Birth Weight, Cord Blood Lipoprotein and Apolipoprotein Levels in Indian Newborns

Objectives: Primordial prevention of chronic disease is of clinical and public health importance. Considering the fetal onset of atherosclerosis, we aimed to determine the cord blood level of lipoproteins and apolipoproteins as well as their correlation with birth weight and gestational age. Methods: This cross-sectional study comprised 100 healthy Indian newborns. Ten ml. of cord blood was collected from placental end of umbilical vein. Serum was separated by centrifugation and ana-lyzed on the same day for lipid profile including total cholesterol (TC), triglycerides (TG), high density lipoprotein- cholesterol (HDL-C), very low density lipoproteincholesterol (VLDL) and low density lipoprotein-cholesterol (LDL-C), apolipoproteins A-I and B (ApoA-I, ApoB). Atherogenic index (AI) was calculated as the ratio of ApoB to ApoA-I. Results: Cord blood of female newborns had higher TC, HDL-C, LDL-C, Apo A-I, Apo B and AI as compared to male newborns, whereas TG and VLDL-C were higher in male than in female newborns. Significant positive correlation was observed between cord blood Apo A-I and HDL-C (r= 0.45, p<0.01), and between cord blood Apo-B and LDL-C (r= 0.44, p<0.01). Non-significant inverse correlation was observed between Apo A-I and ApoB with gestational age. There was a significant inverse correlation be-tween TG and gestational age (r= -0.197, p <0.05). Positive non-significant correlation was observed between AI and birth weight (r=0.046, p>0.05). Conclusions: These findings are another confirmatory evidence for the association of prenatal factors with cord blood lipid pro-file, and can serve as starting point for studying lipid transport system changes during early life

eIF4A inhibitors suppress cell-cycle feedback response and acquired resistance to CDK4/6 inhibition in cancer

CDK4/6 inhibitors are FDA-approved drugs for estrogen receptor-positive (ER+) breast cancer and are being evaluated to treat other tumor types, including KRAS-mutant non-small cell lung cancer (NSCLC). However, their clinical utility is often limited by drug resistance. Here, we sought to better understand the resistant mechanisms and help devise potential strategies to overcome this challenge. We show that treatment with CDK4/6 inhibitors in both ER+ breast cancer and KRAS-mutant NSCLC cells induces feedback upregulation of cyclin D1, CDK4, and cyclin E1, mediating drug resistance. We demonstrate that rocaglates, which preferentially target translation of key cell-cycle regulators, effectively suppress this feedback upregulation induced by CDK4/6 inhibition. Consequently, combination treatment of CDK4/6 inhibitor palbociclib with the eukaryotic initiation factor (eIF) 4A inhibitor, CR-1-31-B, is synergistic in suppressing the growth of these cancer cells in vitro and in vivo Furthermore, ER+ breast cancer and KRAS-mutant NSCLC cells that acquired resistance to palbociclib after chronic drug exposure are also highly sensitive to this combination treatment strategy. Our findings reveal a novel strategy using eIF4A inhibitors to suppress cell-cycle feedback response and to overcome resistance to CDK4/6 inhibition in cancer.Accepted manuscrip

5′-Inositol phosphatase SHIP2 recruits Mena to stabilize invadopodia for cancer cell invasion

Invadopodia are specialized membrane protrusions that support degradation of extracellular matrix (ECM) by cancer cells, allowing invasion and metastatic spread. Although early stages of invadopodia assembly have been elucidated, little is known about maturation of invadopodia into structures competent for ECM proteolysis. The localized conversion of phosphatidylinositol(3,4,5)-triphosphate and accumulation of phosphatidylinositol(3,4)-bisphosphate at invadopodia is a key determinant for invadopodia maturation. Here we investigate the role of the 5′-inositol phosphatase, SHIP2, and reveal an unexpected scaffold function of SHIP2 as a prerequisite for invadopodia-mediated ECM degradation. Through biochemical and structure-function analyses, we identify specific interactions between SHIP2 and Mena, an Ena/VASP-family actin regulatory protein. We demonstrate that SHIP2 recruits Mena, but not VASP, to invadopodia and that disruption of SHIP2–Mena interaction in cancer cells leads to attenuated capacity for ECM degradation and invasion in vitro, as well as reduced metastasis in vivo. Together, these findings identify SHIP2 as a key modulator of carcinoma invasiveness and a target for metastatic disease

