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Upper extremity applications of functional neuromuscular stimulation.

Functional electrical stimulation (FES) has been used for increasing muscle strength, decreasing spasticity, and controlling movement of limbs for many years. Most of this work, however, has been done in a research setting. Over the past decade, FES has moved slowly from the laboratory to the clinical world through feasibility studies in groups of patients with spinal cord injuries and strokes. Electrical stimulation has been shown to decrease spastic tone both during and after the stimulation, allowing for better limb positioning, decrease in contracture formation, and in some cases, improvement of voluntary movement. Electrical stimulation as a motor prosthesis is now being provided to small groups of spinal cord-injured patients (primarily C4, C5 and C6 levels) to assist with hand positioning and to produce hand grasp. In these settings, patients have attained greater independence in activities of daily living and in work-related tasks. Distribution of this technology to multiple centers is continuing through a technology transfer program.

Arm↗

A reversibly immortalized human hepatocyte cell line as a source of hepatocyte-based biological support.

The application of hepatocyte transplantation (HTX) is increasingly envisioned for temporary metabolic support during acute liver failure and provision of specific liver functions in inherited liver-based metabolic diseases. Compared with whole liver transplantation, HTX is a technically simple procedure and hepatocytes can be cryopreserved for future use. A major limitation of this form of therapy in humans is the worldwide shortage of human livers for isolating an adequate number of transplantable human hepatocyes when needed. Furthermore, the numbers of donor livers available for hepatocyte isolation is limited by competition for their use in whole organ transplantation. Considering the cost of hepatocyte isolation and the need for immediate preparation of consistent and functional cells, it is unlikely that human hepatocytes can be obtained on such a scale to treat a large number of patients with falling liver functions. The utilization of xenogenic hepatocytes will result in additional concerns regarding transmission of infectious pathogens and immunological and physiological incompatibilities between animals and humans. An attractive alternative to primary human hepatocytes is the use of tightly regulated human hepatocyte cell lines. Such cell lines can provide the advantages of unlimited availability, sterility and uniformity. We describe here methods for creating transplantable human hepatocyte cell lines using currently available cell cultures and gene transfer technology.

Journal Article↗

Rural water safety from the source to the on-farm tap.

For those Canadians who live in metropolitan areas, good quality water for domestic use and consumption is readily available, and perhaps taken for granted. However, for over 4 million Canadians who rely on private water supplies, access to water that is safe for consumption and suitable for domestic use is a very real issue. This is also true in the agriculture and agri-food sector. Many of these private water supplies are in rural areas, where water is taken from surface or ground sources. These supplies may be of naturally poor quality, or may have had their quality affected by municipal, industrial, or agricultural activities. Options to protect and enhance the quality of private water supplies include source protection using best management practices (BMPs), source enhancement, and water treatment using innovative small-scale systems. With funding from the Canada-Saskatchewan Agri-Food Innovation Fund, Agriculture and Agri-Food Canada has conducted applied research into effective and affordable BMPs (remote livestock watering, low-drift nozzles for spraying farm chemicals, methods to reduce agro-chemical runoff or leaching, etc.). Source enhancement strategies were studied using aeration for farm ponds or preventive maintenance procedures for ground water wells. Various water treatment technologies were adapted to the small-scale needs of farms, including coagulation, biological sand and biological carbon filtration, membrane filtration using microfilters, nanofilters or reverse osmosis processes, and disinfection systems using chlorination or ultra violet light. Each research project included a technology transfer component, to ensure that the knowledge gained from the research was available to those that needed the information, and to help decision makers address rural water quality problems.

Agriculture↗

Harvesting the genomic promise: recombineering sequences for phenotypes.

