Abstract: The N-methyl-D-aspartate (NMDA)-dependent pathway is the major intracellular signaling pathway implemented in both short- and long-term memory formation in the hippocampus which is the most studied brain structure because of its well documented role in learning and memory. However, little is known about the effects of RF-EMR exposure on NMDA receptor signaling pathway including activation of protein kinases, notably Ca2+/calmodulin-dependent protein kinase II alpha (CaMKIIα). The aim of the present study was to investigate the effects of acute and chronic 900 MHz RF-EMR exposure on both passive avoidance behaviour and hippocampal levels of CaMKIIα and its phosphorylated form (pCaMKIIα). Rats were divided into the following groups: Sham rats, and rats exposed to 900 MHz RF-EMR for 2 h/day for 1 week (acute group) or 10 weeks (chronic group), respectively. Passive avoidance task was used as a behavioural method. The hippocampal levels of selected kinases were measured using Western Blotting technique. The results of passive avoidance task showed that both acute and chronic exposure to 900 MHz RF-EMR can impair passive avoidance behaviour with minor effects on chronic group of rats. The analysis of western blot data of selected protein kinases demonstrated that hippocampal levels of CaMKIIα and pCaMKIIα were significantly higher in chronic group of rats as compared to acute groups. Taken together, these findings demonstrated that different duration times (1 week vs 10 weeks) of 900 MHz RF-EMR exposure have different effects on both passive avoidance behaviour of rats and hippocampal levels of selected protein kinases.
Abstract: Several embryonic cellular mechanism including cell
cycle, growth and apoptosis are regulated by phosphatidylinositol-3-
kinase (PI3K)/Akt signaling pathway. The goal of present study is to
determine the effects of annatto (Bixa orellana)-derived δ-tocotrienol
(δ-TCT) on the regulations of PI3K/Akt genes in murine morula.
Twenty four 6-8 week old (23-25g) female balb/c mice were
randomly divided into four groups (G1-G4; n=6). Those groups were
subjected to the following treatments for 7 consecutive days: G1
(control) received tocopherol stripped corn oil, G2 was given 60
mg/kg/day of δ-TCT mixture (contains 90% delta & 10% gamma
isomers), G3 was given 60 mg/kg/day of pure δ-TCT (>98% purity)
and G4 received 60 mg/kg/day α-TOC. On Day 8, females were
superovulated with 5 IU Pregnant Mare’s Serum Gonadotropin
(PMSG) for 48 hours followed with 5 IU human Chorionic
Gonadotropin (hCG) before mated with males at the ratio of 1:1.
Females were sacrificed by cervical dislocation for embryo collection
48 hours post-coitum. About fifty morulas from each group were
used in the gene expression analyses using Affymetrix QuantiGene
Plex 2.0 Assay. Present data showed a significant increase (p
Abstract: Protein kinases participate in a myriad of cellular
processes of major biomedical interest. The in vivo substrate
specificity of these enzymes is a process determined by several
factors, and despite several years of research on the topic, is still
far from being totally understood. In the present work, we have
quantified the contributions to the kinase substrate specificity of
i) the phosphorylation sites and their surrounding residues in the
sequence and of ii) the association of kinases to adaptor or scaffold
proteins. We have used position-specific scoring matrices (PSSMs),
to represent the stretches of sequences phosphorylated by 93 families
of kinases. We have found negative correlations between the number
of sequences from which a PSSM is generated and the statistical
significance and the performance of that PSSM. Using a subset
of 22 statistically significant PSSMs, we have identified specificity
determinant residues (SDRs) for 86% of the corresponding kinase
families. Our results suggest that different SDRs can function as
positive or negative elements of substrate recognition by the different
families of kinases. Additionally, we have found that human proteins
with known function as adaptors or scaffolds (kAS) tend to interact
with a significantly large fraction of the substrates of the kinases to
which they associate. Based on this characteristic we have identified
a set of 279 potential adaptors/scaffolds (pAS) for human kinases,
which is enriched in Pfam domains and functional terms tightly
related to the proposed function. Moreover, our results show that
for 74.6% of the kinase–pAS association found, the pAS colocalize
with the substrates of the kinases they are associated to. Finally, we
have found evidence suggesting that the association of kinases to
adaptors and scaffolds, may contribute significantly to diminish the
in vivo substrate crossed-specificity of protein kinases. In general, our
results indicate the relevance of several SDRs for both the positive
and negative selection of phosphorylation sites by kinase families and
also suggest that the association of kinases to pAS proteins may be
an important factor for the localization of the enzymes with their set
of substrates.
Abstract: Heme oxygenase-1 (HO-1), an enzyme degrading heme to carbon monoxide, iron, and biliverdin, has been recognized as playing a crucial role in cellular defense against stressful conditions, not only related to heme release. In the present study, the effects of TNF-a on the expression of heme oxygenase-1 (HO-1) in human aortic endothelial cells (HAECs) as well as the related mechanisms were investigated. 10 ng/mL TNF-α treatment significantly increased HO-1 expression after 6h, then a further increase at 12h and declined at 24h. Treatment with 2 ng/mL of TNF-a after 12 h resulted in a significant increase in HO-1 expression, which peaked at 10 ng/mL, then declined at 20 ng/mL. TNF-α induced HO-1 expression and then HO-1 expression reduced vascular cell adhesion molecule-1 (VCAM-1) expression. Phosphorylation studies of ERK1/2, JNK, and p38, three subgroups of mitogen-activated protein kinases (MAPKs) demonstrated TNF-α-induced ERK1/2, JNK, and p38 phosphorylation. The increase in HO-1 expression in response to TNF-α treatment was affected by pretreatment with SP600125 (a JNK inhibitor) and SB203580 (a p38 inhibitor), not with PD98059 (an ERK1/2 inhibitor). The expression of HO-1 was stronger in aortas of TNF-α-treated apo-E deficient mice when compared with control mice. These results suggest that low dose of TNF-α treatment notably induced HO-1 expression was mediated through JNK/p38 phosphorylation and may have a protective potential in cardiovascular diseases and inflammatory response through the regulation of HO-1 expression.