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The radial protein concentration profile in the interstitial space of the rat ileal mesentery.

The radial distribution of the protein concentration in the interstitium between arteriolar and venular vessels of the ileal mesentery of the rat was examined. Protein mass was determined by means of uv ultramicrospectrophotometry (UMS) and the relative volume distribution by means of fluorescence microscopy (FM) using the Na fluorescein and FITC-dextran (10,000 mol wt). UMS revealed gradients for protein mass from the vessels out into the interstitial space. FM showed a uniform distribution of fluorescence in the interstitium between the vessels. A gradient for protein mass without a gradient for volume distribution signifies the presence of a concentration gradient for protein in the interstitial space. The protein concentration across the arteriolar wall drops from 5.4 +/- 0.24 (SD) to 2.6 +/- 0.65% and across the venular wall from 5.4 +/- 0.24 to 3.3 +/- 0.43%. From the perivascular site the protein concentration declines exponentially reaching a minimum average interstitial concentration of 1.6 +/- 0.56%. Minimal protein concentration occurred at a point 37 +/- 6.4% of the 295 +/- 37 micron distance from the arteriolar to the venular vessels. In view of this distribution, it is unlikely that lymph or direct samples of interstitial fluid are representative of the perivascular protein concentration.

Animals↗

Effect of ArF-193 nm excimer laser on human dentinal tubules. A scanning electron microscopic study.

The purpose of this study was to evaluate the effects of the ArF-193 nm excimer laser on the dentinal tubules of extracted human teeth under a scanning electron microscope. Fifteen 3 mm thick slices were cut with an electric saw at the cementoenamel junction from 15 extracted human teeth. A diamond bur was used to remove the cementum layer and expose the dentinal tubules. Each slice was scored by a permanent marker into four equal quadrants. The ArF excimer laser was applied for 5 seconds on three of the quadrants with fluences that ranged from 0.2 J/cm2 to 15 J/cm2 and pulse repetition of 25 Hz. The untouched quadrant served as a control. The specimens were mounted on stubs, sputter coated by gold, and examined by a scanning electron microscope. The effects of the ArF excimer laser irradiation varied. Laser fluences of 0.2, 0.5, and 1.0 J/cm2 had no effect. Although fluence of 15 J/cm2 caused significant removal of peritubular dentin, melting and resolidification of the dentinal smear layer was also observed under the scanning electron microscope with a laser fluence of 5 J/cm2.

Argon↗

Apparent relationships between toxins elaborated by the cyanobacterium Trichodesmium erythraeum and those present in the flesh of the narrow-barred Spanish mackerel Scomberomorus commersoni.

The marine cyanobacterium Trichodesmium erythraeum contains toxic water-soluble material that produces signs in mice similar to those produced by water-soluble extracts of the flesh of a specimen of pelagic fish Scomberomorus commersoni from a batch that had been implicated in a poisoning resembling ciguatera. Extracts of water-soluble material from both the cyanobacterium and the fish contained toxins that were chromatographically indistinguishable. A peptide and an alkaloid were detected in partially purified extracts of the water-soluble material. In addition to this material toxic lipid-soluble material was present in some batches of T. erythraeum. Elution of this material with 9:1 chloroform:methanol using column chromatography produced material that was chromatographically indistinguishable from ciguatoxin-like material from S. commersoni and produced signs in mice similar to those produced by this material. Elution of the lipid-soluble material with 97:3 chloroform:methanol yielded a toxin resembling in its chromatographic and toxic properties a scaritoxin-like substance from S. commersoni. Other toxins with Rf values lying between that of the ciguatoxin-like material and that of the scaritoxin-like material were also detected in extracts of T. erythraeum. It is postulated that T. erythraeum is the progenitor of major toxins carried by some ciguateric fish and that water-soluble toxins released into the ambient sea water by T. erythraeum may constitute a health hazard for humans.

Animals↗

Microporous hydrophilic polyurethane vascular grafts as substitutes in the abdominal aorta of dogs.

