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PubMed2002

Particle transport in unsaturated fractured chalk under arid conditions.

A series of field and laboratory experiments were conducted to study the mechanisms of particle detachment and transport from fractures in vadose chalk. Experiments of intermittent flow events along fracture surfaces were carried out in the laboratory. In the field, water was percolated from land surface via a discrete fracture into a compartmental sampler installed inside a horizontal corehole located I m below the surface. The mass, size distribution, and composition of the particles drained from the fracture voids were examined along with flow rates and salt dissolution. Two boreholes penetrating the underlying saturated zone were sampled and analyzed for colloidal concentration and composition. Most of the particle and solute release at the drained effluents occurred during the first several hours of flow, but erratic pulses of particles were still observed after long periods of time. Most of the detached particles had a mean diameter of >2 microm, while the mobile colloidal phase in the groundwater had a mean diameter of approximately 1 microm. Mineralogical composition of the groundwater colloids and the particles detached from the upper vadose fracture were similar. Laboratory observations demonstrated the importance of the existence of a coating layer, made of weathered particles and salts, on particle detachment. The results of this study suggest that: (1) particle detachment causes flow-rate variability in the unsaturated fracture; (2) the mechanisms of particle detachment and salt dissolution within the fracture are linked: and (3) although most of the detached particles are large and likely to accumulate inside fractures, some colloidal particles also eroded from the fracture void and are likely to be transported to the groundwater.

Calcium Carbonate↗
PubMed2003

Isotopes as tracers in a contaminated fractured chalk aquitard.

Clusters of industrial plants often generate contaminant plumes with several potential sources. Prevention of further pollution and designing suitable remedial measures require identification of the contributing source among all potential ones and the sorting of currently active sources from historical ones. In the study area, an industrial complex in the Negev desert, Israel, contaminants could not serve as indicators for the contamination sources because of their extensive spatial distribution across the site. However, stable isotopes of oxygen, hydrogen and sulfur, as well as tritium, proved to be efficient tools for this task. The isotopic characterization of the potential end members provided the criteria for constraining a contaminating source when several alternative sources appeared viable. The isotopic fractionation of oxygen and hydrogen isotopes could be tied to the various disposal phases of the industrial wastewater. The three case studies presented here confirm the important role of isotopes as tracers in contaminated sites.

Environmental Monitoring↗
PubMed1986

Partial purification and characterization of a procollagen C-proteinase from the culture medium of mouse fibroblasts.

A procollagen C-proteinase was purified about 100-fold from the medium of cultured mouse fibroblasts by a combination of ammonium sulfate precipitation, gel-filtration, and affinity chromatography on a column of Sepharose coupled to the carboxyl propeptide of type I procollagen. The purified enzyme did not exhibit other proteolytic activities, and it cleaved type I, II and III procollagens to produce the corresponding pN alpha chains and carboxyl propeptides as the only products. Amino acid sequencing of the first 14-18 residues at the N-terminus of the carboxyl propeptides generated by the enzyme from human pro alpha 1(I), pro alpha 2(I) and pro alpha 1(III) chains showed that the cleavage occurred at the physiological site, i.e. at the specific Ala-Asp bond in the pro alpha 1(I) and pro alpha 2(I) chains, and at the specific Gly-Asp bond in the pro alpha 1(III) chain. The pH optimum of the enzyme is 8.5 and its molecular weight as estimated by gel-filtration is about 125,000 daltons. The enzyme is inhibited by metal-chelators, various amines, dithiothreitol, N-ethylmaleimide and serum, but it is insensitive to pepstatin, leupeptin and serine proteases inhibitors. The enzyme differs from the C-proteinase described by Njieha et al. (Biochemistry 21:757-764, 1982), and the catheptic activities reported by Davidson et al. (Eur. J. Biochem 100:551-558, 1979) and Helseth and Veis (Proc. Natl. Acad. Sci. USA 81:3302-3306, 1984). The specificity of the enzyme is offered as evidence for a unique, C-proteinase, and its recovery from culture medium supports an extracellular location for procollagen processing.

Amino Acid Sequence↗
PubMed1986

Evidence for a protein that enhances the activity of type I procollagen C-proteinase.

