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Biomedical subjects

N R Baker

Publications and source records attributed to N R Baker.

At least 19 recordsLinked to original sources

The role of herbicides in the erosion of salt marshes in eastern England.

Laboratory studies and field trials were conducted to investigate the role of herbicides on saltmarsh vegetation, and their possible significance to saltmarsh erosion. Herbicide concentrations within the ranges present in the aquatic environment were found to reduce the photosynthetic efficiency and growth of both epipelic diatoms and higher saltmarsh plants in the laboratory and in situ. The addition of sublethal concentrations of herbicides resulted in decreased growth rates and photosynthetic efficiency of diatoms and photosynthetic efficiency of higher plants. Sediment stability also decreased due to a reduction in diatom EPS production. There was qualitative evidence that diatoms migrated deeper into the sediment when the surface was exposed to simazine, reducing surface sediment stability by the absence of a cohesive biofilm. Sediment loads on leaves severely reduced photosynthesis in Limonium vulgare. This, coupled with reduced carbon assimilation from the effects of herbicides, could have large negative consequences for plant productivity and over winter survival of saltmarsh plants. The data support the hypothesis that sublethal herbicide concentrations could be playing a role in the increased erosion of salt marshes that has occurred over the past 40 years.

Analysis of Variance↗

Protection against Pseudomonas aeruginosa chronic lung infection in mice by genetic immunization against outer membrane protein F (OprF) of P. aeruginosa.

The Pseudomonas aeruginosa major constitutive outer membrane porin protein OprF, which has previously been shown to be a protective antigen, was targeted as a DNA vaccine candidate. The oprF gene was cloned into plasmid vector pVR1020, and the plasmid vaccines were delivered to mice by biolistic (gene gun) intradermal inoculation. Antibody titers in antisera from immunized mice were determined by enzyme-linked immunosorbent assay, and the elicited antibodies were shown to be specifically reactive to OprF by immunoblotting. The immunoglobulin G (IgG) immune response was predominantly of the IgG1 isotype. Sera from DNA vaccine-immunized mice had significantly greater opsonic activity in opsonophagocytic assays than did sera from control mice. Following the initial immunization and two consecutive boosts, each at 2-week intervals, protection was demonstrated in a mouse model of chronic pulmonary infection by P. aeruginosa. Eight days postchallenge, both lungs were removed and examined. A significant reduction in the presence of severe macroscopic lesions, as well as in the number of bacteria present in the lungs, was seen. Based on these findings, genetic immunization with oprF has potential for development as a vaccine to protect humans against infection by P. aeruginosa.

Animals↗

High resolution imaging of photosynthetic activities of tissues, cells and chloroplasts in leaves.

Through imaging of chlorophyll fluorescence, it is possible to produce parameterized fluorescence images that estimate the operating quantum efficiency of photosystem II (PSII) photochemistry and which can be used to reveal heterogeneous patterns of photosynthetic performance within leaves. The operating quantum efficiency of PSII photochemistry is dependent upon the effective absorption cross-section of the light-harvesting system of PSII and the photochemical capacity of PSII. The effective absorption cross-section is decreased by the process of down-regulation, which is widely thought to operate within the pigment matrices of PSII and which results in non-photochemical quenching of chlorophyll fluorescence. The photochemical capacity is non-linearly related to the proportion of PSII centres in the 'open' state and results in photochemical quenching of chlorophyll fluorescence. Examples of heterogeneity of the operating quantum efficiency of PSII photochemistry during the induction of photosynthesis in maize leaves and in the chloroplast populations of stomatal guard cells of a leaf of Tradescantia albifora are presented, together with analyses of the factors determining this heterogeneity. A comparison of the operating quantum efficiency of PSII photochemistry within guard cells and adjacent mesophyll cells of Commelina communis is also made, before and after stomatal closure through a change in ambient humidity.

Chlorophyll↗

Primary sites of ozone-induced perturbations of photosynthesis in leaves: identification and characterization in Phaseolus vulgaris using high resolution chlorophyll fluorescence imaging.

