Biomedical subjects

M Samiei

Publications and source records attributed to M Samiei.

6 recordsLinked to original sources

PubMed1993

Activation of myelin basic protein and S6 peptide kinases in phorbol ester- and PAF-treated sheep platelets.

The involvement of myelin basic protein (MBP) kinases and ribosomal S6 peptide kinases in sheep platelet signal transduction was investigated. Treatment of platelets with 200 nM 12-O-tetradecanoylphorbol-13-acetate (PMA) led to 5-fold stimulations of cytosolic MBP and S6 peptide kinase activities within 1 min. Immunoblotting analysis of phenyl-Superose-fractionated cytosol from PMA-treated platelets with a panel of mitogen-activated protein (MAP) kinase anti-peptide antibodies revealed that one of the activated MBP kinases was p42mapk. This MAP kinase isoform was also stimulated to a lesser extent (approximately 2-fold) when platelets were exposed to 200 microM platelet-activating factor (PAF) for 3 min. The pathways of PAF-activation of p42mapk also involved a protein kinase C-independent route, since the staurosporin analog compound 3 reduced PAF-induced activation by approximately 30% under conditions in which it inhibited PMA-activation of p42mapk by approximately 80%. Another MAP kinase isoform of 44 kDa, most probably p44erk1, was also detected in platelet cytosol, but it was only marginally modulated in response to PMA or PAF. The predominant PMA- and PAF-activated MBP kinase detected after MonoQ fractionation of platelet cytosol did not appear to correspond to a MAP kinase. MonoQ chromatography of platelet cytosol also resolved two PMA- and PAF-activated S6 peptide kinases, which appeared to coelute on phenyl-Sepharose. Western blotting analysis of the MonoQ fractions with antibodies raised against peptide sequences in the S6 kinases p90rsk and p70S6K revealed immunoreactive proteins of approximately 75 kDa and approximately 95 kDa that coincided with the first S6 peptide kinase peak. These proteins probably corresponded to the 502 and 525 amino-acid-length forms of p70S6K. Only the second peak of S6 peptide kinase activity from MonoQ was appreciably stimulated in response to PAF-treatment of platelets, and this was largely abolished by compound 3. It is more likely that the novel MBP and S6 peptide kinases described here, rather than p42mapk and p70S6K, play a significant role in PAF signal transduction in the platelet.

Amino Acid Sequence↗

PubMed1993

Networking with mitogen-activated protein kinases.

Mitogen activated protein (MAP) kinases and their target ribosomal protein S6 (RSK) kinases have been recognized as shared components in the intracellular signaling pathways of many diverse cytokines. Recent studies have extended this protein kinase cascade by identifying the major activator of vertebrate MAP kinases as a serine/threonine/tyrosine-protein kinase called MEK, which is related to yeast mating factor-regulated protein kinases encoded by the STE7 and byr1 genes. MEK, in turn, may be activated following its phosphorylation on serine by either of the kinases encoded by proto-oncogenes raf1 or mos, as well as by p78mekk, which is related to the yeast STE11 and byr2 gene products. Isoforms of all of these protein kinases may specifically combine to assemble distinct modules for intracellular signal transmission. However, the fundamental architecture of these protein kinase cascades has been highly conserved during eukaryotic evolution.

Animals↗

PubMed1991

Platelet-activating factor- and thrombin-induced stimulation of p34cdc2-cyclin histone H1 kinase activity in platelets.

Numerous studies of cell cycle control in dividing cells have pointed to the central role of a 34-kDa histone H1 kinase (p34cdc2) complexed with regulatory subunits known as cyclins. We now report that p34cdc2-cyclin may also participate in signal transduction in nonproliferating, terminally differentiated cells, in this instance during sheep platelet activation. Immunological evidence for the presence of a p34cdc2 cognant in sheep platelet cytosol was obtained with antipeptide antibodies raised against peptide sequences in the conserved PSTAIRE and C-terminus regions of murine cdc2. The immunoreactive 32-kDa protein was adsorbed onto p13suc1-Sepharose, which selectively binds p34cdc2. A 58-kDa protein that also bound to p13suc1-Sepharose was identified as cyclin A on the basis of its size and immunoreactivity with two different anticyclin peptide antibodies. The p34cdc2-cyclin A complex was regulated during platelet activation. Its histone H1 phosphorylating activity was stimulated 2-fold in p13suc1-Sepharose extracts from platelets that had been exposed to platelet-activating factor or thrombin for 1 min prior to harvesting. Our findings imply that the p34cdc2-cyclin complex may serve alternative functions besides control of cell division.

Amino Acid Sequence↗

PubMed1991

Characterization of calcium-independent forms of protein kinase C-beta in phorbol ester-treated rabbit platelets.

