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PubMed2006

Optical and chemical methods of metal ash analysis and tin recovery.

'Metal ash' presents a waste disposal problem in most of the developing countries as the industries employ obsolete technologies. In this paper we describe analysis of tin ash, zinc ash and aluminium ash by means of optical methods, such as X-ray diffraction (XRD), inductively coupled plasma mass spectrometry (ICP-MS), electron probe micro analysis (EPMA), scanning electron microscopy (SEM) and chemical methods. The results of tin ash obtained by XRD method matched well with the cassiterite, a naturally occurring mineral of tin. ICP-MS studies reveal the presence of a large number of tracer metals, which may cause pollution by tertiary dispersion and this aspect is discussed. Conversely, the data generated by chemical methods are limited. However, the methods are simple and cost-effective. Then, they can easily be adopted by low-budget industries. Simple and cost-effective process to recover tin from tin ash is described. It is based on heating tin ash with sodium cyanide to about 900 degrees C to separate tin component from the metal ash. The process recovers good quality tin and offers a very high yield. The process can be scaled up to small pilot plant.

Environmental Monitoring↗
PubMed2004

Distribution of caveolin-1 in bovine lens and redistribution in cultured bovine lens epithelial cells upon confluence.

The distribution of caveolin-1 in the lens and lens epithelial cells was determined to assess possible roles in cholesterol trafficking, cell to cell communication and signal transduction. Bovine lenses and cultured bovine lens epithelial cells (BLEC) were divided into subcellular fractions and the distribution of proteins recognized by three different caveolin-1 antibodies determined. The immunolocalization of caveolin-1 in the lens epithelium and in subconfluent and confluent cultured BLEC was probed by fluorescence microscopy and laser scanning confocal microscopy. EGF induced phosphorylation of caveolin-1 was detected by Western blotting with an anti-phosphotyrosine antibody to immunoprecipitated caveolin-1 from BLEC and human cancer cells. Monomeric caveolin-1 of about 26 kDa was detected in the epithelial cell membrane of cultured BLEC and fresh epithelia and in the plasma membrane fraction of lens cortical fiber cells. Caveolin-1 of cultured BLEC redistributed from the cytoplasm to plasma membrane as the cells proceeded from subconfluent to confluent states. The apparent abundance of caveolin-1 in cortical fiber cell plasma membrane is consistent with possible roles in distribution of lens membrane cholesterol and membrane structure. The presence of caveolin-1 in the plasma membrane of epithelial cells at - but not before - confluency is consistent with a role of caveolin-1 in cell to cell communications. EGF stimulated phosphorylation of caveolin-1 in human A431 cells but not lens cells.

Animals↗
PubMed2007

Babesia bovis: subcellular localization of host erythrocyte membrane components during their asexual growth.

In the present study, the subcellular localization of the host red blood cell (RBC) membrane components, the alpha2-3-linked sialic acid (SA) residues and the lipid bilayer, was observed during the asexual growth of Babesia bovis using two erythrocyte probes, the SA-specific lectin (MALII) and the lipophilic fluorescent (PKH2) probes, respectively. In confocal laser scanning microscopy with MALII, the SA residues on the surface of parasitized RBCs appeared to accumulate into the intracellular parasites as the parasites matured as well as to remain on the surface of extracellular parasites. Furthermore, when PKH2-labeled RBCs were infected with B. bovis, PKH2 signals were also observed around both the intracellular and the extracellular parasites, similarly to the results of MALII. These results indicated that the components derived from the host erythrocyte membrane are incorporated into the intracellular parasites during their asexual growth within the parasitized RBC, suggesting the possible formation of a parasitophorous vacuole-based network or a parasite surface coat.

Animals↗
PubMed2004

Mutagenicity of cadmium in mammalian cells: implication of oxidative DNA damage.

