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At least 1,135 records · Page 63Linked to original sources
PubMed2006

Cytosolic triacylglycerol biosynthetic pathway in oilseeds. Molecular cloning and expression of peanut cytosolic diacylglycerol acyltransferase.

Triacylglycerols (TAGs) are the most important storage form of energy for eukaryotic cells. TAG biosynthetic activity was identified in the cytosolic fraction of developing peanut (Arachis hypogaea) cotyledons. This activity was NaF insensitive and acyl-coenzyme A (CoA) dependent. Acyl-CoA:diacylglycerol acyltransferase (DGAT) catalyzes the final step in TAG biosynthesis that acylates diacylglycerol to TAG. Soluble DGAT was identified from immature peanuts and purified by conventional column chromatographic procedures. The enzyme has a molecular mass of 41 +/- 1.0 kD. Based on the partial peptide sequence, a degenerate probe was used to obtain the full-length cDNA. The isolated gene shared less than 10% identity with the previously identified DGAT1 and 2 families, but has 13% identity with the bacterial bifunctional wax ester/DGAT. To differentiate the unrelated families, we designate the peanut gene as AhDGAT. Expression of peanut cDNA in Escherichia coli resulted in the formation of labeled TAG and wax ester from [14C]acetate. The recombinant E. coli showed high levels of DGAT activity but no wax ester synthase activity. TAGs were localized in transformed cells with Nile blue A and oil red O staining. The recombinant and native DGAT was specific for 1,2-diacylglycerol and did not utilize hexadecanol, glycerol-3-phosphate, monoacylglycerol, lysophosphatidic acid, and lysophosphatidylcholine. Oleoyl-CoA was the preferred acyl donor as compared to palmitoyl- and stearoyl-CoAs. These data suggest that the cytosol is one of the sites for TAG biosynthesis in oilseeds. The identified pathway may present opportunities of bioengineering oil-yielding plants for increased oil production.

Amino Acid Sequence↗
PubMed2001

Crystallization and preliminary crystallographic studies of antibacterial polypeptide LCI expressed in Escherichia coli.

LCI is a type of novel antibacterial polypeptide secreted by a Bacillus subtilis strain. It consists of 47 residues with a molecular weight of 5468 Da. Using bioengineering, LCI was expressed in Escherichia coli DH5alpha with recombinant plasmid pBVAB16. It was crystallized using PEG 4000 as a precipitant. The crystal belongs to space group P6(2)22 or P6(4)22, with unit-cell parameters a = b = 29.30, c = 187.09 A, and diffracts to 2.44 A. A set of diffraction data to 2.8 A was collected.

Anti-Bacterial Agents↗
PubMedDate not supplied

Healthcare technology, economics, and policy: an evolving balance.

Economic and policy issues increasingly influence healthcare technology solutions as we wrestle with keeping healthcare quality high while keeping costs under control. The healthcare system is also undergoing rapid change brought about by managed care, shifting care to team providers in an outpatient or home environment. The development of health technologies under new market arrangements needs to be examined so that quality of care and cost are not at cross roads. As these issues continue to evolve in the public forum, it is critical for engineers and contributors to technology in healthcare to understand the dynamics that influence technology need and and adoption in the healthcare market. We hope that this special issue will provide a basis for initial understanding of these issues and stimulate further involvement by the bioengineering community at large, for the improvement of healthcare across all of society.

Biomedical Engineering↗
PubMed2004

Implementation of an optical method for the real-time determination of uniaxial strain and vessel mechanics.

The determination of the mechanical properties of vascular growth substrates has seen increasing interest in the bioengineering field. Mechanical features such as rupture strength, compliance characteristics, and viscoelastic properties of vascular grafts are important for their design and are indicative of their success in vivo. Thus a simple inexpensive measurement technique for these parameters would be useful. In this report we describe the implementation of an optical method for the measurement of vessel distention under a transluminal pressure gradient. It is based on the concept of laser light occlusion and allows for real time noncontact diameter measurements of hollow vessels < or = 2 cm. We demonstrate precise and reproducible measurements of diameter changes of less than 10 microm and, further, with the simultaneous determination of both strain and luminal pressure, were able to determine the elastic modulus of commercially available polymeric vessels. Comparison of the manufacturer specifications and our own measurement of the elastic modulus of these vessels, validate the effectiveness of our system. The advantages of this technique are its relative low cost, ease of implementation, high resolution, and flexibility stemming from its modular setup.

Animals↗
PubMed2003

Using a system-on-a-chip implantable device to filter circulating infected cells in blood or lymph.

