Transplantable neural progenitor populations derived from Rhesus monkey embryonic stem cells

Cell-based therapies using embryonic stem cells (ESCs) in the treatment of neural disease will require the generation of homogenous donor neural progenitor (NP) populations. Here we describe an efficient culture system containing hepatocyte growth factor (HGF) and G5 supplement for the production of highly enriched (88.3% +/- 8.1%)populations of NPs from rhesus monkey ESCs. Additional purification resulted in NP preparations that were 98% nestin positive. Moreover, NPs, as monolayers or neurospheres, could be maintained for prolonged periods of time in media containing HGF+G5 or G5 alone. In vitro differentiation and in vivo transplantation assays showed that NPs could differentiate into neurons, astrocytes, and oligodendrocytes. The kinds and quantities of differentiated cells derived from NPs were closely correlated with their niches in vivo. Glial differentiation was predominant in periventricular areas, whereas cells migrating into the cortex were mostly neurons. Cell counts showed that 2 months after transplantation, approximately 25% of transplanted NPs survived and 65% - 80% of the surviving transplanted cells migrated along the ventricular wall or in a radial fashion. Subcloning demonstrated that several clonal lines derived from NPs expressed nestin and differentiated into three neural lineages in vitro and in rat brains in vivo. In contrast, some subcloned lines showed restricted differentiation both in vitro and in vivo in rat brains. These observations set the stage for obtaining highly enriched NPs and evaluating the efficacy of NP-based transplantation therapy in the nonhuman primate and will provide a platform for probing the molecular mechanisms that control neural induction.

Neural progenitors derived from monkey embryonic stem cells in a simple monoculture system

A simple monoculture system. combined with a chemically defined medium containing hepatocyte growth factor (HGF) and G5 supplement, was used to induce rhesus monkey embryonic stem cells (rESC) directly into neuroepithelial (NE) cells. Under these conditio

肝细胞生长因子促进猕猴胚胎干细胞来源的神经前体细胞的增殖

肝细胞生长因子（HGF）是一个多效应因子，在神经系统中具有重要作用， 但是对于HGF在早期神经系统发育（特别是哺乳动物）中的具体作用还不明确， 这方面的研究还很少。我们早前的研究发现采用HGF和G5 supplement结合EB法 可诱导猕猴胚胎干细胞（rESCs）定向分化成高纯度（88.3± 8.1%）的可移植的 神经前体细胞，但是HGF在整个分化过程中的具体作用及HGF与其它因子的关 系还不清楚。 本研究改进先前研究体系，用单层培养诱导体系代替EB法诱导体系，用bFGF 代替G5 supplement。即采用单层培养的方式，分别用同时含bFGF和HGF、只含 HGF或bFGF以及两者都没有添加的成分确定的分化液诱导rESCs向神经细胞分 化。并检测HGF对rESCs来源的神经前体细胞的增殖速度的影响，旨在进一步研 究HGF在整个分化过程中的具体作用及HGF与bFGF的关系。主要结论如下：1） 采用单层培养法，同时添加HGF和bFGF可诱导rESCs在两周内定向分化为高纯度 （>85%）的神经前体细胞，从而建立了一种更为简单的诱导rESCs分化成神经细 胞的方法；2）不同分化条件下都得到了相似比例的神经前体细胞，表明外源性 的HGF在诱导rESC向神经前体细胞转变的过程中对于神经细胞命运的决定并不 起作用；3）HGF能有效地促进rESCs来源的神经前体细胞的增殖，并且与bFGF 具有协同作用。

肝细胞生长因子促进猕猴胚胎干细胞来源的神经前体细胞的增殖

肝细胞牛长因子(hepatoeyte growth factor,HGF)足一个多效应因子,在神经系统中具有重要作用.早前的研究发现采用HGF和G5 supplement结合EB(embryoid body)法可诱导猕猴胍胎干细胞(rhesus embryonic stem cells,rESCs)定向分化成高纯度的可移植的神经前体细胞(neural progenitors),但对于HGF在整个诱导分化过程中的具体作用及机制还不清楚.本研究改进先前研究体系,采用单层培养法,同时添加HGF和bFGF(basic fibroblast growth factor,碱性成纤维细胞生长因子)诱导rESCs在两周内定向分化为高纯度[(81.66±4.37)%]的神经前体细胞,并且单独添加HGF或bFGF以及两者都没有添加的条件下也得到了相似比例的神经前体细胞,表明外源性的HGF在诱导rESCs向神经前体细胞转变的过程中对十神经细胞命运的决定并不起作用;进一步研究发现HGF能有效地促进神经前体细胞的增殖,并且与bFGF具有协同作用.总之,本研究建立了一种更为简单的诱导rESCs分化成神经细胞的方法,发现外源性的HGF在rESCs向神经前体细胞分化的过程中并没有神经诱导的作用,但能与bFGF协同作用促进rESCs来源的神经前体细胞的增殖.

