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PubMed2005

Farm animal genomics and informatics: an update.

Farm animal genomics is of interest to a wide audience of researchers because of the utility derived from understanding how genomics and proteomics function in various organisms. Applications such as xenotransplantation, increased livestock productivity, bioengineering new materials, products and even fabrics are several reasons for thriving farm animal genome activity. Currently mined in rapidly growing data warehouses, completed genomes of chicken, fish and cows are available but are largely stored in decentralized data repositories. In this paper, we provide an informatics primer on farm animal bioinformatics and genome project resources which drive attention to the most recent advances in the field. We hope to provide individuals in biotechnology and in the farming industry with information on resources and updates concerning farm animal genome projects.

Animals↗
PubMed2001

A critical overview of acceptance testing using various measured indices.

It is a requirement of the new European directive on health protection of individuals against the dangers of ionising radiation in relation to medical exposure (97/43/EURATOM), that acceptance testing is carried out on all radiological equipment before its first use for clinical purposes. This directive must be implemented into member state law. In accordance with this requirement, thirteen digital fluorography systems were acceptance tested by the Department of Medical Physics and Bioengineering, St James's Hospital, Dublin. The equipment tested was in six Irish hospitals and included nine systems intended for interventional use, two of which were dedicated cardiac catheterisation laboratories and two of which were dedicated vascular laboratories. Testing was performed in line with current published international guidelines and standards and involved the measurement and analysis of various indices including X ray tube and generator performance, operating dose levels, image quality and electrical, mechanical and radiation safety aspects. The results are presented and compared with international standards and recommendations. Problems were identified with all systems tested with approximately 70% of all systems demonstrating significant problems. This study emphasises the importance of acceptance testing radiological equipment.

European Union↗
PubMed2004

His-404 and His-405 are essential for enzyme catalytic activities of a bacterial indole-3-acetyl-L-aspartic acid hydrolase.

Bacterial indole-3-acetyl-l-aspartic acid (IAA-Asp) hydrolase has shown very high substrate specificity compared with similar IAA-amino acid hydrolase enzymes found in Arabidopsis thaliana. The IAA-Asp hydrolase also exhibits, relative to the Arabidopsis thaliana-derived enzymes, a very high Vmax (fast reaction rate) and a higher Km (lower substrate affinity). These two characteristics indicate that there are fundamental differences in the catalytic activity between this bacterial enzyme and the Arabidopsis enzymes. By employing a computer simulation approach, a catalytic residue, His-385, from a non-sequence-related zinc-dependent exopeptidase of Pseudomonas was found to structurally match His-405 of IAA-Asp hydrolase. The His-405 residue is conserved in all related sequences of bacteria and Arabidopsis. Point mutation experiments of this His-405 to seven different amino acids resulted in complete elimination of enzyme activity. However, point mutation on the neighboring His-404 to eight other residues resulted in reduction, to various degrees, of enzyme activity. Amino acid substitutions for His-404 also showed that this residue influenced the minor activity of the IAA-Asp hydrolase for the substrates IAA-Gly, IAA-Ala, IAA-Ser, IAA-Glu and IAA-Asn. These results show the value and potential of structural modeling for predicting target residues for further study and for directing bioengineering of enzyme structure and function.

Amino Acid Sequence↗
PubMed2004

Characterization and electrotransformation of Lactobacillus crispatus isolated from chicken crop and intestine.

Lactic acid bacteria originating in the intestine have recently undergone intensive study for their potential probiotic properties. Here partial 16S rRNA gene sequencing of 8 Lactobacillus strains proved them to be Lactobacillus crispatus. Fatty acid analysis confirmed strains being closely related. These strains and type strain ATCC33820 were characterized for genetic engineering potential, thus determining aerobic growth, erythromycin sensitivity, and glycine tolerance. Out of 5 plasmids, a 2.9-kb plasmid (pLEB579) was successfully introduced into 4 chicken-originated wild-type L. crispatus strains. Transformation frequency was approximately 30 transformants per microgram of DNA, the first reported electrotransformation into chicken-originated L. crispatus. In spite of its low frequency, transformation enables bioengineering of these strains to improve the probiotic function in feed adsorption, chicken health, and food safety.