This paper describes a system on a chip (SoC) that makes use of nanoscale cellular adhesion mechanisms in an integrated electronic microsystem to filter infected cells from blood or lymph. An example of a human immunodeficiency virus-specific SoC is explored in depth. Such systems work in vivo, and blood and lymph are filtered on a continuous basis. With the intelligence on the chip, captured cells can be identified and lyzed, expelled, or otherwise acted upon. These types of systems transfer the burden of research from traditional chemotherapy to bioengineering and system design.

Animals↗
PubMed2004

Channel-forming membrane proteins as molecular sensors.

Membrane channels are typically around or less than 1 nm in diameter and a description of the flow through them requires a molecular approach called nanofluidic. The ion current through channels is extremely sensitive to pore sizes. It is tempting to use the ion current to probe conformational changes of the channel or, for a fixed channel conformation, the current can be used to follow binding of molecules to the pore surfaces. Here we show the sensitivity of this method. It is possible to observe the passage of single isolated molecules through the channel and it is possible to discriminate between different passing molecules. Bioengineering allows us to modify channel surfaces and the affinity to different host molecules. Combining engineered proteins with the appropriated detection technique will allow a new type of molecular sensor.

Ampicillin↗
PubMed2000

Conformational stability changes of the amino terminal domain of enzyme I of the Escherichia coli phosphoenolpyruvate: sugar phosphotransferase system produced by substituting alanine or glutamate for the active-site histidine 189: implications for phosphorylation effects.

The amino terminal domain of enzyme I (residues 1-258 + Arg; EIN) and full length enzyme I (575 residues; EI) harboring active-site mutations (H189E, expected to have properties of phosphorylated forms, and H189A) have been produced by protein bioengineering. Differential scanning calorimetry (DSC) and temperature-induced changes in ellipticity at 222 nm for monomeric wild-type and mutant EIN proteins indicate two-state unfolding. For EIN proteins in 10 mM K-phosphate (and 100 mM KCl) at pH 7.5, deltaH approximately 140 +/- 10 (160) kcal mol(-1) and deltaCp approximately 2.7 (3.3) kcal K(-1) mol(-1). Transition temperatures (Tm) are 57 (59), 55 (58), and 53 (56) degrees C for wild-type, H189A, and H189E forms of EIN, respectively. The order of conformational stability for dephospho-His189, phospho-His189, and H189 substitutions of EIN at pH 7.5 is: His > Ala > Glu > His-PO3(2-) due to differences in conformational entropy. Although H189E mutants have decreased Tm values for overall unfolding the amino terminal domain, a small segment of structure (3 to 12%) is stabilized (Tm approximately 66-68 degrees C). This possibly arises from an ion pair interaction between the gamma-carboxyl of Glu189 and the epsilon-amino group of Lys69 in the docking region for the histidine-containing phosphocarrier protein HPr. However, the binding of HPr to wild-type and active-site mutants of EIN and EI is temperature-independent (entropically controlled) with about the same affinity constant at pH 7.5: K(A)' = 3 +/- 1 x 10(5) M(-1) for EIN and approximately 1.2 x 10(5) M(-1) for EI.

Alanine↗
PubMed1997

Fas signal transduction triggers either proliferation or apoptosis in human fibroblasts.

Although shown to be highly expressed by the epidermis in inflammatory skin disease, the ability of the Fas protein to trigger apoptosis in the distinct cell subpopulations of cutaneous tissue, particularly with regard to receptor density and the degree of crosslinking, has not been fully characterized. We therefore determined the effect of Fas cross-linking in primary human dermal fibroblasts at both high and low levels of Fas receptor expression. First, we examined the effects of the anti-Fas monoclonal antibody, CH-11, on fibroblasts expressing low basal levels of Fas. In these cells Fas aggregation stimulated proliferation by 160 +/- 10% over untreated controls. In contrast, the same concentration of CH-11 had an inhibitory effect on epidermal keratinocyte growth. Because Fas is upregulated in inflamed skin, we next examined the effects of Fas cross-linking on fibroblasts expressing augmented levels ofFas. Fibroblasts were either transfected with plasmids for overexpression of full length or bioengineered Fas receptors or were transduced with a retroviral Fas expression vector. In these cells Fas oligomerization triggered the morphologic changes indicative of apoptosis regardless of whether or not the Fas-signaling domain was tethered to the plasma membrane. These studies indicate that Fas oligomerization in dermal fibroblasts may initiate dual signaling programs, either proliferation or apoptosis, and that the chosen outcome may depend upon the magnitude of Fas aggregation.

Adult↗
PubMed2004

Circadian and ultradian (12 h) variations of skin blood flow and barrier function in non-irritated and irritated skin-effect of topical corticosteroids.

