnormalities in enamel formation in various phenotypes. Many mutations in the <i>FAM83H</i> gene have been identified to result in autosomal dominant hypocalcified amelogenesis imperfecta in different populations. However, the structure and function of FAM83H and its pathological mechanism have yet to be further explored. Evolutionary analysis is an alternative for revealing residues or motifs that are important for protein function. In the present study, we chose 50 vertebrate species in public databases representative of approximately 230 million years of evolution, including 1 amphibian, 2 fishes, 7 sauropsidas and 40 mammals, and we performed evolutionary analysis on the FAM83H protein. By sequence alignment, conserved residues and motifs were indicated, and the loss of important residues and motifs of five special species (Malayan pangolin, platypus, minke whale, nine-banded armadillo and aardvark) was discovered. A phylogenetic time tree showed the <i>FAM83H</i> divergent process. Positive selection sites in the C-terminus suggested that the C-terminus of <i>FAM83H</i> played certain adaptive roles during evolution. The results confirmed some important motifs reported in previous findings and identified some new highly conserved residues and motifs that need further investigation. The results suggest that the C-terminus of FAM83H contain key conserved regions critical to enamel formation and calcification.</p></div

SERPING1, GBP1, PLA1A and LTF gene mRNA expression.

<p>(*:p<0.01;**:P<0.05).</p

Differentially expressed genes among these three groups were involved in the inflammation signal pathway.

<p>Red and blue represents the up- and down- regulated genes respectively.</p

The results of selection tests.

<p>The results of selection tests.</p

Alignment of FAM83H protein sequences among human and the five special species.

<p>Conserved amino acids in vertebrates excluding the five special species are indicated in the yellow box; conserved amino acids in mammals excluding the five special species are highlighted in the pink fill and in the yellow box; substitutions of conserved sites in the five special species are indicated in green, bold and italic font; phosphorylation sites in human FAM83H are presented in blue, bold and italic font; positive selection residues are shown in red, bold and italic font with a red border in the significant selection site.</p

The phylogenetic time tree for <i>FAM83H</i> in 40 mammals.

<p>The time tree was computed using the Reltime method (GTR+G model) involving 40 mammalian <i>FAM83H</i> coding sequences under one calibration constraint. The platypus was selected as an outgroup. There are a total of 6441 positions with gaps in the final dataset. The phylogenic and divergence timescales were implemented in MEGA 7.0. Euarchontoglires (red) and marsupialia (green) are highlighted.</p

Functional analysis of genes that exhibited altered expression levels.

<p>Functional analysis of genes that exhibited altered expression levels.</p

A new ferulic acid ester from <i>Rhodiola wallichiana</i> var<i>. cholaensis</i> (Crassulaceae)

<p>A new ferulic acid ester, 6-feruloyloxyhexanoic acid (<b>1</b>), was isolated along with 10 known ones (<b>2</b>–<b>11),</b> from the concentrated water extract of <i>Rhodiola wallichiana</i> var<i>. cholaensis.</i> Their chemical structures were elucidated on the basis of extensive spectroscopic methods including Two-dimensional nuclear magnetic resonance (2D NMR) experiments. Compound <b>3</b> was isolated from this plant for the first time. The protective effects against H₂O₂-induced myocardial cell injury in cultured H9c2 cells were also evaluated. Compounds <b>1</b>, <b>5</b> and <b>7</b>–<b>11</b> provided significant protective effects on H₂O₂-induced H9c2 cells injury at the concentration of 25 μg/mL. And the protective effects of compound <b>1</b> was also investigated by the oxygen–glucose deprivation/reperfusion (OGD/R) tests.</p

A new flavonol glycoside from the florets of <i>Carthamus tinctorius</i> L.

<div><p>One new flavonol glycoside, 6-hydroxykaempferol-3-<i>O</i>-β-D-glucoside-7-<i>O</i>-β-D-glucuronide (<b>1</b>), together with eight known flavonoids and three known quinochalcones, was isolated from the florets of <i>Carthamus tinctorius</i> L. Their structures were determined by extensive spectroscopic analyses. Their cardioprotective effects against H₂O₂-induced apoptosis in H9c2 cells were also evaluated; compounds <b>1</b>, <b>2</b>, <b>4</b>–<b>5</b>, <b>7</b>–<b>10</b> and <b>12</b> provided significant protective effects on H₂O₂-induced H9c2 cells at the concentration of 25 μg/mL.</p></div