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At least 1,333 records · Page 74Linked to original sources
PubMed1999

Probing the microenvironment of intracellular bacterial pathogens.

The identification of bacterial genes regulated in response to the intracellular environment is crucial to the understanding of host-pathogen interactions. Several techniques have been developed to identify and characterize bacterial genes that are induced during the intracellular infection and, potentially, may play a role in pathogenesis. This review discusses the strategies that have been utilized to examine differential gene expression by bacterial pathogens during the intracellular infection. Furthermore, a number of the differentially expressed genes are described.

Animals↗
PubMed2002

Engineering cellular microenvironments to improve cell-based drug testing.

Recent progress in the biology of cell adhesion is enabling cell culture models to better reproduce in vivo functions. Cues from adhesion to extracellular matrix and neighboring cells are important regulators of cell behaviors. The recent adaptation of semiconductor tools to spatially organize cells and their adhesions has enhanced our ability to engineer cell functions ex vivo. By using these tools to create more in vivo-like cultures, cell-based drug discovery and target validation could be improved. This review explores the biological advances made by these microfabrication tools and discusses how they could enable high-throughput cell-based assays.

Animals↗
PubMed1999

Effect of stocking density on ewes' milk yield, udder health and microenvironment.

Three groups of 12 lactating Comisana ewes were housed in separate rooms of the same building that contained straw-bedded pens with total areas of 12, 18 and 24 m2. The ewes were assigned to groups with different stocking densities: high (HSD, 1 m2/animal), medium (MSD, 1.5 m2/animal) or low (LSD, 2 m2/animal). The LSD treatment resulted in lower air concentrations of total microorganisms and coliform bacteria than the other two. Ewes in the LSD group gave greater yields of milk, total protein and fat than those in the other two groups. LSD treatment increased milk casein content and improved clotting time and clot firmness. LSD also resulted in ewes giving milk with lower somatic cell counts and smaller concentrations of mesophiles, psychrotrophs and faecal coliforms than MSD and HSD. Subclinical mastitis occurred in three ewes of the MSD groups and four ewes of the HSD group, while no cases were recorded in the LSD group. These results indicate that stocking density is a critical factor in dairy sheep housing and suggest that a space allocation < 2 m2/animal may adversely affect the performance and health of the lactating ewe.

Air Microbiology↗
PubMed1995

The microenvironment created by non-blocking embryos in aggregates may rescue blocking embryos via cell-embryo adherent contacts.

Under our culture conditions, mouse embryos from the BALB/c inbred mouse strain develop successfully in culture only from the late 2-cell stage onwards (so-called 2-cell block), whether or not EDTA is added to the culture medium. (CBA x C57BL) F2 embryos do not exhibit a 2-cell block. Medium conditioned by culture of non-blocking embryos from the 2-cell to the 8-cell stage did not improve the development of blocking embryos, nor did co-culture of blocking and non-blocking embryos, with or without conditioned medium. On the other hand phytohaemagglutinin (PHA)-assisted aggregation of an early 2-cell BALB/c embryo with five surrounding non-blocking F2 embryos (2-cell or 8-cell) or five BALB/c 8-cell embryos allowed the early 2-cell BALB/c embryos to develop into blastocysts within 72 h. Aggregation of blocking BALB/c 2-cell embryos with each other had no 'rescue' effect. When blocking and non-blocking 2-cell embryos were aggregated together, an integrated blastocyst was formed; but when the early 2-cell BALB/c embryos were aggregated with non-blocking 8-cell embryos, the blocking embryos formed a separate small blastocyst, which nonetheless retained adherent contact with the non-blocking embryos throughout the culture period. Ultrastructural analysis showed that 2-cell embryos aggregated with the aid of PHA form close adherent cell contacts up to several micrometres in length.

Animals↗
PubMed1994

Site-directed mutagenesis, fluorescence, and two-dimensional NMR studies on microenvironments of effector region aromatic residues of human c-Ha-Ras protein.