Gel-filtration separated type I procollagen C-proteinase from a glycoprotein that enhanced the enzyme activity by approximately 4-fold. The enhancer was purified by affinity chromatography on a column of Sepharose coupled to the carboxyl propeptide of type I procollagen. Sodium-dodecyl-sulfate- polyacrylamide gel electrophoresis of the affinity-purified enhancer revealed two active major protein bands with molecular weights of 36 and 34 kdal. Both proteins were glycosylated, as shown by binding to concanavalin-A. The enhancer is extremely heat stable (100 degrees C, 15 min) but its activity is totally abolished by treatment with trypsin or bacterial elastase. The enhancer does not alter the digestion intermediates or final products of the enzymatic reaction but it changes the kinetic properties of the reaction, increasing the apparent Km and Vmax values 16- and 20-fold, respectively. It is suggested that the enhancer might play a regulatory role in procollagen processing.

Animals↗
PubMed1996

Characterization of normal and malignant human hepatocytes by Raman microspectroscopy.

Raman microspectroscopy was used to characterize normal and malignant hepatocytes in both cultured cells and human liver tissues. Consistent spectral changes were observed, including intensity increases at 1040 and 1083 cm(-1) with malignancy. A loss of intensity at 1241 cm(-1) was also observed in cancer cells, but was obscured in tissues by the overlap of a 1253 cm(-1) band, thought to originate from heme proteins. Normal liver tissue also differed from both the malignant tumor and its accompanying cirrhotic tissue at 1182 cm(-1). These results demonstrate the potential usefulness of Raman spectroscopy in clinical diagnosis, and investigations into the source of the observed spectral changes will provide information on the underlying mechanisms of carcinogenesis.

Carcinoma, Hepatocellular↗
PubMed1994

Modulation of the fibrinolytic system by major peripheral ischemia.

PURPOSE: A rat model was developed to investigate the effects of acute peripheral ischemia on the components of the fibrinolytic system. METHODS: Laparotomy was performed and ischemia was introduced by total aortic clamping at a subrenal position. Control animals underwent sham laparotomy alone. Plasma and tissue samples were collected for analysis at 30, 60, 90, and 120 minutes after operation. RESULTS: Functional assays of rat plasma revealed a dramatic and transient increase in tissue-type plasminogen activator (tPA) activity within 30 minutes of the onset of ischemia. A simultaneous decline in plasminogen activator inhibitor activity was observed. Immunohistochemical analysis suggested this initial increase in tPA activity resulted primarily from the release of stored tPA from ischemic vascular tissues. Northern blot analysis revealed that both tPA and plasminogen activator inhibitor-1 messenger RNA levels were elevated at 60 to 120 minutes in well-perfused tissues distant from the ischemic insult. CONCLUSIONS: Collectively, these data demonstrate that acute peripheral ischemia results in a rapid and transient increase in plasma fibrinolytic activity, concomitant with the early release of stored tPA from ischemic vascular tissues. In addition, peripheral ischemia appears to stimulate both tPA and plasminogen activator inhibitor-1 gene expression in well-perfused tissues at later time points, consistent with the existence of humoral mediators.

Adipose Tissue↗
PubMed1996

A point scoring system for the clinical diagnosis of Buerger's disease.

INTRODUCTION: The literature on thromboangiitis obliterans (TAO, Buerger's disease) suffers from the lack of a unified method of establishing the diagnosis of the disease. The aim of this study was to test the application of a point scoring system (PSS) in the diagnosis of TAO. METHOD: Points are awarded for young age at onset, foot claudication, upper extremity involvement, superficial vein thrombosis and vasospastic phenomena. Atypical features detract points, and the resultant score classifies the diagnosis of TAO as being of low, medium or high probability. One hundred and seven patients diagnosed and classified according to our previous accepted criteria for TAO, were independently reclassified by the PSS, and the results of the two diagnostic processes were compared. RESULTS: Of the 107 patients diagnosed by our old criteria (OC) the diagnosis of TAO was rejected by the PSS in 20 patients. Of the remaining 87 patients the degree of certainty in the diagnosis (PSS vs. OC) was lower in 31 equal in 47 and higher only in nine. CONCLUSION: The proposed PSS is more discriminating than our OC. If this type of scoring system can be agreed upon the certainty in the diagnosis of TAO would be increased. Data collection would be improved, and the cause of clinical and basic research would be advanced.

Adult↗
PubMed1981

Resonance Raman spectra of cytochromes c and b in Paracoccus denitrificans membranes: evidence for heme--heme interactions.

