Using the Chick Embryo Brain as a Model for In Vivo and Ex Vivo Analyses of Human Glioblastoma Cell Behavior.

The chick embryo has been an ideal model system for the study of vertebrate development, particularly for experimental manipulations. Use of the chick embryo has been extended for studying the formation of human glioblastoma (GBM) brain tumors in vivo and the invasiveness of tumor cells into surrounding brain tissue. GBM tumors can be formed by injection of a suspension of fluorescently labeled cells into the E5 midbrain (optic tectum) ventricle in ovo. Depending on the GBM cells, compact tumors randomly form in the ventricle and within the brain wall, and groups of cells invade the brain wall tissue. Thick tissue sections (350 µm) of fixed E15 tecta with tumors can be immunostained to reveal that invading cells often migrate along blood vessels when analyzed by 3D reconstruction of confocal z-stack images. Live E15 midbrain and forebrain slices (250-350 µm) can be cultured on membrane inserts, where fluorescently labeled GBM cells can be introduced into non-random locations to provide ex vivo co-cultures to analyze cell invasion, which also can occur along blood vessels, over a period of about 1 week. These ex vivo co-cultures can be monitored by widefield or confocal fluorescence time-lapse microscopy to observe live cell behavior. Co-cultured slices then can be fixed, immunostained, and analyzed by confocal microscopy to determine whether or not the invasion occurred along blood vessels or axons. Additionally, the co-culture system can be used for investigating potential cell-cell interactions by placing aggregates of different cell types and colors in different precise locations and observing cell movements. Drug treatments can be performed on ex vivo cultures, whereas these treatments are not compatible with the in ovo system. These two complementary approaches allow for detailed and precise analyses of human GBM cell behavior and tumor formation in a highly manipulatable vertebrate brain environment.

Numerical assessment of hemodynamic perspectives of a left ventricular assist device and subsequent proposal for improvisation.

Due to the unavailability of donors, the use of left ventricular assist devices has emerged to be a reliable line of alternative treatment for heart failure. However, ventricular assist devices (VAD) have been associated with several postoperative complications such as thrombosis, hemolysis, etc. Despite considerable improvements in technology, blood trauma due to high shear stress generation has been a major concern that is largely related to the geometrical feature of the VAD. This study aims to establish the design process of a centrifugal pump by considering several variations in the geometrical feature of a base design using the commercial solver ANSYS-CFX. To capture the uncertain behavior of blood as fluid, Newtonian, as well as non-Newtonian (Bird-Carreau model), models are used for flow field prediction. To assess the possibility of blood damage maximum wall shear stress and hemolysis index have been estimated for each operating point. The results of the simulations yield an optimized design of the pump based on parameters like pressure head generation, maximum shear stress, hydraulic efficiency, and hemolysis index. Further, the design methodology and the steps of development discussed in the paper can serve as a guideline for developing small centrifugal pumps handling blood.

Polyunsaturated Fatty Acids from Astrocytes Activate PPARγ Signaling in Cancer Cells to Promote Brain Metastasis.

Brain metastasis, the most lethal form of melanoma and carcinoma, is the consequence of favorable interactions between the invading cancer cells and the brain cells. Peroxisome proliferator-activated receptor γ (PPARγ) has ambiguous functions in cancer development, and its relevance in advanced brain metastasis remains unclear. Here, we demonstrate that astrocytes, the unique brain glial cells, activate PPARγ in brain metastatic cancer cells. PPARγ activation enhances cell proliferation and metastatic outgrowth in the brain. Mechanistically, astrocytes have a high content of polyunsaturated fatty acids that act as "donors" of PPARγ activators to the invading cancer cells. In clinical samples, PPARγ signaling is significantly higher in brain metastatic lesions. Notably, systemic administration of PPARγ antagonists significantly reduces brain metastatic burden in vivo. Our study clarifies a prometastatic role for PPARγ signaling in cancer metastasis in the lipid-rich brain microenvironment and argues for the use of PPARγ blockade to treat brain metastasis. SIGNIFICANCE: Brain-tropic cancer cells take advantage of the lipid-rich brain microenvironment to facilitate their proliferation by activating PPARγ signaling. This protumor effect of PPARγ in advanced brain metastases is in contrast to its antitumor function in carcinogenesis and early metastatic steps, indicating that PPARγ has diverse functions at different stages of cancer development.This article is highlighted in the In This Issue feature, p. 1631.

