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Biomedical subjects

J R Lu

Publications and source records attributed to J R Lu.

At least 19 recordsLinked to original sources
PubMed2006

Neutron reflection study of a water-soluble biocompatible diblock copolymer adsorbed at the air/water interface: the effects of pH and polymer concentration.

The effect of varying both the solution pH and copolymer concentration on the structure of layers of poly[2-(methacryloyloxy)ethyl phosphorylcholine-block-2-(dimethylamino)ethyl methacrylate] copolymer (denoted as MPC(30)-DMA(60), M(n) = 18,000) adsorbed at the air/water interface is studied using surface tension and specular neutron reflection. The surface structure of the adsorbed diblock copolymer is represented by a dense layer of 10-15 A on the air side, accompanied by a loose layer of 20-30 A extending into the aqueous phase. Although the uniform layer model generally provided a reasonable description of the adsorbed copolymer chains, some deviations were observed. A more detailed analysis showed that the distribution of the copolymer across the interface required a minimum of three layers to take into account the structural inhomogeneities. Refinement of the structural distributions involved the combined fitting of partially deuterated copolymer in null reflecting water and D(2)O and the fully hydrogenated copolymer in D(2)O, leading to a substantial improvement in the reliability of the structural profiles obtained. The data analysis showed an increase in surface excess at higher copolymer concentrations and at more alkaline pH. However, the copolymer layer was fully immersed in water under all conditions studied. Because the surface excess showed a steady increase across the cmc over the high pH range, we speculate that copolymer adsorption above the cmc involves the formation of surface micellar aggregates under these conditions.

Adsorption↗
PubMed2006

The structure of zwitterionic phosphocholine surfactant monolayers.

The structure of a zwitterionic phosphocholine (PC) surfactant monolayer adsorbed on the surface of water has been determined using neutron reflectivity in combination with H/D isotopic substitution. The most significant results of this study are the level of hydration of the PC headgroup and the lack of dehydration with increasing temperature and salt addition. The fraction of the alkyl chain (f(c)) immersed in water for all three chain isomers studied was found to be around 0.15, suggesting that the PC headgroup geometries influenced not only the headgroup hydration but also the degree of immersion of the alkyl chain in water. At the critical micelle concentration (CMC), the number of water molecules associated with the PC headgroup in C(m)PC (m = 12, 14, 16) was on order of 15. This value was significantly greater than that obtained for nonionic and ionic surfactants with similar limiting area per molecule at the CMC (A(cmc)). However, the fraction of the chain immersed in water for the ionic and nonionic surfactants was much greater. This suggests that the unique surface biocompatibility of PC surfactants arises from their strong affinity for water, and the relatively low fraction of mixing with the alkyl chain arises from the higher structural order within the PC monolayer. As surface coverage decreased, the number of water molecules associated with each PC headgroup increased, but f(c) remained constant for all the surfactants. This observation was consistent with the small variation in the thickness of the headgroup region, and the entire layer changed little with surfactant concentration. This is attributed to the role of PC headgroup geometries to maintain the conformational order within the layer as packing density varies. Further structural analysis based on a kinematic approach showed that, as the chain length was increased from C12 to C14 to C16 at the CMC, the angle of tilt for the alkyl chain increased from 40 degrees to 48 degrees to 53 degrees , respectively, whereas the thickness of the whole layer and that of the PC head region was largely constant. The almost vertical projection of the PC headgroup from these single alkyl chain surfactants is in sharp contrast to its strongly tilted conformation, as reported for dichain phospholipids such as dipalmitoyl glycerol phosphocholine (DPPC).

Ions↗
PubMed2005

Adsorption of single chain zwitterionic phosphocholine surfactants: effects of length of alkyl chain and head group linker.