Resonance Raman (RR) scattering from cytochromes b and c in the bacterium Paracoccus denitrificans was recorded by exciting with the 568.2-, 530.9-, and 520.8-nm lines of a Kr+ laser. The main features of the spectra were similar to those of the analogous cytochrome b-c1 complex derived from pigeon breast mitochondria. Differences in the 1300-cm-1 region were interpreted in terms of marker bands for heme type and spectral coupling of the hemes on the membranes. It is difficult to explain the results without invoking sharing of electronic and vibrational wave functions among the hemes. This conclusion documents the potential to study the physics of electron transport in functioning membrane by monitoring the RR spectra.

Bacterial Proteins↗
PubMed1979

Photoreductive titration of the resonance Raman spectra of cytochrome oxidase in whole mitochondria.

A photoreductive titration of the resonance Raman (RR) spectra of cytochrome c oxidase in whole mitochondria was recorded by exploiting the preferential enhancement of the Raman signals of reduced cytochrome oxidase excited at 441.6 nm. When the sample was cooled to about--10 degrees C, it was possible to slow down the photoreductive effect of the laser and to record RR spectra at various states of reduction. Compared to the earliest recorded scan (most oxidized), the dithionite-reduced sample shows the appearance of new bands at 216, 363, 560, and 1665 cm-1. At intermediate stages of photoreduction, the 216- and 560-cm-1 bands appear before the 363- and 1665-cm-1 bands; photoreduction induces full intensity in the former bands, whereas the latter bands are photoreduced to 50% of the dithionite-reduced intensity. The relative intensities of a doublet at 1609--1623 cm-1 are affected by reduction: the band at 1609 cm-1 is weaker in the earlier scans; in later scans this band has grown to equal intensity with the 1623-cm-1 band. We conclude that this reductive titration of the RR spectrum of cytochrome c oxidase reflects three states in its reduction. The behavior of the doublet at 1609--1623 cm-1 suggests that the two hemes are nonequivalent but interacting. The band at 216 cm-1 may be indicative of an iron-copper interaction that is affected by the presence of external ligands.

Animals↗
PubMed1978

Resonance Raman spectra of whole mitochondria.

The resonance Raman spectra of reduced cytochromes b and c and cytochrome oxidase in whole mitochondria have been recorded without any instrument modifications. The contributions of the individual cytochromes have been identified by comparison with the characteristic features observed in partially purified preparations including: (i) the strong dependence of the intensity patterns on excitation wavelength relative to the peak positions of the alpha, beta, and gamma absorption bands of the cytochromes; and (ii) the presence of marker bands for heme type. Since the Raman spectra can be used as an intrinsic indicator of interaction between hemes, the ability to record spectra in intact mitochondria opens the possibility to study heme-heme interactions in the functioning membrane in situ.

Animals↗
PubMed1997

Redefinition of the carbohydrate specificity of Erythrina corallodendron lectin based on solid-phase binding assays and molecular modeling of native and recombinant forms obtained by site-directed mutagenesis.

Binding of the N-acetyllactosamine-specific lectin from Erythrina corallodendron (ECorL) to four glycosphingolipids has been tested using the microtiter well assay. The role of several amino acids in the binding site region was studied by combining binding assays and molecular modeling for native and recombinant forms of the lectin. Seven single-point mutants at positions 106 (Y106A), 108 (Y108A, T), 218 (A218G), and 219 (Q219A, N or E) were investigated. A comparison with more than 30 known sequences of legume lectins showed that ECorL is unique in displaying a tyrosine residue or a structural equivalent at position 106. Analyses of the binding results obtained for mutants at positions 106 and 108 using molecular modeling point to complex conformational dependencies between these and several other residues around the binding site. Gln 219 was found to have a large conformational flexibility, which, paradoxically, favors the binding of N-acetyllactosamine-containing glycosphingolipids. Particularly significant is the fact that ECorL exhibits a higher affinity for Fuc alpha2Gal beta4GlcNAc beta-terminated glycosphingolipids than N-acetyllactosamine-terminated ones, in accordance with molecular modeling revealing a perfect fit of the alpha2-linked fucose in a cavity extending from the Gal beta4 binding pocket. These findings lead to a redefinition of the specificity of this lectin, where the affinity for the terminal Fuc alpha2Gal beta4GlcNAc beta trisaccharide should be considered in the first place. The possible biological significance of this specificity remains to be investigated.

Adsorption↗
PubMed2005

Biodegradation of 2,4,6-tribromophenol during transport in fractured chalk.

