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PubMed1995

A sensitive staining method for NORs.

A sensitive staining method for nucleolar organizer regions (NORs) is described, using blue toning of AgNORs. NORs are loops of DNA which are transcribed into ribosomal RNA. NORs can be demonstrated by staining with silver nitrate, since NOR-associated proteins are argyrophilic, producing structures termed AgNORs. Normal blood lymphocytes were stained with both methods. The number and resolution of NORs increased 2-3 times by blue toning (30 mmol/l FeCl3, 11 mmol/l potassium hexacyanoferrate(III), and 33 mmol/l oxalic acid) compared with silver staining. A significant difference in the number of NORs was noticed between silver-stained and blue-toned cells (P < 0.001). The blue toning technique thus appears to be more sensitive in detecting NORs than the AgNOR method and may prove a useful alternative for applications in histopathology.

Chlorides↗
PubMed2005

Adsorption of 2-aminobenzothiazole on colloidal silver particles: an experimental and theoretical surface-enhanced Raman scattering study.

Surface-enhanced Raman scattering (SERS) spectra of the biologically important 2-aminobenzothiazole (2-ABT) molecule adsorbed on silver hydrosols are compared with its FTIR spectrum and normal Raman spectroscopy (NRS) spectrum in the bulk and in solution. The optimized structural parameters and the computed vibrational wavenumbers of the compound have been estimated from ab initio (Hatree-Fock) and density functional calculations. Some vibrational modes of the molecule have been reassigned. Concentration-dependent SERS spectra of the molecule reveal the existence of two types of vertically adsorbed species on colloidal silver particles, whose relative population varies with the adsorbate concentrations. The adsorption geometry and structural parameters of one type of adsorbed species are related to the NRS spectrum of the chemically prepared and theoretically modeled 2-ABT-Ag(I) coordination compound.

Journal Article↗
PubMed1984

A silver and gold technique for axons and axon-bundles in formalin-fixed central and peripheral nervous tissue.

We have modified the reduced silver procedure of Liesegang and added a gold treatment. The technique has been successfully and routinely used to impregnate both peripheral and central nervous tissues of mice of different ages, and central nervous tissue of man. It was applied on conventionally (formalin) fixed and cryostat-cut material. In the periphery, nerves, individual axons, and sensory and motor endings stained a dark purple. In the brain, both bundles of axons and individual fine axonal branches were stained. When counterstained by a Nissl method, an excellent overall image of CNS structure is obtained.

Age Factors↗
PubMed1996

Polyfluorinated Tris(pyrazolyl)borates. Syntheses and Spectroscopic and Structural Characterization of Group 1 and Group 11 Metal Complexes of [HB(3,5-(CF(3))(2)Pz)(3)](-) and [HB(3-(CF(3))Pz)(3)](-).

