An Improved In Vivo Methodology to Visualise Tumour Induced Changes in Vasculature Using the Chick Chorionic Allantoic Membrane Assay

Background/Aim: Decreasing the vascularity of a tumour has proven to be an effective strategy to suppress tumour growth and metastasis. Anti-angiogenic therapies have revolutionized the treatment of advanced-stage cancers, however there is still demand for further improvement. This necessitates new experimental models that will allow researchers to reliably study aspects of angiogenesis. The aim of this study was to demonstrate an in vivo technique in which the highly vascular and accessible chorioallantoic membrane (CAM) of the chick embryo is used to study tumour-induced changes in the macro and microvessels. Materials and Methods: Two cancer cell lines (human melanoma (C8161) and human prostate cancer (PC3)) were selected as model cells. Human dermal fibroblasts were used as a control. One million cells were labelled with green fluorescent protein and implanted on the CAM of the chick embryo at embryonic development day (EDD) 7 and angiogenesis was evaluated at EDDs 10, 12 and 14. A fluorescently-tagged lectin (lens culinaris agglutinin (LCA)) was injected intravenously into the chick embryo to label endothelial cells. The LCA is known to label the luminal surface of endothelial cells, or dextrans, in the CAM vasculature. Macrovessels were imaged by a hand-held digital microscope and images were processed for quantification. Microvessels were evaluated by confocal microscopy. Tumour invasion was assessed by histological and optical sectioning. Results: Tumour cells (C8161 and PC3) produced quantifiable increases in the total area covered by blood vessels, compared to fibroblasts when assessed by digital microscopy. Tumour invasion could be demonstrated by both histological and optical sectioning. The most significant changes in tumour vasculature observed were in the microvascular structures adjacent to the tumour cells, which showed an increase in the endothelial cell coverage. Additionally, tumour intravasation and tumour thrombus formation could be detected in the areas adjacent to tumour cells. The fragility of tumour blood vessels could be demonstrated when tumour cells seeded on a synthetic scaffold were grown on CAM. Conclusion: We report on a modification to a well-studied CAM in vivo assay, which can be effectively used to study tumour induced changes in macro and microvasculature

Resolved-sideband Raman cooling to the ground state of an optical lattice

We trap neutral Cs atoms in a two-dimensional optical lattice and cool them close to the zero-point of motion by resolved-sideband Raman cooling. Sideband cooling occurs via transitions between the vibrational manifolds associated with a pair of magnetic sublevels and the required Raman coupling is provided by the lattice potential itself. We obtain mean vibrational excitations \bar{n}_x \approx \bar{n}_y \approx 0.01, corresponding to a population \sim 98% in the vibrational ground state. Atoms in the ground state of an optical lattice provide a new system in which to explore quantum state control and subrecoil laser coolingComment: PDF file, 13 pages including 3 figure

Recent concepts in biodegradable polymers for tissue engineering paradigms: a critical review

Tissue engineering and regenerative medicine are emerging as future approaches for the treatment of acute and chronic diseases. Numerous clinical conditions exist today and include congenital disorders, trauma, infection, inflammation, and cancer, in which hard and soft tissue damage, organ failure and loss are still not treated effectively. Researchers are constantly developing new biomaterials and tissue-engineered technologies to stimulate tissue regeneration. Various emerging approaches according to organ, tissue, disease and disorder are identified. Irrespective, engineered biomaterials are required to regenerate and ultimately reproduce the original physiological, biological, chemical, and mechanical properties. Biodegradable materials have been used extensively as regenerative therapies. The selection, design, biological and physicochemical properties of these materials are important and must be considered for stimulating tissue growth. In this review, we critique recently developed biodegradable materials for tissue regeneration of some targeted organs e.g., skin, nerves, blood vessels, heart, cornea, trachea, dental/oral structure and bones