Cadmium and cadmium compounds are well established human carcinogens and are ubiquitously present in the environment. The carcinogenic mechanism(s) of cadmium remains largely unknown since direct mutagenic effect is weak in bacterial and in standard mammalian cell mutation assays. In this study, we show that when evaluated using the human-hamster hybrid A(L) cell mutation assay in which both intragenic and multilocus deletions can readily be detected, CdCl(2) is a strong mutagen that induces predominantly large deletion mutations. Concurrent treatment of A(L) cells with the oxyradical scavenger dimethyl sulfoxide significantly reduced the number of cadmium-induced mutations. In contrast, pre-treatment of cells with buthionine sulfoximine that depletes intracellular glutathione, increased cytotoxicity and mutagenicity of cadmium. These results demonstrate that reactive oxygen species mediate cadmium induced mutations in A(L) cells. With laser scanning confocal microscopy and the fluorescent probe 5-(and-6)-chloromethyl-2',7'-dichlorodihydrofluorescein diacetate, we demonstrated that cadmium induced a dose and time dependent formation of intracellular oxyradicals. Using immunoperoxidase staining coupled with a monoclonal antibody-specific for 8-OHdG adducts in DNA, we demonstrated that cadmium induced a dose dependent increase of 8-OHdG adducts, which accumulated with prolonged exposure. Furthermore, we showed that at low concentration, cadmium, attenuated removal of hydrogen peroxide induced 8-OHdG adducts. Thus, the carcinogenicity of cadmium can, in part, be explained by its mutagenic activity, which is mediated by reactive oxygen species induced DNA damage and by its interference with the repair of oxidative DNA damage.

Animals↗
PubMed1999

Three-dimensional measurement of cerebral microvascular plasma perfusion, glial fibrillary acidic protein and microtubule associated protein-2 immunoreactivity after embolic stroke in rats: a double fluorescent labeled laser-scanning confocal microscopic study.

Early astroglial response to post-ischemic microvascular hypoperfusion may contribute to progressive cerebral microcirculatory impairment and ischemic neuronal injury. Using laser-scanning confocal microscopy and three fluorescent probes, we measured in three-dimensions cerebral microvascular plasma perfusion, astrocytic reactivity, and neuronal injury assessed by fluorescein isothiocyanate (FITC)-dextran, GFAP immunoreactivity, and microtubule associated protein-2 (MAP2) immunoreactivity, respectively, in rats subjected to 2 h of middle cerebral artery occlusion. Three-dimensional quantitative analysis revealed that 2 h of embolic ischemia resulted in a significant (P<0.05) reduction of cerebral microvascular plasma perfusion in the ipsilateral cortex and subcortex. Tissue within the ipsilateral cortex and subcortex with low plasma perfusion exhibited a significant (P<0.05) increase in GFAP immunoreactivity compared with the homologous contralateral tissue. Three-dimensional re-constructed images showed that prominent GFAP immunoreactive astrocytes surrounded large vessels with decreased plasma perfusion in downstream capillaries in the ipsilateral MCA territory when compared to the vessels in the contralateral homologous tissue. Triple fluorescence probe-stained sections showed that tissue with decreased plasma perfusion and with increased GFAP immunoreactivity was accompanied by a reduction of MAP2 immunoreactivity. The present study demonstrates that an impairment of microvascular perfusion induces an early increase in GFAP immunoreactivity, and reactive astrocytes may contribute to a further reduction of cerebral microvascular plasma perfusion. The three-dimensional quantitative imaging analysis used in the present study provides a means to investigate parenchymal cellular responses to changes of cerebral microvascular plasma perfusion after MCA occlusion.

Animals↗
PubMed1994

Dietary xylitol prevents ovariectomy induced changes of bone inorganic fraction in rats.