The fluorinated tris(pyrazolyl)borate ligands [HB(3,5-(CF(3))(2)Pz)(3)](-) and [HB(3-(CF(3))Pz)(3)](-) (where Pz = pyrazolyl) have been synthesized as their sodium salts from the corresponding pyrazoles and NaBH(4) in high yield. These sodium complexes and the related [HB(3,5-(CF(3))(2)Pz)(3)]K(DMAC) were used as ligand transfer agents in the preparation of the copper and silver complexes [HB(3,5-(CF(3))(2)Pz)(3)]Cu(DMAC), [HB(3,5-(CF(3))(2)Pz)(3)]CuPPh(3), [HB(3,5-(CF(3))(2)Pz)(3)]AgPPh(3), and [HB(3-(CF(3))Pz)(3)]AgPPh(3). Metal complexes of the fluorinated [HB(3,5-(CF(3))(2)Pz)(3)](-) ligand have highly electrophilic metal sites relative to their hydrocarbon analogs. This is evident from the formation of stable adducts with neutral oxygen donors such as H(2)O, dimethylacetamide, or thf. Furthermore, the metal compounds derived from fluorinated ligands show fairly long-range coupling between fluorines of the trifluoromethyl groups and the hydrogen, silver, or phosphorus. The solid state structures show that the fluorines are in close proximity to these nuclei, thus suggesting a possible through-space coupling mechanism. Crystal structures of the sodium adducts exhibit significant metal-fluorine interactions. The treatment of [HB(3,5-(CF(3))(2)Pz)(3)]Na(H(2)O) with Et(4)NBr led to [Et(4)N][HB(3,5-(CF(3))(2)Pz)(3)], which contains a well-separated [Et(4)N](+) cation and the [HB(3,5-(CF(3))(2)Pz)(3)](-) anion in the solid state. Crystal data with Mo Kalpha (lambda = 0.710 73 Å) at 193 K: [HB(3,5-(CF(3))(2)Pz)(3)]Na(H(2)O), C(15)H(6)BF(18)N(6)NaO, a = 7.992(2) Å, b = 15.049(2) Å, c = 9.934(2) Å, beta = 101.16(2) degrees, monoclinic, P2(1)/m, Z = 2; [{HB(3-(CF(3))Pz)(3)}Na(thf)](2), C(32)H(30)B(2)F(18)N(12)Na(2)O(2), a = 9.063(3) Å, b = 10.183(2) Å, c = 12.129(2) Å, alpha = 94.61(1) degrees, beta = 101.16(2) degrees, gamma = 95.66(2) degrees, triclinic, &Pmacr;1, Z = 1; [HB(3,5-(CF(3))(2)Pz)(3)]Cu(DMAC), C(19)H(13)BCuF(18)N(7)O, a = 15.124(4) Å, b = 8.833(2) Å, c = 21.637(6) Å, beta = 105.291(14) degrees, monoclinic, P2(1)/n, Z = 4; [HB(3,5-(CF(3))(2)Pz)(3)]CuPPh(3), C(33)H(19)BCuF(18)N(6)P, a = 9.1671(8) Å, b = 14.908(2) Å, c = 26.764(3) Å, beta = 94.891(1) degrees, monoclinic, P2(1)/c, Z = 4; [HB(3,5-(CF(3))(2)Pz)(3)]AgPPh(3).0.5C(6)H(14), C(36)H(26)AgBF(18)N(6)P, a = 13.929(2) Å, b = 16.498(2) Å, c = 18.752(2) Å, beta = 111.439(6) degrees, monoclinic, P2(1)/c, Z = 4; [Et(4)N][HB(3,5-(CF(3))(2)Pz)(3)], C(23)H(24)BF(18)N(7), a = 10.155(2) Å, b = 18.580(4) Å, c = 16.875(5) Å, beta = 99.01(2) degrees, monoclinic, P2(1)/n, Z = 4.

Journal Article↗
PubMed2004

High-frequency plant regeneration through callus initiation from mature embryos of maize ( Zea Mays L.).

An efficient maize regeneration system was developed using mature embryos. Embryos were removed from surface-sterilized mature seeds and sliced into halves. They were used as explants to initiate callus on induction medium supplemented with 4.0 mg l(-1) 2,4-dichlorophenoxyacetic acid (2,4-D). The induction frequency of primary calli was over 90% for all inbred lines tested. The primary calli were then transferred onto subculture medium supplemented with 2.0 mg l(-1) 2,4-D. Following two biweekly subcultures, embryogenic calli were formed. Inclusion of a low concentration (0.2 mg l(-1)) of 6-benzylaminopurine (BA) in the subculture medium significantly promoted the formation of embryogenic callus. The addition of silver nitrate (10 mg l(-1)) also supported an increased frequency of embryogenesis. The embryogenic callus readily formed plantlets on regeneration medium supplemented with 0.5 mg l(-1) BA. The regenerated plantlets were transferred to half-strength Murashige and Skoog medium supplemented with 0.6 mg l(-1) indole-3-butyric acid to develop healthy roots. The regenerated plantlets were successful on transfer to soil and set seed. Using this system, plantlets were regenerated from seven elite maize inbred lines. The frequency of forming green shoots ranged from 19.8% to 32.4%. This efficient regeneration system provides a solid basis for genetic transformation of maize.

2,4-Dichlorophenoxyacetic Acid↗
PubMed1991

Liver units in three dimensions: I. Organization of argyrophilic connective tissue skeleton in porcine liver with particular reference to the "compound hepatic lobule".

