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SoxRS-mediated regulation of chemotrophic sulfur oxidation in Paracoccus pantotrophus.

Paracoccus pantotrophus GB17 requires thiosulfate for induction of the sulfur-oxidizing (Sox) enzyme system. The soxRS genes are divergently oriented to the soxVWXYZA-H genes. soxR predicts a transcriptional regulator of the ArsR family and soxS a periplasmic thioredoxin. The homogenate mutant GBOmegaS carrying a disruption of soxS by the Omega-kanamycin-resistance-encoding interposon expressed a low thiosulfate-oxidizing activity under heterotrophic and mixotrophic growth conditions. This activity was repressed by complementation with soxR, suggesting that SoxR acts as a repressor and SoxS is essential for full expression. Sequence analysis uncovered operator characteristics in the intergenic regions soxS-soxV and soxW-soxX. In each region a transcription start site was identified by primer extension analysis. Both regions were cloned into the vector pRI1 and transferred to P. pantotrophus. Strains harbouring pRI1 with soxS-soxV or soxW-soxX expressed the sox genes under heterotrophic conditions at a low rate, indicating repressor titration. Sequence analysis of SoxR suggested a helix-turn-helix (HTH) motif at position 87-108 and uncovered an invariant Cys-80 and a cysteine residue at the C-terminus. SoxR was overproduced in Escherichia coli with an N-terminal His6-tag and purified to near homogeneity. Electrophoretic gel mobility shift assays with SoxR retarded the soxS-soxV region as a single band while the soxW-soxX region revealed at least two protein-DNA complexes. These data demonstrated binding of SoxR to the relevant DNA. This is believed to be the first report of regulation of chemotrophic sulfur oxidation at the molecular level.

Amino Acid Sequence↗

Regulation of the gravitropic response and ethylene biosynthesis in gravistimulated snapdragon spikes by calcium chelators and ethylene inhibitors.

The possible involvement of Ca2+ as a second messenger in snapdragon (Antirrhinum majus L.) shoot gravitropism, as well as the role of ethylene in this bending response, were analyzed in terms of stem curvature and gravity-induced asymmetric ethylene production rates, ethylene-related metabolites, and invertase activity across the stem. Application of Ca2+ chelators (ethylenediaminetetraacetic acid, trans-1,2-cyclohexane dinitro-N,N,N',N'-tetraacetic acid, 1,2-bis(2-aminophenoxy)ethane-N,N,N',N',-tetraacetic acid) or a Ca2+ antagonist (LaCl3) to the spikes caused a significant loss of their gravitropic response following horizontal placement. Conversely, the Ca2+ ionophore A23187 or the agonist Bay K-8644 increased gravibending. Longitudinally halved stem sections had significantly higher amounts of ethylene, 1-aminocyclopropane-1-carboxylic acid, and 1-(malonylamino) cyclopropane-1-carboxylic acid compared with vertical controls, with the extra production arising exclusively from the lower half of the stem. trans-1,2-cyclohexane dinitro-N,N,N',N'-tetraacetic acid pretreatment completely abolished the gravity-induced ethylene gradient across the stem, thereby leading to a significant reduction of the curvature. Similarly, reduction of the ethylene produced in the gravistimulated with CoCl2 or inhibition of its action by silver thiosulfate or 2,5-norbornadiene significantly inhibited the subsequent gravibending. Silver thiosulfate and CoCl2 also abolished the gravity-induced gradient of invertase activity across the stem, which is associated with the asymmetric stem elongation. These results suggest that cytosolic Ca2+ may regulate auxin action in snapdragon spikes, manifested as increased ethylene production, which is, in turn, intimately correlated with stem bending. Therefore, both hormones seem to play significant roles in induction and progress of the gravibending of snapdragon spikes.

Amino Acids↗

Role of ethylene in the senescence of isolated hibiscus petals.

Senescence of petals isolated from flowers of Hibiscus rosa-sinensis L. (cv Pink Versicolor) was associated with increased ethylene production. Exposure to ethylene (10 microliters per liter) accelerated the onset of senescence, as indicated by petal in-rolling, and stimulated ethylene production. Senescence was also hastened by basal application of 1-aminocyclopropane-1-carboxylic acid (ACC). Aminooxyacetic acid, an inhibitor of ethylene biosynthesis, effectively inhibited ethylene production by petals and delayed petal in-rolling. In marked contrast to these results with mature petals, immature petals isolated from flowers the day before flower opening did not respond to ethylene in terms of an increase in ethylene production or petal in-rolling. Furthermore, treatment with silver thiosulfate the day before flower opening effectively prevented petal senescence, while silver thiosulfate treatment on the morning of flower opening was ineffective. Application of ACC to both immature and mature petals greatly stimulated ethylene production indicating the presence of an active ethylene-forming enzyme in both tissues. Immature petals contained less free ACC than mature, presenescent petals and appeared to possess a more active system for converting ACC into its conjugated form. Thus, while the nature of the lack of responsiveness of immature petals to ethylene is unknown, ethylene production in hibiscus petals appears to be regulated by the control over ACC availability.

Journal Article↗

Molecular cloning and characterization of senescence-related genes from carnation flower petals.

