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PubMed2005

Radiotargeted gene therapy.

The radiotargeted gene therapy approach to localizing radionuclides at tumor sites involves inducing tumor cells to synthesize a membrane-expressed receptor with a high affinity for injected radiolabeled ligands. A second strategy involves transduction of the sodium iodide symporter (NIS) and free radionuclide therapy. Using the first strategy, induction of high levels of human somatostatin receptor subtype 2 expression and selective tumor uptake, imaging, or growth inhibition with radiolabeled somatostatin analogs has been achieved in human tumor xenograft models. Therapy studies have been performed on several tumor xenograft models with various radionuclides using the NIS radiotargeted gene therapy approach. The use of gene transfer technology to induce expression of high-affinity membrane receptors or transporters can enhance the specificity and extent of radioligand or radionuclide localization in tumors, and the use of radionuclides with appropriate emissions can deliver radiation-absorbed cytotoxic doses across several cell diameters to compensate for limited transduction efficiency. Clinical studies are needed to determine the most promising of these new therapeutic approaches.

Animals↗
PubMed2005

Gene therapy for lympho-hematopoietic disorders.

Inherited lympho-hematopoietic disorders are considered to be some of the most amenable targets for development of gene therapy because of their defined molecular biology and pathophysiology and the potential for corrected cells to exhibit profound growth and survival advantages. Recently, several clinical studies have shown that conventional gene transfer technology can produce major beneficial therapeutic effects.

Genetic Therapy↗
PubMed2004

[The potential role of nuclear matrix attachment regions (MARs) in regulation of gene expression].

Gene transfer technology is being used to enhance agronomic performance or improve quality traits in a wide variety of crop species. However, it is sometimes severely handicapped by difficulty in obtaining material in which transgene expression is predictable and stable over many generations. Because integration seemed to occur randomly in the plant genome, it was thought that some transgenes would be integrated in a relatively uncondensed, transcriptionally active chromatin environment, while others in a condensed, transcriptionally inert chromatin structure. Nuclear matrix attachment regions (MARs) are defined as DNA sequences that bind preferentially to the proteins of the nuclear matrix. They typically are localized at the borders of gene domains, implicating them in the formation of individual loops of higher order chromatin structure and transcription regulation. When MARs are positioned on either side of a transgene their presence usually results in higher and more stable espression in transgenic plants, most likely by minimizing gene silencing. In this review, we focus mainly on novel findings and our observations concerning the function of MARs in transcription regulation. Our objective is not only to summarize the current data and present several possible models to explain MAR effects on the transcription regulation, but also to point out some open questions involving the utilization of MARs in constructing high efficient expression vectors.

Chromatin↗
PubMed2005

Effectiveness of liposomes to transfect livestock fibroblasts.

The development of an efficient transfection system in livestock cells is an important step towards investigating gene transfer and the functioning and production of transgenic animals. Important factors involved in cationic liposome mediated gene transfer were evaluated through in vitro transfection of bovine, caprine and ovine fibroblast cells. Transfection of plasmid DNA complexes of different commercially available liposomes (Lipofectamine, Lipofectin, Cellfectin and DMRIE-C; Gibco-BRL, USA) was evaluated utilizing the following parameters: DNA/liposome ratio, cell density, DNA conformation, and the effect of transfection time on the efficiency of bovine fibroblasts to express a reporter gene. The effects and concentrations of liposomes were also evaluated in caprine and ovine fibroblasts. Lipofectamine alone and Lipofectamine with Plus reagent induced high-frequency expression of beta-galactosidase and neo genes in all cells evaluated (47 and 88.3%, respectively). Regarding phenotype, chromosomal stability was similar in transfected and non-transfected cells. The parameters set in this study will establish a foundation for utilizing transfected fibroblast cells to generate transgenic animals through nuclear transfer technology and gene function studies.

Animals↗
PubMed2005

Immune deficiency enhances expression of recombinant human antibody in mice after nonviral in vivo gene transfer.