The past decade has witnessed the construction of linkage and physical maps defining quantitative trait loci (QTL) in various domesticated species. Targeted chromosomal regions are being further characterized through the construction of bacterial artificial chromosome (BAC) contigs in order to isolate and characterize genes contributing towards phenotypic variation. Whole-genome BAC contigs are also being constructed that will serve as the tiling path for genomic sequencing. Harvesting this genetic information for biological gain requires either genetic selection or the production of genetically modified animals. This later approach when coupled with nuclear transfer technology (NT) provides "clones" of genetically modified animals. However, to date, the production of genetically modified animals has been limited to either microinjection of small gene constructs into embryos with random insertion or complex gene constructs designed to knock-out targeted gene expression. Neither of these approaches provides for introducing directed genetic manipulation allowing for allelic substitution [knock-in], subsequent analyses of gene expression, and cloning. An alternative approach utilizing genomic sequence information and recombineering to direct gene targeting of specific porcine BACs is presented here.

Animals↗

Family-based interventions for substance use and misuse prevention.

Because "substance abuse" is a "family disease" of lifestyle, including both genetic and family environmental causes, effective family strengthening prevention programs should be included in all comprehensive substance abuse prevention activities. This article presents reviews of causal models of substance use and evidence-based practices. National searches by the authors suggest that there is sufficient research evidence to support broad dissemination of five highly effective family strengthening approaches (e.g., behavioral parent training, family skills training, in-home family support, brief family therapy, and family education). Additionally, family approaches have average effect sizes two to nine time larger than child-only prevention approaches. Comprehensive prevention programs combining both approaches produced much larger effect sizes. The Strengthening Families Program (SFP) is the only one of these programs that has been replicated with positive results by independent researchers with different cultural groups and with different ages of children. Few research-based programs have been adopted by practitioners, partly because of technology transfer issues. Overall, research on ways to improve dissemination, marketing, training, and funding is needed to improve adoption of effective prevention programs.

Adolescent↗

The status of health services research on adjudicated drug-abusing juveniles: selected findings and remaining questions.

This article focuses on the factors that affect the availability, accessibility, and opportunity of effective treatment for the adjudicated adolescent drug user The social and developmental context of "drug abuse" and its treatment in adolescents, and particularly in adjudicated adolescents, is reviewed and compared with that in adults. Selected health services research findings on this population are discussed in terms of the effectiveness of delivering treatment services; the organization, management, and financing of services; and adoption of best practices (technology transfer).

Adolescent↗

Functional complementation of a genetic deficiency with human artificial chromosomes.

We have shown functional complementation of a genetic deficiency in human cultured cells, using artificial chromosomes derived from cloned human genomic fragments. A 404-kb human-artificial-chromosome (HAC) vector, consisting of 220 kb of alphoid DNA from the centromere of chromosome 17, human telomeres, and the hypoxanthine guanine phosphoribosyltransferase (HPRT) genomic locus, was transferred to HPRT-deficient HT1080 fibrosarcoma cells. We generated several cell lines with low-copy-number, megabase-sized HACs containing a functional centromere and one or possibly several copies of the HPRT1 gene complementing the metabolic deficiency. The HACs consisted of alternating alphoid and nonalphoid DNA segments derived only from the input DNA (within the sensitivity limits of FISH detection), and the largest continuous alphoid segment was 158-250 kb. The study of both the structure and mitotic stability of these HACs offers insights into the mechanisms of centromere formation in synthetic chromosomes and will further the development of this human-gene-transfer technology.

Blotting, Southern↗

What are today's orphaned vaccines?

Development costs for new biological agents are increasing, and the time span from laboratory research to introduction of a product on the world market is becoming ever longer. Complex regulatory requirements add barriers and additional costs to early introduction abroad. This results in reluctance by manufacturers to undertake development of a vaccine that will be used for a tropical disease in only the public sector of a poor country. The chances of recovery of huge investment costs before patents expire are not good, unless such a new vaccine can also be sold at high cost in North America and Europe. These are some of the reasons that we still do not have a modern Japanese encephalitis vaccine or products against malaria and dengue fever. Many tropical countries must find a way to develop their own vaccine production facilities. Innovative help for technology transfer will have to be forthcoming, or many new life-saving products will never bridge the gap between research unit and production.

Developed Countries↗

Nuclear cloning, stem cells, and genomic reprogramming.

