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At least 487 records · Page 27Linked to original sources
PubMed2000

Technological considerations related to the up-scaling of protein microencapsulation by spray-drying.

Research and development of therapeutics and vaccines based on biodegradable polymers are intensive and one of the most promising fields in controlled drug delivery. However, new applications necessitate successful technology transfer and industrial scale-ups. In an endeavour to produce clinical samples of a single-administration tetanus vaccine based on poly(lactide-co-glycolide) microspheres, we report on technological parameters that are of importance in the up-scaling of the spray-drying process. The results show that an up-scaling of the encapsulation of protein vaccines or drug by spray-drying is feasible, but that additives, the type of polymer solvent, the polymer concentration, the w/o ratio and the product collection method influence process and product quality.

Biocompatible Materials↗
PubMed2005

Physical methods for gene transfer: improving the kinetics of gene delivery into cells.

One factor critical to successful gene therapy is the development of efficient delivery systems. Although advances in gene transfer technology, including viral and non-viral vectors, have been made, an ideal vector system has not yet been constructed. This review describes the basic principles behind various physical methods for gene transfer and assesses the advantages and performance of such approaches, compared to other transfection systems. In particular, the kinetics and efficiency of gene delivery, the toxicity, in vivo feasibility, and targeting ability of different physical methodologies are discussed and evaluated.

Animals↗
PubMedDate not supplied

Laser printing of pluripotent embryonal carcinoma cells.

A technique by which to print patterns and multilayers of scaffolding and living cells could be used in tissue engineering to fabricate tissue constructs with cells, materials, and chemical diversity at the micron scale. We describe here studies using a laser forward transfer technology to print single-layer patterns of pluripotent murine embryonal carcinoma cells. This report focuses on verifying cell viability and functionality as well as the ability to differentiate cells after laser transfer. We find that when cells are printed onto model tissue scaffolding such as a layer of hydrogel, greater than 95% of the cells survive the transfer process and remain viable. In addition, alkaline comet assays were performed on transferred cells, showing minimal single-strand DNA damage from potential ultraviolet-cell interaction. We also find that laser-transferred cells express microtubular associated protein 2 after retinoic acid stimulus and myosin heavy chain protein after dimethyl sulfoxide stimulus, indicating successful neural and muscular pathway differentiation. These studies provide a foundation so that laser printing may next be used to build heterogeneous multilayer cellular structures, enabling cell growth and differentiation in heterogeneous three-dimensional environments to be uniquely studied.

Animals↗
PubMed2000

Functional analysis of cardiovascular renin-angiotensin system using a gain or loss of function approach.

The study of the effect of autocrine-paracrine vasoactive modulators on cardiovascular biology is very difficult in vivo, because in vivo studies are limited. In particular, characterization of the role of components of the renin-angiotensin system (RAS) in vivo is limited by the difficulty in manipulating individual components of the RAS as well as by methodological limitations in studying the function of a local RAS in the absence of any contribution by the circulatory system. Recent progress in in vivo gene transfer technologies has provided us with the opportunity to study cellular responses to the manipulation of the individual components (i.e., by overexpression or inhibition). Many researchers have recently developed various in vivo gene transfer techniques for cardiovascular applications. Using in vivo gene transfer approaches, the roles of various tissues in the RAS, such as cardiac angiotensin, have been identified. Such an approach may increase our understanding of the biology and pathobiology of the autocrine-paracrine system. This review discusses the potential utility of in vivo gene transfer methods.

Angiotensins↗
PubMed2001

Deep intrauterine insemination and embryo transfer in pigs.

A new method for non-surgical deep intrauterine catheterization of pigs, without sedation of the sow, is described. Insemination results obtained with this method using fresh spermatozoa demonstrate that, in comparison to conventional artificial insemination (AI) (3 x 10(9) spermatozoa in 80-100 ml), a 20-60-fold reduction in the number of spermatozoa inseminated and at least a 8-10-fold reduction in the dose volume can be used without affecting fertility if spermatozoa are deposited deep (middle or upper) into one of the uterine horns. Results from deep intrauterine insemination with frozen-thawed spermatozoa and flow-sorted spermatozoa are also presented and the effect of deep intrauterine insemination on sperm transport is discussed. In addition, a brief description of the advances made in non-surgical embryo transfer technology is reported.