A cDNA encoding human antibody against hepatitis B virus was expressed in normal and severe combined immune deficiency (SCID) mice to clarify whether or not host immune status affects circulating levels of the recombinant human antibody (RhAb) after nonviral in vivo gene transfer. For transferring genes, either electroporation (EP) or hydrodynamics-based transfection (HD) was employed. The former was applied to the leg muscle to express the gene, while the latter primarily targeted foreign gene expression in the liver. The expressed RhAb was secreted into the blood circulation, and its existence was assayed by ELISA. Prior to the investigation of host immune status, suitable forms of plasmid expression vectors and types of electrodes were determined in normal mice. Results showed that the vector encoding both the light and heavy chains driven by the CMV promoter had the highest plasma RhAb concentrations, and a pair of pincette-type electrodes conferred the best performance. In both EP and HD, the SCID state showed an increased and prolonged RhAb production in the blood circulation due probably to suppressed recognition of RhAb as a foreign protein to the host animal. The difference in gene transfer methods demonstrated a characteristic pattern: an early and sharp rise followed by a relatively rapid decrease in HD, in contrast to a gradual rise followed by a plateau level maintained in EP. As a result, with the same amount of gene transferred, the plasma RhAb concentrations for the first 7 or 8 weeks were higher in HD than EP, while the reverse was true for the latter period. Multiple gene transfer contributed to maintaining and prolonging high RhAb concentrations in plasma by both methods with similar characteristic patterns accompanying the respective gene transfer method. These results suggest the importance of host immunological potency for maintaining plasma RhAb concentrations if these gene transfer technologies are used for clinical and therapeutic purposes.

Animals↗
PubMed2005

The impact of computing technology on pharmaceutical and biotech research.

Pharmaceutical and biotechnology companies continue to make investments in research techniques including those using high-performance computing technology. Much of this research is fueled by the need to analyze the ever-growing store of genomic and proteomic data. However, many research and development investments, including techniques to mine newly produced genomic data, have not provided the anticipated improvements in productivity. This raises the questions of whether advancements expected from next-generation computing technology can translate into tangible benefits for pharmaceutical and biotechnology researchers, and whether researchers can capitalize on an increased understanding of biological systems using tools made more readily available in high-performance computing. The author provides some background on the source of the advancements in the computing industry and offers examples from other scientific applications that point to potential benefits in life science research.

Biotechnology↗
PubMed2006

Autocrine and intracrine signaling for cardiogenesis in embryonic stem cells: a clue for the development of novel differentiating agents.

Cardiogenesis, one of the earliest and most complex morphogenetic events in the embryo, is not fully understood at the molecular level and is typically a low-yield process. Affording a high throughput of cardiogenesis from a suitable population of pluripotent cells is therefore a major assignment in the perspective of a stem cell therapy for heart failure. Analysis of cardiac lineage commitment in mouse embryonic stem cells and in vivo models of cardiac differentiation revealed that a number of crucial growth factors are released from precursor cells, acting in an autocrine fashion on specific plasma membrane receptors to prime a cardiogenic decision. Nevertheless, it is increasingly becoming evident that cell nuclei harbor the potential for intrinsic signal transduction pathways. The term "intracrine" has been proposed for growth regulatory peptides that have been shown to act within their cell of synthesis at the level of the nuclear envelope, chromatin, or other subnuclear components. Considerable evidence links known intracrines with transcriptional responses and self-sustaining loops that behave as long-lived signals and impart features characteristic of differentiation, growth regulation and cell memory. This review focuses on a number of autocrine and intracrine systems within the context of cardiac differentiation and emphasizes the identification of cardiogenic mechanisms as a clue for the development of unprecedented differentiating strategies. In this regard, recently synthesized mixed esters of hyaluronan with butyric and retinoic acid primed the expression of cardiogenic genes and elicited a remarkable increase in cardiomyocyte yield in mouse embryonic stem cells. This demonstrates the potential for chemically modifying the gene program of cardiac differentiation without the aid of gene transfer technologies and sets the basis for the design of a novel generation of chemicals suited for the organization of targeted lineage patterning in stem cells.

Animals↗
PubMedDate not supplied

Molecular entomology: a new promising tool for malaria control.

The recent biotechnological revolution in Molecular Entomology explores new promising tools for the control of vector borne diseases through genetic manipulation of vectorial competence. The gene transfer technology is hoped to make the malaria vectors incapable of supporting the development of malaria parasite which will ultimately lead to eradication of the parasites and the disease.

Animals↗
PubMed2005

Liposomal drug delivery system from laboratory to clinic.