The generation of adult animals by nuclear cloning from adult donor cells is extremely inefficient, with most clones dying soon after implantation. In contrast, cloning from embryonic stem cell donor nuclei is significanty more efficient than from adult donor cells. However, regardless of donor cell type, all clones that survive to birth and beyond suffer serious phenotypic and gene expression abnormalities. All available evidence is consistent with the notion that the anomalous phenotypes of cloned animals are caused by faulty epigenetic reprogramming of the donor nucleus. Faulty reprogramming appears to be caused by the cloning process itself as well as by the epigenetic state of the donor nucleus. In contrast to reproductive cloning, faulty reprogramming of the donor nucleus does not tend to interfere with the application of nuclear transfer technology for therapeutic purposes (therapeutic cloning).

Animals↗

Patent prosecution in structural proteomics.

This paper presents a brief overview of intellectual property rights and the various areas in proteomics to which intellectual property rights may be applicable. Technology transfer, including licensing and business agreements, are not covered in this paper. Instead, issues and complications related to national and overseas patent prosecution in this relatively new field will be discussed.

Intellectual Property↗

Globalizing biosecurity.

A harmonized international regime that enhances biosecurity is needed to reduce the risk of bioterrorism. Like other security regimes, this will entail mutually reinforcing strands, which need to include: enactment of legally binding control of access to dangerous pathogens, transparency for sanctioned biodefense programs, technology transfer and assistance to developing countries to jointly advance biosafety and biosecurity, global awareness of the dual-use dilemma and the potential misuse of science by terrorists, and development of a global ethic of compliance. To work, this effort must be undertaken collectively, utilizing the international and regional institutions that already have a role to play in providing safety and security. Most notably, it must grow in a top-down manner from the Biological Weapons Convention accord, in which States Parties have agreed to ban the development of biological weapons, and in a bottom-up manner from the scientific and health communities, which are engaged in the research and public health efforts that must be protected against misuse-especially involving the World Health Organization.

Bioterrorism↗

BETA2/NeuroD protein transduction requires cell surface heparan sulfate proteoglycans.

BETA2/NeuroD protein is important for regulating insulin gene transcription and for the terminal differentiation of islet cells, including insulin- and glucagon-producing cells. We reported that BETA2/NeuroD protein can permeate several cell types, including pancreatic islets, because of an arginine- and lysine-rich protein transduction domain (PTD) sequence in its structure. Here we provide genetic and biochemical evidence that cell membrane heparan sulfate proteoglycans are involved in extracellular BETA2/NeuroD internalization. We tested whether soluble glycosaminoglycans (GAGs) could inhibit BETA2/NeuroD internalization. Heparin almost completely prevented BETA2/NeuroD entry, whereas chondroitin sulfate A, B, and C caused only limited inhibition. Moreover, treatment with heparinase III impaired BETA2/NeuroD internalization, whereas treatment with chondroitinase ABC, or with chondroitinase AC, was completely ineffective in inhibiting BETA2/NeuroD internalization. We also examined various mutant cell lines originating from CHOK1 cells and defective in GAG biosynthesis. The observation using mutant cell lines supports the notion that the selective sulfation of heparan sulfate is an important determinant for NeuroD/heparan sulfate recognition. These data indicate that cell surface heparan sulfate proteoglycans are required for BETA2/NeuroD internalization and that BETA2/NeuroD protein transduction could be a safe and valuable strategy for enhancing insulin gene transcription without requiring gene transfer technology.

Animals↗

Cancer initiatives in developing countries.

The coming decades will bring dramatic increases in morbidity and mortality from cancer in the developing world. The burden of cancer is increasing globally, with an expected 20 million new cases per year in 2020, half of which will be in low- and middle-income countries. Despite an already overwhelming burden of health problems, developing countries must somehow address this cancer pandemic and their alarming share of cancer illness. Many large-scale and innovative worldwide initiatives have been launched in the fight against cancer. These programs represent colossal efforts to expand resources for health education among people and healthcare workers in developing countries and to increase awareness of cancer prevention. Many novel evidence-based and culturally-relevant programs are being implemented for screening and early prevention of common cancers. Several key initiatives are supporting clinical research, guideline development, oncology training, and appropriate technology transfer, such as access to electronic medical literature. Treatment programs are expanding access and quality of radiologic and pharmacologic therapies for cancer. These initiatives represent an unprecedented level of and cooperation among international agencies, governmental and nongovernmental organizations, international foundations, scientific societies, local institutions, and industry. This review examines the scope of need in response to the increasing burden of cancer in the developing world and major initiatives that have been created to respond to these needs.