Animals↗
PubMed2005

Cutting through the obstacles and resurrecting the promise of gene therapy.

Gene therapy is a promising biomedical discipline that could potentially lead to new treatments and perhaps long-term curative effects for a plethora of diseases including hereditary disorders, cardiovascular and neurological diseases, cancer, diabetes and even infectious or autoimmune diseases. These diseases affect millions of people worldwide and the development of effective and safe gene-based drugs obviously represent a tremendous market potential. Convincing evidence continues to emerge from clinical trials demonstrating that gene therapy can be effective in patients suffering from a limited number of different diseases. Nevertheless, as with any emerging new biomedical discipline, gene therapy has also faced a number of setbacks, and there have been concerns regarding the safety of some gene delivery approaches, however, these hurdles are not insurmountable. Gene transfer technologies are improving rapidly and have led to the development of new and more efficacious gene delivery approaches with fewer side effects. The success of gene therapy is still highly dependent upon the continuous development of improved gene delivery technologies, the progress of which should hopefully and ultimately cure diseases that are refractory to current treatment paradigms.

Gene Transfer Techniques↗
PubMed2005

Science and ethics of sham surgery: a survey of Parkinson disease clinical researchers.

BACKGROUND: Sham surgery is used in neurosurgical clinical trials in Parkinson disease (PD) but remains controversial. The controversy may be compounded when gene-transfer technologies are tested in sham surgical trials. OBJECTIVE: To determine the perspective of PD clinical researchers on the science and ethics of sham-surgery controls when used to test novel interventions such as gene transfer for PD. DESIGN: Internet survey eliciting both quantitative and qualitative responses. PARTICIPANTS: Investigator members of the Parkinson Study Group. RESULTS: Overall response rate was 103 (61.3%) of 168 researchers. A large majority (97%) of PD clinical researchers believe sham-surgery controls are better than unblinded controls for testing the efficacy of neurosurgical interventions such as gene transfer for PD. Half of the researchers believe an unblinded control efficacy trial would be unethical because it may lead to a falsely positive result. A minority (less than 22%) believe that an invasive sham condition that involves penetration of brain tissue is justified. CONCLUSION: It appears unlikely that the PD clinical research community will perceive future neurosurgical interventions for PD, such as gene-transfer therapies, as truly efficacious unless a sham-control condition is used to test it.

Adult↗
PubMedDate not supplied

The zootechnical applications of biotechnology in animal reproduction: current methods and perspectives.

The development of the four generations of Reproductive Biotechnology, particularly in cattle and since the last world war, represents one of the best examples of the success story of technology transfer. This review will only refer to the first three generations and will not deal with nuclear transfer nor transgenesis. Based on sound so-called "finalised" research, Artificial Insemination first, then in vivo collected embryo transfer and later in vitro fertilised embryo transfer have been implemented worldwide. Each of these Biotechnologies has many advantages and limitations. In addition to the specificity of each of them, one major point is that farmers and breeders may choose either collectively or individually, the best technology to be used in order to achieve the goals they have set for their industry. It is noteworthy that these technologies have been able to match with the economics demands over the last decades and yet are in a very good capacity to respond to the contemporary demand of sustainable development. In this context, there are further advantages such as potentially contributing to maintaining biodiversity or allowing preservation ex situ of genes otherwise threatened to extinction.

Animal Husbandry↗
PubMed2003

The potential for improving the growth and development of cultured farm animal oocytes.

Previous predictions that the technologies for producing genetically engineered large animal embryos containing genes for faster growth rates, leaner carcasses, greater disease resistance and improved lactational performance would be available early in the twenty-first century have been, for the most part, realized. The animal industries have been slow to adopt these technological advances and it cannot be said that any of them are currently having great impact on animal agriculture worldwide. A major reason for this is the inefficiencies of the techniques for superovulation, ovum recovery, in vitro fertilization, nuclear transfer, cloning and embryo transfer. Although improvements in these techniques can be expected, the best hope for increasing the impact of embryo transfer technologies on the animal industries lies in developing ways to mature, harvest, store and fertilize in vitro the large numbers of primordial oocytes present in the ovaries of all farm animals. Although limited progress has been made in the culture of bovine primordial oocytes, it is clear that much more research is needed to achieve success in this important area.