The adsorption of a range of single chain zwitterionic phosphocholine surfactants (C(n)P(m)C) at the air/liquid interface has been studied by a combination of surface tension and neutron reflectivity. The critical micellar concentration (CMC) for C(n)PC (or C(n)P(2)C), where n varied from 12, 14 to 16, was found to be 0.91, 0.14, and 1.2 x 10(-2) mM respectively, and followed the same trend as observed for other zwitterionic and non-ionic surfactants. The area per molecule at the CMC, A(cmc), for C(n)PC was found to remain constant between 50 and 53 A(2), indicating that the increase in the alkyl chain length had little effect on A(cmc) at the interface. The neutron reflection measurement also showed an almost constant layer thickness (tau) of 20+/-2 A from all the alkyl chain deuterated PC surfactants (dC(n)hPC) in null reflecting water (NRW), suggesting that the alkyl chains of the surfactant responded to changes in either chain length or solution concentration by varying their angle of tilt. In contrast, increasing the length of head group linker between P and N atoms in C(12)P(m)C, where m=2, 4, to 6, resulted in a much slower decrease of CMC from 0.91, 0.7, to 0.5 mM, consistent with a different contribution to the free energy of micellization. A(cmc) for C(12)P(m)C did not vary when m was increased from 2 to 4, and this observation together with the thickness of the head group region indicated an almost perpendicular projection of the head group in C(12)P(2)C and C(12)P(4)C. A further increase in m to 6 resulted in an A(cmc) of 70 A(2). This increase in A(cmc) however did not result in any change in either the total layer thickness or the fraction of the head group region submerged in the aqueous subphase, suggesting that the head group in C(12)P(6)C was bent away from the surface normal direction. Both increase in temperature from 25 to 40 degrees C and the addition of 0.1 M NaCl had little effect on the area per molecule or the thickness of C(12)P(m)C surfactant layer, showing that the C(12)P(m)C series behaved like C(n)P(2)C series. The main conclusion from this study is that for all the C(n)P(m)C surfactants studied, change in m or n has little effect on the total thickness, the thickness of the alkyl chain or that of the head group region.

Adsorption↗
PubMed2005

Surface adsorption of zwitterionic surfactants: n-alkyl phosphocholines characterised by surface tensiometry and neutron reflection.

The surface adsorption of n-dodecyl phosphocholine (C12PC) has been characterised by a combined measurement of surface tension and neutron reflectivity. The critical micellar concentration (CMC) was found to be 0.91 mM at 25 degrees C in pure water. At the CMC, the limiting area per molecule (A(cmc)) was found to be 52+/-3 A2 and the surface tension (gamma(cmc)) to be ca. 40.0+/-0.5 mN/m. The parallel study of chain isomer n-hexadecyl phosphocholine (C16PC) showed a decrease of the CMC to 0.012 mM and a drop of gamma(cmc) to 38.1+/-0.5 mN/m. However, A(cmc) for C16PC was found to be 54+/-3 A2, showing that increase in alkyl chain length by four methylene groups has little effect on A(cmc). The almost constant A(cmc) suggested that the limiting area per molecule was determined by the bulky PC head group. It was further found that the surface tension and related key physical parameters did not vary much with temperature, salt addition, solution pH or any combination of these, thus showing that surface adsorption and solution aggregation from PC surfactants is largely similar to the zwitterionic betaine surfactants and is distinctly different from ionic and non-ionic surfactants. The thickness of the adsorbed monolayers measured from both dC12hPC and dC16hPC was found to be 20-22 A at the CMC from neutron reflectivity. Neither A(cmc) nor layer thickness varied with alkyl chain length, indicating that as the alkyl chain length became longer it was further tilted away from the surface normal direction and the layer packing density increased. It was also observed that the thickness of the layer varied little with surfactant concentration, indicating that the average conformational orientation of the alkyl chain remained unchanged against varying surface coverage.

Adsorption↗
PubMed2001

Neutron reflection from a dimyristoylphosphatidylcholine monolayer adsorbed on a hydrophobised silicon support.

Neutron specular reflection has been used to study the structure of a monolayer of dimyristoylphosphatidylcholine (DMPC) deposited using the Langmuir-Blodgett technique onto a silicon oxide substrate. A self-assembled monolayer of octadecyltrichlorosilane with a deuterated alkyl chain (d-OTS) had been previously bonded onto this silicon oxide substrate which rendered it hydrophobic. In the system under study, the alkyl chains of the phospholipid were found to penetrate extensively into the d-OTS layer with the mixed chain region (d-OTS and DMPC) having a total thickness of 30.5 A. This mixed region was divided into two halves for analysis; the 'lower half' (nearest to the substrate surface) was found to comprise anchored d-OTS chains mixed with the lipid chains in the volume ratio approx. 0.60:0.35. The corresponding volume ratio in the 'upper half' of this region was determined to be approx. 0.50:0.40. The thicknesses of these regions were found to be 17.9 A (incorporating approx. 6% solvent) and 12.6 A (incorporating approx. 9% solvent) for the lower and upper halves respectively. The DMPC head groups were found to be confined to the most external layer (furthest away from the silicon substrate). This layer was found to have a thickness of 9.4 A and included a small fraction of the lipid alkyl chains with approx. 47% solvent.

Dimyristoylphosphatidylcholine↗
PubMed2001

Association of COOH-terminal-binding protein (CtBP) and MEF2-interacting transcription repressor (MITR) contributes to transcriptional repression of the MEF2 transcription factor.