Minnelide effectively eliminates CD133+ side population in pancreatic cancer

BACKGROUND: Pancreatic Ductal Adenocarcinoma (PDAC) is a devastating disease hallmarked by limited patient survival. Resistance to chemotherapy, a major cause of treatment failure in PDAC patients, is often attributed to Cancer Stem Cells (CSCs). Pancreatic CSCs are a small subset of quiescent cells within a tumor represented by surface markers like CD133. These cells are responsible not only for tumor recurrence, but also poor prognosis based on their “stem-like” characteristics. At present, conventional therapy is directed towards rapidly dividing PDAC cells and thus fails to target the CSC population. METHODS: MIA PaCa-2, S2-013 and AsPC-1 were treated with 12.5 nM triptolide (12 T cells) for 7 days. The surviving cells were recovered briefly in drug-free growth media and then transferred to Cancer Stem cell Media (CSM). As a control, untreated cells were also transferred to CSM media (CSM). The 12 T and CSM cells were tested for stemness properties using RNA and protein markers. Low numbers of CSM and 12 T cells were implanted subcutaneously in athymic nude mice to study their tumorigenic potential. 12 T and CSM cells were sorted for CD133 expression and assayed for their colony forming ability and sphere forming ability. Invasiveness of 12 T cells, CSM and MIA PaCa-2 were compared using Boyden chamber assays. RESULTS: Treated 12 T cells displayed increased expression of the surface marker CD133 and the drug transporter ABCG2 compared to untreated cells (CSM cells). Both 12 T and CSM cells formed subcutaneous tumors in mice confirming their tumor-initiating properties. When tested for invasion, 12 T cells had increased invasiveness compared to CSM cells. CD133(+) cells in both CSM and 12 T showed greater colony and sphere forming ability compared to CD133(−) cells from each group. Consistent with these data, when injected subcutaneously in mice, CD133(−) cells from CSM or 12 T did not form any tumors whereas CD133(+) cells from both groups showed tumor formation at a very low cell number. Despite pre-exposure to triptolide in 12 T CD133(+) cells, treatment of tumors formed by these cells with Minnelide, a triptolide pro-drug, showed significant tumor regression. CONCLUSION: Our results indicated that triptolide enhanced and enriched the “stemness” in the PDAC cell lines at a low dose of 12.5 nM, but also resulted in the regression of tumors derived from these cells. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s12943-015-0470-6) contains supplementary material, which is available to authorized users