The Tyr residues in positions 32 and 40 of human c-Ha-Ras protein were replaced by site-directed mutagenesis (Y32F, Y32W, Y40K, and Y40W) to examine their roles in the signal-transducing activity and the sensitivity to the GTPase activating protein (GAP). The signal-transducing activity of the oncogenic Ras protein in PC12 cells was lost upon mutations Y32F and Y40K, but retained upon mutations Y32W and Y40W. These results suggest that residues 32 and 40 are both required to have aromatic groups and residue 32 is further required to have a hydrogen donor. On the other hand, three mutations (Y32F, Y32W, and Y40W) caused no appreciable reduction in either GAP-binding affinity or GAP sensitivity. By the Y40K mutation, GAP-binding affinity was slightly lowered, while GAP sensitivity was drastically impaired. Therefore, for residues 32 and 40 of Ras, interactions with GAP appear to be different from those with the target of signal transduction in the PC12 cell. As for the Y32W-Ras protein bound with an unhydrolyzable GTP analogue (GMPPNP), the Trp32 fluorescence is appreciably red-shifted, weaker, and more susceptible to KI quenching as compared to that of the GDP-bound form. Two-dimensional NMR spectroscopy with selectively deuterated Ras proteins revealed fewer and weaker nuclear Overhauser effects on the aromatic protons of Trp32 in the GMPPNP-bound form than in the GDP-bound form. This indicates that the side chain of Trp32 is more exposed to the solvent in the GMPPNP-bound form than in the GDP-bound form.(ABSTRACT TRUNCATED AT 250 WORDS)

Amino Acid Sequence↗
PubMed1986

Phospholipid distribution in the microenvironment of the immunoglobulin E-receptor from rat basophilic leukemia cell membrane.

It has been previously found that lipids were required to maintain intact the tetrameric structure of the receptor for immunoglobulin E (IgE) (Fc epsilon R) in detergent solutions [Rivnay, B., Rossi, G., Henkart, M., & Metzger, H. (1984) J. Biol. Chem. 259, 1212-1217, and references cited therein]. Failure of commercially obtained lipids to provide sufficient protection, however, underscored the necessity for development of additional analytical approaches. In order to identify the phospholipid distribution in the intimate natural environment of this receptor, both the plasma membrane vesicles and the ligand-receptor complex (IgE-Fc epsilon R) have been isolated by affinity chromatography. The phospholipids of both preparations were compared. After extensive washing with detergent lipid micelles, IgE-Fc epsilon R retained 0.1-1% of the total phospholipids in the purified plasma membrane. The receptor-bound lipids were shown to contain phosphatidylcholine and sphingomyelin; the content of the latter lipid was enriched 2-5-fold compared with that in the plasma membranes. This pattern was observed with several detergents employed for purification and under a variety of experimental conditions. In light of the general distribution of choline phospholipids in the outer leaflet of plasma membranes, this enrichment may not be a characteristic of this particular receptor exclusively. These observations should be particularly helpful in studies on aggregation-induced functions of the isolated Fc epsilon receptor. In general, the methods employed enable isolation of purified and lipid-protected integral proteins and also provide an appropriate reference source of intact membrane vesicles. These qualities render this approach useful in similar studies of other membrane proteins.

Animals↗
PubMed1989

Characterization of lens alpha-crystallin tryptophan microenvironments by room temperature phosphorescence spectroscopy.

Room temperature phosphorescence techniques were used to study the structural and dynamic features of the tryptophan residues in bovine alpha-crystallin. Upon excitation at 290 nm, the characteristic signature of tryptophan phosphorescence was observed with an emission maximum at 442 +/- 2 nm. The phosphorescence intensity decay was biphasic with lifetimes of 5.4 ms (71%) and 42 ms (29%). Phosphorescence quenching measurements strongly suggest that each component corresponds to one class of tryptophans with the more buried residues having the longer emission lifetime. Three small-molecule quenchers were surveyed, and in order of increasing quenching efficiency: iodide less than nitrite less than acrylamide. A heavy-atom effect was observed in iodide solutions, and an upper limit of 5% was placed on the quantum yield of triplet formation in iodide-free solutions, while the phosphorescence quantum yield was estimated to be approximately 3.2 x 10(-4). The temperature dependence of the phosphorescence lifetime was measured between 5 and 40 degrees C. Arrhenius plots exhibited discontinuities at 26 and 29 degrees C for the short- and long-lived components, respectively, corresponding to abrupt transitions in segmental flexibility. Denaturation studies revealed conformational transitions between 1 and 2 M guanidine hydrochloride, and 4 and 6 M urea. Long-lived phosphorescence lifetimes of 3 and 7 ms were measured in 6 M guanidine hydrochloride and 8 M urea, respectively, suggesting that some structural features are preserved even at very high concentrations of denaturant. Our studies demonstrate the sensitivity of room temperature phosphorescence spectroscopy to the structure of alpha-crystallin, and the applicability of this technique for monitoring conformational changes in lens crystallin proteins.(ABSTRACT TRUNCATED AT 250 WORDS)

Acrylamide↗
PubMed2004

Application of multicriteria decision making methods to air quality in the microenvironments of residential houses in Brisbane, Australia.