Hepatocyte growth factor enhances the generation of high-purity oligodendrocytes from human embryonic stem cells

Generation of homogeneous oligodendrocytes as donor cells is essential for human embryonic stem cell (hESC)-based cell therapy for demylinating diseases. Herein we present a novel method for efficiently obtaining mature oligodendrocytes from hESCs with high purity (79.7 +/- 6.9%), using hepatocyte growth factor (HGF) and G5 supplement(containing insulin, transferrin, selenite, biotin, hydrocortisone, basic fibroblast growth factor and epidermal growth factor) in a four-step method. We induced hESCs into neural progenitors (NP) with HGF (5 ng/ml) and G5 (1 x) supplemented medium in an adherent differentiation system. The purified NPs were amplified in suspension as neurospheres for 1 month, and terminal oligodendrocyte differentiation was then induced by G5 supplement withdrawal and HGF treatment (20 ng/ml). The cells generated displayed typical morphologies of mature oligodendrocytes and expressed oligodendrocyte markers O4 and myelin basic protein (MBP). Our result revealed that HGF significantly enhanced the proliferation of hESC-derived NPs and promoted the differentiation as well as the maturation of oligodendrocytes from NPs. Further studies suggest that HGF/c-Met signaling pathway might play an important role in oligodendrocyte differentiation in our system. Our studies provide a means for generating the clinically relevant cell type and a platform for deciphering the molecular mechanisms that control oligodendrocyte differentiation. (C) 2009 International Society of Differentiation. Published by Elsevier Ltd. All rights reserved.

RAP-A Indigenous supplement : guidelines for the adaptation and implementation of the RAP Program for indigenous adolescents

The RAP-A Indigenous supplement has been designed to provide guidelines for the Adaptation and implementation of the RAP Program for indigenous adolescents. It describes a variety of adaptations that have been made to RAP-A to make it more suitable for indigenous teenagers.

A forced titration study of the antioxidant and immunomodulatory effects of Ambrotose AO supplement

Background Oxidative stress plays a role in acute and chronic inflammatory disease and antioxidant supplementation has demonstrated beneficial effects in the treatment of these conditions. This study was designed to determine the optimal dose of an antioxidant supplement in healthy volunteers to inform a Phase 3 clinical trial. Methods The study was designed as a combined Phase 1 and 2 open label, forced titration dose response study in healthy volunteers (n = 21) to determine both acute safety and efficacy. Participants received a dietary supplement in a forced titration over five weeks commencing with a no treatment baseline through 1, 2, 4 and 8 capsules. The primary outcome measurement was ex vivo changes in serum oxygen radical absorbance capacity (ORAC). The secondary outcome measures were undertaken as an exploratory investigation of immune function. Results A significant increase in antioxidant activity (serum ORAC) was observed between baseline (no capsules) and the highest dose of 8 capsules per day (p = 0.040) representing a change of 36.6%. A quadratic function for dose levels was fitted in order to estimate a dose response curve for estimating the optimal dose. The quadratic component of the curve was significant (p = 0.047), with predicted serum ORAC scores increasing from the zero dose to a maximum at a predicted dose of 4.7 capsules per day and decreasing for higher doses. Among the secondary outcome measures, a significant dose effect was observed on phagocytosis of granulocytes, and a significant increase was also observed on Cox 2 expression. Conclusion This study suggests that Ambrotose AO® capsules appear to be safe and most effective at a dosage of 4 capsules/day. It is important that this study is not over interpreted; it aimed to find an optimal dose to assess the dietary supplement using a more rigorous clinical trial design. The study achieved this aim and demonstrated that the dietary supplement has the potential to increase antioxidant activity. The most significant limitation of this study was that it was open label Phase 1/Phase 2 trial and is subject to potential bias that is reduced with the use of randomization and blinding. To confirm the benefits of this dietary supplement these effects now need to be demonstrated in a Phase 3 randomised controlled trial (RCT).

Supplement : Efficient weak second-order stochastic Runge-Kutta methods for non-commutative Stratonvich stochastic differential equations

This paper gives a modification of a class of stochastic Runge–Kutta methods proposed in a paper by Komori (2007). The slight modification can reduce the computational costs of the methods significantly.

Errata supplement for QUT thesis 'Heavy vehicle suspensions : testing and analysis'

Errata supplement to QUT thesis: 'Heavy vehicle suspensions : testing and analysis'

Assessing the impact of a food supplement on the nutritional status and body composition of HIV-infected Zambian women on ARVs

Background Zambia is a sub-Saharan country with one of the highest prevalence rates of HIV, currently estimated at 14%. Poor nutritional status due to both protein-energy and micronutrient malnutrition has worsened this situation. In an attempt to address this combined problem, the government has instigated a number of strategies, including the provision of antiretroviral (ARV) treatment coupled with the promotion of good nutrition. High-energy protein supplement (HEPS) is particularly promoted; however, the impact of this food supplement on the nutritional status of people living with HIV/AIDS (PLHA) beyond weight gain has not been assessed. Techniques for the assessment of nutritional status utilising objective measures of body composition are not commonly available in Zambia. The aim of this study is therefore to assess the impact of a food supplement on nutritional status using a comprehensive anthropometric protocol including measures of skinfold thickness and circumferences, plus the criterion deuterium dilution technique to assess total body water (TBW) and derive fat-free mass (FFM) and fat mass (FM). Methods/Design This community-based controlled and longitudinal study aims to recruit 200 HIV-infected females commencing ARV treatment at two clinics in Lusaka, Zambia. Data will be collected at four time points: baseline, 4-month, 8-month and 12-month follow-up visits. Outcome measures to be assessed include body height and weight, body mass index (BMI), body composition, CD4, viral load and micronutrient status. Discussion This protocol describes a study that will provide a longitudinal assessment of the impact of a food supplement on the nutritional status of HIV-infected females initiating ARVs using a range of anthropometric and body composition assessment techniques.