Liver units were investigated in pig livers by means of histologic serial tracing, physical model building, and computer-aided three-dimensional imaging. Observations of the argyrophilic connective tissue skeleton were based mainly on the celloidin-embedded serial sections treated with silver impregnation. The parenchymal mass that clothed the initial segments of hepatic venous radicles was demarcated by fibrous septa which formed isolable units with two basic patterns: the simple hepatic lobule (SHL) and the compound hepatic lobule (CHL). Both lobule types presented regular limiting structures circumscribing each unit. Three-dimensional studies revealed that 25% of the lobules in a section belonged to the SHL type and 75% to the CHL type, the latter being predominant among the surface lobules. When considered in only two dimensions, however, the SHL-like lobules constituted the majority. Polygonal analysis disclosed that the pentagonal lobule was the most typical, instead of the "hexagonal" or "classic" lobule. The CHLs represented a multiaxial unit containing a system of venous tributaries in accordance with intralobular septation, whereas the SHLs were found with one axial vessel having a dendritic tendency at the incipient end; some SHLs were drained eccentrically by separate vessels into a sublobular vein. It was observed that, in dividing CHLs, whereas particular sinosoids were transformed into portal twigs, other sinusoids were changed into central venous tributaries. Fibrous deposition occurred along the septal-line sinusoids, bringing into view the septum-initiating plane. Fibroconnective tissue was supplied from the portal area and central (sublobular) adventitia, where portal triad structures and adventitial arterioles, respectively, were included. The findings of the present study facilitate the understanding of several characters of the lobules that have been reported previously, or occasionally postulated, such as the portal-central bridging tendency, the intralobular arterioles or ductules, the translobular artery or portal vein, the "portal-portal" or "portal-central" anastomoses, and the apposition of pericentral zone close to periportal zone. Based on differences in argyrophilia of sinusoidal reticulum, in proportion of lobule types, and in vasculature, the anatomic heterogeneity of liver unit was demonstrable in zonality, regionality, and locality.

Animals↗
PubMed2003

1, 2-dihydroisoquinoline-N-acetic acid derivatives as new carriers for specific brain delivery I: synthesis and estimation of oxidation kinetics using multivariate calibration method.

In order to overcome the slow oxidation of 1, 2-dihydro-N-alkylisoquinoline, 1, 2-dihydroisoquinoline-N-acetic acid derivatives (3b-d) were designed and synthesized as new chemical delivery systems (CDS) for the brain. Molecular orbital calculations for the suggested derivatives revealed that these carriers are stable against oxidation. However, hydrolysis to their corresponding anions will accelerate the rate of oxidation, and accordingly increase the efficiency of brain specific delivery. A multivariate calibration method for in vitro determination of the oxidation rate for the suggested brain specific chemical delivery system is described. The method is based on measurement of individual rates of oxidation of prepared 1, 2-dihydroisoquinolines, using silver ions to provide the corresponding quaternary forms. Components of binary mixtures formed through the oxidation step (composed of the dihydro-compound and its quaternary form), showed a considerable degree of spectral overlapping - more than 90% in all cases. Resolution of these binary mixtures under investigation has been accomplished mainly using classical least squares analysis. Kinetic oxidation data of the tested compounds revealed that these new CDSs have a reasonable oxidation rate for efficient brain delivery.

Acetic Acid↗
PubMed2004

Low-temperature sealed tube combustion of gaseous, liquid and solid organic compounds for 13C/12C and 14C analysis.

A new, low-temperature sealed tube technique for combustion of organic carbon prior to subsequent off-line isotope analysis is proposed. Complete oxidation is achieved with potassium peroxodisulfate and silver permanganate as oxidants at temperatures not exceeding 500 degrees C. The combustion of gaseous (methane), solid (cane sugar, vanilla, N-thiazolyl-2-sulfamide, ascorbic acid, phenanthrene, thiourea, polyethylenefilm, tetrafluoropolyethylene, polyetheretherketone, graphite, and Suwannee River Fulvic Acid), and liquid (tetrachloroethene, toluene, and oil) model compounds and international standards was tested. A 24 h combustion at 500 degrees C was sufficient for complete oxidation in all cases. The time required for complete oxidation of Suwannee River Fulvic Acid, typical of refractory freshwater dissolved organic carbon, as a function of combustion temperature was 2 h at 500 degrees C, 6 h at 400 degrees C, and 24 h at 300 degrees C. Preparation of saline solution parallels of cane sugar, vanilla, N-thiazolyl-2-sulfanilamide, and ascorbic acid gave consistent results. For reproducible delta13C analyses using a Thermoquest MAT 252 MS, a minimum of 5 microg C had to be combusted. Reliable 14C results, measured at an accelerator mass spectrometer facility, were obtained from coal and from cane sugar combusted for 24 h at 500 degrees C by the proposed method.

