Activation of Akt at T308 and S473 in alcohol, tobacco and HPV-induced HNSCC:is there evidence to support a prognostic or diagnostic role?

BACKGROUND: Tobacco, alcohol and HPV infection are associated with increased risk of HNSCC. However, little is known about the underlying signaling events influencing risk. We aimed to investigate the relationship between these risk factors and Akt phosphorylation, to determine prognostic value. METHOD: VEGF-positive HNSCC biopsies, with known HPV status, were analyzed by immunohistochemistry (IHC) for Akt, phosphorylated at residues S473 and T308. Comparisons between the tissues were carried out using a Mann–Whitney U test. Associations between the variables and continuous immunohistochemical parameters were evaluated with general linear models. Patient characteristics and pAkt IHC score were analyzed for possible association with overall survival by Cox proportional hazard models. RESULTS: Immunohistochemistry revealed that cancer patients had significantly higher levels of pAkt T308 than S473 (P < 0.001). Smoking and alcohol were found to be independent risk factors for Akt phosphorylation at T308 (P = 0.022 and 0.027, respectively). Patients with tumors positive for HPV or pAkt S473 had a poorer prognosis (P = 0.005, and 0.004, respectively). Patients who were heavy drinkers were 49 times more likely to die than non-drinkers (P = 0.003). Patients with low pAkt T308 were more likely to be HPV positive (P = 0.028). Non-drinkers were also found to have lower levels of pAkt T308 and were more likely to have tumors positive for HPV than heavy drinkers (P = 0.044 and 0.007, respectively). CONCLUSION: This study suggests different mechanisms of carcinogenesis are initiated by smoking, alcohol and HPV. Our data propose higher phosphorylation of Akt at T308 as a reliable biomarker for smoking and alcohol induced HNSCC progression and higher phosphorylation of Akt at S473 as a prognostic factor for HNSCC

Review: Electric field driven pumping in microfluidic device

Pumping of fluids with precise control is one of the key components in a microfluidic device. The electric field has been used as one of the most popular and efficient nonmechanical pumping mechanism to transport fluids in microchannels from the very early stage of microfluidic technology development. This review presents fundamental physics and theories of the different microscale phenomena that arise due to the application of an electric field in fluids, which can be applied for pumping of fluids in microdevices. Specific mechanisms considered in this report are electroosmosis, AC electroosmosis, AC electrothermal, induced charge electroosmosis, traveling wave dielectrophoresis, and liquid dielectrophoresis. Each phenomenon is discussed systematically with theoretical rigor and role of relevant key parameters are identified for pumping in microdevices. We specifically discussed the electric field driven body force term for each phenomenon using generalized Maxwell stress tensor as well as simplified effective dipole moment based method. Both experimental and theoretical works by several researchers are highlighted in this article for each electric field driven pumping mechanism. The detailed understanding of these phenomena and relevant key parameters are critical for better utilization, modulation, and selection of appropriate phenomenon for efficient pumping in a specific microfluidic application

The alternate GNB3 splice variant, Gβ3s, exhibits an altered signalling response to EGF stimulation, which leads to enhanced cell migration

It has recently been reported that the duplication of the GNB3 gene has been shown to be directly linked to an obesity phenotype, both in humans and also in a humanised mouse model. Moreover, the common human GNB3 c.825C&gt;T polymorphism (rs5443) causes this ubiquitously expressed gene to be aberrantly spliced approximately 50% of the time leading to the production of both a normal Gβ3 protein and a truncated, possibly less stable subunit, known as Gβ3s. The presence of the GNB3 825T allele has previously been shown to be associated with predisposition to hypertension, obesity, various cancers, Alzheimers, age related cognitive function, erectile dysfunction as well as a marker for pharmacogenetic drug action. Great controversy, however, currently exists as to whether these phenotypes associated with the 825T allele are a) mainly due to the presence of the smaller, possibly more active, Gβ3s subunit or b) merely down to the haploinsufficiency of the normal GNB3 transcript, due to its frequent aberrant splicing. In order to try and address these two conflicting hypothesis, we report on the identification and characterisation of signalling alterations unique to the presence of Gβ3s protein subunit. Moreover we also show the physiological consequences associated with altered signalling, directly induced by the Gβ3s subunit. For this, we used both an EBV transformed lymphoblast cell line homozygote for GNB3 825T/825T (TT) and a stable Gβ3s expressing recombinant COS-7 clone. In both of these cell lines that express the Gβ3s subunit, we found enhanced cytosolic calcium influx upon stimulation with EGF, TGFα and VEGF ligands, as compared to “normal” GNB3 controls with the 825C/825C (CC) genotype. This aberrant calcium influx also led to an increase in ERK, but not AKT1, phosphorylation. Despite the lack of AKT1 activation, we paradoxically observed a significant increase in phosphorylation of its downstream substrates, namely mTOR and p70S6k (KS6B2). Moreover we observed a decrease in phospho FoxO3a only in Gβ3s expressing cells, but not in the “normal” GNB3 (CC) control cell line. The presence of the Gβ3s subunit also appeared to alter the distinct localisation patterns of both Foxo3a and AKT1, while also increasing the colocalisation of mTOR and p70S6K. Subsequent growth factor stimulation studies revealed that EGF treatment, of Gβ3s expressing cells, appeared to cause a significant decrease in cAMP levels, which, in turn resulted in both enhanced caveolin-1a phosphorylation, and an increase in actin stress fibre formation. The identification of these distinct Gβ3s specific signalling alterations were indicative of a more aggressive migratory phenotype. This led us to further investigate and confirm that the presence of the Gβ3s subunit also appears to cause significantly enhanced migration and robust scratch wound healing kinetics, as compared to cells harbouring only the normal copy of the gene. These data therefore present convincing evidence that the Gβ3s subunit is stable, functional and its presence can significantly alter signalling pathways, in different cell types

Chemical and biological investigations of Delonix regia (Bojer ex Hook.) Raf.