The skin is the organ that receives the greatest exposure to light and shows a high-amplitude circadian rhythm in epidermal cell proliferation. We have previously demonstrated that the skin barrier function has a significant circadian rhythm. Corticosteroids (CS) are the most commonly used topical treatment in dermatology. Time-dependent differences in their efficacy and side-effects would be of considerable interest. The aims of the current study were to examine time-dependent cycles in the effect of topical CS application in healthy and irritated skin on skin blood flow and its relationship to barrier function. Twenty clinically healthy, diurnally active subjects were examined at eight and nine time points over a 24 or 28 h span respectively, using non-invasive skin bioengineering techniques of laser Doppler imaging, a transepidermal water loss (TEWL) device and a skin thermometer in a 28 h session. The results of this current study demonstrate circadian and ultradian (12 h) variations in skin blood flow. A significant correlation was found between skin temperature and skin blood flow but not with TEWL. Circadian and ultradian rhythms are maintained during treatment with high-potency and mid-potency CS in healthy skin. These rhythms persist during stratum corneum disruption with and without CS application.

Administration, Topical↗
PubMed2004

An in vivo model of wound healing in genetically modified skin-humanized mice.

Cutaneous wound-healing disorders are a major health problem that requires the development of innovative treatments. Whithin this context, the search for reliable human wound-healing models that allow us to address both mechanistic and therapeutic matters is warranted. In this study, we have developed a novel invivo wound-healing model in a genetically modified human context. Our model is based on the regeneration of human skin on the back of nude mice by transplantation of a cultured bioengineered skin equivalent previously designed in our laboratory. In this setting, human keratinocytes in the epidermal compartment were genetically modified with a retroviral vector encoding the enhanced green fluorescent protein (EGFP). After stable engraftment of the EGFP skin was achieved (9-12 wk after grafting), a small circular full thickness wound was performed on this mature human skin. A wide variety of parameters involved in wound healing were monitored, including tissue architecture, cell proliferation, epidermal differentiation, dermal remodelling, and basement membrane regeneration. Wounded gene-targeted skin-humanized mice re-capitulated native skin wound-healing features. In addition, when keratinocyte growth factor (KGF), a growth factor that has been shown to improve wound healing, was added to wounds during 3 d, the re-epithelialization was significantly accelerated. The present wound-healing model system provides a suitable in vivo tool to test gene transfer strategies for human skin repair. It also serves as a complementary platform for studies in genetically modified mice and as a model to evaluate pharmaceutical therapeutic approaches for impaired wound healing.

Animals↗
PubMed2004

Thermal dependence of enzyme function and inhibition; implications for, herbicide efficacy and tolerance.

Environmental temperature is a critical factor in the lives of almost all organisms. Plants experience periods of thermal stress related to seasonal patterns of temperature and periodic water deficits. Within the range of non-lethal temperatures, there are a number of thermal effects on metabolism that are a result of the thermal dependence of enzymes. The thermal dependence of enzyme kinetic parameters was used to predict that the efficacy of the herbicide pyrithiobac on Palmer amaranth would be reduced at temperatures outside a 20-34 degrees C thermal application range. This prediction is validated in a controlled environment study described in this paper. Palmer amaranth was grown for 16 days in growth chambers with 34/18 degrees C day/night temperature regime. Pyrithiobac was applied to plants at 18, 27 or 40 degrees C. After 1 h at the application temperatures the plants were returned to the 34/18 degrees C regime for 14 days and post-application biomass accumulation (efficacy) was determined. Dry weight accumulation, as a percentage of untreated controls, was 25, 2.5 and 70% for 18, 27 and 40 degrees C application temperatures. Pyrithiobac efficacy was highest for the application within the thermal application range and significantly reduced at temperatures above and below. The validation of the earlier prediction suggests that temperature-related kinetic limitations on herbicide efficacy may also occur in plants with bioengineered herbicide resistance based on herbicide metabolism. The theoretical aspects of such thermal limitations on herbicide resistance mechanisms are discussed.

Journal Article↗
PubMed2003

Skin strain and its influence on systemic exposure to a glycol ether in offset printing workers.

