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Biomedical subjects

R K Wang

Publications and source records attributed to R K Wang.

At least 19 recordsLinked to original sources

Effect of laser and environmental parameters on reducing microbial contamination of stainless steel surfaces with Nd:YAG laser irradiation.

AIMS: The effect of laser (pulse repetition frequency, pulse energy and exposure time) and environmental parameters (pH, NaCl concentration and wet or dry samples) of Nd:YAG laser decontamination of stainless steel inoculated with Escherichia coli, Staphylococcus aureus and Listeria monocytogenes was investigated. METHODS AND RESULTS: Stainless steel discs were inoculated with the bacterial samples and exposed to laser energy densities to about 900 J cm(-2). These inactivation curves allowed selection of laser parameters for two-level multifactorial designed experiments, the results of which allowed comparisons to be made between effects of individual and combined parameters on the laser inactivation efficiency. Escherichia coli was inactivated most effectively as a wet film with L. monocytogenes and S. aureus showing similar response. For the multifactorial experiments all laser parameters were significant and were smallest for S. aureus as a wet film. CONCLUSIONS: pH and NaCl concentration had little effect on the efficacy of laser inactivation but dry or wet states and all laser parameters were significant. SIGNIFICANCE AND IMPACT OF THE STUDY: Such systems may prove to be applicable in industrial processes where stainless steel may be contaminated with acidic solutions or salt, e.g. in the food industry with laser inactivation seeming to be independent of these parameters. Parameters have been identified that allow optimization of the treatment process.

Analysis of Variance↗

Doppler optical coherence tomography for measuring flow in engineered tissue.

The engineering of human tissue represents a major paradigm shift in clinical medicine. Early embodiments of tissue engineering are currently being taken forward to the clinic by production methods that are essentially extensions of laboratory manual procedures. However, to achieve the status of routine large-scale clinical practice, automation and scale-out processes are required. This in turn will require the development of reliable on-line monitoring and control systems. This paper examines one demand of crucial importance, namely the real time in vitro monitoring of the flow characteristics through growing tissue since this has a complex interrelationship. Doppler optical coherence tomography (DOCT) is a recently developed imaging technique for studying the rheological properties of tissues in vivo. Capable of non-invasive imaging in real time with high resolution, it is potentially ideal for the continuous monitoring of engineered tissues in vitro. As a base line, the current status of DOCT in vivo is therefore reviewed. This paper also reports the first preliminary use of DOCT in tissue engineering. The application described involves the imaging of a fully developed laminar flow through a combined tissue fabrication/bioreactor with a tissue-engineered construct (substitute blood vessel) in situ.

Bioreactors↗

The potential of optical coherence tomography in the engineering of living tissue.

The better repair of human tissue is an urgent medical goal and in order to achieve a safe outcome there is a parallel need for sensitive, non-invasive methods of assessing the quality of the engineered tissues and organs prior to surgical implantation. Optical coherence tomography (OCT) can potentially fulfil this role. The current status of OCT as an advanced imaging tool in clinical medicine, developmental biology and material science is reviewed and the parallels to the engineering of living tissue and organs are discussed. Preliminary data are also presented for a tissue engineering bioreactor with in situ OCT imaging. The data suggest that OCT can be utilized as a real time, non-destructive, non-invasive tool to critically monitor the morphology of tissue-engineered constructs during their fabrication and growth.

Animals↗

Vertex/propagator model for least-scattered photons traversing a turbid medium.

The least-scattered photons that arrive at a detector through highly scattering tissues have the potential to image internal structures, functions, and status with high imaging resolution. In contrast, optical diffusing tomography is based on the use of the late-arriving photons, which have been diffusely scattered, leading to very low imaging resolution. A good model of the early-arriving photons, i.e., the least-scattered photons, may have a significant effect on the development of imaging algorithms and a further understanding of imaging mechanisms within current high-resolution optical-imaging techniques. We describe a vertex/propagator approach that attempts to find the probabilities for least-scattered photons traversing a scattering medium, based on analytical expressions for photon histories. The basic mathematical derivations for the model are outlined, and the results are discussed and found to be in very good agreement with those from the Monte Carlo simulations.