High resolution imaging of chlorophyll a fluorescence was used to identify the sites at which ozone initially induces perturbations of photosynthesis in leaves of Phaseolus vulgaris. Leaves were exposed to 250 and 500 nmol mol(-1) ozone at a photosynthetically active photon flux density of 300 micromol m(-2) s(-1) for 3 h. Images of fluorescence parameters indicated that large decreases in both the maximum and operating quantum efficiencies of photosystem II had occurred in cells adjacent to stomata in the upper, but not lower, leaf surfaces. However, this treatment did not produce any significant changes in the maximum or operating quantum efficiencies of photosystem II in the leaves when estimated from fluorescence parameters measured with a conventional, integrating fluorometer. The localized decreases in photosystem II photochemical efficiencies were accompanied by an increase in the minimal fluorescence level, which is indicative of photoinactivation of photosystem II complexes and a decrease in stomatal conductance. Perturbations of photochemical efficiencies were not observed in cells associated with all of the stomata on the upper leaf surface or within cells distant from the upper leaf surface. It is concluded that ozone penetrates the leaf through stomata and initially damages only cells close to stomatal pores.

Chlorophyll↗

Growth in elevated CO(2) can both increase and decrease photochemistry and photoinhibition of photosynthesis in a predictable manner. Dactylis glomerata grown in two levels of nitrogen nutrition.

Biochemically based models of C(3) photosynthesis can be used to predict that when photosynthesis is limited by the amount of Rubisco, increasing atmospheric CO(2) partial pressure (pCO(2)) will increase light-saturated linear electron flow through photosystem II (J(t)). This is because the stimulation of electron flow to the photosynthetic carbon reduction cycle (J(c)) will be greater than the competitive suppression of electron flow to the photorespiratory carbon oxidation cycle (J(o)). Where elevated pCO(2) increases J(t), then the ratio of absorbed energy dissipated photochemically to that dissipated non-photochemically will rise. These predictions were tested on Dactylis glomerata grown in fully controlled environments, at either ambient (35 Pa) or elevated (65 Pa) pCO(2), and at two levels of nitrogen nutrition. As was predicted, for D. glomerata grown in high nitrogen, J(t) was significantly higher in plants grown and measured at elevated pCO(2) than for plants grown and measured at ambient pCO(2). This was due to a significant increase in J(c) exceeding any suppression of J(o). This increase in photochemistry at elevated pCO(2) protected against photoinhibition at high light. For plants grown at low nitrogen, J(t) was significantly lower in plants grown and measured at elevated pCO(2) than for plants grown and measured at ambient pCO(2). Elevated pCO(2) again suppressed J(o); however growth in elevated pCO(2) resulted in an acclimatory decrease in leaf Rubisco content that removed any stimulation of J(c). Consistent with decreased photochemistry, for leaves grown at low nitrogen, the recovery from a 3-h photoinhibitory treatment was slower at elevated pCO(2).

Acclimatization↗

An evaluation of the potential triggers of photoinactivation of photosystem II in the context of a Stern-Volmer model for downregulation and the reversible radical pair equilibrium model.

Photoinactivation of photosystem II (PS II) is a light-dependent process that frequently leads to break-down and replacement of the D1 polypeptide. Photoinhibition occurs when the rate of photoinactivation is greater than the rate at which D1 is replaced and results in a decrease in the maximum efficiency of PS II photochemistry. Downregulation, which increases non-radiative decay within PS II, also decreases the maximum efficiency of PS II photochemistry and plays an important role in protecting against photoinhibition by reducing the yield of photoinactivation. The yield of photoinactivation has been shown to be relatively insensitive to photosynthetically active photon flux density (PPFD). Formation of the P680 radical (P680+), through charge separation at PS II, generation of triplet-state P680 (3P680*), through intersystem crossing and charge recombination, and double reduction of the primary stable electron acceptor of PS II (the plastoquinone, Q(A)) are all potentially critical steps in the triggering of photoinactivation. In this paper, these processes are assessed using fluorescence data from attached leaves of higher plant species, in the context of a Stern-Volmer model for downregulation and the reversible radical pair equilibrium model. It is shown that the yield of P680+ is very sensitive to PPFD and that downregulation has very little effect on its production. Consequently, it is unlikely to be the trigger for photoinactivation. The yields of 3P680* generated through charge recombination or intersystem crossing are both less sensitive to PPFD than the yield of P680+ and are both decreased by down regulation. The yield of doubly reduced Q(A) increases with incident photon flux density at low levels, but is relatively insensitive at moderate to high levels, and is greatly decreased by downregulation. Consequently, 3P680* and doubly reduced Q(A) are both viable as triggers of photoinactivation.

