Mitochondrial dysfunction enhances the migration of vascular smooth muscles cells via suppression of Akt phosphorylation

Background

Atherosclerosis is one of the major complications of diabetes, which may result from insulin resistance via mitochondrial dysfunction. Although a strong association between insulin resistance and cardiovascular disease has been suggested, it is not clear yet whether stress-inducing factors damage mitochondria and insulin signaling pathway in cardiovascular tissues.

Methods

We investigated whether stress-induced mitochondrial dysfunction might alter the insulin/Akt signaling pathway in A10 rat vascular smooth muscle cells (VSMC).

Results

The treatment of oxidized low density lipoprotein (oxLDL) decreased ATP contents, mitochondrial respiration activity, mRNA expressions of OXPHOS subunits and IRS-1/2 and insulin-mediated phosphorylations of Akt and AMP-activated protein kinase (AMPK). Similarly, dideoxycytidine (ddC), the mtDNA replication inhibitor, or rotenone, OXPHOS complex I inhibitor, inhibited the insulin-mediated pAkt while increased pAMPK regardless of insulin. Reciprocally, an inhibitor of Akt, triciribine (TCN), decreased cellular ATP contents. Overexpression of Akt dominant positive reversed the oxLDL- or ddC-mediated ATP decrease but AMPK activator did not. Akt activation also normalized the aberrant VSMC migration induced by ddC.

Conclusions

Defective insulin signaling and mitochondrial function may collectively contribute to developing cardiovascular disease.

General significance

Akt may be a possible therapeutic target for treating insulin resistance-associated atherosclerosis.     

Activation and expression of urokinase-type plasminogen activator are modulated by freezing/thawing process through activation of redox signal pathway in primary porcine endometrial cells

Plasminogen activators (PAs) play a pivotal role in a variety of uterine physiologies, such as endometrial function, trophoblast invasion, and implantation process, but its alteration in expression or activity during cryopreservation of primary uterine cells has received little attention. In this study, we investigated whether PA expression and activity were modulated in first passage primary porcine uterus endometrial epithelium cells (PUEECs) treated with or without a freezing-thawing procedure. Western blotting and zymographic analysis showed that uPA expression and activity increased significantly in frozen-thawed PUEECs in a passage-dependent manner as compared to freshly prepared control cells. Moreover, intracellular reactive oxygen species (ROS) were increased by freezing-thawing and longer culturing, and were more prominent in frozen-thawed PUEECs than in control cells. However, the increase in both uPA expression and activity was greatly reduced or alleviated by treatment with either ROS scavenger N-acetylcysteine or extracellular signal-regulated kinase (ERK) inhibitor PD98059. These results suggest that ROS/ERK-mediated uPA activation may be an important factor in cryo-damage of primary uterine cells.     

Molecular identification of Ammodytes (PISCES, Ammodytidae) larvae, with ontogenetic evidence on separating populations

A total of ten larval Ammodytes were collected from the Yellow (5 specimens) and Bering Seas (5), and a 638 base pair sequence from their mitochondrial DNA COI genes was analyzed for species identification. Sequences were compared with those of adult Ammodytes personatus from the East (4), the Yellow (2) and the East China Seas (2), and Japan (3). Average genetic distance was 0.064 between the Yellow Sea larvae and the Bering Sea larvae, but was 0.0043 between the Yellow Sea larvae and all the adult A. personatus except two individuals from the East Sea. NJ-tree showed that five Yellow Sea larvae were closely clustered with adult A. personatus, except for two individuals. The five Bering Sea larvae were located distantly from adult A. personatus, and are thought to be A. hexapterus. We found morphological differences among two populations of A. personatus larvae and A. hexapterus larvae in morphometric characters and pigmentation. Our results indicated that the East Sea population of A. personatus is closer to A. hexapterus than to the Yellow Sea population of A. personatus, suggesting the East Sea population may be part of separate species.     