Carbon↗
PubMed2000

Waste water bacterial isolates resistant to heavy metals and antibiotics.

Sewage water of Casablanca, an industrial city in Morocco, was studied for microorganisms resistant to heavy metals. Isolates were purified and collected on agar slants to be screened for resistance to heavy metals, including mercury in vitro. The strains that showed high resistance to heavy metals were also studied for their resistance to antibiotics and aromatic hydrocarbons. Results indicated that the strains most resistant to all tested products belonged to Ps. fluorescens, Ps. aeruginosa, Klebsiella pneumoniae, Proteus mirabilis, and Staphylococcus sp. These strains exhibit high minimal inhibitory concentrations for heavy metals such as cadmium (2 mm) or mercury (1.2 mm). Growth of Ps. fluorescens and Klebsiella pneumoniae in the presence of heavy metals was also determined, and the growth curves indicated that mercury, copper, and zinc present a slight inhibitory action, while cadmium and silver could have a potent inhibitory action on growth compared with the controls. These studies also investigated growth in media containing aromatic compounds as the sole source of carbon. The results demonstrate that these strains could be good candidates for remediation of some heavy metals and aromatic compounds in heavily polluted sites.

Anti-Bacterial Agents↗
PubMed1994

A new technique for the direct demonstration of overlapping cutaneous innervation territories of peptidergic C-fibre afferents of rat hindlimb nerves.

A new technique based on the phenomenon of vascular labelling has been devised for the direct visualisation of overlapping innervation territories of cutaneous nerves. The saphenous, peroneal and sural nerves on one side in anaesthetised rats were exposed, cut centrally and successively stimulated antidromically to induce a neurogenic inflammatory response after an intravenous injection of either a 1% colloidal silver solution or a suspension of 3% Monastral Blue B. Light microscopic examination of transparent preparations of the dorsal hindpaw skin revealed labelled blood vessels of different colours which represented cutaneous territories served by different nerves. Blood vessels labelled with both substances were regarded as areas of overlapping innervation. Such areas were typically localised along the border of adjacent innervation territories. In addition, distinct areas exhibiting double-labelled blood vessels were regularly encountered in regions separate from this border zone. Areas of interest were drawn with the aid of a camera lucida and measured by means of a computerised system. The results indicate a significant, although topographically variable, degree of overlap of these cutaneous innervation areas. This new technique offers a possibility to explore the importance of normally existing overlap in the reinnervation of a denervated skin area by collateral nerve sprouting.

Animals↗
PubMed2001

Analysis of diacyl peroxides by Ag+ coordination ionspray tandem mass spectrometry: free radical pathways of complex decomposition.

Organic peroxides have significance in organic synthesis and biological processes. Characterization of these compounds with weak O-O bonds is sometimes difficult due to their thermal instability and sensitivity to acid or base. Coordination of diacyl peroxides with AgBF4 provides a means for analysis of these compounds by coordination ionspray tandem mass spectrometry (CIS-MS/MS). Precursor ion (Q1) scans of acetyl benzoyl peroxide give two Ag+ adducts, [M + Ag + solvent]+ and [M + Ag + M]+. These silver ion adducts can be selectively dissociated (CID) to give unique structural information about the analyte. Decomposition of the [M + Ag + solvent]+ adduct generates fragmentation products due to apparent homolytic cleavage of the O-O bond followed by decarboxylation of the resultant radicals. The bis-diacylperoxide complex, [M + Ag + M]+ gives CID pathways that involve homolysis of the (O-O bond and free radical cross-coupling of the two diacyl peroxides coordinated to the silver ion, i.e. formation of dibenzoyl peroxide, phenyl benzoate, and biphenyl from acetyl benzoyl peroxide. The observation of free radical CID modes is uncommon in mass spectrometry but these pathways are consistent with well-known solution and gas phase processes for peroxide compounds. The proposed fragmentation pathways have been supported by experiments with (18)O and deuterated substrates. This technique can be applied to analyze diacyl peroxides with different substituents as well.