The class II histone deacetylases (HDACs) 4, 5, and 7 share a common structural organization, with a carboxyl-terminal catalytic domain and an amino-terminal extension that mediates interactions with members of the myocyte enhancer factor-2 (MEF2) family of transcription factors. Association of these HDACs with MEF2 factors represses transcription of MEF2 target genes. MEF2-interacting transcription repressor (MITR) shares homology with the amino-terminal extensions of class II HDACs and also acts as a transcriptional repressor, but lacks a histone deacetylase catalytic domain. This suggests that MITR represses transcription by recruiting other corepressors. We show that the amino-terminal regions of MITR and class II HDACs interact with the transcriptional corepressor, COOH-terminal-binding protein (CtBP), through a CtBP-binding motif (P-X-D-L-R) conserved in MITR and HDACs 4, 5, and 7. Mutation of this sequence in MITR abolishes interaction with CtBP and impairs, but does not eliminate, the ability of MITR to inhibit MEF2-dependent transcription. The residual repressive activity of MITR mutants that fail to bind CtBP can be attributed to association with other HDAC family members. These findings reveal CtBP-dependent and -independent mechanisms for transcriptional repression by MITR and show that MITR represses MEF2 activity through recruitment of multicomponent corepressor complexes that include CtBP and HDACs.

Alcohol Oxidoreductases↗
PubMed2001

Beta-casein adsorption at the hydrophobized silicon oxide-aqueous solution interface and the effect of added electrolyte.

The effect of the presence of NaCl, CaCl(2), or MgCl(2) at the same ionic strength on the structure of beta-casein layers adsorbed on hydrophobic surfaces has been investigated by neutron reflectivity measurements. The data were fitted to a four-layer model. The volume fraction versus distance profiles have a similar shape whether beta-casein is adsorbed from NaCl, CaCl(2), and MgCl(2) of the same ionic strength or whether the protein concentration is lowered 10 times. In particular at larger distances from the surface, the volume fraction values are low and similar. However, close to the hydrophobic surface the volume fraction of protein decreases in the order CaCl(2) > MgCl(2) > NaCl. We have also used a specific proteolytic enzyme, endoproteinase Asp-N, which cleaves off the hydrophilic part of beta-casein, as a tool to reveal the interfacial structure of the protein. For all the different types of added electrolytes, endoproteinase Asp N only affects the outermost beta-casein layer. Subsequent addition of beta-casein in all cases led to large increases in amounts adsorbed and in the thickness of the outer layers.

Adsorption↗
PubMed2001

beta-Casein adsorption at the silicon oxide--aqueous solution interface.

Neutron reflectometry was used to investigate the time-dependent beta-casein adsorption at the silica-aqueous solution interface. The transient and steady-state structural characteristics of the adsorbed layer were determined from reflectivity curves, fitted to three-layer and two-layer models. The results show that the beta-casein adsorption to silica is very slow. The adsorption process involves the formation of an inner dense protein layer with a mean thickness of about 30 A onto which a more hydrated outer layer is self-associated. The surface excess and the total layer thickness of the asymmetric bilayer were, after 5 h adsorption time, estimated to be about 6.5 mg/m2 and 105 A, respectively. The adsorption behavior observed on silica contrasts with that previously reported for hydrophobic substrates, where beta-casein adsorption is much more rapid and the final surface excess is less than half of that observed for silica. Rinsing the silica surface with protein-free buffer resulted in a substantial desorption; much more pronounced than observed for hydrophobic substrates. This behavior suggests a weak adsorption affinity for a fraction of the adsorbed casein molecules; most likely the outer self-associated casein molecules in the adsorbed bilayer. The comparative desorption from hydrophobic surfaces was shown to be marginal. The difference between the layer structures adopted on hydrophobic and hydrophilic surfaces is also mirrored in the effects that the addition of a specific proteolytic enzyme (endoproteinase Asp-N) has on the adsorbed layer properties. The rinsing and endoproteinase cleavage processes result together in more than 80% reduction of the originally adsorbed mass at the silica surface. Only a thin but dense adsorbed layer remains after these treatments. The corresponding reduction reported for the hydrophobic adsorbent system was only about 20%. It is concluded that beta-casein adsorption on silica results in the formation of an asymmetric surface bound bilayer that stands in strong contrast to the monolayer structure formed at hydrophobic surfaces. This finding support the previous results obtained by using ellipsometry. The study also shows that neutron reflection, despite its limitations in time resolution, can be used for studying dynamic interfacial phenomena in protein systems.

Adsorption↗
PubMed2001

Application of digital image analysis and flow cytometry to enumerate marine viruses stained with SYBR gold.