Abortion, Veterinary↗
PubMed1999

Expression of recombinant NMDA receptors in hippocampal neurons by adenoviral-mediated gene transfer.

N-methyl-d-aspartate (NMDA) receptors have attracted a great deal of attention because they are intimately involved in brain development, synaptic plasticity and a variety of neurological disorders. The ability to artificially alter the properties of NMDA receptors in central nervous system (CNS) neurons would be useful for elucidating the physiological roles of these receptors. It would also raise the possibility of gene therapy of neurological diseases caused by malfunction of NMDA receptors. In this study, we constructed three recombinant adenoviruses encoding rat NMDA receptor subunit cDNAs, NMDAR1 (NR1), NMDAR2B (NR2B) and mutant NR1(N598R) in which the asparagine (N) site of the wild-type NR1 was replaced with arginine (R) by site-directed mutagenesis. PC12 cells co-infected with recombinant adenoviruses bearing NR1 and NR2B cDNAs expressed conventional NMDA receptors that were permeable to Ca2+ and sensitive to Mg2+, whereas those with viruses bearing NR1(N598R) and NR2B cDNAs expressed Ca2+-impermeable and Mg2+-insensitive receptors. When rat hippocampal neurons in culture were infected with NR1(N598R) and NR2B viruses, both Ca2+ permeability and Mg2+ sensitivity of NMDA receptors were markedly reduced in the infected neurons. Excitatory postsynaptic currents (EPSCs) mediated by NMDA receptors also became much less sensitive to Mg2+. Thus, the NR1(N598R)/NR2B receptors were more dominant than the native NMDA receptors in the infected neurons, and the former receptors introduced by the adenoviral vectors functioned as postsynaptic receptors. These results indicate that the functional properties of postsynaptic NMDA receptors can be manipulated by gene transfer technology using adenoviral vectors.

Adenoviridae↗
PubMed2001

Transgenics in crops.

With rapid world population growth and declining availability of fresh water and arable land, a new technology is urgently needed to enhance agricultural productivity. Recent discoveries in the field of crop transgenics clearly demonstrate the great potential of this technology for increasing food production and improving food quality while preserving the environment for future generations. In this review, we briefly discuss some of the recent achievements in crop improvement that have been made using gene transfer technology.

Agriculture↗
PubMed2004

Prospects for gene therapy of haemophilia.

That gene therapy offers the promise of a cure for haemophilia was apparent more than a decade ago. After years of failure, substantial progress in the efficiency of gene transfer technology has recently resulted in impressive success in animal models with haemophilia. However, fears of the risks intrinsic to such therapy have been raised by the fate of two children cured of immune deficiency by gene transfer who have, however, subsequently developed leukaemia as a result of insertional mutagenesis. The purpose of this review is to outline the current status of gene therapy in light of recent successes and tragedies and to consider the prospects for curing haemophilia in the short-to-medium term.

Adenoviridae↗
PubMed2000

[Gene transfer].

Mammalian cells in culture can be modified by gene transfer and these procedures are routinely used in experimental biology. Yet, efficient approaches to modify certain complex cell populations, stem cells or cells organized within a tissue are still lacking. The hurdles to gene transfer can be listed by describing the pathway of a nucleic acid molecule, from the external medium towards the cell nucleus where its encoded information will be expressed. The requirements include the necessity to compact the size of the macromolecule, to overcome electrostatic repulsion, to cross a series of membranes and to establish itself permanently. Viruses have evolved to achieve these goals and understanding their strategies for cell invasion allows to design vectors for gene transfer. Chemicals or biochemicals able to bind DNA, as well as physical methods inducing changes in the structure of membranes can also be useful for gene transfer. The outcome of gene transfer technologies and their improvement pave the way to inovative medical applications and provide powerful tools for exploring the living world.

Animals↗
PubMed2005

[The risk of congenital malformations and genomic imprinting defects in assisted reproductive technologies and nuclear transfer cloning].