Animals↗
PubMed1983

Abnormal patterns of normal eye movements in schizophrenia.

The multilevel control of eye movements for the researcher in the schizophrenic disease process includes the saccadic trajectory, the dual mode tracking system, and schematically directed eye movements in fixation, in scanpaths, and other looking strategies, and in reading. Coordination of eye movements with head movement in gaze and sensory consequences and concomitants of eye movements round out the picture. Recent studies with precise bioengineering instrumentation have defined two abnormal patterns of eye movements--saccadic intrusions and saccadic "smooth" pursuit. Although these signs are not pathognomonic, they raise interesting questions concerning the relationship of these eye movements to the schizophrenic disease process.

Attention↗
PubMed2003

Generation of live fry from intraperitoneally transplanted primordial germ cells in rainbow trout.

Germ cell transplantation has tremendous applications in transgenic animal production, assisted reproductive technology, and germline stem cell research. Here, we report for the first time the production of individuals from intraperitoneally transplanted primordial germ cells (PGCs) in animals. To trace the behavior of exogenous PGCs in recipients, PGCs visualized by a green fluorescent protein gene were used as donors. The PGCs prepared from the genital ridges of hatching embryos were transplanted into recipients at various developmental stages. The PGCs injected into the peritoneal cavities of hatching embryos had the ability to migrate toward, and to colonize, the genital ridges of recipient embryos. Furthermore, donor-derived PGCs proliferated and differentiated into mature eggs and sperm in the allogenic gonads; the resulting gametes produced live fry, showing the donor-derived phenotype, through fertilization. Combined with in vitro culture, genetic modification, and cryopreservation of PGCs, this technique provides new approaches for fish bioengineering.

Age Factors↗
PubMed2005

Green fluorescent protein labeling of primordial germ cells using a nontransgenic method and its application for germ cell transplantation in salmonidae.

Transplanting primordial germ cells (PGCs) has a number of potential applications in fish bioengineering. Previously, we established a system to visualize live PGCs in the rainbow trout by introducing the green fluorescent protein (Gfp) gene driven by rainbow trout vasa gene regulatory regions. However, for PGC transplantation to be practically useful in aquaculture, visualization of PGCs using a nontransgenic technique is required. In this study, we demonstrate a method for labeling PGCs from various fish species by introducing chimeric RNAs composed of the Gfp coding region and vasa gene 3'-untranslated regions (UTRs); these sequences play a critical role in stabilizing mRNA in zebrafish PGCs. The GFP chimeric RNAs, including vasa 3'-UTR RNAs from rainbow trout, Nibe croaker, and zebrafish, were microinjected into the cytoplasm of fertilized eggs of several Salmonidae species. All the resulting embryos showed specific labeling in PGCs after the somatogenesis stage, which continued to be visible for at least 50 days. To apply this technique to PGC transplantation, PGCs labeled with chimeric RNA were microinjected into the peritoneal cavity of newly hatched salmonid embryos. The GFP labeling was sufficiently long-lived for the initial stage of donor PGC behavior to be followed in the recipient embryos. Importantly, donor PGCs from brown trout and masu salmon were incorporated into xenogeneic genital ridges in recipient rainbow trout. This nontransgenic method for labeling fish PGCs should be extremely useful for applications of PGC transplantation where the resulting progeny are to be released into the environment, such as PGC cryopreservation for fish stocks and surrogate brood stock technology.

3' Untranslated Regions↗
PubMed2003

Innervated human corneal equivalents as in vitro models for nerve-target cell interactions.

A sensory nerve supply is crucial for optimal tissue function. However, the mechanisms for successful innervation and the signaling pathways between nerves and their target tissue are not fully understood. Engineered tissue substitutes can provide controllable environments in which to study tissue innervation. We have therefore engineered human corneal substitutes that promote nerve in-growth in a pattern similar to in vivo re-innervation. We demonstrate that these nerves (a) are morphologically equivalent to natural corneal nerves; (b) make appropriate contact with target cells; (c) can generate action potentials; (d) respond to chemical and physical stimuli; and (e) play an important role in the overall functioning of the bioengineered tissue. This model can be used for studying the more general topics of nerve ingrowth or regeneration and the interaction between nerves and their target cells and, more specifically, the role of nerves in corneal function. This model could also be used as an in vitro alternative to animals for safety and efficacy testing of chemicals and drugs.