Thirty-six 3-month-old female Wistar rats were labelled with a single intraperitoneal tetracycline injection. Twenty-four animals were subsequently ovariectomized, while the control group of 12 animals underwent sham operations. All animals received the basal Ewos R3 diet and half of the ovariectomized animals (n = 12) were given an additional 5% dietary xylitol supplementation. Three months later, following the collection of blood and urine, the animals were killed by decapitation. The tibiae were detached and prepared for chemical and other studies. The weight and density of the tibiae were measured. The right tibiae were dried and pulverized for chemical analysis of calcium, phosphorus and citric acid. The total inorganic fraction was determined by ashing the powdered bone. The left tibiae were cross-sectioned at the tibio-fibular junctions for the measurement of the width of periosteally formed bone, which was identified by tetracycline fluorescence. The examination of mineral content of bone was performed by scanning electron microscopy, using an electron probe microanalytic technique. The results indicate that the supplementation of the diet with 5% xylitol had a protective effect against the loss of bone mineral after ovariectomy in the rat. This was clearly seen in tibial density and in the inorganic fraction of the bone, and in the concentrations of bone Ca and phosphorus. Ovariectomy caused a doubling in periosteal bone formation relative to the controls, whereas the growth of the periosteally formed bone was somewhat reduced following xylitol supplementation as compared with ovariectomy alone.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed2002

Nucleus behavior during the closed mitosis of Tritrichomonas foetus.

We present observations on the fine structure and the division process of the nucleus in the protist Tritrichomonas foetus, parasite of the urogenital tract of cattle. The nucleus was followed by immunofluorescence and electron microscopy during interphase and mitosis. Conventional karyotyping coupled to image processing and bright field Panotic staining were used to follow nucleus modifications, chromosome number and condensation pattern along the whole cell cycle. Confocal laser scanning microscopy (CLSM) using DNA fluorescent probes, followed by image processing in the SURF-Driver program, produced three-dimensional reconstruction data of the mitotic nucleus under each phase of the division process. Immunocytochemistry in thin-sections revealed the chromosome spatial arrangement after bromodeoxyuridine incorporation and immunogold labeling using anti-DNA monoclonal antibodies. Our results indicate that: (1) the nucleus assumes different size and shapes along mitosis: it appears oval in interphase, becoming lobed or concave in prophase, then undergoing torsion and constriction, displaying an 'S' shape (metaphase). Next, it becomes elongated and it is finally separated in two nuclei at the transition of anaphase to telophase; (2) T. foetus nucleus harbors five chromosomes; (3) chromosomes become condensed in a pre-mitotic phase; (4) the nucleolus persists during the mitosis.

Anaphase↗
PubMed1997

The uptake of particulates by an urban woodland: site description and particulate composition.

Woodlands may improve local air quality by increasing the uptake rates of gaseous, particulate and aerosol pollutants from the atmosphere and can also act as relatively permanent sinks for some pollutants. Rough Wood, Walsall was selected for a study of the material which accumulates on tree foliage because of its location in a densely populated urban area, and its proximity to a motorway with high traffic flow (the M6) and to other pollutant sources. Methods were developed for leaf washing to allow determination of the quantity of dust and the identification of the dust particles present on oak leaves. Elemental analysis of particles was also undertaken using scanning electron microscopy coupled with electron probe microanalysis. A large proportion of particles were organic in origin. Of the inorganic particles, the majority contained silicon and aluminium in varying proportions suggesting that they were soil derived. Some particles were clearly identified as the products of combustion, and sea or road salt was present on leaf surfaces. Some particles contained copper, tin and titanium which may reflect the proximity of Rough Wood to local metal workings. The number of particles counted on leaf surfaces decreased as distance from the motorway increased.

Journal Article↗
PubMed1997

Simultaneous topographic and fluorescence imagings of recombinant bacterial cells containing a green fluorescent protein gene detected by a scanning near-field optical/atomic force microscope.

