Actin Inhibition Increases Megakaryocyte Proplatelet Formation through an Apoptosis-Dependent Mechanism

Background

Megakaryocytes assemble and release platelets through the extension of proplatelet processes, which are cytoplasmic extensions that extrude from the megakaryocyte and form platelets at their tips. Proplatelet formation and platelet release are complex processes that require a combination of structural rearrangements. While the signals that trigger the initiation of proplatelet formation process are not completely understood, it has been shown that inhibition of cytoskeletal signaling in mature megakaryocytes induces proplatelet formation. Megakaryocyte apoptosis may also be involved in initiation of proplatelet extension, although this is controversial. This study inquires whether the proplatelet production induced by cytoskeletal signaling inhibition is dependent on activation of apoptosis.

Methods

Megakaryocytes derived from human umbilical cord blood CD34+ cells were treated with the actin polymerization inhibitor latrunculin and their ploidy and proplatelet formation were quantitated. Apoptosis activation was analyzed by flow cytometry and luminescence assays. Caspase activity was inhibited by two compounds, ZVAD and QVD. Expression levels of pro-survival and pro-apoptosis genes were measured by quantitative RT-PCR. Protein levels of Bcl-XL, Bax and Bak were measured by western blot. Cell ultrastructure was analyzed by electron microscopy.

Results

Actin inhibition resulted in increased ploidy and increased proplatelet formation in cultured umbilical cord blood-derived megakaryocytes. Actin inhibition activated apoptosis in the cultured cells. The effects of actin inhibition on proplatelet formation were blocked by caspase inhibition. Increased expression of both pro-apoptotic and pro-survival genes was observed. Pro-survival protein (Bcl-x_L) levels were increased compared to levels of pro-apoptotic proteins Bak and Bax. Despite apoptosis being activated, the megakaryocytes underwent minimal ultrastructural changes during actin inhibition.

Conclusions

We report a correlation between increased proplatelet formation and activation of apoptosis, and that the increase in proplatelet formation in response to actin inhibition is caspase dependent. These findings support a role for apoptosis in proplatelet formation in this model.     

Investigating Polycyclic Aromatic Hydrocarbons Profiles in Higher Plants Using Statistical Models

Thirty-six higher plants sampled from Olomoro, Irri, Uzere, and Oginni exploration sites in the Niger Delta region of Nigeria were subjected to GC/MS analysis to assess the occurrence, distribution and profiles of polycyclic aromatic hydrocarbons (PAHs) contained in them. The Σ₂₈PAHs ranged from 335 to 3094 ng/g. The results of the nonparametric regression models showed that PAHs concentration in a plant cannot be used in isolation to deduce the total PAHs concentration in soils hosting the plant since PAHs concentration in a plant is influenced by the presence (or absence) of other plants in that location. A combination of Factor analysis (FA) and principal component analysis (PCA) were used to recognize PAHs concentration patterns among the plants in the studied locations and individual PAHs compounds. Woody annuals and perennial plants formed similar patterns in Oginni and Irri locations. Three main clusters were formed by all the compounds with naphthalene and 2-methylnaphthalene standing as outliers in all the four locations.

Supplemental materials are available for this article. Go to the publisher's online edition of International Journal of Phytoremediation to view the supplemental file.     

Biomarkers for early detection of high risk cancers: From gliomas to nasopharyngeal carcinoma

Nasopharyngeal carcinoma (NpC) is a malignant disease associated with Epstein-Barr virus infection, and often diagnosed at an advanced stage. This significantly curtails patient survival. We hypothesize that a panel of biomarkers can be assembled to assess NpC incidence, early detection, and tumor progression during therapeutic intervention. Our thesis rests on a model of successfully predicting high-risk gliomas by means of a carefully crafted panel of molecular mitotic biomarkers (i.e., securin, survivin and MCM2). The strategy we propose holds strong promise for prevention and cure of NpC. The approach we propose seeks to identify certain biomarkers from viral materials, patient tissues and assessment of related diseases, whose signatures, taken together, will be endowed with some degree of congruency, or sense of a coordinated language (i.e., “votes”). Biomarker “voting” will then permit to outline a broad coordinated molecular map for the molecular and epigenetic characterization of each individual patient''s NpC tumor. We will draw on the process of contrasting biomarkers in health and disease, which rests on the auto-proteomic concept particularly relevant in high-risk cancer individuals, such as is the case for NpC. In brief we defend, current advances in human proteome profiling proffers the possibility of having individual baseline proteomic profiles using local body fluids (e.g., saliva, nasal secretions, sputum) or systemic fluids (e.g., plasma, serum, cerebrospinal fluid) to unravel a personalized molecular map for high-risk NpC individuals. Regular check-up will monitor for new or impending manifestations of NpC, and provide a secure assessment of incidence and early detection.