U radu je opisana izolacija pet sastojaka petroleterske i diklormetanske frakcije metanolnog ekstrakta kore biljke Delonix regia: lupeol (1), epilupeol (2), β-sitosterol (3), stigmasterol (4) i p-metoksibenzaldehid (5). Nadalje, testirano je antimikrobno djelovanje različitih ekstrakata difuzijskom metodom na disku (15 μg mm2). Zone inhibicije za sastojke topljive u petroleteru, tetraklormetanu i diklormetanu bile su 914 mm, 1113 mm, odnosno 920 mm, dok je zona inhibicije standarda kanamicina bila 2025 mm. U biološkom pokusu smrtnosti morskih kozica najveću toksičnost pokazali su spojevi topljivi u tetraklormetanu (LC50 = 0,83 μg mL1), dok je topljivost sastojaka topljivih u petroleteru i diklormetanu bila LC50 14,94, odnosno 3,29 μg mL1, a standarda vinkristin sulfata 0,812 μg mL1. Ovo je prvo izvješće o izolaciji sastojaka, antimikrobnom djelovanju i citotoksičnosti biljke D. regia.In this study five compounds, lupeol (1), epilupeol (2), β-sitosterol (3), stigmasterol (4) and p-methoxybenzaldehyde (5) were isolated from the petroleum ether and dichloromethane fractions of a methanolic extract of the stem bark of Delonix regia. Antimicrobial screening of the different extracts (15 μg mm2) was conducted by disc diffusion method. The zones of inhibition demonstrated by the petroleum ether, carbon tetrachloride and dichloromethane fractions ranged from 914 mm, 1113 mm and 920 mm, respectively, compared to kanamycin standard with the zone of inhibition of 2025 mm. In brine shrimp lethality bioassay, the carbon tetrachloride soluble materials demonstrated the highest toxicity with LC50 of 0.83 μg mL1, while petroleum ether and dichloromethane soluble partitionates of the methanolic extract revealed LC50 of 14.94 and 3.29 μg mL1, respectively, in comparison with standard vincristine sulphate with LC50 of 0.812 μg mL1. This is the first report on compounds separation from D. regia, their antimicrobial activity and cytotoxicity

Synthesis, Characterization and Bioactivities of Some Novel Oxovanadium(IV) Glycinato Complexes

The novel oxovanadium(IV) complexes, [VIVO(GlyH)(Gly)]+ClO4 - .H2O (1), [VIVO(GlyH)(Gly)]+NO3 - .H2O (2), [VIVO(GlyH)(Gly)]+CH3COO- .H2O (3) were synthesized and characterized by FT-IR, UV-Vis and 1H NMR spectroscopic measurements. The cumulative spectroscopic assessment envisaged that, the complexes adopt a square pyramidal structure, in which the two glycine ligands coordinate to vanadium(IV) center in bidentate fashions conforming a homoleptic structure. The amino nitrogen and a carboxylato oxygen atom coordinate the vanadium(IV) center from both sides making a five members chelate by each side. All the complexes are stable in amorphous state and in aerobic and anaerobic solution. Significantly, all the complexes have the antifungal activities against Aspergillus niger and Penicillium notatum but ineffective against Candida tropicalis. No antibacterial activity was observed for the complexes against tested bacteria and unfortunately, they were found cytotoxic against brine shrimp bioassay

TANGENT BUNDLES OF LP-SASAKIAN MANIFOLD ENDOWED WITH GENERALIZED SYMMETRIC METRIC CONNECTION

The aim of the present work is to study and establish conditions for anLP-Sasakian manifold on the tangent bundle $TM$. An LP-Sasakian manifold with the generalized symmetric metric connection on $TM$ is investigated. Next, the curvature tensor and the Ricci tensor of an LP-Sasakian manifold with respect to the generalized symmetric metric connection on $TM$ are calculated. Moreover, the projective curvature tensor with respect to the generalized symmetric metric connection on $TM$ is studied and showed that $TM$ is not $\hat{\xi}^C$-projectively flat. In particular, if $\alpha=0$ and $\beta=1$ then $TM$ is $\hat{\xi}^C$-projectively flat

Synthesis and Functional Evaluation of DNA-Assembled Polyamidoamine Dendrimer Clusters for Cancer Cell-Specific Targeting

SummaryWe sought to produce dendrimers conjugated to different biofunctional moieties (fluorescein [FITC] and folic acid [FA]), and then link them together using complementary DNA oligonucleotides to produce clustered molecules that target cancer cells that overexpress the high-affinity folate receptor. Amine-terminated, generation 5 polyamidoamine (G5 PAMAM) dendrimers are first partially acetylated and then conjugated with FITC or FA, followed by the covalent attachment of complementary, 5′-phosphate-modified 34-base-long oligonucleotides. Hybridization of these oligonucleotide conjugates led to the self-assembly of the FITC- and FA-conjugated dendrimers. In vitro studies of the DNA-linked dendrimer clusters indicated specific binding to KB cells expressing the folate receptor. Confocal microscopy also showed the internalization of the dendrimer cluster. These results demonstrate the ability to design and produce supramolecular arrays of dendrimers using oligonucleotide bridges. This will also allow for further development of DNA-linked dendrimer clusters as imaging agents and therapeutics