Nutritional support in children with end-stage liver disease : a randomized crossover trial of a branched-chain amino acid supplement

Malnutrition is common in children with end-stage liver disease (ESLD) awaiting orthotopic liver transplantation (OLT), and nutritional support is assuming an important role in preoperative management. To evaluate preoperative nutritional therapy, 19 children (median age 1.25 y) with ESLD awaiting OLT were prospectively studied. Two high-energy, isoenergetic and isonitrogenous nutritional formulations delivered nasogastrically were compared: a branched-chain amino acid (BCAA)-enriched semielemental formulation and a matched standard semielemental formulation. Twelve of 19 patients completed a randomized controlled study before OLT and 10 of 19 completed a full crossover study. Improvements in weight and height occurred during the BCAA supplements, with no statistical change on the standard formulation. Significant increases in total body potassium, midupper arm circumference, and subscapular skinfold thickness occurred during the BCAA supplements, whereas no significant changes occurred during the standard formulation period. Significantly fewer albumin infusions were required during the BCAA supplement. These findings suggest that BCAA-enriched formulas have advantages over standard semielemental formulas in improving nutritional status in children with ESLD. and are deserving of wider application and study.

Variability in Voluntary Intake of a Molasses-based Supplement by Cows and Calves

Supplements are often fed to ruminants in extensive grazing situations to provide minerals and nitrogen likely to be deficient in pasture. However a large proportion of animals offered such supplements may not consume any supplement, while among consumer animals the variability in supplement intake may be high (Wheeler et al., 1980; Dixon et al., 1998). An experiment examined the distribution of intake of a molasses-based supplement containing phosphorus and urea in a breeder herd. A herd of mixed-age breeder cows, calves, heifers and bulls were offered ad libitum a molasses-based supplement containing 13% urea and 17% phosphoric acid. After 2 weeks lithium-labelled supplement (2 mg Li/kg LW) was offered on one day to measure individual intakes of supplement. The molasses was offered in three 560 mm diameter feeders placed together near the water point.

A Guide to Disability Statistics from the National Health Interview Survey – Disability Supplement

The National Health Interview Survey - Disability supplement (NHIS-D) provides information that can be used to understand myriad topics related to health and disability. The survey provides comprehensive information on multiple disability conceptualizations that can be identified using information about health conditions (both physical and mental), activity limitations, and service receipt (e.g. SSI, SSDI, Vocational Rehabilitation). This provides flexibility for researchers in defining populations of interest. This paper provides a description of the data available in the NHIS-D and information on how the data can be used to better understand the lives of people with disabilities.

Evaluation of a remote drafting system for regulating sheep access to supplement

Remote drafting technology now available for sheep makes possible targeted supplementation of individuals within a grazing flock. This system was evaluated by using 68 Merino wethers grazing dry-season, native Mitchell grass pasture (predominantly Astrebla spp.) as a group and receiving access to lupin grain through a remote drafter 0, 1, 2, 4 or 7 days/week for 8 weeks. The sole paddock watering point was separately fenced and access was via a one-way flow gate. Sheep exited the watering point through a remote drafter operated by solar power and were drafted by radio frequency identification (RFID) tag, according to treatment, either back into the paddock or into a common supplement yard where lupins were provided ad libitum in a self-feeder. Sheep were drafted into the supplement yard on only their first time through the drafter during the prescribed 24-h period and exited the supplement yard via one-way flow gates in their own time. The remote drafter operated with a high accuracy, with only 2.1% incorrect drafts recorded during the experimental period out of a total of 7027 sheep passes through the remote drafter. The actual number of accesses to supplement for each treatment group, in order, were generally less than that intended, i.e. 0.02, 0.69, 1.98, 3.35 and 6.04 days/week. Deviations from the intended number of accesses to supplement were mainly due to sheep not coming through to water on their allocated day of treatment access, although some instances were due to incorrect drafts. There was a non-linear response in growth rate to increased frequency of access to lupins with the growth rate response plateauing at similar to 3 actual accesses per week, corresponding to a growth rate of 72.5 g/head. day. This experiment has demonstrated the application of the remote drafting supplementation system for the first time under grazing conditions and with the drafter operated completely from solar power. The experiment demonstrates a growth response to increasing frequency of access to supplement and provides a starting point with which to begin to develop feeding strategies to achieve sheep weight-change targets.