Action Potentials↗
PubMed2004

Gene and cell-based therapies for heart disease.

Heart disease remains the prevalent cause of premature death and accounts for a significant proportion of all hospital admissions. Recent developments in understanding the molecular mechanisms of myocardial disease have led to the identification of new therapeutic targets, and the availability of vectors with enhanced myocardial tropism offers the opportunity for the design of gene therapies for both protection and rescue of the myocardium. Genetic therapies have been devised to treat complex diseases such as myocardial ischemia, heart failure, and inherited myopathies in various animal models. Some of these experimental therapies have made a successful transition to clinical trial and are being considered for use in human patients. The recent isolation of endothelial and cardiomyocyte precursor cells from adult bone marrow may permit the design of strategies for repair of the damaged heart. Cell-based therapies may have potential application in neovascularization and regeneration of ischemic and infarcted myocardium, in blood vessel reconstruction, and in bioengineering of artificial organs and prostheses. We expect that advances in the field will lead to the development of safer and more efficient vectors. The advent of genomic screening technology should allow the identification of novel therapeutic targets and facilitate the detection of disease-causing polymorphisms that may lead to the design of individualized gene and cell-based therapies.

Coronary Artery Disease↗
PubMed2005

Functional human corneal endothelial cell sheets harvested from temperature-responsive culture surfaces.

This study reports a new method for fabricating bioengineered human corneal endothelial cell sheets suitable for ocular surgery and repair. We have initially cultured human corneal endothelial cells on type IV collagen-coated dishes and, after several passages, expanded cells were then seeded onto novel temperature-responsive culture dishes. Four weeks after reaching confluence, these cultured endothelial cells were harvested as intact monolayer cell sheets by simple temperature reduction without enzymatic treatment. Scanning electron microscopy indicated that these cells were primarily hexagonal with numerous microvilli and cilia, similar to the native corneal endothelium. The Na+, K+-ATPase pump sites were located at the cell borders as in vivo. Moreover, cell densities and numbers of pump sites were identical to those of in vivo human corneal endothelium under optimized conditions. A 3H-ouabain binding analysis demonstrated a linear proportionality for cell pump density between confluent cell densities of 575 cells/mm2 and 3070 cells/mm2. We also confirmed Na+, K+-ATPase activity in the sheets in vitro. Xenograft transplantation results showed that the fabricated sheets retain their function of maintaining proper stromal hydration in vivo. We have established a regimen to culture and proliferate human corneal endothelial cells and fabricate endothelial sheets ex vivo morphologically and functionally similar to the native corneal endothelium. Our results support the value of harvested cell sheets for clinical applications in ocular reconstructive surgery in patients with ocular endothelial decompensation.

Cell Culture Techniques↗
PubMed2006

Polysurgery of cell sheet grafts overcomes diffusion limits to produce thick, vascularized myocardial tissues.

Recently, the field of tissue engineering has progressed rapidly, but poor vascularization remains a major obstacle in bioengineering cell-dense tissues, limiting the viable size of constructs due to hypoxia, nutrient insufficiency, and waste accumulation. Therefore, new technologies for fabricating functional tissues with a well-organized vasculature are required. In the present study, neonatal rat cardiomyocytes were harvested as intact sheets from temperature-responsive culture dishes and stacked into cell-dense myocardial tissues. However, the thickness limit for layered cell sheets in subcutaneous tissue was approximately 80 microm (3 layers). To overcome this limitation, repeated transplantation of triple-layer grafts was performed at 1, 2, or 3 day intervals. The two overlaid grafts completely synchronized and the whole tissues survived without necrosis in the 1 or 2 day interval cases. Multistep transplantation also created approximately 1 mm thick myocardium with a well-organized microvascular network. Furthermore, functional multilayer grafts fabricated over a surgically connectable artery and vein revealed complete graft perfusion via the vessels and ectopic transplantation of the grafts was successfully performed using direct vessel anastomoses. These cultured cell sheet integration methods overcome long-standing barriers to producing thick, vascularized tissues, revealing a possible solution for the clinical repair of various damaged organs, including the impaired myocardium.