Endoscopic third ventriculostomy for tumor-related hydrocephalus in a pediatric population.

OBJECT: Endoscopic third ventriculostomy (ETV) has become a common alternative for managing hydrocephalus in select patients. Nevertheless, there is still controversy regarding the indications for ETV as the primary procedure, given its variable success rates. The purpose of this study is to review the authors' experience with ETV for a variety of patients. METHODS: A total of 43 children underwent ETV between July 1992 and June 2003. Their medical records, operative reports, and imaging studies, when available, were retrospectively reviewed with regard to outcome, complications, and patency rate. Treatment failure was defined as the need to place a shunt within 4 weeks of performing ETV in the patient. There were 20 male and 23 female patients with a mean age of 9.6 years (range 8 weeks-21 years). The overall success rate was 69.8%, and the mean follow-up duration was 24.6 months. Six patients underwent eight repeated ETVs at a mean interval of 25 months, with a patency rate of 62.5% after the second procedure. Only two surgeries were aborted for anatomical reasons. The highest success rates (100% in each instance) were achieved for obstructive hydrocephalus resulting from midbrain/tectal tumor (four patients) and pineal tumor (three patients). CONCLUSIONS: The ETV procedure is an effective management tool for obstructive hydrocephalus in children. It should be considered the primary procedure, rather than ventriculoperitoneal shunts, in carefully selected children. The success rate is dependent on the origin of the hydrocephalus.

Antibacterial efficacy of acridine derivatives conjugated with gold nanoparticles.

Adsorption of acridine derivatives viz. 9-aminoacridine hydrochloride hydrate (9AA-HCl), acridine yellow (AY), acridine orange (AO), and proflavine (Pro) on citrate stabilized gold nanoparticle surface were studied using different analytical techniques like UV-vis absorption spectroscopy, Fourier transform infrared spectroscopy (FT-IR), and transmission electron microscopy (TEM). The amine moiety of acridine derivative binds strongly to the gold nanoparticles as confirmed by spectroscopic studies. The plasmon band observed for the wine red colloidal gold at 525 nm in the UV-vis spectrum is characteristic of gold nanoparticles. However, with the addition of acridine derivatives the intensity of the absorption band at 525 nm decreases and a new peak emerges at red-end region - a signature of formation of gold-drug complex. The TEM images show the average size of citrate stabilized gold nanoparticles as 15-20 nm, which becomes larger in the presence of various drugs due to aggregation. From the thermogravimetric analyses (TGA) we have measured the number of drug molecules attached per gold nanoparticle (AuNP). These gold nanoparticles are very important as drug delivery vehicles and for clinical applications it is necessary to understand their activity in vivo. The antibacterial efficacy of drugs coated gold nanoparticles were studied against various strains of Gram positive and Gram negative bacteria. Among the four drugs, 9AA-HCl and AO showed antibacterial activity and for both of them the AuNP conjugated drug showed better antibacterial efficacy than the bare drug. Because of the high penetrating power and large surface area of Au(0), a single gold nanoparticle can adsorb multiple drug molecules, hence this total entity acts as a single group against the bacteria.

Viscoelastic behavior of human lamin A proteins in the context of dilated cardiomyopathy.

Lamins are intermediate filament proteins of type V constituting a nuclear lamina or filamentous meshwork which lines the nucleoplasmic side of the inner nuclear membrane. This protein mesh provides a supporting scaffold for the nuclear envelope and tethers interphase chromosome to the nuclear periphery. Mutations of mainly A-type lamins are found to be causative for at least 11 human diseases collectively termed as laminopathies majority of which are characterised by aberrant nuclei with altered structural rigidity, deformability and poor mechanotransduction behaviour. But the investigation of viscoelastic behavior of lamin A continues to elude the field. In order to address this problem, we hereby present the very first report on viscoelastic properties of wild type human lamin A and some of its mutants linked with Dilated cardiomyopathy (DCM) using quantitative rheological measurements. We observed a dramatic strain-softening effect on lamin A network as an outcome of the strain amplitude sweep measurements which could arise from the large compliance of the quasi-cross-links in the network or that of the lamin A rods. In addition, the drastic stiffening of the differential elastic moduli on superposition of rotational and oscillatory shear stress reflect the increase in the stiffness of the laterally associated lamin A rods. These findings present a preliminary insight into distinct biomechanical properties of wild type lamin A protein and its mutants which in turn revealed interesting differences.