Journal Article↗

Bactericidal action of high-power Nd:YAG laser light on Escherichia coli in saline suspension.

Infra-red light (1064 nm) from a high-power Nd:YAG laser caused more than 90% loss of viability of Escherichia coli during exposures that raised the temperature of PBS suspensions of the bacteria to 50 C in a thermocouple-equipped cuvette. In contrast, there was minimal loss of viability after heating the same suspensions to 50 degrees C in a water-bath, or in a PCR thermal cycler. The mechanism of laser killing at 50 degrees C was explored by differential scanning calorimetry, by laser treatment of transparent and turbid bacterial suspensions, and by optical absorbency studies of E. coli suspensions at 1064 nm. Taken together, the data suggested that the bactericidal action of Nd:YAG laser light at 50 degrees C was due partly to thermal heating and partly to an additional, as yet undefined, mechanism. Scanning electron microscopy revealed localized areas of surface damage on laser-exposed E. coli cells.

Colony Count, Microbial↗

[Antagonizing effect of sodium ferulate on the changes of hepatic antioxidative function induced by ethanol in mice].

Effects of ethanol at different dosages on hepatic antioxidative and detoxicating functions and the antagonizing effect of sodium ferulate have been investigated in mice. The data showed that ethanol (11.4 g.kg-1, ig) could induce the increase of hepatic glutathione peroxidase (GSH-Px) activity and decreases of hepatic glutathione reductase (GSH-Re), superoxide dismutase (SOD) and glutathione S-transferase (GST) activities and reduce glutathione (GSH) content. At the same time, serum GST activity was increased. Pretreatment with sodium ferulate (100 mg.kg-1 ig, qd x 10 d) completely reversed these changes induced by ig ethanol in mice, indicating that sodium ferulate could protect mice from ethanol-induced acute hepatotoxicity. The hepato-protective mechanism of sodium ferulate may be related to intensification of the function of glutathione oxidative-reductive enzymes, enhancement of SOD activity and promotion of glutathione conjugation. The results also indicate that the serum GST level is a sensitive indicator in ethanol-induced hepatotoxicity.

Animals↗

Expression of murine beta 7, alpha 4, and beta 1 integrin genes by rodent mast cells.

The screening of a rat mast cell cDNA library with a probe selected to recognize those genes preferentially or exclusively expressed by mast cells identified a rat gene sequence, RF-17, that shared homology with the beta-integrins. This integrin was expressed in rat tissues enriched for mast cells and T cells. The rat RF-17 sequence was used to isolate the murine homologue from a spleen cDNA library. The murine gene encodes a protein of 806 amino acids that is the probable homologue to the human beta 7 chain. Transcripts specific for the murine gene are found in the thymus, spleen, and lung. To attempt to identify the gene product for this new integrin chain, we examined the murine T cell line TK-1, which expresses a novel integrin heterodimer, lymphocyte Peyer's patch high endothelial venule adhesion molecule (LPAM-1), of a known alpha 4 chain and an unknown beta P chain, for expression of murine beta 7 (RF-17). This cell line expresses high levels of RF-17 transcripts, suggesting that beta P is encoded by the beta 7 gene. Bone marrow cells induced to differentiate into mast cells via IL-3 express the beta 7 gene as well as the genes encoding the murine integrin alpha 4 and beta 1 proteins. Surface staining analysis indicates that these cells express an alpha 4-containing integrin complex throughout the differentiation process. These data suggest that the Peyer's patch homing LPAM-1 receptor expressed by a subset of T cells consists of the beta 7 gene product and the alpha 4 chain, and that this integrin chain complex is also found on the surface of maturing mast cells. The presence of beta 1 transcripts also suggests that these maturing mast cells possess the LPAM-2 integrin complex (alpha 4/beta 1) as well. The experimental strategy described in this manuscript has, thus, identified a novel murine beta-integrin chain that is expressed by rodent T cells and mast cells.