Sub-Poissonian statistics in order-to-chaos transition

We study the phenomena at the overlap of quantum chaos and nonclassical statistics for the time-dependent model of nonlinear oscillator. It is shown in the framework of Mandel Q-parameter and Wigner function that the statistics of oscillatory excitation number is drastically changed in order-to chaos transition. The essential improvement of sub-Poissonian statistics in comparison with an analogous one for the standard model of driven anharmonic oscillator is observed for the regular operational regime. It is shown that in the chaotic regime the system exhibits the range of sub- and super-Poissonian statistics which alternate one to other depending on time intervals. Unusual dependence of the variance of oscillatory number on the external noise level for the chaotic dynamics is observed.Comment: 9 pages, RevTeX, 14 figure

Dichotomy of Tyrosine Hydroxylase and Dopamine Regulation between Somatodendritic and Terminal Field Areas of Nigrostriatal and Mesoaccumbens Pathways

Measures of dopamine-regulating proteins in somatodendritic regions are often used only as static indicators of neuron viability, overlooking the possible impact of somatodendritic dopamine (DA) signaling on behavior and the potential autonomy of DA regulation between somatodendritic and terminal field compartments. DA reuptake capacity is less in somatodendritic regions, possibly placing a greater burden on de novo DA biosynthesis within this compartment to maintain DA signaling. Therefore, regulation of tyrosine hydroxylase (TH) activity may be particularly critical for somatodendritic DA signaling. Phosphorylation of TH at ser31 or ser40 can increase activity, but their impact on L-DOPA biosynthesis in vivo is unknown. Thus, determining their relationship with L-DOPA tissue content could reveal a mechanism by which DA signaling is normally maintained. In Brown-Norway Fischer 344 F1 hybrid rats, we quantified TH phosphorylation versus L-DOPA accumulation. After inhibition of aromatic acid decarboxylase, L-DOPA tissue content per recovered TH protein was greatest in NAc, matched by differences in ser31, but not ser40, phosphorylation. The L-DOPA per catecholamine and DA turnover ratios were significantly greater in SN and VTA, suggesting greater reliance on de novo DA biosynthesis therein. These compartmental differences reflected an overall autonomy of DA regulation, as seen by decreased DA content in SN and VTA, but not in striatum or NAc, following short-term DA biosynthesis inhibition from local infusion of the TH inhibitor α-methyl-p-tyrosine, as well as in the long-term process of aging. Such data suggest ser31 phosphorylation plays a significant role in regulating TH activity in vivo, particularly in somatodendritic regions, which may have a greater reliance on de novo DA biosynthesis. Thus, to the extent that somatodendritic DA release affects behavior, TH regulation in the midbrain may be critical for DA bioavailability to influence behavior

Phase diffusion as a model for coherent suppression of tunneling in the presence of noise

We study the stabilization of coherent suppression of tunneling in a driven double-well system subject to random periodic $\delta-$function ``kicks''. We model dissipation due to this stochastic process as a phase diffusion process for an effective two-level system and derive a corresponding set of Bloch equations with phase damping terms that agree with the periodically kicked system at discrete times. We demonstrate that the ability of noise to localize the system on either side of the double-well potenital arises from overdamping of the phase of oscillation and not from any cooperative effect between the noise and the driving field. The model is investigated with a square wave drive, which has qualitatively similar features to the widely studied cosinusoidal drive, but has the additional advantage of allowing one to derive exact analytic expressions.Comment: 17 pages, 4 figures, submitted to Phys. Rev.

State determination in continuous measurement

The possibility of determining the state of a quantum system after a continuous measurement of position is discussed in the framework of quantum trajectory theory. Initial lack of knowledge of the system and external noises are accounted for by considering the evolution of conditioned density matrices under a stochastic master equation. It is shown that after a finite time the state of the system is a pure state and can be inferred from the measurement record alone. The relation to emerging possibilities for the continuous experimental observation of single quanta, as for example in cavity quantum electrodynamics, is discussed.Comment: 12 pages, 4 figures, Revte

The impact of fatty acids biosynthesis on the risk of cardiovascular diseases in Europeans and East Asians:A Mendelian randomization study