A novel nucleic acid stain, SYBR Gold, was used to stain marine viral particles in various types of samples. Viral particles stained with SYBR Gold yielded bright and stable fluorescent signals that could be detected by a cooled charge-coupled device camera or by flow cytometry. The fluorescent signal strength of SYBR Gold-stained viruses was about twice that of SYBR Green I-stained viruses. Digital images of SYBR Gold-stained viral particles were processed to enumerate the concentration of viral particles by using digital image analysis software. Estimates of viral concentration based on digitized images were 1.3 times higher than those based on direct counting by epifluorescence microscopy. Direct epifluorescence counts of SYBR Gold-stained viral particles were in turn about 1.34 times higher than those estimated by the transmission electron microscope method. Bacteriophage lysates stained with SYBR Gold formed a distinct population in flow cytometric signatures. Flow cytometric analysis revealed at least four viral subpopulations for a Lake Erie sample and two subpopulations for a Georgia coastal sample. Flow cytometry-based viral counts for various types of samples averaged 1.1 times higher than direct epifluorescence microscopic counts. The potential application of digital image analysis and flow cytometry for rapid and accurate measurement of viral abundance in aquatic environments is discussed.

Cyanobacteria↗
PubMed2000

Surfactant layers at the air/water interface: structure and composition.

The use of neutron reflectometry to study the structure and composition of surfactant layers adsorbed at the air/water interface is reviewed. A critical assessment of the results from this new technique is made by comparing them with the information available from all other techniques capable of investigating this interface.

Adsorption↗
PubMed1999

The reduced adsorption of lysozyme at the phosphorylcholine incorporated polymer/aqueous solution interface studied by spectroscopic ellipsometry.

Coating hydrogel polymers onto solid substrates can reduce the adsorption of proteins onto these surfaces, but the extent of the reduction in protein adsorption is strongly dependent on how the surface layer is coated. We have examined the effect of coating conditions on the structure of thin polymer films formed from a number of poly(methacrylate)-based hydrogel polymers via the dip-coating method. We show in this work how the polarity of the solvent, the speed of lifting, and the annealing temperature affect the thickness and uniformity of ultrathin phosphorylcholine (PC)-incorporated polymer films coated on the surface of native oxide on silicon and the subsequent interaction of these coated surfaces with lysozyme molecules. Our results show that the uniformity of the polymer film, and thus the smoothness of the outer film surface, influence the extent of reduction in protein adsorption. We suggest that the reduction in lysozyme adsorption is the result of a layer of PC groups on the surface of the polymer film. The improvement of the smoothness of the film results in the formation of a close-packed PC layer on the outer surface of the polymer film, leaving few defects or cavities on which protein molecules can bind.

Adsorption↗
PubMed1999

FOG-2, a heart- and brain-enriched cofactor for GATA transcription factors.

Members of the GATA family of zinc finger transcription factors have been shown to play important roles in the control of gene expression in a variety of cell types. GATA-1, -2, and -3 are expressed primarily in hematopoietic cell lineages and are required for proliferation and differentiation of multiple hematopoietic cell types, whereas GATA-4, -5, and -6 are expressed in the heart, where they activate cardiac muscle structural genes. Friend of GATA-1 (FOG) is a multitype zinc finger protein that interacts with GATA-1 and serves as a cofactor for GATA-1-mediated transcription. FOG is coexpressed with GATA-1 in developing erythroid and megakaryocyte cell lineages and cooperates with GATA-1 to control erythropoiesis. We describe a novel FOG-related factor, FOG-2, that is expressed predominantly in the developing and adult heart, brain, and testis. FOG-2 interacts with GATA factors, and interaction of GATA-4 and FOG-2 results in either synergistic activation or repression of GATA-dependent cardiac promoters, depending on the specific promoter and the cell type in which they are tested. The properties of FOG-2 suggest its involvement in the control of cardiac and neural gene expression by GATA transcription factors.

Amino Acid Sequence↗
PubMed1998

A calcineurin-dependent transcriptional pathway for cardiac hypertrophy.

In response to numerous pathologic stimuli, the myocardium undergoes a hypertrophic response characterized by increased myocardial cell size and activation of fetal cardiac genes. We show that cardiac hypertrophy is induced by the calcium-dependent phosphatase calcineurin, which dephosphorylates the transcription factor NF-AT3, enabling it to translocate to the nucleus. NF-AT3 interacts with the cardiac zinc finger transcription factor GATA4, resulting in synergistic activation of cardiac transcription. Transgenic mice that express activated forms of calcineurin or NF-AT3 in the heart develop cardiac hypertrophy and heart failure that mimic human heart disease. Pharmacologic inhibition of calcineurin activity blocks hypertrophy in vivo and in vitro. These results define a novel hypertrophic signaling pathway and suggest pharmacologic approaches to prevent cardiac hypertrophy and heart failure.