Amino Acid Sequence↗

Caloric restriction affects liver microsomal monooxygenases differentially in aging male rats.

Caloric restriction (CR) extends life span and retards the onset of physiological changes and pathologies associated with aging, but the underlying mechanisms remain unresolved. This study demonstrates that CR postpones the documented age-related declines in and/or enhances the activity and microsomal concentration of several liver monooxygenases in male rats, i.e., NADPH cytochrome P-450 reductase, total cytochromes P-450. However, the relative concentration of cytochrome P-450b+C did not exhibit statistically significant changes, whereas another isozyme, the male specific P-450h, declined significantly in both ad libitum-fed and CR rats as a function of increasing age. While CR appears to retard age-associated changes in certain liver enzymes, this effect is by no means universal. The hepatic monooxygenases constitute a well-characterized enzyme system in which to examine the perturbation of the aging process by CR.

Aging↗

Effects of age and gender on in vitro properties of human liver microsomal monooxygenases.

Aging in humans is associated with marked declines in the disposition of numerous drugs and other xenobiotics that require hepatic biotransformation before elimination. Considerable pharmacokinetic evidence in humans, coupled with data on in vitro liver microsomal monooxygenase functions generated in inbred male rodent models, has implicated impaired liver phase I drug metabolism (i.e., diminished efficacy of microsomal monooxygenases) in reduced drug clearance in the elderly. This study (1) assessed the in vitro activities and amounts of liver microsomal monooxygenases as a function of donor age and gender in healthy humans and (2) provides the most extensive and comprehensive data to date demonstrating the absence of significant age- and gender-dependent differences in the activities and contents of human liver monooxygenases.

Adolescent↗

Mucosal immune response to cholera toxin in ageing rats. I. Antibody and antibody-containing cell response.

Although ageing is accompanied by systemic immunodeficiencies, the status of the mucosal immune system in the elderly remains unresolved. The gastrointestinal mucosal immune response was evaluated in young, mature and old male rats subjected to intra-intestinal immunization with cholera toxin (CTx). Five days following secondary immunization, the alpha-CTx-IgA titre in the bile of immunized rats was markedly reduced, i.e. the values measured in young rats were approximately five-fold higher than those of old animals. alpha-CTx-IgA levels in non-immunized rats were negligible and age-related shifts in other antibody titres (alpha-CTx IgG and IgM) were not significant. The antibody response to CTx was not reflected in the total IgA content of the samples. The number of alpha-CTx antibody-containing cells (ACCs) in the small intestinal lamina propria was significantly reduced in old immunized rats in comparison with the young or mature animals. These data suggest that ageing compromises both non-immune cell (antibody transport by hepatocytes) and immune cell (number of ACCs in the gut wall) functions in response to cholera toxin immunization in this animal model.

Aging↗

Age-dependent alterations in the physicochemical properties of rat liver microsomes.