Down-Regulation↗

Blindness following a diabetic foot infection: a variant to the 'eye-foot syndrome'?

AIMS: The 'eye-foot syndrome' was initially described by Walsh et al. to highlight the important association of foot lesions in patients with diabetic retinopathy. We present a case of a 58-year-old patient with Type 2 diabetes mellitus who developed blindness following endogenous staphylococcal endophthalmitis from an infected foot ulcer. RESULTS: Our case describes the link between the eye and the foot but is somewhat different to the association as described by Walsh et al. Endogenous endophthalmitis is rare with diabetic patients being especially at risk, and we report the first case of endogenous staphylococcal endophthalmitis related to a diabetic foot lesion. CONCLUSIONS: Our case illustrates several important issues in the management of diabetic patients admitted to hospital with infection; the need to thoroughly examine the feet to ascertain any foot lesions and any underlying peripheral vascular disease or peripheral neuropathy, to treat aggressively any infected foot lesions to prevent serious complications of septicaemia and to consider rare conditions like endogenous endophthalmitis in any diabetic patient presenting with acute visual impairment and septicaemia.

Blindness↗

Effects of drought on photosynthesis in Mediterranean plants grown under enhanced UV-B radiation.

The effects of drought on the photosynthetic characteristics of three Mediterranean plants (olive, Olea europea L.; rosemary, Rosmarinus officinalis L.; lavender, Lavandula stoechas L.) exposed to elevated UV-B irradiation in a glasshouse were investigated over a period of weeks. Drought conditions were imposed on 2-year-old plants by withholding water. During the onset of water stress, analyses of the response of net carbon assimilation of leaves to their intercellular CO2 concentration were used to examine the potential limitations imposed by stomata, carboxylation velocity and capacity for regeneration of ribulose 1,5-bisphosphate on photosynthesis. Measurements of chlorophyll fluorescence were used to determine changes in the efficiency of light utilization for electron transport, the occurrence of photoinhibition of photosystem II photochemistry and the possibility of stomatal patchiness across leaves. The first stages of water stress produced decreases in the light-saturated rate of CO2 assimilation which were accompanied by decreases in the maximum carboxylation velocity and the capacity for regeneration of ribulose 1,5-bisphosphate in the absence of any significant photodamage to photosystem II. Leaves of rosemary and lavender were more sensitive than those of olive during the first stages of the drought treatment and also exhibited increases in stomatal limitation. With increasing water stress, significant decreases in the maximum quantum efficiency of photosystem II photochemistry occurred in lavender and rosemary, and stomatal limitation was increased in olive. No indication of any heterogeneity of photosynthesis was found in any leaves. Drought treatment significantly decreased leaf area in all species, an important factor in drought-induced decreases in photosynthetic productivity. Exposure of plants to elevated UV-B radiation (0.47 W m(-2)) prior to and during the drought treatment had no significant effects on the growth or photosynthetic activities of the plants. Consequently, it is predicted that increasing UV-B due to future stratospheric ozone depletion is unlikely to have any significant impact on the photosynthetic productivity of olive, lavender and rosemary in the field.

Carbon Dioxide↗

Analysis of immunization with DNA encoding Pseudomonas aeruginosa exotoxin A.