Under workplace conditions, it is difficult to prove the influence of skin lesions on skin penetration by chemical substances. The aim of the present study was to show whether systemic exposure to glycol ether increases due to lesions of the skin in printing workers. 28 male printers, exposed to 2-(2-butoxyethoxy)ethanol (BEE), were interviewed about the workplace exposure by a standardized questionnaire. The systemic exposure in printers was determined by biological monitoring of the main metabolite of BEE butoxyethoxyacetic acid (BEAA) in urine. Furthermore, clinical examination of the skin, transepidermal water loss, capacitance and skin surface pH measurements were carried out. Erythema and scaliness were the most important factors showing an effect on dermal absorption. The mean urinary BEAA excretions for printers with skin lesions on the hands were higher (20.62 mg/l for scaliness and 14.40 mg/l for erythema) compared to that for printers without detectable skin lesions (12.08 mg/l for scaliness and 13.03 mg/l for erythema). Bioengineering measurements to predict skin strain and percutaneous absorption were only supportive. We were able to show that by using a multiple spectrum of methods an enhancement of percutaneous absorption of BEE could be demonstrated in workers with skin lesions.

Adult↗
PubMed2005

Long-term repetitive sodium lauryl sulfate-induced irritation of the skin: an in vivo study.

Skin may adapt to topical irritants through accommodation. This study focuses on long-term exposure to irritants and attempts to demonstrate accommodation. Sodium lauryl sulfate (SLS) induced irritant contact dermatitis at 3 concentrations (0.025% to 0.075%). Distilled water, acetone and an empty chamber served as controls. Experimental compounds were applied to forearms of 7 healthy volunteers for 24 hr before replacing by a fresh chamber for 6 non-consecutive weeks over 103 days. Possible accommodation was quantified by visual scoring (erythema and dryness) and by bioengineering parameters: transepidermal water loss (TEWL), capacitance, chromametry and laser Doppler flowmetry (LDF). Significant erythema, dryness, elevated TEWL, skin colour reflectance and LDF values occurred during the exposure periods. Upon repeat exposure, an immediate and augmented response in erythema, TEWL, skin colour reflectance and LDF developed. However, irritant skin changes were not sustained. Irritation parameters return to baseline after cessation of exposure. There was no evidence of sustained irritation or accommodation after the last exposure. Study findings do not document sustained accommodation or adaptive hyposensitivity after long-term repetitive irritant exposure under these test conditions. Alternative models should be developed to prove or disprove the accommodation hypothesis.

Adaptation, Physiological↗
PubMed2005

Age and skin structure and function, a quantitative approach (I): blood flow, pH, thickness, and ultrasound echogenicity.

BACKGROUND/PURPOSE: The aging process has been studied with fervor recently, given our shifting demographics. Since age's effects are so manifest in skin's appearance, structure, mechanics, and barrier function, it is not surprising that much effort has been placed in research to better understand them. Quantitative measurements permitted by bioengineering have allowed us to objectively and precisely study aging skin. These overviews piece together the immense amounts of information that have emerged from recent technological advances in dermatological research in order to develop a unified understanding of the quantitative effects of age on the skin. METHODS: We performed a literature on age-related changes in blood flow, pH, skin thickness, and ultrasound imaging data, searching Pub-med, Em-Base, Science Citation Index, and the UCSF dermatological library's collection of books on the topic of aging skin. RESULTS: Despite the many tools and techniques available for quantitative analysis of skin, age studies are often conflicting, especially in the areas of blood flow and skin thickness. Trends indicate that blood flow may decrease with age, especially in sites exposed to the environment. pH apparently varies little until the age of 70, after which it declines. Skin thickness data are difficult to interpret; while the stratum corneum is generally accepted to maintain its thickness during aging, dermal, epidermal, and whole skin thickness changes are controversial. Ultrasound reveals the appearance of a subepidermal low echogenic band that thickens with age, especially in environmentally exposed areas. Some studies also indicate the presence of an echogenic band in the lower dermis which thins with increased age. However, the whole dermis appears to become more echogenic in elderly people. CONCLUSION: Much remains to be done if we are to reach consensus on the effects of age on skin structure and function. Future studies would be benefited by increased standardization of skin sites tested, methodology, and increased sample size.

Aging↗
PubMed2006

Age and skin structure and function, a quantitative approach (II): protein, glycosaminoglycan, water, and lipid content and structure.