This paper reports the first application of the multicriteria decision making methods, PROMETHEE and GAIA, to indoor and outdoor air quality data. Fourteen residential houses in a suburb of Brisbane, Australia were investigated for 21 air quality-influencing criteria, which included the characteristics of the houses as well as the concentrations of volatile organic compounds, fungi, bacteria, submicrometer, and supermicrometer particles in their indoor and outdoor air samples. Ranking information necessary to select one house in preference to all others and to assess the parameters influencing the differentiation of the houses was found with the aid of PROMETHEE and GAIA. There was no correlation between the rank order of each house and the health complaints of its occupants. Patterns in GAIA plots show that indoor air quality in these houses is strongly dependent on the characteristics of the houses (construction material, distance of the house from a major road, and the presence of an in-built garage). Marked similarities were observed in the patterns obtained when GAIA and factor analysis were applied to the data. This underscores the potential of PROMETHEE and GAIA to provide information that can assist source apportionment and elucidation of effective remedial measures for indoor air pollution.

Air Pollution, Indoor↗
PubMed2005

The microenvironment effect on the generation of reactive oxygen species by Pd-bacteriopheophorbide.

Generation of reactive oxygen species (ROS) is the hallmark of important biological processes and photodynamic therapy (PDT), where ROS production results from in situ illumination of certain dyes. Here we test the hypothesis that the yield, fate, and efficacy of the species evolved highly depend on the dye's environment. We show that Pd-bacteriopheophorbide (Pd-Bpheid), a useful reagent for vascular targeted PDT (VTP) of solid tumors, which has recently entered into phase II clinical trials under the code name WST09 (trade name TOOKAD), forms appreciable amounts of hydroxyl radicals, superoxide radicals, and probably hydrogen peroxide in aqueous medium but not in organic solvents where singlet oxygen almost exclusively forms. Evidence is provided by pico- and nanosecond time-resolved spectroscopies, ESR spectroscopy with spin-traps, time-resolved singlet oxygen phosphorescence, and chemical product analysis. The quantum yield for singlet oxygen formation falls from approximately 1 in organic solvents to approximately 0.5 in membrane-like systems (micelles or liposomes), where superoxide and hydroxyl radicals form at a minimal quantum yield of 0.1%. Analysis of photochemical products suggests that the formation of oxygen radicals involves both electron and proton transfer from (3)Pd-Bpheid at the membrane/water interface to a colliding oxygen molecule, consequently forming superoxide, then hydrogen peroxide, and finally hydroxyl radicals, with no need for metal catalysis. The ability of bacteriochlorophyll (Bchl) derivatives to form such radicals upon excitation at the near infrared (NIR) domain opens new avenues in PDT and research of redox regulation in animals and plants.

Bacteriochlorophylls↗
PubMed2006

Comparison of the lipase activity in hydrolysis and acyl transfer reactions of two latex plant extracts from babaco (Vasconcellea x Heilbornii Cv.) and Plumeria rubra: Effect of the Aqueous microenvironment.

The enzymatic properties of Plumeria rubra latex have been evaluated for the first time, showing a high activity in both hydrolysis and synthesis reactions, and compared to the biocatalytic behavior of babaco (Vasconcellea x Heilbornii cv.) latex. Both biocatalysts have been optimized by studying the various parameters that influence reaction kinetics. The optimum temperatures for hydrolysis reactions were 50 and 55 degrees C for babaco and Plumeria, respectively. The optimum pH for babaco latex was 7, whereas for Plumeria latex, two optimal pH values (4 and 7) were observed. With regard to esterification and acyl transfer reactions such as alcoholysis and interesterification, the influence of thermodynamic water activity on reaction yields was determined and correlated with water sorption and desorption isotherms. When babaco latex is used as a biocatalyst, optimal synthesis reaction yields are obtained when the enzymatic extract is stabilized at a water activity value of 0.38, which corresponds to a water content of 5.7%. This optimal level of hydration is located on the linear portion of the biocatalyst's sorption isotherm, where the water molecules exhibit high-energy interactions with the protein network. In synthesis reactions (esterification, alcoholysis, and interesterification) biocatalyzed by Plumeria latex, correlation between best reaction yields and water activity cannot be done. Indeed, the sorption isotherm plot has an atypical shape, indicating that water might be trapped in the latex matrix and, consequently, that the water content of the biocatalyst is highly dependent on the hydration history of the latex.

Acylation↗