Polyurethanes are emerging as promising biomaterials. A microporous vascular graft fabricated from Mitrathane, a new polyetherurethane urea, appeared to be particularly interesting according to in vitro evaluation and was tested in vivo as an infrarenal aortic substitute (i.d. 5 mm) in 24 dogs. After implantation for scheduled periods ranging from 4 h to 6 mnth, morphology evaluation and healing analysis of the grafts were performed. At harvesting, 18 grafts were patent and 6 were thrombosed. The thrombosed grafts had been implanted for 4 h (1 graft), 1 mnth (2 grafts) and 6 mnth (3 grafts). No macroscopic deposits other than red mural thrombi were observed. At 1 mnth complete external encapsulation was observed in 2 grafts. The capsule then contracted and became thinner; over the long term it shrank, became translucent and incomplete at 6 mnth. Yellow and brown stains appeared on many of the grafts. Characterization of the brown stains showed them to be iron (Fe3+) and the yellow stains were associated with the deposition of a bile pigment (bilirubin).

Animals↗

Separation and analysis of the major forms of plasma fibronectin.

Human plasma fibronectin exists in circulation in multiple molecular forms that are distinguishable by SDS-polyacrylamide gel electrophoresis (zone I, approx. 450 kDa dimers; zone II, 190-235 kDa; Zone III, 146-175 kDa). (Chen, A.B., Amrani, D.L. and Mosesson, M.W. (1977) Biochim. Biophys. Acta 493, 310-322). We report here on investigations of plasma fibronectin that had been purified from the 'heparin-precipitable fraction' of plasma by DEAE-cellulose chromatography using buffers containing a chaotropic salt (KSCN). Zone I fibronectin and zone II fibronectin were subsequently separated by Sepharose CL-6B chromatography in the presence of 0.3 M KSCN. Electrophoresis of reduced zone I fibronectin dimers showed the presence of three types of subunits (i.e., 220 kDa, 215 kDa, 207 kDa), evidently all having the same NH2-terminal sequence. Subunits of this size were also found in reduced zone II fibronectin, as well as another polypeptide of 190 kDa, the latter amounting to under 5% of the total. Unreduced zone I fibronectin was resolved by gel electrophoresis into a doublet. The upper component amounted to approx. 90% of the total and was comprised of 220 kDa and/or 215 kDa subunits; the lower component contained 207 kDa plus a 220 kDa or 215 kDa subunit. Scanning transmission electron microscopy indicated that under physiologic conditions zone II fibronectin molecules, like those in zone I, exist as pleiomorphic, loosely folded structures (approx. 16 X 8-12 nm) that are somewhat smaller than dimeric zone I molecules (approx. 24 X 16 nm). Circular dichroic spectral analyses suggests that both types have similarly folded local domains. Affinity chromatography experiments revealed a relative decrease in the binding of zone II fibronectin to gelatin but no difference from zone I fibronectin with respect to heparin or fibrin binding.

Amino Acids↗

Prophage distribution in coryneform bacteria.

Four temperate bacteriophages of corynebacteria were isolated after UV induction. Phages phi 304L and phi 304S were both induced from Corynebacterium glutamicum ATCC 13058, ATCC 21488, ATCC 21649 and ATCC 21650 strains, and have no known sensitive host. Phages phi 15 and phi 16 were both induced from ATCC 14020 and ATCC 21792. Phage phi 15 formed turbid plaques on Corynebacterium sp. ATCC 21857 and on C. glutamicum ATCC 13058, ATCC 21488, ATCC 21649 and ATCC 21650. Phage phi 16 produced turbid plaques only on C. glutamicum ATCC 21792 cured of prophage phi 16. All these phages belong to the Siphoviridae family. Their genomes consist of a double-stranded DNA with cohesive ends and share no homology with each other. Prophages phi 16, phi 304L and phi 304S were integrated into their respective host chromosomes, whereas prophage phi 15 seemed to persist free in the cell. Cross-hybridizations between phage DNAs and total cellular DNA obtained from 20 strains belonging to the genera Corynebacterium and Brevibacterium did not show the presence of these prophages in strains other than their respective hosts.

Bacteriophages↗

UV-B-laser-induced DNA damage in lymphocytes observed by single-cell gel electrophoresis.