Aged wild-type littermates and APPswe+PS1/ΔE9 mice present similar deficits in associative learning and spatial memory independent of amyloid load

APPswe+PS1/ΔE9 transgenic (Tg) mice with Aβ plaque formation in neocortex and hippocampus were evaluated in tests measuring exploratory activity, anxiety, and memory ability using open field test (OFT), Y-maze, contextual fear conditioning (CFC), and Morris water maze (MWM). Wild type (WT) and Tg mice over eight months old showed same locomotion activity and anxiety level in novel stimulation, open field, and Y-maze contexts. In other experiments that measured associative memory and spatial memory in Tg mice and their littermates, the subjects also presented similar deficiencies in memory acquisition. These two aged groups showed abnormal freezing level variance especially in CFC test. In comparison to that in non-transgenic 8-week-old mice group, the acquisition of spatial memory in MWM task was impaired in aged WT and bigenic Tg mice. Taken together, aged wild-type littermates and Tg mice present similar deficits in associative learning and spatial memory independent of amyloid plaques.     

Generation of transgenic chickens that produce bioactive human granulocyte-colony stimulating factor

We report here the generation of transgenic chickens that produce human granulocyte-colony stimulating factor (hG-CSF) using replication-defective Moloney murine leukemia virus (MoMLV)-based vectors packaged with vesicular stomatitis virus G glycoprotein (VSV-G). The recombinant retrovirus was injected beneath the blastoderm of nonincubated chicken embryos (stage X). Out of 140 injected eggs, 17 chicks hatched after 21 days of incubation and all hatched chicks were found to express vector-encoded hG-GSF gene. The biological activity of the recombinant hG-CSF was significantly higher than its commercially derived E. coli-derived counterpart. Successful germline transmission of the transgene was also confirmed in G(1) transgenic chicks produced from the cross of Go transgenic roosters with nontransgenic hens, but most of the G(1) progeny were dead within 1 month of hatching.     

Seedling Growth and Heavy Metal Accumulation of Candidate Woody Species for Revegetating Korean Mine Spoils

Selecting plant species that can overcome harsh soil and microclimatic conditions and speed the recovery of degraded minelands remains a worldwide restoration challenge. This study evaluated the potential of three woody species and various organic and inorganic fertilization treatments for revegetating abandoned metalliferous mines in Korea. We compared survival, growth, and heavy metal uptake of species common to Korean minelands in two spoil types and a reference forest soil. Substrate type and fertilization both influenced seedling growth and metal concentrations substantially, but they had little effect on seedling survival. Fertilization increased the growth of all three species when grown in mine spoils but influenced the growth of seedlings grown in forest soil only marginally. Initial seedling survival and growth indicate that the study species can tolerate the heavy metal concentrations and other soil constraints of metalliferous spoil types. We estimate that plants can stabilize 2–22% of various heavy metals contained in spoil materials into plant biomass during 20 years of plantation growth. Combined with the erosion control and site amelioration benefits of mineland reforestation, stabilization of heavy metals in forest biomass justifies this treatment on abandoned Korean metalliferous mines.     

Leaf traits and their interrelationship of 23 plant species in southeast of Keerqin Sandy Lands,China

Six leaf traits,i.e.,fresh mass (FM),dry mass (DM),leaf dry matter content (DMC),area (AR),specific leaf area (SLA) and thickness (TH) from 23 plant species in the southeastern Keerqin Sandy Lands,China were measured.The results show that leaf traits of herbs were more diversified than those of shrubs and trees and aver-age SLA tended towards a decreasing trend from herbs to shrubs to trees.On the contrary,DMC and DM show an upward trend from herbs to shrubs to trees.No apparent difference was found in TH.Except for DM and TH,there were significant variations in SLA and DMC among three different growth forms.Moreover,a significant correla-tion was found between SLA and DMC.It is concluded that SLA and DMC could be used to predict species posi-tion along a resource use gradient.