Cardiac Stem Cells for Myocardial Regeneration: They Are Not Alone

Heart failure is the number one killer worldwide with ~50% of patients dying within 5 years of prognosis. The discovery of stem cells, which are capable of repairing the damaged portion of the heart, has created a field of cardiac regenerative medicine, which explores various types of stem cells, either autologous or endogenous, in the hope of finding the “holy grail” stem cell candidate to slow down and reverse the disease progression. However, there are many challenges that need to be overcome in the search of such a cell candidate. The ideal cells have to survive the harsh infarcted environment, retain their phenotype upon administration, and engraft and be activated to initiate repair and regeneration in vivo. Early bench and bedside experiments mostly focused on bone marrow-derived cells; however, heart regeneration requires multiple coordinations and interactions between various cell types and the extracellular matrix to form new cardiomyocytes and vasculature. There is an observed trend that when more than one cell is coadministered and cotransplanted into infarcted animal models the degree of regeneration is enhanced, when compared to single-cell administration. This review focuses on stem cell candidates, which have also been tested in human trials, and summarizes findings that explore the interactions between various stem cells in heart regenerative therapy

Cryopreservation of Neurospheres Derived from Human Glioblastoma Multiforme

Cancer stem cells have been shown to initiate and sustain tumor growth. In many instances, clinical material is limited, compounded by a lack of methods to preserve such cells at convenient time points. Although brain tumor-initiating cells grown in a spheroid manner have been shown to maintain their integrity through serial transplantation in immune-compromised animals, practically, it is not always possible to have access to animals of suitable ages to continuously maintain these cells. We therefore explored vitrification as a cryopreservation technique for brain tumor-initiating cells. Tumor neurospheres were derived from five patients with glioblastoma multiforme (GBM). Cryopreservation in 90% serum and 10% dimethyl sulfoxide yielded greatest viability and could be explored in future studies. Vitrification yielded cells that maintained self-renewal and multipotentiality properties. Karyotypic analyses confirmed the presence of GBM hallmarks. Upon implantation into NOD/SCID mice, our vitrified cells reformed glioma masses that could be serially transplanted. Transcriptome analysis showed that the vitrified and nonvitrified samples in either the stem-like or differentiated states clustered together, providing evidence that vitrification does not change the genotype of frozen cells. Upon induction of differentiation, the transcriptomes of vitrified cells associated with the original primary tumors, indicating that tumor stem-like cells are a genetically distinct population from the differentiated mass, underscoring the importance of working with the relevant tumor-initiating population. Our results demonstrate that vitrification of brain tumor-initiating cells preserves the biological phenotype and genetic profiles of the cells. This should facilitate the establishment of a repository of tumor-initiating cells for subsequent experimental designs

SDS-PAGE-Based Quantitative Assay for Screening of Kidney Stone Disease

Kidney stone disease is a common health problem in industrialised nations. We developed a SDS-PAGE-based method to quantify Tamm Horsfall glycoprotein (THP) for screening of kidney stone disease. Urinary proteins were extracted by using ammonium sulphate precipitation at 0.27 g salt/mL urine. The resulted pellet was dissolved in TSE buffer. Ten microliters of the urinary proteins extract was loaded and separated on 10% SDS-PAGE under reducing condition. THP migrated as single band in SDS-PAGE. The assay reproducibility and repeatability were 4.8% CV and 2.6% CV, respectively. A total of 117 healthy subjects and 58 stone patients were tested using this assay, and a distinct cut-off (P < 0.05) at 5.6 μg/mL THP concentration was used to distinguish stone patients from healthy subjects. The sensitivity and specificity of the method were 92.3% and 83.3%, respectively

Web services integration on the fly

In a net-centric environment, data, tools and people operate in a distributed network. A key research question is whether a software framework can become so usable and intelligent that integration of web services can be done on-the-fly as self-integration. Given data, software agents and supporting software infrastructure, web services integration on the fly means that human coding is not required to integrate web services into a Web Service Architecture. This thesis explores a generic, flexible, scalable, usable and intelligent web services architecture framework that enables sharing and integration of data and tools on the fly. This software framework is a key enabler for systems of systems architecture in a net-centric environment. The envisioned Web Service Architecture Intelligent Framework (WSAIF) is applied to the Modeling, Virtual Environments and Simulation (MOVES) domain. Specifically, the framework is applied to provide the capability to search and retrieve visualization models and their matching behavior models in a collaborative environment. This thesis elaborates on the design, implementation, deployment and test results of web services for the Scenario Authoring and Visualization for Advanced Graphical Environments (SAVAGE) archive, which is a set of web-based 3D graphics models plus corresponding agent-behaviour models. SAVAGE web services can perform both "find" and "get" operations for models in the archives. SAVAGE web services operations can be composed to form business processes. These business processes can be expressed using modeling techniques such as Web Service Business Process Execution Language (WSBPEL). Future capabilities include semantic activities using Web Ontology Language for Services (OWL-S). The study and comparison of various modeling techniques that enable integration, orchestration and adaptation of composable web services is mentioned. The design and implementation approach matches industry best practices for information architectures. The modeling techniques are essential to and will eventually be used in WSAIF Orchestration and Adaptation components. This thesis further explores how WSAIF software agents, modeling data and supporting software infrastructure can someday enable web services integration on the fly and concludes with recommendations for future work.http://archive.org/details/webservicesinteg109453758Singapore DSO National Laboratories (civilian).Approved for public release; distribution is unlimited.Approved for public release; distribution is unlimited

Qualification and application of an ELISA for the determination of Tamm Horsfall Protein (THP) in human urine and its use for screening of Kidney Stone Disease

<p>Kidney stone disease affects 1 - 20% of the general population. At present, the diagnosis of a stone is done using radiography method when noticeable symptoms appeared. We developed a non-invasive quantitative assay for urinary THP, namely ELISA; whereby our previous study and other reports had shown the usefulness of THP as biomarker for kidney stone disease. Since urine is biological fluid that is easily obtainable, this method could be used as a screening assay for kidney stone prior to confirmation with radiography. The ELISA gave assay linearity r² &#62; 0.999 within the range of 109 ng/mL to 945 ng/mL THP. Assay precisions were &#60; 4% (C.V.) for repeatability and &#60; 5% (C.V.) for reproducibility. Assay accuracy range from 97.7% to 101.2% at the various THP concentrations tested. Assay specificity and sensitivity were 80% and 86%, respectively. The cut-off points at <i>P</i> &#60; 0.05 were 37.0 and 41.2 &#956;g/mL for male and female, respectively. The assay is cost effective and rapid whereby the cost for assaying each urine sample in duplicate is approximately USD0.35 and within 5 hours, 37 samples can be assayed alongside full range of standards and 3 QC samples in each plate. Furthermore, sample preparation is relatively easy where urine sample was diluted 10 times in TEA buffer. The usability of the ELISA method for diagnosis of kidney stone disease is evaluated with 117 healthy subjects and 58 stone formers.</p