The promising arena of DNA-based vaccines has led us to investigate possible candidates for immunization against bacterial pathogens. One such target is the opportunistic pathogen Pseudomonas aeruginosa which produces exotoxin A (PE), a well-characterized virulence factor encoded by the toxA gene. In its native protein form, PE is highly cytotoxic for susceptible eukaryotic cells through ADP-ribosylation of elongation factor-2 following internalization and processing of the toxin. To study the biologic and immunological effects of PE following in situ expression, we have constructed eukaryotic plasmid expression vectors containing either the wild-type or a mutated, non-cytotoxic toxA gene. In vitro analysis by transfection of UM449 cells suggests that expression of the wild-type toxA gene is lethal for transfected cells whereas transfection with a mutated toxA gene results in the production of inactive PE which can be readily detected by immunoblot analysis of cell lysates. To investigate the effects resulting from the intracellular expression of potentially cytotoxic gene products in DNA vaccine constructs, we immunized mice with both the wild-type and mutant toxA plasmid constructs and analyzed the resulting humoral and cellular immune responses. Immunization with the mutated toxA gene results in production of neutralizing antibodies against native PE and potentiates a T(H)1-type response, whereas only a minimal humoral response can be detected in mice immunized with wild-type toxA. DNA-based vaccination with the non-cytotoxic toxA(mut) gene confers complete protection against challenge with the wild-type PE. Therefore, genetic immunization with genes encoding potentially cytotoxic gene products raises concern with regard to the selection of feasible gene targets for DNA vaccine development.

ADP Ribose Transferases↗

Studies on new phosphodiesterase inhibitors. I. Synthesis of 1-(2,3-epoxypropoxy)-2(4)-fluorobenzenes and 1-(2-hydroxy-3-morpholinopropoxy and piperazino)fluorobenzene derivatives.

A series of 1-(2,3-epoxypropoxy)-2(4)-fluorobenzenes and their corresponding 1-(2-hydroxy-3-morpholinopropoxy and piperazino)fluorobenzene derivatives (8a--f) were synthesised via a short synthetic route in good chemical yields and were evaluated for inotropic and chronotropic activity in isolated guinea-pig atria preparation. The compounds act as potential phosphodiesterase (PDE) inhibitors with desirable biological activity and have considerable promise in heart therapy.

Animals↗

Analysis of Light-Induced Depressions of Photosynthesis in Leaves of a Wheat Crop during the Winter.

The photosynthetic performances of individual leaves of a wheat (Triticum aestivum cv Bezostaya) crop were assessed daily and throughout individual days during the winter when temperature and light levels were fluctuating. Measurements of chlorophyll fluorescence induction and the maximum quantum yield of O(2) evolution were made on individual leaves. Depressions in the ratio of variable to maximal fluorescence (F(v)/F(m)) were correlated with low temperatures and high light levels throughout the winter and during the course of individual days. Depressions in F(v)/F(m) observed in the field during the day were not accompanied by any significant change in the ability of photosystem II complexes to bind 3-(3,4-dichlorophenyl)-1-dimethyl urea, indicating that the depressions in F(v)/F(m) were not attributable to photodamage to the D1 protein of the photosystem II reaction center. Decreases in F(v)/F(m) were associated with increases in the rate of dissipation of excitation energy by radiationless decay processes and decreases in the quantum efficiency of CO(2) assimilation, indicative of a rapidly reversible light-induced "downregulation" of photosynthesis. No major changes were observed in the maximum quantum efficiency of O(2) evolution of leaves throughout periods of fluctuating temperature and light, because light-induced depressions in photosynthetic efficiency recovered within the time required to make these measurements.

Journal Article↗

Modifications to Thylakoid Composition during Development of Maize Leaves at Low Growth Temperatures.