The main objective of drug delivery systems is to deliver a drug effectively, specifically to the site of action and to achieve greater efficacy and minimise the toxic effects compared to conventional drugs. Amongst various carrier systems, liposomes have generated a great interest because of their versatility. Liposomes are vesicular concentric bilayered structures, which are biocompatible, biodegradable and nonimmumnogenic. They can control the delivery of drugs by targeting the drug to the site of action or by site avoidance drug delivery or by prolonged circulation of drugs. Amphotericin B (Amp B) remains the drug of choice in most systemic mycoses and also as a second line treatment for Kala azar. However, its toxic effects often limit its use. Although the liposome delivery system has been tried for several drugs, only a few have been used in patients due to the slow development of necessary large-scale pharmaceutical procedures. This paper reviews the development of the technique for liposomal Amphotericin B (L-Amp-LRC-1, Fungisome) drug delivery system in our laboratory in collaboration with the department of Biochemistry, Delhi University in India and proving the safety and efficacy of this preparation in clinical practice. It also attempts to compare the efficacy and benefits of our product for Indian patients with those of similar products and it includes facts from the publications that flowed from our work. As compared to conventional Amp B, Fungisome is infused over a much shorter period requiring a smaller volume and no premedication. It was found to be safe in patients who had developed serious unacceptable toxicity with conventional Amp B. In renal transplant patients, Fungisome did not produce any nephrotoxicity. Fungisome is effective in fungal infections resistant to fluconazole, conventional Amp B and in virgin and resistant cases of visceral leishmaniasis. The cost of any drug is of great significance, especially in India. We have therefore devoted a section of our review to the relative costs of our product and those of other commercially available products. This patient-worthy formulation is safe, efficacious and cheaper than the commercially available formulation of liposomal amphotericin B. The product has been patented and technology transferred to a pharmaceutical company for marketing. Results of postmarketing study also document safety and efficacy as observed in premarketing studies. A brief review of this work is provided here.

Amphotericin B↗
PubMed2006

[Outsourced real-time PCR diagnosis of cutaneous leishmaniasis in the outbreak region of Constatine, Algeria].

Taking into account the re-emergence of leishmaniasis in the world, the geographic variability of its epidemiology and the growing numbers of travellers, a pilot study on the diagnosis of cutaneous leishmaniasis was undertaken in Constatine, one of the outbreak regions in eastern Algeria. A total of 143 specimens were collected on blotters and tested by real-time PCR. Results were compared with those of direct examination. Diagnosis was positive for leishmaniasis in 81% of cases using PCR versus 48% of cases using microscopy. Real-time PCR showed a significant quantitative difference between patients for whom microscopic diagnosis was positive and those for whom direct examination was negative. The results presented in this study demonstrated the effectiveness and sensitivity of PCR in the diagnosis of cutaneous leishmaniasis from blotter specimens. This technique enabled in-field collection of specimens from each patient and provided prompt results. North-South cooperation based on the use of simple means for transmission of specimens for molecular diagnosis allowed creation of an effective partnership for daily diagnosis and promoted exchange between investigators in preparation for technology transfer.

Adolescent↗
PubMed2006

Translating research findings into practice: example of treatment services for adolescents in managed care.

An important question in the alcoholism treatment field is how research findings can be translated into real-world clinical practice. Researchers have developed a new research-practice integration (RPI) model that can both drive the formulation of studies and new research questions and promote improvements in treatment quality. The hallmark of this model is a collaborative relationship between the key stakeholders in both alcohol and other drug (AOD) treatment and research, including health plan administrators and clinicians, treatment program administrators, psychiatry and primary care departments, patients and their families, purchasers, and researchers. The issue of technology transfer is especially relevant in the realm of adolescent AOD treatment. The implementation and feasibility of the RPI model are illustrated by a case study of a managed health care plan's treatment services for adolescents with AOD dependence. In this setting, key research findings are being used to shape the plan's adolescent health services.

Adolescent↗
PubMed2006

Challenges and options for animal and public health services in the next two decades.

Trade in livestock and livestock products makes up approximately one sixth of global agriculture trade. This trade is demand driven, primarily by growing human populations, changing economies, and consumer preferences in developing countries. Different rates of population growth, economic growth, urbanisation, environmental sustainability, and technology transfer will determine which countries will reap the greatest benefits. Global trends in demand and supply for food, not terrorism, will drive the future of animal and public health service delivery. To benefit the greatest number of people and countries, animal and public health services should support policies that temper growing disparities among rich and poor countries, city and rural populations, and the sexes. Economic growth is critical to overcoming disparities between countries and best supported by integrated animal health, public health, labour, and foreign policies. Opportunities for job growth will be the greatest along the value added chain of food production and will require significant investments in science- (risk-) based education.