Angiotensin II↗
PubMed1995

Pollution by PCDDS and PCDFS in sediment from freshwater fish culture ponds near incineration sites for metal reclamation in Wan-Li, Taiwan, Republic of China.

Sediment samples from nine culture ponds for freshwater fish at and near waste incineration sites for metal reclamation in Wan-Li, southern Taiwan, Republic of China were analyzed for PCDDs and PCDFs by HRGC-HRMS. Four of nine samples were heavily polluted by PCDDs and PCDFs, indicating the latter to be dominant. The total TEQ concentrations of PCDDs and PCDFs in the four samples were in the range of 257 to 12200 pg/g dry weight. In addition, the levels corresponded to those of surface soil samples from the same waste combustion areas.

Animals↗
PubMed1995

[Hypoxia induced increase of MDA and echinocytes from erythrocytes in rabbit's blood with special reference to inhibition of the increase by MPEG-SOD].

When rabbits acutely exposed to hypobaric hypoxia (at imitative altitude 5,000 m, PaO2 = 6.7 kPa), the percentage of the activity of superoxide dismutase (SOD in erythrocyte; EC 1.15.1.1) decreased respectively to 86%, 76%, 81%, 84%, 55%, 81%, 84% and 95% for a period of 0.25, 0.5, 1, 2, 3, 4, 5 and 24 h hypoxia while MDA (malondialdehyde) in erythrocyte and plasma increased significantly during hypoxia periods of 1, 2, 3, 5 and 24 h (n = 5, P < 0.05, P < 0.01 vs. normoxia group). The increase of MDA in plasma could be prevented by vitamin E (V. E, i.p, 1 mg daily for 3 consecative days before hypoxia), but that in erythrocyte did not show significant change probably because the injected V.E. without vehicle cannot penetrate through the erythrocyte membrane effectively. The echinocytes from erythrocytes increased significantly after hypoxia for 24 h (n = 5, P < 0.05 vs. normoxia group). However, by injecting MPEG-SOD (monomethoxypolythlene glycol-SOD) intravenously 30 min before hypoxia, the increase of MDA (both of erythrocyte and plasma) and echinocytes (from erythrocytes) were both inhibited obviously during hypoxia for 24 h (n = 5, P < 0.05, P < 0.01 vs. hypoxia group). It is inferred that the increase of MDA and echinocytes from erythrocytes may be due to lipid peroxidation by superoxide free radicals (O2-.) generated in erythrocytes through Fenton pattern Haber-Weiss reaction during hypobaric hypoxia.

Altitude↗
PubMed1992

[Morphologic and biochemical characteristics of viridans streptococci isolated from dental plaque].

The purposes of this study were to give the characteristics of viridans streptococci isolated from dental plaque, and the relationships with dental caries. We stained 69 patients' teeth on supragingival plaques (Plaque Index = 1) with the Pfiffer's reagent or FD&C RED#28 dye, then isolated 80 strains of oral streptococci from 69 patients. Cultures were grown in TSB, MS, and MSB media. A total of 80 fresh-isolated plaque streptococci, including strains of S. salivarius, S. sanguis, S. mitis, and S. mutans were identified on the basis of morphologic, dark field, SEM, and biochemical characteristics. All of the oral plaque streptococci were to be found in significant percentages in dental plaque. Patients who had caries harboured S. mutans more often (37/40) than patients without caries (7/29). S. mutans comprised the greatest percentage (37/69) of microorganisms encountered in caries' plaque, and there was an association between high counts of S. mutans and the presence of dental caries. Cultural and biochemical methods were used for the isolation and identification of the different species of S. mutans. The result indicated that biotype I were the most predominant species (50%) in dental plaque. Thus, it is possible to isolate, screen and identify unknown strains through the morphologic, darkfield, SEM, and biochemical characteristics of viridans streptococci isolated from human dental plaque. By means of the present methods for identification, we hold considerable promises for contributing to the prevention and diagnosis of dental diseases.

Adolescent↗
PubMed1989

Identification of the types of adenovirus isolates with monoclonal antibodies.

The types of 59 isolates of adenovirus (Adv) which were isolated between winter, 1984 and spring, 1986 were identified by type-specific monoclonal antibodies (McAb) against Adv types 3 and 7. The results showed that the method could not only identify the types of Adv, but also discover the changes of their subtypes with time. The tedious neutralization test can be replaced by this simple method.

Adenoviruses, Human↗