The effects of reductions in growth temperature on the development of thylakoids of maize (Zea mays var LG11) leaves are examined. Thylakoids isolated from mesophyll cells of leaves grown at 17 degrees and 14 degrees C, compared with 25 degrees C, exhibited a decreased accumulation of many polypeptides, which was accompanied by a loss of activity of photosystems (PS) I and II. Probing the polypeptide profiles with a range of antibodies specific for thylakoid proteins demonstrated that a number of polypeptides encoded by the chloroplast genome failed to accumulate at low temperatures. Although thylakoid protein synthesis was reduced severely at 14 degrees C compared with 25 degrees C, major synthesis of both chloroplast and nuclear encoded polypeptides was detected. It is suggested that the lack of accumulation of some thylakoid proteins at low temperatures may be due to an inability to stabilize the proteins in the membranes. A number of thylakoid polypeptides were found to appear as the growth temperature was decreased. Analyses of pigments and polypeptides demonstrated that decreases in the photosystem reaction center core complexes occur relative to the light harvesting complex associated with PS II at reduced growth temperatures. Differential effects on the development of PSI and PSII were also observed, with PSII activity being preferentially reduced. Reductions in PSII content and activity occurred in parallel with decreases in the quantum yield and light-saturated rate of CO(2) assimilation. Fractionation of thylakoid pigment-protein complexes showed that the ratio of monomeric:oligomeric form of the light harvesting complex associated with PSII increased at low growth temperature, which is consistent with a chill-induced modification of thylakoid organization. Many, but not all, of the characteristic changes in thylakoid protein metabolism, which were observed when leaves were grown at low temperatures in controlled environments, were identified in leaves of a field maize crop during the early growing season when low temperatures were experienced by the crop. Chill-induced perturbations of thylakoid development can occur in the field in temperate regions and may have implications for the photosynthetic productivity of the crop.

Journal Article↗

The Sequence of Change within the Photosynthetic Apparatus of Wheat following Short-Term Exposure to Ozone.

The basis of inhibition of photosynthesis by single acute O(3) exposures was investigated in vivo using analyses based on leaf gas exchange measurements. The fully expanded second leaves of wheat plants (Triticum aestivum L. cv Avalon) were fumigated with either 200 or 400 nanomoles per mole O(3) for between 4 and 16 hours. This reduced significantly the light-saturated rate of CO(2) uptake and was accompanied by a parallel decrease in stomatal conductance. However, the stomatal limitation, estimated from the relationship between CO(2) uptake and the internal CO(2) concentration, only increased significantly during the first 8 hours of exposure to 400 nanomoles per mole O(3); no significant increase occurred for any of the other treatments. Analysis of the response of CO(2) uptake to the internal CO(2) concentration implied that the predominant factor responsible for the reduction in light-saturated CO(2) uptake was a decrease in the efficiency of carboxylation. This was 58 and 21% of the control value after 16 hours at 200 and 400 nanomoles per mole O(3), respectively. At saturating concentrations of CO(2), photosynthesis was inhibited by no more than 22% after 16 hours, indicating that the capacity for regeneration of ribulose bisphosphate was less susceptible to O(3). Ozone fumigations also had a less pronounced effect on light-limited photosynthesis. The maximum quantum yield of CO(2) uptake and the quantum yield of oxygen evolution showed no significant decline after 16 hours with 200 nanomoles per mole O(3), requiring 8 hours at 400 nanomoles per mole O(3) before a significant reduction occurred. The photochemical efficiency of photosystem II estimated from the ratio of variable to maximum chlorophyll fluorescence and the atrazine-binding capacity of isolated thylakoids demonstrated that photochemical reactions were not responsible for the initial inhibition of CO(2) uptake. The results suggest that the apparent carboxylation efficiency appears to be the initial cause of decline in photosynthesis in vivo following acute O(3) fumigation.

Journal Article↗

Pseudomonas aeruginosa exoenzyme S is an adhesion.

Exoenzyme S from Pseudomonas aeruginosa has been studied as an adhesion for glycosphingolipids and buccal cells. Binding of exoenzyme S to gangliotriosylceramide (GalNAc beta 1-4Gal beta 1-4Glc beta 1-1Cer), gangliotetraosylceramide (Gal beta 1-3 GalNAcT beta 1-4 Gal beta 1-4Glc beta 1-1Cer), and lactosylceramide (Gal beta 1-4Glc beta 1-1Cer) separated on thin-layer chromatograms was observed. Binding curves for exoenzyme S with dilutions of gangliotetraosylceramide immobilized on plastic plates were similar to previously reported results for the intact bacteria. Binding of exoenzyme S to sialylated counterparts of these glycosphingolipids was not seen, indicating that the addition of a sialic acid residue interferes with binding. Exoenzyme S and monoclonal antibody to exoenzyme S inhibit the binding of P. aeruginosa to buccal cells. The presence of exoenzyme S on the surface of P. aeruginosa was detected by immunogold labeling of bacteria with antibodies to exoenzyme S. Results of these studies led us to conclude that exoenzyme S is an important adhesion of P. aeruginosa.