Temperature sensing in plants

It is now well established that cold-triggered calcium influx mediates cold-induced gene expression and development of freezing tolerance (cold acclimation). In this thesis, cold signaling events both upstream and downstream of calcium influx were examined.First, it was shown that the studies on calcium mediation of cold acclimation in alfalfa cell suspension cultures could be applied to intact seedlings of Arabidopsis. Calcium chelators and channel blockers caused a strong reduction in the cold-induced accumulation of kin1 and kin2 transcripts, suggesting that calcium influx was an essential event during cold signaling and that the source of calcium for this influx was largely the calcium-rich cell wall. Evidence suggesting the involvement of calcium-dependent protein kinases (CDPKs) was also obtained.Second, the nature of events upstream of calcium influx was explored. For this study, transgenic Brassica napus seedlings possessing both the endogenous cold-inducible BN115 gene and the coding part of beta-glucuronidase (GUS) gene placed under the control of the BN115 promoter were used. Thus cold-activation of the BN115 promoter drove the expression of both BN115 at the transcriptional level and the GUS enzyme activity at the translational level. Cold-activation of BN115 was inhibited by chemicals which cause membrane fluidization, cytoskeletal stabilization and inhibition of Ca2+ influx, and mimicked at 25°C by chemicals causing membrane rigidification, cytoskeletal destabilization and Ca2+ influx. Inhibitors of protein and lipid kinases prevented cold-activation of BN115, but inhibition of protein phosphatases activated BN115 at 25°C.Third, given the increasing importance of mitogen-activated protein kinases (MAPKs) in signal transduction, the nature of molecular mechanisms that lead to cold-activation of a previously reported MAPK, SAMK, was investigated. During this study, the first plant MAPK activated by heat shock was discovered and named HAMK (Heat-shock-activated MAPK). It was shown that cold-activation of SAMK is mediated by cold-induced membrane rigidification, whereas the heat shock-activation of HAMK occurs through heat shock-induced membrane fluidization. Whereas activation of both SAMK and HAMK is blocked by an actin microfilament stabilizer, it is mimicked at 25°C by chemical destabilizers of microtubules or actin microfilaments. All of these events are inhibited by blocking the influx of extracellular Ca 2+. Cold-activation of SAMK and heat-activation of HAMK was prevented by treatment of cells with inhibitors of CDPKs. Thus, cold and heat shock are sensed by structural changes in the plasma membrane, which transduces the signal via cytoskeletal rearrangements to the opening of calcium channels, leading to Ca2+ influx, activation of CDPKs and activation of distinct MAPK cascades

Mechanism of Action of the Anti-cancer Agent, Triptolide

Triptolide, a dipertene triepoxide isolated from the roots of the Chinese herb Tripterygium wilfordii Hook F., is a promising anti-cancer agent. While its role as a promoter of cell death, both in vivo and in vitro, in various cancers is well established, the mechanism by which it induces cell death in cancer cells is not well understood, and has therefore been the subject of intense interest in the past decade. Studies to date have shown that triptolide acts in a pleiotropic fashion, resulting in decrease of HSP70 expression, affecting calcium release, causing lysosomal membrane depolarization, inhibiting NFκB activity, iNOS and Cox-2 expression, as well as acting as a transcription inhibitor and an anti-angiogenesis factor. In this review, we discuss the possible modes of action of triptolide in various cancers, as well as a novel compound derived from triptolide currently being prepared for Phase I clinical trials

Crk Associates with a Multimolecular Paxillin/GIT2/β-PIX Complex and Promotes Rac-dependent Relocalization of Paxillin to Focal Contacts

We have previously demonstrated that the CrkII and CrkL adapter proteins are required for the spreading of epithelial colonies and the breakdown of adherens junctions in response to hepatocyte growth factor. When overexpressed, CrkII and CrkL promote lamellipodia formation, cell spreading, and the loss of epithelial adherens junctions in the absence of hepatocyte growth factor. The exact mechanism by which Crk proteins elicit these changes is unclear. We show that the overexpression of CrkII or CrkL, but not Src homology 2 or amino-terminal Src homology 3 domain mutant Crk proteins, promotes the relocalization of Paxillin to focal contacts throughout the cell and within lamellipodia in a Rac-dependent manner. In stable cell lines overexpressing CrkII, enhanced lamellipodia formation and cell spreading correlate with an increased association of CrkII with Paxillin, GIT2 (an ARF-GAP) and β-PIX (a Rac1 exchange factor). Mutants of Paxillin that fail to associate with Crk or GIT2, or do not target to focal adhesions inhibit Crk-dependent cell spreading and lamellipodia formation. We conclude from these studies that the association of Crk with Paxillin is important for the spreading of epithelial colonies, by influencing the recruitment of Paxillin to focal complexes and promoting the enhanced assembly of Paxillin/GIT2/β-PIX complexes