Recent studies show that assisted reproductive technologies (ART), whether in vitro fertilization (IVF) or intra-cytoplasmatic sperm injection (ICSI) or applied to cloning by somatic cell nuclear transfer (SCNT) are associated to a higher risk of congenital malformations and errors in deprogramming, maintenance or reprogramming genomic imprinting in humans and animals. IVF and ICSI are also associated to an increased admission to neonatal intensive care units and more need for health care resources in infancy. A mutagenic effect of a chemical used in SCNT has been reported and gene depression was found in bovine embryos obtained by IVF or SCNT. The causes of these anomalies could be pathological conditions for which ART is applied, a direct effect of technologies on the zygotes or embryos, avoidance for zygotes or embryos of the oviduct path that is needed to elicit necessary immunity or genomic programming processes, or adaptive selective steps acquired during thousands of millions of generations in evolution. The knowledge of evolution is emphasized as essential in the scientific ethical analysis.

Animals↗
PubMed2002

Recent progress and problems in animal cloning.

It is remarkable that mammalian somatic cell nuclei can form whole individuals if they are transferred to enucleated oocytes. Advancements in nuclear transfer technology can now be applied for genetic improvement and increase of farm animals, rescue of endangered species, and assisted reproduction and tissue engineering in humans. Since July 1998, more than 200 calves have been produced by nuclear transfer of somatic cell nuclei in Japan, but half of them were stillborn or died within several months of parturition. Morphologic abnormalities have also been observed in cloned calves and embryonic stem cell-derived mice. In this review, we discuss the present situation and problems with animal cloning and the possibility for its application to human medicine.

Animals↗
PubMed2000

Drug discovery: a historical perspective.

Driven by chemistry but increasingly guided by pharmacology and the clinical sciences, drug research has contributed more to the progress of medicine during the past century than any other scientific factor. The advent of molecular biology and, in particular, of genomic sciences is having a deep impact on drug discovery. Recombinant proteins and monoclonal antibodies have greatly enriched our therapeutic armamentarium. Genome sciences, combined with bioinformatic tools, allow us to dissect the genetic basis of multifactorial diseases and to determine the most suitable points of attack for future medicines, thereby increasing the number of treatment options. The dramatic increase in the complexity of drug research is enforcing changes in the institutional basis of this interdisciplinary endeavor. The biotech industry is establishing itself as the discovery arm of the pharmaceutical industry. In bridging the gap between academia and large pharmaceutical companies, the biotech firms have been effective instruments of technology transfer.

Chemistry, Pharmaceutical↗
PubMed2006

Emerging vectors and targeting methods for nonviral gene therapy.

Until recently, nonviral vectors were outside the mainstream of gene transfer technology. Recent problems in clinical trials using viral vectors renewed interest in these methods. The clinical usefulness of nonviral methods is still hindered by their relatively low gene delivery/transgene expression efficiencies. Vectors must navigate a series of obstacles before the therapeutic gene can be expressed. This review considers these barriers and the properties of components of nonviral vectors that are essential for nucleic acid transfer. Although developments of new physical methods (hydrodynamic delivery, ultrasound, electroporation) have made a significant impact on gene transfer efficiency, various chemical carriers (lipids and polymers) have been shown to achieve high-level gene delivery and functional expression. Success of nonviral gene targeting will depend not only on the efficacy, but also safety of this methodology, and this aspect is also discussed. Understanding problems associated with nonviral targeting can also help in designing better viral vectors. In fact, interplay between viral and nonviral technologies should lead to a continued refinement of both methodologies.

Animals↗
PubMed2004

New angles in mycology: studies in directional growth and directional motility.

Mycology is changing as an era of extensive genome sequencing comes of age and provides vital information that enables questions to be addressed about fungi in all the major taxonomic groups. As technology transfer facilitates what was once only possible for a very small number of model species, it becomes possible to explore the biology and biodiversity of fungi as a whole. The availability of genome sequence information and reverse genetic technologies allows hypotheses that emerge from biological observations to be tested. Genomic and post-genomic technologies will underline the importance of fungi as excellent models for the study of fundamental biological phenomena. Two enduring areas of research in my own laboratory are described that are now being extended using post-genomic approaches. These projects relate to how fungal hyphae extend and guide their tips and secondly how plant pathogenic oomycete zoospores are guided on their journey to the plant surface.

Fungi↗