ADP Ribose Transferases↗

Relationship between the Quantum Efficiencies of Photosystems I and II in Pea Leaves.

The irradiance dependence of the efficiencies of photosystems I and II were measured for two pea (Pisum sativum [L.]) varieties grown under cold conditions and one pea variety grown under warm conditions. The efficiencies of both photosystems declined with increasing irradiance for all plants, and the quantum efficiency of photosystem I electron transport was closely correlated with the quantum efficiency of photosystem II electron transport. In contrast to the consistent pattern shown by efficiency of the photosystems, the redox state of photosystem II (as estimated from the photochemical quenching coefficient of chlorophyll fluorescence) exhibited relationships with both irradiance and the reduction of P-700 that varied with growth environment and genotype. This variability is considered in the context of the modulation of photosystem II quantum efficiency by both photochemical and nonphotochemical quenching of excitation energy.

Journal Article↗

Adherence of Pseudomonas aeruginosa to tracheal epithelium.

Adherence of mucoid and nonmucoid strains of Pseudomonas aeruginosa to tracheal epithelium was studied with a perfused-trachea model. The species specificity of adherence was studied by infecting tracheas from hamsters, guinea pigs, or mice. Perfused tracheas from hamsters were infected with strains of P. aeruginosa in the presence of various sugars, lectins, cations, or charged polymers. Adherence of mucoid strains of P. aeruginosa was greatest for guinea pigs; that for hamsters and mice was approximately the same. Nonmucoid strains did not adhere well to epithelium from any of the species tested. N-Acetylglucosamine, galactose, and N-acetylneuraminic acid were the best inhibitors of adherence of mucoid strains of P. aeruginosa. Phaseolus vulgaris agglutinin and Arachis hypogaea agglutinin enhanced adherence of mucoid strains. Adherence of mucoid strains was also enhanced by the presence of Ca2+ in the incubation medium. Poly-L-lysine, poly-L-aspartic acid, and polyglycine inhibited adherence of a mucoid strain by 96, 86, and 52%, respectively. In general, the adherence of nonmucoid strains was not affected. The results indicate that carbohydrates are involved in the interaction of mucoid strains of P. aeruginosa with tracheal cells and that divalent cations may enhance this interaction. The lectin data show that lectins can interact with the mucoid organisms and the host and suggest that lectins may play a role in the adhesion process.

Animals↗

Purification and cytotoxic potential of myeloperoxidase in cystic fibrosis sputum.

The neutrophil myeloperoxidase-H2O2-halide enzyme system produces hypochlorous acid and chlorinated amine compounds capable of killing a variety of target cells. In the present study we hypothesized that the myeloperoxidase enzyme system is one mechanism for airway epithelial damage in patients with cystic fibrosis (CF). Enzyme linked immunosorbent assay detected high antigenic levels of myeloperoxidase in sputum samples of seven patients with CF. Myeloperoxidase was purified to homogeneity from CF sputum and from blood neutrophils by a three-step technique involving dialysis, gel filtration, and ion-exchange chromatography. CF sputum myeloperoxidase and neutrophil myeloperoxidase appeared identical by acid gel electrophoresis and Ouchterlony experiments. CF sputum myeloperoxidase also contained approximately the same enzymatic activity as neutrophil myeloperoxidase. The myeloperoxidase enzyme system was tested for its cytotoxic potential in a tracheal ring culture system. Myeloperoxidase-induced cytotoxicity for airway epithelium was confirmed by light microscopy and radiolabelling experiments. These findings suggest a possible role for neutrophil myeloperoxidase in CF lung disease.

Animals↗