Despite early interest, the evidence linking fatty acids to cardiovascular diseases (CVDs) remains controversial. We used Mendelian randomization to explore the involvement of polyunsaturated (PUFA) and monounsaturated (MUFA) fatty acids biosynthesis in the etiology of several CVD endpoints in up to 1 153 768 European (maximum 123 668 cases) and 212 453 East Asian (maximum 29 319 cases) ancestry individuals. As instruments, we selected single nucleotide polymorphisms mapping to genes with well-known roles in PUFA (i.e. FADS1/2 and ELOVL2) and MUFA (i.e. SCD) biosynthesis. Our findings suggest that higher PUFA biosynthesis rate (proxied by rs174576 near FADS1/2) is related to higher odds of multiple CVDs, particularly ischemic stroke, peripheral artery disease and venous thromboembolism, whereas higher MUFA biosynthesis rate (proxied by rs603424 near SCD) is related to lower odds of coronary artery disease among Europeans. Results were unclear for East Asians as most effect estimates were imprecise. By triangulating multiple approaches (i.e. uni-/multi-variable Mendelian randomization, a phenome-wide scan, genetic colocalization and within-sibling analyses), our results are compatible with higher low-density lipoprotein (LDL) cholesterol (and possibly glucose) being a downstream effect of higher PUFA biosynthesis rate. Our findings indicate that PUFA and MUFA biosynthesis are involved in the etiology of CVDs and suggest LDL cholesterol as a potential mediating trait between PUFA biosynthesis and CVDs risk

TNF-α increases human melanoma cell invasion and migration in vitro: the role of proteolytic enzymes

Inflammatory mediators have been reported to promote malignant cell growth, invasion and metastatic potential. More specifically, we have recently reported that tumour necrosis factor alpha (TNF-a) increases melanoma cell attachment to extracellular matrix (ECM) substrates and invasion through fibronectin. In this study, we extend these investigations asking specifically whether the TNF-a effect on cell invasion and migration involves activation of proteolytic enzymes. We examined the effect of TNF-a on melanoma expression/activation of type IV gelatinases matrix metalloproteinases 2 and 9 (MMPs -2 and -9) and general proteolytic enzymes. Stimulation with TNF-a significantly increased both melanoma cell migration at 24 h ( þ 21%) and invasion through fibronectin ( þ 35%) but did not upregulate/activate the expression of latent MMP-2 constitutively produced by these cells and did not upregulate their general protease activity. However, the increased cell migration and invasion through fibronectin observed following stimulation with TNF-a were inhibited by the general protease inhibitor a2 macroglobulin. These findings suggest that the promigratory and proinvasive effect of TNF-a on this melanoma cell line may be mediated to some extent by induction of localised cell membrane-bound degradative enzyme activity, which is not readily detected in biochemical assays

The polygenic nature of telomere length and the anti-ageing properties of lithium

Telomere length is a promising biomarker for age-related disease and a potential anti-ageing drug target. Here, we study the genetic architecture of telomere length and the repositioning potential of lithium as an anti-ageing medication. LD score regression applied to the largest telomere length genome-wide association study to-date, revealed SNP-chip heritability estimates of 7.29%, with polygenic risk scoring capturing 4.4% of the variance in telomere length in an independent cohort (p = 6.17 × 10-5). Gene-enrichment analysis identified 13 genes associated with telomere length, with the most significant being the leucine rich repeat gene, LRRC34 (p = 3.69 × 10-18). In the context of lithium, we confirm that chronic use in a sample of 384 bipolar disorder patients is associated with longer telomeres (p = 0.03). As complementary evidence, we studied three orthologs of telomere length regulators in a Caenorhabditis elegans model of lithium-induced extended longevity and found all transcripts to be affected post-treatment (p  0.05). Consequently, this suggests that lithium may be catalysing the activity of endogenous mechanisms that promote telomere lengthening, whereby its efficacy eventually becomes limited by each individual's inherent telomere maintenance capabilities. Our work indicates a potential use of polygenic risk scoring for the prediction of adult telomere length and consequently lithium's anti-ageing efficacy