A scanning near-field optical/atomic force microscope (SNOAM) system was applied for simultaneous topographic and fluorescence imaging of biological samples in air and liquid. The SNOAM uses a bent optical fiber simultaneously as a dynamic mode atomic force microscopy cantilever and as a scanning near-field optical microscopy probe. Optical resolution of this system was about 50-100 nm in fluorescence mode for fluorescent latex beads on a quartz glass plate. Green fluorescent protein (GFP) is a convenient indicator of transformation and should allow cells to be separated by fluorescence-activated cell sorting. The gene coding to GFP was cloned in recombinant Escherichia coli. The SNOAM system used 458- or 488-nm irradiation from a multiline Ar ion laser for excitation of GFP, since a native GFP has been known to give a maximum at 395 nm and a broad absorption spectrum until 500 nm. Topographic and fluorescence images of recombinant E. coli were obtained simultaneously with a high spatial resolution which was apparently better than that of a conventional confocal microscope. A nanoscopic GFP fluorescence spectrum was obtained by positioning the optical fiber probe above the bright area of the E. coli cells. Comparing topographic and fluorescence images, it can be seen that individual E. coli cells expressed different fluorescence intensities. Fluorescence obtained by SNOAM indicated that GFP oxidation possibly occurred near the cell surface. A SNOAM system also indicated the possibility of precise imaging of native cells in liquid.

Escherichia coli↗
PubMed2001

Structural elucidation of microporous and mesoporous catalysts and molecular sieves by high-resolution electron microscopy.

Twenty years ago, one of us embarked (Bursill, L. A.; Lodge, E. A.; Thomas, J. M. Zeolitic structures as revealed by high-resolution electron microscopy. Nature 1980, 286, 111-113) on the study of zeolites (renowned for their electron-beam sensitivity) by high-resolution transmission electron microscopy (HRTEM). In the ensuing years, high-resolution imaging aided by optical diffractometry has yielded details of the open framework structures of a number of new aluminosilicate and aluminophosphate molecular sieves and catalysts. The nature of intergrowth and recurrently twinned structures, as well as new types of structural imperfection in hitherto uncharacterized materials, has also been elucidated. With continued improvements in instrumental development, encompassing higher accelerating voltages, better objective lenses and vacua, computational advances, and the arrival of slow-scan CCD detectors, electron crystallographic methods and HRTEM imaging now enable the ab initio three-dimensional structures of micro- and mesoporous solids, with their occluded structure-directing organic species, to be determined. High-resolution scanning transmission electron microscopy using subnanometric probes provides supplementary structural and ultramicro analytical information and electron spectroscopic imaging (at the attogram level). In its high-angle annular dark-field mode, it is capable of locating and determining the composition of individual nanoparticle catalysts (consisting of just a few atoms) supported on porous hosts.

Journal Article↗
PubMed2003

Observation of the marcus inverted region of electron transfer reactions at a liquid/liquid interface.

Scanning electrochemical microscopy was used to probe the influence of a driving force on the heterogeneous electron transfer (ET) processes at the externally polarized water/1,2-dichloroethane interface. Being a part of the driving force, the Galvani potential difference at the interface, Deltaowphi, can be quantitatively controlled in a wide range, allowing the precise measurements of the rate constants of the ET reactions. Two opposite systems were chosen in this work. One was 5,10,15,20-tetraphenyl 21H,23H-porphyrin zinc (ZnPor, O)/Fe(CN)64- (W), and the other was TCNQ (O)/Fe(CN)63- (W). For both systems studied, the relations between the rate constant and the Deltaowphi were of parabolic shape; that is, the rate constants increased initially with the Deltaowphi until reaching a maximum and then decreased steadily as the Deltaowphi increased further. This is in accordance with the prediction of the Marcus theory. To our knowledge, this is the first report that the Marcus inverted region can be observed electrochemically at an unmodified liquid/liquid interface with only one redox couple at each phase. The effect of the concentrations of the organic supporting electrolyte has also been discussed in detail.

Journal Article↗
PubMed2003

Understanding and tuning the epitaxy of large aromatic adsorbates by molecular design.