BACKGROUND/PURPOSE: The aging process has been studied with fervor recently, given our shifting demographics. As age's effects are so manifest in the skin's appearance, structure, mechanics, and barrier function, it is not surprising that much effort has been made in research to better understand them. Quantitative measurements permitted by bioengineering have allowed us to objectively and precisely study aging skin. These overviews piece together the immense amounts of information that have emerged from recent technological advances in dermatological research in order to develop a unified understanding of the quantitative effects of age on skin. METHODS: We performed a literature search on age-related changes in protein, glycosaminoglycan (GAG), water, and lipid content and structure, searching Pub-med, Em-Base, Science Citation Index, and the UCSF dermatological library's collection of books on the topic of aging skin. RESULTS: Collagen becomes sparser and less soluble in intrinsically aged skin, but is thickened and more soluble in extrinsically aged areas. Elastin is degraded slowly and accumulates damage with intrinsic aging; also, increased synthesis of abnormally structured elastin occurs in photoexposed areas. This leads to an age-related accumulation of aberrant elastoic material, clumped in the papillary dermis. Generally, age leads to increased folding and decreased interaction of proteins with water. Also, despite increased GAGs in aged skin, these are abnormally deposited on the elastoic material and cannot interact properly with water. Hence, in aged skin, water is found in the tetrahedron form, bound to itself rather than other molecules. Lipid content appears to decrease with age, although the proportion of different lipid classes seems to remain fairly constant. CONCLUSION: Much work remains to be carried out to reach a consensus on the effects of age on skin structure and function. Future studies would be benefited by increased standardization of skin sites tested, methodology, and increased sample sizes.

Body Water↗
PubMed2006

Assessment of anti-inflammatory activity of Poria cocos in sodium lauryl sulphate-induced irritant contact dermatitis.

OBJECTIVE: In the present study, we evaluated the anti-inflammatory activity of Poria cocos (PoCo) on experimentally induced irritant contact dermatitis (ICD) in a repeated sodium lauryl sulphate (SLS) irritation model. METHODS: The anti-irritative effect of PoCo was evaluated with a visual score and quantified by non-invasive bioengineering methods, namely chromametry and transepidermal water loss. Three concentrations of PoCo in base cream DAC (amphiphilic emollient; German pharmacopoeia) were tested in a 4-day repetitive irritation test using SLS. RESULTS: A statistically significant anti-inflammatory activity was observed for PoCo by all three methods when applied in parallel to the induction period of ICD. Application of PoCo after induction of ICD once a day for 5 days, starting just at the end of 4 days, was without any effect. CONCLUSION: An anti-inflammatory efficacy of PoCo on the elicitation phase of the ICD induced by repeated SLS test could be observed and quantified by three independent, non-invasive biophysical assessment parameters. This effect can be explained by its influence on pro-inflammatory enzymes, namely phospholipase A2.

Adult↗
PubMedDate not supplied

Optimizing wound treatment through health care structuring and professional education.

Considerable developments have occurred in wound healing and care over the last few decades. Innovative technologies such as use of growth factors, bioengineered skin, topical negative pressure, heat, oxygen, and others have entered the clinic. These techniques have been advantageous, but even the most advanced and sophisticated product requires proper wound care and wound bed preparation in order to function optimally. It has been shown that the organization of a wound team, not the change in products, increases the healing of problem wounds (MTV Report, Denmark, 2003). Efficacy studies are generally performed in a few centers with investigators already experienced in the field. This probably leads to an optimal efficacy of the product. Problems arise when the product is marketed and is used by clinicians not specially educated in wound care. If no treatment plan and education is performed, product efficacy and cost-effectiveness decrease dramatically. This is clearly a major problem for patients, but also for communities and, in the end, for companies' sale of the product. Structuring of treatment and education is for these reasons the largest present challenge in the area of wound healing. The optimal way to treat nonhealing wounds is based on a multidisciplinary concept that has been implemented in a national health care system as a recognized expert function. It should consist of a specific treatment plan including modern wound care products and well-educated personnel. This structure is similar to other areas of the medical profession, but has to a lesser degree been established in the treatment of problem wounds. A growing understanding of the importance of this area is, however, shown by the increasing number of working groups focusing on organization and education in wound care. Presently the European Wound Management Association, the World Union of Wound Healing Societies, and The Danish Wound Healing Society have created such types of working groups.

Cooperative Behavior↗
PubMed2005

Atelocollagen sponge and recombinant basic fibroblast growth factor combination therapy for resistant wounds with deep cavities.

Recent advances in bioengineering have introduced materials that enhance wound healing. Even with such new tools, some deep ulcers surrounded by avascular tissues, including bone, tendon, and fascia, are resistant to various therapies and easily form deep cavities with loss of subcutaneous tissue. Atelocollagen sponges have been used as an artificial dermis to cover full-thickness skin defects. Topical recombinant human basic fibroblast growth factor has been introduced as a growth factor to induce fibroblast proliferation in skin ulcers. We applied these materials in combination in two patients with deep resistant wounds: one with a cavity reaching the mediastinum through a divided sternum and one with deep necrotic wounds caused by electric burns. These wounds did not respond to the topical basic fibroblast growth factor alone. In contrast, the combination therapy closed the wounds rapidly without further surgical treatment. This combination therapy is a potent treatment for resistant wounds with deep cavities.

Aged↗