Aging results in a significant decline in liver drug metabolism which is largely attributable to changes in the microsomal mixed function oxidase system. For example, the mixed function oxidase system in the livers of senescent rats is characterized by: (1) a reduced cytochrome P-450 content; (2) a decline in the specific activity of NADPH-cytochrome c (P-450) reductase; and (3) a slower rate of ethylmorphine N-demethylation in comparison to young adult animals. Since several factors intrinsic to the microsomes may influence the efficacy of the mixed function oxidase system, e.g. the phospholipid and cholesterol contents, the saturation index of the fatty acids and the fluidity of the membranes, we conducted a physicochemical analysis of liver microsomes isolated from young adult (3-4 months), mature (12-16 months) and senescent (25-27 months) male Fischer rats. Although the microsomal cholesterol content did not change appreciably between maturity and senescence, there was a marked decline in the total phospholipid content. This resulted in a significant increase in the cholesterol/phospholipid ratio, 0.49 to 0.65 between 16 and 27 months of age. The age-related changes in the total phospholipid content were largely reflected in each of the major fractions, i.e. phosphatidylcholine, phosphatidylinositol and phosphatidylethanolamine + phosphatidylserine. Small increases in the relative percentages of highly unsaturated fatty acid species were offset by similar decreases in the more frequent and more saturated species as a function of increased age. As a result, the net change in the fatty acid saturation index was probably minimal. However, the increase in the cholesterol/phospholipid ratio most likely contributes to the significant decline in the order parameter of microsomes isolated from old rats which, in turn, may impair the functional capacity of the hepatic mixed function oxidase system.

Aging↗

Age-dependent alterations in rat liver microsomal NADPH-cytochrome c (P-450) reductase: a qualitative and quantitative analysis.

The hepatic drug-metabolizing capacities of rodents and man exhibit age-dependent declines. The extensive use of medication in geriatric patients demonstrates the need to characterize the mechanism(s) responsible for this reduced liver function. An analysis of microsomal NADPH-cytochrome c (P-450) reductase in young adult (3 months), mature (9 months) and senescent (27 months) male Fischer 344 rats revealed specific age-related qualitative and quantitative changes in this enzyme. The specific activity of the purified enzyme from young animals was two-fold higher than that recovered from the other age groups. In addition, there was: (1) no change in molecular weight, (2) alterations in heat inactivation profiles, (3) an apparent reduction in substrate affinity, and (4) a two-fold loss of enzyme activity per unit of immunoprecipitable material as a function of animal age. Our data suggest that this important liver drug-metabolizing enzyme undergoes post-translational modifications in its conformation which are reflected in the above parameters and which, ultimately, affect its efficacy.

Age Factors↗

Age-dependent changes in rat liver microsomal NADPH cytochrome C (P-450) reductase: a kinetic analysis.

Rat liver microsomal NADPH cytochrome c (P-450) reductase (EC 1.6.2.4) exhibits several marked age-dependent changes, including a decline in specific activity, reduced inducibility and enhanced thermostability, which are indicative of an alteration in the quality of this enzyme. The present study examined the kinetic profile of the microsomal-bound enzyme in an effort to further define the effects of aging on the hepatic mixed function oxidase system. Intact microsomes isolated from young adult (3 months), mature (16 months) and senescent (27 months) rats were subjected to an extensive double reciprocal kinetic analysis employing NADPH and cytochrome c as substrates. The Km values obtained with the initial substrate (NADPH) remained unchanged with animal age, whereas there was a decline in this parameter for the artificial acceptor substrate, cytochrome c. The Vmax values for both substrates were reduced as a function of increasing age, perhaps reflecting a concomitant decline in the relative amount(s) of efficient reductase in the microsomes.

Aging↗

Effects of aging and phenobarbital on the rat liver microsomal drug-metabolizing system.

Significant declines in the non-induced activities of liver microsomal drug-metabolizing enzymes and in the amount of cytochrome P-450 occur between maturity (16 months) and senescence (27 months) in male Fischer 344 rats, whereas there are essentially no differences between very young (1 month) and mature animals. Several hepatic responses to chronic phenobarbital administration also demonstrate marked age-dependent changes. The livers of young and mature animals exhibit: (1) greater hepatomegaly; (2) faster rates of induction and post-induction recovery of microsomal mixed function oxidase enzyme activities and hemoprotein concentration; and (3) higher maximally induced levels of these components in comparison to senescent rats. When considered with information from previous studies, the present data suggest that the age-related decline in liver drug metabolism may be due to qualitative and/or quantitative changes in the structural and/or functional components of the hepatic microsomal mixed function oxidase system.

Aging↗