If the rich functionality of organic molecules is to be exploited in devices such as light-emitting diodes or field-effect transistors, interface properties of organic materials with various (metallic and insulating) substrates must be tailored carefully. In many cases, this calls for well-ordered interfaces. Organic epitaxy-that is, the growth of molecular films with a commensurate structural relationship to their crystalline substrates--relies on successful recognition of preferred epitaxial sites. For some large pi-conjugated molecules ('molecular platelets') this works surprisingly well, even if the substrate exhibits no template structure into which the molecules can lock. Here we present an explanation for site recognition in non-templated organic epitaxy, and thus resolve a long-standing puzzle. We propose that this form of site recognition relies on the existence of a local molecular reaction centre in the extended pi-electron system of the molecule. Its activity can be controlled by appropriate side groups and--in a certain regime--may also be probed by molecularly sensitized scanning tunnelling microscopy. Our results open the possibility of engineering epitaxial interfaces, as well as other interfacial nanostructures for which specific site recognition is essential.

Journal Article↗
PubMed2005

Evaluation of hexagonal boron nitride as a new tablet lubricant.

In this study, hexagonal boron nitride (HBN) was evaluated as a new lubricant for pharmaceutical tablet manufacturing. The other conventional lubricants such as magnesium stearate (MGST), stearic acid (STAC), and glyceryl behenate (COMP) were also tested along with HBN. Tablets were manufactured on an instrumented single-station tablet press to monitor and quantify the lower punch ejection force (LPEF). The force ratio, tablet crushing strength, disintegration time, and thickness were measured. The lubricant film formation and lubricant distribution in tablets were studied using the scanning electron microscopy (SEM) and electron probe micro analyzer (EPMA). Based on the force ratio, a good lubrication was obtained at 1% for MGST and HBN; in contrast, STAC and COMP did not show a good lubrication. After 1%, all lubricants performed well. MGST was found to be the most effective lubricant based on LPEF-lubricant concentration profile. HBN provided a 50% decrease in LPEF at 2% lubricant concentration and was rated as an effective tablet lubricant. HBN was better than either STAC or COMP. Unlike MGST, HBN had no significant prolongation effect on tablet disintegration times.

Boron Compounds↗
PubMed2006

Electron transport via local polarons at interface atoms.

Electronic transport is profoundly modified in the presence of strong electron-vibration coupling. We show that in certain situations, the electron flow takes place only when vibrations are excited. By controlling the segregation of boron in semiconducting Si(111)-square root 3 x square root 3 R 30 degrees surfaces, we create a type of adatom with a dangling-bond state that is electronically decoupled from any other electronic state. However, probing this state with scanning tunnelling microscopy at 5 K yields high currents. These findings are rationalized by ab-initio calculations that show the formation of a local polaron in the transport process.

Journal Article↗
PubMedDate not supplied

Near-field scanning optical microscopy studies of L-alpha-dipalmitoylphosphatidylcholine monolayers at the air-liquid interface.

The phase structure in L-alpha-dipalmitoylphosphatidylcholine-20 mol% fluorescent 1,1'-dioctadecyl-3,3,3',3'-tetramethylindocarbocyanine perchlorate Langmuir monolayers dispersed on a 2 M sucrose solution subphase is studied with near-field scanning optical microscopy (NSOM). Cantilevered NSOM probes operating in a tapping-mode feedback or an optical interferometric feedback mode are capable of tracking the air-sucrose solution interface. At the micrometre scale, the NSOM fluorescence images reveal lipid domain features similar to those observed previously in supported Langmuir-Blodgett (LB) monolayers. At the submicrometre scale, the small nanometric lipid islands seen in LB films are not observed at the air-sucrose interface. This supports a mechanism in which domain formation in LB films can be induced by means of the transfer process onto the solid support. Progress towards extending these studies to films at the air-water interface using the optical interferometric feedback method is also discussed.