Animals↗
PubMed2002

Development and evaluation of a graphical anesthesia drug display.

BACKGROUND: Usable real-time displays of intravenous anesthetic concentrations and effects could significantly enhance intraoperative clinical decision-making. Pharmacokinetic models are available to estimate past, present, and future drug effect-site concentrations, and pharmacodynamic models are available to predict the drug's associated physiologic effects. METHODS: An interdisciplinary research team (bioengineering, architecture, anesthesiology, computer engineering, and cognitive psychology) developed a graphic display that presents the real-time effect-site concentrations, normalized to the drugs' EC(95), of intravenous drugs. Graphical metaphors were created to show the drugs' pharmacodynamics. To evaluate the effect of the display on the management of total intravenous anesthesia, 15 anesthesiologists participated in a computer-based simulation study. The participants cared for patients during two experimental conditions: with and without the drug display. RESULTS: With the drug display, clinicians administered more bolus doses of remifentanil during anesthesia maintenance. There was a significantly lower variation in the predicted effect-site concentrations for remifentanil and propofol, and effect-site concentrations were maintained closer to the drugs' EC(95). There was no significant difference in the simulated patient heart rate and blood pressure with respect to experimental condition. The perceived performance for the participants was increased with the drug display, whereas mental demand, effort, and frustration level were reduced. In a post-simulation questionnaire, participants rated the display to be a useful addition to anesthesia monitoring. CONCLUSIONS: The drug display altered simulated clinical practice. These results, which will inform the next iteration of designs and evaluations, suggest promise for this approach to drug data visualization.

Anesthesia, Intravenous↗
PubMed1979

John Davies Reese and the Reese dermatome.

John Davies Reese was a unique "loner" whose expertise lay in the field of bioengineering. This led to his development of the calibrated dermatome that bears his name. A bachelor, Reese devoted all his spare time to perfecting dermatape, glue, and the instrument itself.

History, 19th Century↗
PubMed2000

Cost-efficacy of cultured epidermal autografts in massive pediatric burns.

OBJECTIVE: To assess the efficacy of cultured epidermal autografts (CEA) for closure of burn wounds in pediatric burn patients with full-thickness burns of more than 90% total body surface area. SUMMARY BACKGROUND DATA: Paucity of donor sites in massive burns makes the use of expanded skin of paramount importance. CEA techniques have been used in burned patients with differing and controversial results. The true impact and the efficacy of such techniques in massive burns remain uncertain. METHODS: Patients with full-thickness burns of more than 90% body surface area treated between May 1988 and May 1998 were studied. Patients grafted with CEA were compared with patients grafted with conventional meshed autografts. Rates of death and complications, length of hospital stay (LOS), hospital cost, acute readmissions for reconstruction, and quality of scars were studied as outcome measures. RESULTS: Patients treated with CEA had a better quality of burn scars but incurred a longer LOS and higher hospital costs. Both groups had comparable readmissions for open wounds, but patients treated with CEA required more reconstructive procedures during the first 2 years after the injury. The incidence of sepsis and pneumonia in both groups was comparable. CONCLUSIONS: Conventional meshed autografts are superior to CEA for containing hospital cost, diminishing LOS, and decreasing the number of readmissions for reconstruction of contractures. However, the use of CEA provides better scar quality such that perhaps future research should focus on bioengineered dermal templates to promote take and diminish long-term fragility.

Burns↗
PubMed1991

Effect of cancer therapy on host response and immunobiology.

The advances in bioengineering and hybridoma technology and the acquisition of basic knowledge about human antitumor defenses have facilitated the development of biologic therapies as a fourth cancer treatment approach. It is becoming important, therefore, to reconsider how conventional therapies affect host responses. It is apparent from the work of many laboratories that both the biologic agents and certain anticancer agents can cause augmentation of antitumor host defenses. These augmenting effects are generally seen at less than maximally tolerated doses. In contrast, at maximally tolerated doses these agents may induce long-lasting untoward effects, some of which (eg, long-term immunodepression) seem incongruent with effective host response-based therapies. This review examines developments in these areas over the past year and suggests that the maximal benefit may not be derived from the combination of chemotherapeutic agents with biologic therapies unless the optimal conditions for their administration in such combinations are redefined.