Human lymphocytes from a cell culture were irradiated with 12 mJ pulses (524 J m-2) of an XeCl excimer laser (308 nm). DNA strand breaks were analysed with the "comet assay", a single-cell method. The DNA damage of individual cells could be investigated depending on the energy and the irradiated volume. DNA damage could be detected already after a single laser pulse of 12 mJ. Since the irradiation time is short compared with the DNA repair time, DNA damage is solely dependent on the total number of photons. A quantitative evaluation of the number of damaged cells vs. number of photons can be best explained by a detection threshold model. The calculated threshold is 1.25 x 10(8) photons cell-1. From this and the principal sensitivity of the comet assay, one can estimate that the upper limit for DNA damaging efficiency is 400,000 photons.

Cells, Cultured↗

Interfacing myoglobin to graphite electrode with an electrodeposited nanoporous ZnO film.

A biocompatible, nanoporous ZnO film was prepared on graphite electrode by the simple electrodeposition method. Based on the film's strong adsorption ability and friendly microenvironment, it can be used as a good matrix to immobilize myoglobin (Mb) through simple adsorption. Moreover, the entrapped Mb realized direct electron transfer with the electrode and displayed an elegant catalytic activity toward the reduction of hydrogen peroxide, nitrite, and trichloroacetic acid, by which the mediator-free biosensors could be fabricated. Atomic force microscopy, UV-Vis spectra, electrochemical impedance spectroscopy, and cyclic voltammetry, etc. were used to characterize the nanoporous ZnO film and Mb-modified ZnO film. Compared to other methods, electrodeposition of porous material supplied a more simple and convenient approach to prepare biocompatible materials for biosensors.

Adsorption↗

Trifunctional norrisolide probes for the study of Golgi vesiculation.

Inspired by the effect of norrisolide on the Golgi complex, we synthesized norrisolide probes that contain: the perhydroindane core of the parent natural product for Golgi localization, a crosslinking unit (aryl azide or epoxide) for covalent binding to the target, and a tag (biotin or iodine) for subsequent target purification. We found that biotin-containing probes 14, 20 and 24 induced inefficient Golgi vesiculation. However, the iodinated probe 25 induced extensive and irreversible Golgi fragmentation. This probe can be used for the isolation of the cellular target of norrisolide.

Animals↗

Photodegradation of Diquat and Paraquat in aqueous solutions by titanium dioxide: evolution of degradation reactions and characterisation of intermediates.

The titanium dioxide assisted photodegradation of Diquat and Paraquat herbicides solutions has been the subject of the present investigation, considering its direct application in the treatment of contaminated waters and soils. To have a better understanding of the photodegradation process, different types of TiO2, commercial and 'home prepared' Ti(1-x)FexO2 (x = 0% and 4%), were used as catalysts, using an UV light as radiation source. The degradation reactions were followed by UV spectroscopy and the intermediates and reaction products were characterised by electrospray ionisation mass spectrometry (ESIMS) combined with collision-induced dissociation (CID) and tandem mass spectrometry (MS/MS). The present study shows that, for photocatalytic degradation of Diquat and Paraquat solutions, a basic pH can be determinant, as well as the type of catalyst. The type of catalyst can also strongly influence the degradation pattern of the herbicide. Regarding complete degradation, we were able to show that Diquat is more persistent than Paraquat. During the photocatalytic processes, several intermediate and reaction products are sequentially formed, to which structures are proposed.

Catalysis↗

On-chip electrochromatography using sol-gel immobilized stationary phase with UV absorbance detection.

A chromatography column on a chip was fabricated by immobilizing reversed-phase stationary phase particles (5 microm, C4) using sol-gel technology. Channels were fabricated in quartz using photolithography and wet etching. Localization of the stationary phase was achieved by immobilizing the stationary phase at the desired location in the separation channel prior to bonding of the cover plate. Cross channel design was employed for gated injection. An optical fiber setup was developed for carrying out on-chip UV absorbance detection. The effective optical path length was theoretically determined for the trapezoidal shaped channel and the result was shown to match closely with the experimentally determined value. The effect of applied voltage on velocity was evaluated using thiourea as an unretained marker. Separation performance of the stationary phase was demonstrated by separation of three peptides (Trp-Ala, Leu-Trp and Trp-Trp) under isocratic chromatographic conditions.