Chromatography, Thin Layer↗
PubMed1997

3-Nitropropionic acid neurotoxicity: visualization by silver staining and implications for use as an animal model of Huntington's disease.

The neuronal damage produced by the mitochondrial toxin 3-nitropropionic acid (3NPA) has been suggested to replicate much of the neuropathology seen in Huntington's disease (HD) and therefore might be used in an animal model. We investigated the susceptibility to this toxin of different neuronal populations in addition to the commonly studied caudate putamen by injecting 3NPA into seven different brain regions as well as systemically. After different survival times, rats were intracardially perfused, brain sections were processed with the Gallyas silver technique, and impregnated neurons were mapped with a computerized microscope. Intracerebral administration of 3NPA resulted in a lesion, the center of which was devoid of tissue while the area was surrounded by a halo of Golgi-like impregnated neurons. In addition to local damage, rats receiving microinjections into the frontal cortex, caudate putamen, basal forebrain, and amygdala displayed argyrophilic neurons in the thalamus corresponding to the topography of thalamofugal neurons projecting to the individual injection sites. On the other hand, negligible secondary damage was seen after injections into the internal capsule, thalamus, or substantia nigra, implicating that thalamofugal axons are especially vulnerable to the local effect of this toxin. Two and a half days after systemic administration of 3NPA, a diffuse argyrophilic neuronal reaction was seen in the dorsolateral part of the caudate putamen, together with a more regionally selective staining of neurons in different cortical areas and the hippocampus. These morphopathological changes were also accompanied by motor deficits. The affected neurons in the cortical regions were primarily in those layers (V and VI) and areas (medial prefrontal, caudal insular/periphinal, and ventral temporal) that do not project toward the lesioned striatal area; therefore, the cortical pathology may represent another primary site of action of the toxin. Among the affected neurons in the hippocampal complex were pyramidal neurons in the CA1 region as well as various neurons in the CA3 region and dentate hilar area. These studies suggest that a combination of 3NPA administration and a sensitive silver-impregnation method may unravel the potential site of primary neuronal damage in this animal model. Furthermore, these findings may contribute to the understanding of how the disease progresses in HD from the originally affected neuronal population(s) by the recruitment of closely related systems and pathways.

Animals↗
PubMed1995

Effects of aluminum chloride on the nucleus and nucleolus in root tip cells of Hordeum vulgare.

The cytotoxic effects of aluminum chloride at different concentrations (1 x 10(-4)-5 x 10(-2) M) on the nucleus and the nucleolus in root tip cells of Hordeum vulgare were investigated using the carbol fuchsin staining method and the silver staining technique separately. Results showed that aluminum chloride could induce nuclear aberrations comprising elongated, irregular, ruptured and fractured nuclei and micronucleus formation. After the treatment with aluminum, it was observed that the nucleolar material was extruded from the nucleus into the cytoplasm. The nucleolar material in the cytoplasm was disintegrated, forming silver-stained particulate material, which occurred in both the inner root meristem cells and the root cap cells. The possible mechanism of the aluminum toxicity on the nucleus and nucleolus is briefly discussed.

Aluminum Chloride↗
PubMed1996

Analysis of triacylglycerols by silver-ion high-performance liquid chromatography-atmospheric pressure chemical ionization mass spectrometry.