Antineoplastic Agents↗
PubMed2001

Fetal electrocardiogram waveform analysis in labour.

The fetal electrocardiogram recorded during labour has so far been used for heart rate recording only. Extensive experimental, bioengineering and clinical research for more than 25 years on changes in the morphology of the electrocardiogram have provided more detailed information on fetal reactions to the stress and strain of labour. ST waveform analysis has been shown to provide new parameters that have the ability to monitor intrapartum hypoxia, thereby not only reducing the risk of the baby being born with hypoxia, but also significantly reducing the number of operative interventions.

Acid-Base Imbalance↗
PubMedDate not supplied

Forces, moments, and accelerations acting on an unrestrained dummy during simulations of three wheelchair accidents.

To determine the magnitude and distribution of the forces, moments, and accelerations acting on an individual sitting in a wheelchair during three possible accidents occurring while negotiating a sidewalk curb, experimental trials were performed in a bioengineering laboratory using a 50th percentile Hybrid III dummy seated in a standard wheelchair. A ramp was designed with an adjustable incline to allow the wheelchair to reach the edge of a sidewalk height platform at the desired forward speed of 2.5 miles per hour (comfortable walking speed). The wheelchair velocity was monitored via an optical pickup. Three types of accidents were simulated: (1) a wheelchair hitting straight into a curb; (2) a wheelchair falling forward straight off a curb; (3) a wheelchair falling diagonally off a curb. Each experiment was repeated three times. Each run was photographed using high-speed cameras and videotaped from three perspectives: frontal, lateral, and overhead. The results were averaged and compared with published injury Assessment Values (IAV) and Head Injury Criteria (HIC). Of significance were the following results. In the straight into a curb experiments, the wheelchair remained upright and the dummy seated. Low magnitude forces (23-73 N), moments (1-12 Nm), and accelerations (0.2-1 G) were recorded at the neck and head. The HIC was low at 0.3. These results were of no clinical significance. In the straight off a curb experiments, properly attached footrests prevented the wheelchair from toppling over but did not prevent the dummy from falling off the wheelchair. Forces (187-4,176 N), moments (3-178 Nm), and accelerations (131-206 G) of great magnitude were recorded at the head and neck when the dummy fell off the wheelchair. These values were above IAV. The HIC was 960. In the diagonally off a curb experiments, both the wheelchair and the dummy fell sideways. High-magnitude forces (274-2,313 N), moments (4-110 Nm), and acceleration (140-236 G) were recorded in the head and neck regions. The HIC was 975. These values were close to IAV and may signify potential serious injuries.

Acceleration↗
PubMedDate not supplied

Scaffold precoating with human autologous extracellular matrix for improved cell attachment in cardiovascular tissue engineering.

Cell attachment to a scaffold is a precondition for the development of bioengineered valves and vascular substitutes. This attachment is generally facilitated by the use of precoating factors, but some can cause toxic or immunologic side effects. Autologous extracellular matrix (ECM) is used as a precoating factor in our study. Ascending aortic tissue was cultured to obtain human myofibroblasts. Autologous ECM was extracted from the same aortic tissue. Poly(glycolic acid) (PGA) scaffolds were precoated with autologous ECM, human serum, or poly-L-lysine; the control group was pretreated with phosphate buffered saline (PBS). Myofibroblasts were seeded onto each scaffold, and the cell attachment was assayed and compared. Compared with the control group, precoating with human serum, poly-L-lysine, and ECM increased number of attached cells by 24%, 53%, and 48%, respectively. Differences between precoating groups were significant (p < 0.01), except for ECM versus poly-L-lysine. Scanning electron microscopy also demonstrated the high degree of cell attachment to the PGA fibers on scaffolds precoated with ECM and poly-L-lysine. Precoating polymeric scaffold with autologous human extracellular matrix is a very effective method of improving cell attachment in cardiovascular tissue engineering without the potential risk of immunologic reactions.

Aorta↗