Developing Countries↗

Role of lesser developed nations in rabies research.

The contributions of lesser developed nations to rabies research can be significant in several areas. The type of health system that supports complex veterinary-public health programs, the operational research that is required in these countries, the follow-up epidemiologic surveillance, and the involvement in technology transfer concerning new rabies vaccines and rapid and simple diagnostic procedures are all crucial to the success of rabies prevention in developing countries. Results of a relevant research project carried out in the Kathmandu Valley in Nepal, India, are discussed.

Developing Countries↗

CAM and cell fate targeting: molecular and energetic insights into cell growth and differentiation.

Evidence-based medicine is switching from the analysis of single diseases at a time toward an integrated assessment of a diseased person. Complementary and alternative medicine (CAM) offers multiple holistic approaches, including osteopathy, homeopathy, chiropractic, acupuncture, herbal and energy medicine and meditation, all potentially impacting on major human diseases. It is now becoming evident that acupuncture can modify the expression of different endorphin genes and the expression of genes encoding for crucial transcription factors in cellular homeostasis. Extremely low frequency magnetic fields have been found to prime the commitment to a myocardial lineage in mouse embryonic stem cells, suggesting that magnetic energy may direct stem cell differentiation into specific cellular phenotypes without the aid of gene transfer technologies. This finding may pave the way to novel approaches in tissue engineering and regeneration. Different ginseng extracts have been shown to modulate growth and differentiation in pluripotent cells and to exert wound-healing and antitumor effects through opposing activities on the vascular system, prompting the hypothesis that ancient compounds may be the target for new logics in cell therapy. These observations and the subtle entanglement among different CAM systems suggest that CAM modalities may deeply affect both the signaling and transcriptional level of cellular homeostasis. Such a perception holds promises for a new era in CAM, prompting reproducible documentation of biological responses to CAM-related strategies and compounds. To this end, functional genomics and proteomics and the comprehension of the cell signaling networks may substantially contribute to the development of a molecular evidence-based CAM.

Journal Article↗

Embryonic stem-cell gametes: the new frontier in human reproduction.

As infertility increases and gamete donations decline, an alternate source of sex cells may prove valuable for research and infertility treatment. This article examines the social and scientific value of gametes derived from the differentiation of established human embryonic stem (ES)-cell lines (ES-cell-derived gametes) and customized gametes created using nuclear transfer technologies to contain a haploid set of genes creating children genetically related to parent(s). ES-cell-derived gametes may be valuable as a resource for biomedical research, instruction and training in assisted reproductive technologies and perhaps for creating children. The creation of children by ES-cell-derived and customized gametes may not result in psychological harm to children but customized gametes may lead to physical harm to children or an accumulation of gene mutations in a population. Although the creation of new types of children using ES-cell gametes provides more reproductive choices to both fertile and infertile individuals, the risk or physical harm to children from customized gametes may be so severe that the scope of reproductive liberty must be limited. Further scientific and ethical analysis of the creation of children by ES-cell gametes is required.

Cell Differentiation↗

Human gene therapy: why draw a line?

Despite widespread agreement that it would be ethical to use somatic cell gene therapy to correct serious diseases, there is still uneasiness on the part of the public about this procedure. The basis for this concern lies less with the procedure's clinical risks than with fear that genetic engineering could lead to changes in human nature. Legitimate concerns about the potential for misuse of gene transfer technology justify drawing a moral line that includes corrective germline therapy but excludes enhancement interventions in both somatic and germline contexts.

Beneficence↗