Triacylglycerols of the seed oils rich in alpha- and/or gamma-linolenic acid moieties were separated by silver-ion high-performance liquid chromatography (HPLC) followed by on-line atmospheric pressure chemical ionization-mass spectrometric (APCI-MS) detection. Mass spectra of most triacylglycerols exhibited abundant [M + H]+ and [M - RCO2]+ ions, which defined the molecular weight and the molecular association of fatty acyl residues of a triacylglycerol, respectively. Silver ions formed weaker complexes with triacylglycerols containing gamma-linolenic acid than with those containing alpha-linolenic acid, i.e., the elution order of molecules was XYT gamma > XYT alpha, XT gamma T gamma > XT gamma T alpha > XT alpha T alpha, and T gamma T gamma T gamma > T gamma T gamma T alpha > T gamma T alpha T alpha > T alpha T alpha T alpha, where T alpha = alpha-linolenic acid, T gamma = gamma-linolenic acid, and X, Y = fatty acids different from linolenic acid. Furthermore, silver-ion HPLC resulted in partial separation within equally unsaturated triacylglycerols according to differences in the combined number of acyl carbons. Regioisomeric forms of triacylglycerols were not determined from the seed oil samples, although differences were measured with reference compounds in the relative abundances of [M - RCO2]+ ions formed by a loss of a fatty acyl residue from the sn-2 position and the sn-1/3 positions. Silver-ion HPLC/APCI-MS provided valuable information for structure elucidation of seed oil triacylglycerols: 43 molecular species were identified from cloudberry seed oil, 39 from evening primrose oil, 79 from borage oil, 44 from alpine currant, and 56 from black currant seed oils. The quantitation requires to be studied further, especially in those cases where several molecular weight species of triacylglycerols eluted in a single chromatographic peak.

Chromatography, High Pressure Liquid↗
PubMed1991

Acute toxicity tests using rotifers. IV. Effects of cyst age, temperature, and salinity on the sensitivity of Brachionus calyciflorus.

Several aspects of the response to toxicants using a standardized toxicity test with the freshwater rotifer Brachionus calyciflorus are described. Test animals are obtained by hatching cysts which produce animals of similar age and physiological condition. The acute toxicity of 28 compounds is described with 24-hr LC50's. The LC50's span five orders of magnitude, from silver at 0.008 mg.liter-1 to benzene at more than 1000 mg.liter-1. Control mortality in 84 tests averaged 2% with a standard deviation of 3%, indicating very consistent test sensitivity. Only once in 84 trials did a test fail because of excessive control mortality, yielding a failure rate of 1.2%. Cyst age from 0 to 18 months had no effect on the sensitivity of neonates to reference toxicants. Both high and low temperatures increased rotifer sensitivity to reference toxicants. Copper sensitivity was greater at 10, 25, and 30 degrees C compared with results at 20 degrees C. Likewise, sodium pentachlorophenol toxicity was greater at 10 and 30 degrees C compared with results at 20 degrees C. Survivorship curves at 25 degrees C of neonates under control conditions indicated that mortality begins at about 30 hr. This places a practical limit on toxicant exposure for the assay of 24 hr. B. calyciflorus cysts hatch at salinities up to 5 ppt and acute toxicity tests using pentachlorophenol at this salinity yielded LC50's about one-half those of standard freshwater. B. calyciflorus is preferred over Brachionus plicatilis for toxicity tests in salinities up to 5 ppt because it is consistently more sensitive.

Animals↗
PubMed2004

Novel approaches to the syntheses of N-substituted S-glycosyl-sulfenamides.

Bis(tetra-O-acetyl-beta-D-glucopyranosyl)disulfide reacts, under silver ion activation, with primary and secondary aliphatic as well as aromatic amines to furnish the title compounds in moderate to good yields. The same derivatives could also be obtained from (tetra-O-acetyl)-beta-D-glucopyranosyl methanethiolsulfonate 1 by nucleophilic substitution with amines. It was shown that the polarization of the S-S-bond in 1 is enhanced by Ag+ so as to allow reaction with sterically hindered amines as well.

Glycosides↗
PubMed2003

A novel, highly electrical conducting, single-component molecular material: [Ag2(ophen)2] (Hophen = 1H-[1,10]phenanthrolin-2-one).

This communication describes a highly conducting, single-component molecular material [Ag2(ophen)2] (Hophen = 1H-[1,10]phenanthrolin-2-one) based on very strong off-set pi-pi stacking interactions with an unusually short ring-to-ring distance of ca. 3.15 A between adjacent molecules and silver(I)-pi interactions with the Ag...C contact of 3.082 A between adjacent molecules. The title compound, having a room-temperature conductivity of 14 S cm-1, is structurally different from both the known conducting single-component molecular materials of transition-metal complexes with sulfur-containing pi-delocalized dithiolene ligands and the known conducting multicomponent organosilver(I) materials.

Journal Article↗

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