Animal Husbandry↗
PubMed2006

[Views on the financing of biomedical scientific research; the report 'Erop of eronder' (Sink or swim) from the KNAW (Royal Netherlands Academy of Arts and Sciences)].

In a recent report, entitled 'Erop of eronder. Financiering van (bio)medisch wetenschappelijk onderzoek' [Sink or swim. The financing of (bio)medical scientific research], the Royal Netherlands Academy of Arts and Sciences has reviewed various aspects of the current financing system. The recommendations concern a funding process with emphasis on open programmes focusing on researcher-initiated research, creating an earmarked budget for matching obligations, creating technology transfer centres offering broad expertise for the validation ofthe scientific results, and giving priority to the establishment of a European Research Council that encourages excellence in fundamental research within the European research area.

Biomedical Research↗
PubMed1991

[Smoking and alcohol drinking behavior among Japanese adolescents--results from "Japan Know Your Body Study"].

The "Japan Know Your Body Study" (JKYB) is an example of collaborative activities of technology transfer carried out in Japan in several phases. The first phase is a needs assessment and data collection survey and it was administered during 1989. A survey was conducted throughout 9 prefectures in Japan with participation by 49 schools. Subjects were students in elementary, junior and senior high schools who ranged in age from 6 to 18 years. Participants, 12,892 in total, completed a survey on knowledge, attitude and behavior regarding cigarette smoking, alcohol drinking, diet, exercise and so on. The main results concerning cigarette smoking and alcohol drinking behavior were as follows: 1) The percentage of monthly male smokers rose remarkably from 8% in the third grade of junior high school to 37% in the third grade of senior high school, while that of female smokers showed a slow rise from 3% to 15%. The sex difference for smoking behavior was clear among senior high school students. 2) Smoking rates increased in proportion to the numbers of parents, siblings and friends who smoked. Above all, smoking behavior of friends was strongly related. Among senior high school students, for example, 50% of males and 46% of females who had two or more smoking friends were smokers, as compared to 5% of males and 2% of females with no smoking friends. 3) The percentage of monthly drinkers rose at a steady pace from 13% of males and 7% of females in the fifth grade of elementary schools to 59% of males and 50% of females in the third grade of senior high schools. The sex difference for alcohol drinking behavior among senior high school students, while statistically significant, was not as pronounced as that for smoking behavior. These results suggest that tobacco- and alcohol-use prevention programs should be introduced in elementary schools and those programs should include components relating to the social influences of parents, siblings and friends.

Adolescent↗
PubMed1991

Pharmacological characteristics of high-affinity serotonin uptake systems established through gene transfer.

Inactivation of a neurotransmitter, after its stimulated release, via high-affinity uptake mechanisms is an essential regulatory step of neurotransmission in both the central and peripheral nervous systems. To initiate explorations of the molecular mechanisms and the underlying biochemical architecture of high-affinity neurotransmitter uptake systems, we have used gene transfer technology to establish and identify novel cellular models that express these systems. Human genomic DNA was transfected into mouse L-M fibroblasts and two independently arising, clonal cell lines (L-S1 and L-S2) have been identified as expressing high-affinity serotonin (5-HT) uptake systems. The 5-HT uptake characteristics of L-S1 and L-S2 are essentially comparable (in terms of Na+ dependence, temperature sensitivity, imipramine antagonizability, kinetic saturability and high affinities) and those of L-S1 have been reported previously. Furthermore, competition studies utilizing catecholamine neurotransmitters and their amino acid precursors demonstrated that these systems are highly specific for 5-HT. Several known inhibitors of high-affinity 5-HT uptake systems (including amitriptyline, desipramine, fluoxetine, imipramine, nortriptyline, tryptamine, 5-methoxytryptamine and N-acetyl 5-methoxytryptamine) were assessed in terms of their respective potencies to inhibit 5-[3H]HT uptake by L-S1 and L-S2 cells. For L-S1 cells, the rank order of inhibitor potencies is imipramine greater than amitriptyline greater than fluoxetine greater than desipramine = nortriptyline greater than tryptamine greater than 5-methoxytryptamine greater than N-acetyl-5-methoxytryptamine. For L-S2, the rank order is similar to that of L-S1 except that fluoxetine is more potent than amitriptyline.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