The subcellular distribution, size, and activation state of protein kinase C (PKC) were studied after short term exposure of rabbit platelets to a saturating dose of 12-O-tetradecanoylphorbol 13-acetate (TPA). Cytosolic and Nonidet P-40-solubilized particulate extracts prepared from TPA-treated platelets were subjected to analytical column chromatography on Mono Q, hydroxylapatite, and Superose 6/12. PKC activity was assayed according to the ability of the enzyme to phosphorylate (i) histone H1 in the presence of the activators calcium, diacylglycerol, and phosphatidylserine; (ii) histone H1 after proteolytic activation of PKC with trypsin; and (iii) protamine in the absence of calcium and lipid. Within 1 min of TPA treatment of platelets, greater than 95% of the PKC activity was particulate associated, as assessed by all three methods. The particulate PKC activity from 1-min TPA-treated cells eluted from Mono Q with approximately 0.35 M NaCl (peak I), and it was highly dependent upon Ca2+ and lipid for optimal histone H1 phosphorylation. With longer exposure times of platelets to TPA, the disappearance of the Mono Q peak I form of PKC was correlated with the production of new PKC species that were released from Mono Q with approximately 0.4 M NaCl (peak II), approximately 0.5 M NaCl (peak III), and approximately 0.6 M NaCl (peak IV). These last forms of PKC were still lipid activated but exhibited little Ca2+ dependence. The Mono Q peak III form displayed a particularly high level of histone H1 phosphorylating activity in the absence of lipid and Ca2+. All of these forms behaved as approximately 65-kDa proteins on Superose 6/12, but on sodium dodecyl sulfate-polyacrylamide gels, Western blotting with anti-PKC-beta antibodies revealed immunoreactive polypeptides of approximately 79 kDa (Mono Q peaks I, II, and IV) and approximately 100-kDa (Mono Q peak III). Hydroxylapatite column chromatography permitted partial resolution of the Mono Q peaks I and II forms, which were eluted within a concentration range of potassium phosphate (100-150 mM) which was typical of the beta isozyme of PKC. Treatment of the Mono Q peak III and IV PKC forms with alkaline phosphatase resulted in the production of the peak I form, which implicated protein phosphorylation in the interconversion of the various PKC forms.

Alkaline Phosphatase↗

PubMed1991

Clonal selection within metastatic SP1 mouse mammary tumors is independent of metastatic potential.

Our previous studies using randomly integrated plasmid DNA as unique clonotypic markers of SPI mouse mammary tumor cells transplanted into syngeneic CBA/J or nude mice demonstrated reproducible selection and eventual overgrowth of the primary transplant tumors by genotypically distinct metastatic subclones. Two independent metastatic SPI clones, neo5 and ras1, were shown to exhibit "clonal dominance" relative to the non-metastatic SPI tumor-cell population. These results suggested that the capacity for preferential growth within the tumors may be related to cellular properties associated with metastatic ability. To investigate the clonal interactions of metastatic SPI clones present within the same tumor mass, we have analyzed tumors composed of paired mixtures of neo5 and ras1. The tumors were monitored for the relative proportion of each clone by Southern blot analysis. The ras1 clone was found to dominate over the neo5 clone in the majority of tumors examined, even when present as 1% of the mixed inoculum. This represents a 20- to 50-fold enrichment of ras1, while the proportion of neo5 within the tumors was reduced at least 5-fold. No evidence for selection of either clone was seen during co-culture in vitro. Neo5 and ras1 are indistinguishable with respect to tumorigenic and metastatic potential when inoculated separately into different mice, suggesting that clonal dominance is independent of metastatic ability. Analysis of the metastases resulting from mixed inocula indicates that it is possible for a subpopulation representing less than 1% of the primary tumor mass to give rise to metastases. This also suggests that the process of metastasis within metastatic tumors is independent of clonal dominance.

Adenocarcinoma↗

PubMedDate not supplied

An Internet-based exercise as a component of an overall training program addressing medical aspects of radiation emergency management.

The use of ionizing radiation and radioactive materials continues to increase worldwide in industry, medicine, agriculture, research, electrical power generation, and nuclear weaponry. The risk of terrorism using weapons of mass destruction or simple radiological devices also has increased, leading to heightened concerns. Radiation accidents occur as a consequence of errors in transportation of radionuclides, use of radiation in medical diagnosis and therapy, industrial monitoring and sterilization procedures, and rarely, nuclear power generation. Compared to other industries, a small number of serious radiation accidents have occurred over the last six decades with recent cases in the Republic of Georgia, Peru, Japan, and Thailand. The medical, psychological, and political consequences of such accidents can be considerable. A number of programs designed to train medical responders in the techniques of radiation accident management have been developed and delivered in many countries. The low frequency of serious radiation accidents requires constant re-training, as skills are lost and medical staff turnover occurs. Not all of the training involves drills or exercises in which responders demonstrate learning or communication over the broad spectrum of medical response capabilities. Medical preparedness within the context of a total emergency response program is lacking in many parts of the world, particularly in Central and Eastern Europe and the Newly Independent States. This paper describes an effort to enhance medical preparedness in the context of a total program of international cooperation and conventions facilitated by the International Atomic Energy Agency. The paper concludes that novel application of telecommunications technology as part of a training activity in radiation accident preparedness can help address gaps in training in this field in which preparedness is essential but experience and practical field exercises are lacking.

Attitude of Health Personnel↗