The effect of physicochemical conditions (residence time, oxygen concentrations, and chalk characteristics) on the biodegradation of 2,4,6-tribromophenol (TBP) during transport was investigated in low-permeability fractured-chalk cores. Long-term (approximately 600 d) biodegradation experiments were conducted in two cores (approximately 21 cm diameter, 31 and 44 cm long, respectively), intersected by a natural fracture. TBP was used as a model contaminant and as the sole carbon source for aerobic microbial activity. Bacterial isolates were recovered and identified by both Biolog identification kit and 16S rDNA sequences from batch enrichment cultures. One of the strains, with 98% similarity (based on the 16S rDNA data) to Achromobacter xylosoxidans, was shown to have the ability to degrade TBP in the presence of chalk. The decrease in TBP concentration along the fracture due to biodegradation was not affected by reducing the residence time from 49 to 8 min. In contrast, adding oxygen to the water at the inlet and increasing the flow rates improved TBP removal. Although the matrix pore-size distribution limits microbial activity to the fracture void, the chalk appears to provide an excellent environment for biodegradation activity. Approximately 90% of TBP removal occurred within 10 cm of the TBP source, indicating that in-situ bioremediation can be used to remove organic contaminants in low-permeability fractured rocks if nutrient-delivery pathways within the aquifer are secured.

Biodegradation, Environmental↗
PubMed2005

Chemical transformation of 3-bromo-2,2-bis(bromomethyl)-propanol under basic conditions.

The mechanism of the spontaneous decomposition of 3-bromo-2,2-bis(bromomethyl)propanol (TBNPA) and the kinetics of the reaction of the parent compound and two subsequent products were determined in aqueous solution at temperatures from 30 to 70 degrees C and pH from 7.0 to 9.5. TBNPA is decomposed by a sequence of reactions that form 3,3-bis(bromomethyl)oxetane (BBMO), 3-bromomethyl-3-hydroxymethyloxetane (BMHMO), and 2,6-dioxaspiro[3.3]-heptane (DOH), releasing one bromide ion at each stage. The pseudo-first-order rate constant of the decomposition of TBNPA increases linearlywith the pH. The apparent activation energy of this transformation (98+/-2 KJ/mol) was calculated from the change of the effective second-order rate constant with temperature. The pseudoactivation energies of BBMO and BMHMO were estimated to be 109 and 151 KJ/mol, respectively. Good agreement was found between the rate coefficients derived from changes in the organic molecules concentrations and those determined from the changes in the Br- concentrations. TBNPA is the most abundant semivolatile organic pollutant in the aquitard studied, and together with its byproducts they posess an environmental hazard. TBNPA half-life is estimated to be about 100 years. This implies that high concentrations of TBNPA will persist in the aquifer long after the elimination of all its sources.

Half-Life↗
PubMed2004

Computational studies on the electrocyclizations of 1-amino-1,3,5-hexatrienes.

Electrocyclizations of 1,3,5-hexatrienes containing up to four electron-donating and/or electron withdrawing substituents have been studied computationally using the hybrid density functional, B3LYP. Electron donating substituents at positions C-1 and C-5 decrease activation barriers by 0.3 to 2.3 kcal/mol. Introducing of an electron-withdrawing group, CO(2)Me, at C-4 further decreases the activation energy by 7 kcal/mol. Electron-withdrawing groups (NO(2), SO(2)Ph and C=N(+)Me(2)) at C-2 have a profound effect of 17-25 kcal/mol on the activation energy.

Journal Article↗
PubMed2005

Self-motion perception during locomotor recalibration: more than meets the eye.

Do locomotor aftereffects depend specifically on visual feedback? In 7 experiments, 116 college students were tested, with closed eyes, at stationary running or at walking to a previewed target after adaptation, with closed eyes, to treadmill locomotion. Subjects showed faster inadvertent drift during stationary running and increased distance (overshoot) when walking to a target. Overshoot seemed to saturate (i.e., reach a ceiling) at 17% after as little as 1 min of adaptation. Sidestepping at test reduced overshoot, suggesting motor specificity. But inadvertent drift effects were decreased if the eyes were open and the treadmill was drawn through the environment during adaptation, indicating that these effects involve self-motion perception. Differences in expression of inadvertent drift and of overshoot after adaptation to treadmill locomotion may have been due to different sets of ancillary cues available for the 2 tasks. Self-motion perception is multimodal.

Adaptation, Psychological↗