Chromatography, Micellar Electrokinetic Capillary↗

Sun protection enhancement of titanium dioxide crystals by the use of carnauba wax nanoparticles: the synergistic interaction between organic and inorganic sunscreens at nanoscale.

Carnauba wax is partially composed of cinnamates. The rational combination of cinnamates and titanium dioxide has shown a synergistic effect to improve the sun protection factor (SPF) of cosmetic preparations. However, the mechanism of this interaction has not been fully understood. In this study, an ethanolic extract of the carnauba wax and an ethanolic solution of a typical cinnamate derivative, ethylcinnamate, were prepared and their UV absorption and SPF either alone or in the presence of titanium dioxide were compared. The titanium dioxide crystals and the cinnamates solutions were also distributed into a matrix composed of saturated fatty acids to emulate the structure of the crystallized carnauba wax. SPF, differential scanning calorimetry (DSC) and X-ray studies of these matrices were performed. Additionally, carnauba wax nanosuspensions containing titanium dioxide either in the lipid phase or in the aqueous phase were prepared to evaluate their SPFs and their physical structure. Strong UV absorption was observed in diluted suspensions of titanium dioxide after the addition of cinnamates. The saturated fatty acid matrices probably favored the adsorption of the cinnamates at the surface of titanium dioxide crystals, which was reflected by an increase in the SPF. No modification of the crystal structure of the fatty acid matrices was observed after the addition of cinnamates or titanium dioxide. The distribution of the titanium dioxide inside the lipid phase of the nanosuspensions was more effective to reach higher SPFs than that at the aqueous phase. The close contact between the carnauba wax and the titanium dioxide crystals after the high-pressure homogenization process was confirmed by transmission electron microscopy (TEM).

Calorimetry, Differential Scanning↗

Morphology characterization of layer-by-layer films from PAH/MA-co-DR13: the role of film thickness.

We report on the use of dynamic scale theory and fractal analyses in the study of distinct growth stages of layer-by-layer (LBL) films of poly(allylamine hydrochloride) (PAH) and a side-chain-substituted azobenzene copolymer (Ma-co-DR13). The LBL films were adsorbed on glass substrates and characterized with atomic force microscopy with the Ma-co-DR13 at the top layer. The granular morphology exhibited by the films allowed the observation of the growth process inside and outside the grains. The growth outside the grains was found to follow the Kardar-Parisi-Zhang model, with fractal dimensions of ca. 2.6. One could expect that inside the grains the morphology would be close to a Euclidian surface with fractal dimension of ca. 2 for any growth stage. The latter, however, was observed only for thicker films containing more than 10 bilayers. For thinner films the morphology was well described by a self-affine fractal. Such dependence of the growth behavior with the film thickness is associated with a more complete coverage of adsorption sites in thicker films due to diffusion of polymer molecules.

Azo Compounds↗

Polymer micelles with cross-linked ionic cores for delivery of anticancer drugs.

This work reports the design of polymer micelles with cross-linked ionic cores that display high stability. Block ionomer complexes of poly(ethylene oxide)-b-poly(methacrylic acid) copolymer and divalent metal cations were utilized as micellar templates for the synthesis of the cross-linked micelles. Such micelles represent hydrophilic nanospheres of core-shell morphology. The core comprises a network of the cross-linked polyanions, which is surrounded by the shell of hydrophilic PEO chains. The ionic character of the core provided for pH-dependent swelling/collapse behavior of the nanogels. Cisplatin, a potent chemotherapeutic agent, was incorporated into the ionic core of the micelles with remarkably high efficiency (22% w/w). The drug-loaded micelles were stable in aqueous dispersions exhibiting no aggregation or precipitation for a prolonged period of time. Slow release of platinum complexes was observed in sustained manner from the cisplatin-loaded cross-linked micelles in physiological saline. In vitro studies using human A2780 ovarian carcinoma cells demonstrated that the cross-linked micelles rapidly internalized and delivered cisplatin into cells. These results indicated that polymer micelles with cross-linked ionic cores are promising for further fundamental material studies and practical applications as drug delivery carriers.

Antineoplastic Agents↗