1,2-Dipalmitoylphosphatidylcholine↗
PubMed2000

Simultaneous detection of the establishment of seed-inoculated Pseudomonas fluorescens strain DR54 and native soil bacteria on sugar beet root surfaces using fluorescence antibody and in situ hybridization techniques.

Colonization at sugar beet root surfaces by seedling-inoculated biocontrol strain Pseudomonas fluorescens DR54 and native soil bacteria was followed over a period of 3 weeks using a combination of immunofluorescence (DR54-targeting specific antibody) and fluorescence in situ hybridization (rRNA-targeting Eubacteria EUB338 probe) techniques with confocal laser scanning microscopy. The dual staining protocol allowed cellular activity (ribosomal number) to be recorded in both single cells and microcolonies of strain DR54 during establishment on the root. After 2 days, the population density of strain DR54 reached a constant level at the root basis. From this time, however, high cellular activity was only found in few bacteria located as single cells, whereas all microcolony-forming cells occurring in aggregates were still active. In contrast, a low density of strain DR54 was observed at the root tip, but here many of the bacteria located as single cells were active. The native population of soil bacteria, comprising a diverse assembly of morphologically different forms and size classes, initiated colonization at the root basis only after 2 days of incubation. Hence the dual staining protocol allowed direct microscopic studies of early root colonization by both inoculant and native soil bacteria, including their differentiation into active and non-active cells and into single or microcolony-forming cells.

Journal Article↗
PubMed2000

Combined molecular ecological and confocal laser scanning microscopic analysis of peat bog methanogen populations.

Confocal laser scanning microscopy, using fluorescently labelled oligonucleotide probes targeting the 16S rRNA of different physiological groups of methanogens, was used to identify which methanogenic genera were present and to describe their in situ spatial locations in samples taken at different depths from blanket peat bog cores. Total bacterial DNA was also extracted and purified from the samples and used as template for amplification of 16S rRNA and regions of methyl CoM reductase-encoding genes using the polymerase chain reaction, as well as for oligonucleotide hybridisation experiments. These techniques, used in concert, demonstrated that methanogens of several physiological groups were present in highest numbers in the mid regions of 25 cm deep peat cores. Some discrepancies were apparent in the findings of the microscopic and molecular methods, though these may be partially accounted for by the different sensitivities of the techniques employed. The combined approaches used in this study gave an insight into the diversity and distribution of methanogens in peat environments not possible using molecular ecological methods alone.

DNA, Archaeal↗
PubMed2002

Oxidative stress and inflammatory reaction modulation by white wine.

Wine and olive oil, essential components of the Mediterranean diet, are considered important factors for a healthy life style. Tyrosol (T) and caffeic acid (CA) are found in both extra virgin olive oil and in white wine. Three white wines from the northeast Italy and four white wines from Germany were analyzed for their content of T and CA. These compounds were tested for their antioxidant activity and their capacity to modulate three different cytokines: IL-1 beta, IL-6, and TNF-alpha, which are currently considered to be the major cytokines influencing the acute phase of the inflammatory response. Furthermore, the antioxidant activity of T and CA was analyzed by monitoring the oxidation of a redox-sensitive probe by using laser scanning confocal microscopy. T and CA, applied at nanomolar range, were found to significantly reduce the generation of oxidants induced by azobis-amidinopropanedihydrochloride. Peripheral blood mononuclear cells (PBMC) from healthy volunteers were incubated at 37 degrees C for 12 hours with 100 ng LPS (E. coli and P. maltofilia). Increasing doses of T and CA (150 nM to 300 microM) were added and cell-associated IL-1 beta and TNF-alpha were determined by immunoreactive tests after three freeze-thaw cycles. IL-6 release was also determined in cell surnatants. LPS-stimulated PBMC showed a significant increase in cytokine release, while T and CA, used at nanomolar concentrations, were able to modulate their expression. Taken together, these results suggest a remarkable effect of white wine non-alcoholic compounds on oxidative stress and inflammatory reaction.

Antioxidants↗