The senescence of carnation (Dianthus caryophyllus L.) flower petals is associated with increased production of ethylene which plays an important role in regulating this developmental event. Three senescence-related cDNA clones were isolated from a cDNA library prepared from mRNA isolated from senescing petals. These cDNAs are representative of two classes of mRNAs which increase in abundance in senescing petal tissue. The mRNA for one class is present at low levels during the early stages of development and begins to accumulate in mature petals prior to the increase in ethylene production. The accumulation of this mRNA is reduced, but not eliminated, in petals treated with aminooxyacetic acid, an inhibitor of ethylene biosynthesis, or silver thiosulfate, an ethylene action inhibitor. In contrast, expression of the second class of mRNAs appears to be highly regulated by ethylene. These mRNAs are not detectable prior to the rise in ethylene production and increase in abundance in parallel with the ethylene climacteric. Furthermore, expression of these mRNAs is significantly inhibited by both aminooxyacetic acid and silver thiosulfate. Expression of these mRNAs in vegetative and floral organs was limited to floral tissue, and predominantly to senescing petals.

Journal Article↗

Effect of Ethylene Action Inhibitors upon Wound-Induced Gene Expression in Tomato Pericarp.

The contribution of wound-ethylene to wound-induced gene expression was investigated in unripe tomato pericarp using inhibitors of ethylene action. Wounded unripe tomato pericarp was treated with 2,5-norbornadiene or silver thiosulfate to inhibit specifically the induction of ethylene-dependent mRNA species. Poly(A)(+) RNAs isolated from these tissues after 12 hours of wounding were translated in vitro in a rabbit reticulocyte lysate system and [(35)S]methionine-labeled polypeptides were compared to unwounded controls after separation by one and two-dimensional polyacrylamide gel electrophoresis. Results show that mechanical wounding induces a dramatic shift in gene expression (over 50 mRNA species) but expression of less than 15% of these genes is affected by the treatment with ethylene action inhibitors. A selective decrease in mRNAs coding for a 37 kilodalton doublet and 75 kilodalton polypeptides is observed in 2,5-norbornadiene and silver thiosulfate treated wounded pericarp. Levels of hydroxyproline-rich glycoprotein mRNAs induced in wounded tissue were not influenced by inhibitors of ethylene action.

Journal Article↗

Bis(1,10-phenanthroline-N,N')(thiosulfato-O, S)nickel(II)-water-methanol (1/0.92/1.4) and bis(2,2'-bipyridyl-N, N')(thiosulfato-O,S)nickel(II)-water-methanol (1/2/0.55)

The title compounds, [Ni(S(2)O(3))(C(12)H(8)N(2))(2)].0.92H(2)O.1. 4CH(4)O and [Ni(S(2)O(3))(C(10)H(8)N(2))(2)].2H(2)O.0.55CH(4)O, are monomeric, containing nickel(II) in a distorted octahedral coordination environment provided by the four N atoms of two bidentate bipy or phen groups and one S and one O atom from a chelating thiosulfate anion. The crystals are highly unstable outside their mother liquors and are stabilized in solution by a not fully determined number of water and methanol solvate molecules. The phenanthroline structure includes two independent moieties related by a non-crystallographic inversion center. The thiosulfate anions display the usual S-O lengthening found when the anion acts in a bidentate mode.

Journal Article↗

Bis(1,10-phenanthroline)(thiosulfato)manganese(II) methanol solvate and catena-poly[[diaqua(2,9-dimethyl-1,10-phenanthroline)manganese(II)]-mu-thiosulfato].

The structure of bis(1,10-phenanthroline-kappa(2)N,N')(thiosulfato-kappa(2)O:S)manganese(II) methanol solvate, [Mn(S(2)O(3))(C(12)H(8)N(2))(2)].CH(3)OH, is made up of Mn(2+) centers coordinated to two bidentate phenanthroline (phen) groups and an S,O-chelating thiosulfate anion, forming monomeric entities. The structure of catena-poly[[diaqua(2,9-dimethyl-1,10-phenanthroline-kappa(2)N,N')manganese(II)]-mu-thiosulfato-kappa(2)O:S], [Mn(S(2)O(3))(C(14)H(12)N(2))(H(2)O)(2)](n), is polymeric, consisting of Mn(dmph)(H(2)O)(2) units (dmph is 2,9-dimethyl-1,10-phenanthroline) linked by thiosulfate anions acting in an S,O-chelating manner.

Journal Article↗

catena-poly[[aqua(3,4,7,8-tetramethyl-1,10-phenanthroline-kappa2N,N')cadmium(II)]-mu3-thiosulfato-kappa3S:S:O].

The title compound, [Cd(S(2)O(3))(C(16)H(16)N(2))(H(2)O)](n), presents a polymeric one-dimensional structure running along the P2(1)/c glide direction, with elementary units defined by six-coordinate Cd(II) atoms bonded to three symmetry-related thiosulfate groups, a bidentate tetramethylphenanthroline ligand and one aqua ligand. The bridging thiosulfates bind metal centers through two different sequences, viz. Cd-S-Cd' and Cd'-S'-S'-O'-Cd, defining a closed six-membered ring. Individual chains are held together via pi-pi interactions to generate two-dimensional networks parallel to the (100) plane. These, in turn, are connected by much weaker van der Waals interactions.

Journal Article↗

Relevance of positive patch test reactions in patients attending a dermatology tertiary referral centre.

Our purpose was to determine the proportion of relevant patch test reactions among all patch tested patients in the National Skin Centre (NSC), Singapore, from 1 January 1999 to 31 December 2000. A retrospective study was made of the patch test records of all patients patch tested in the study period. The proportion of relevant patch tests in this population was then determined. The allergens most likely to have present or past relevance were: nickel sulfate (79%), p-phenylenediamine (76%), colophonium (73%), cobalt chloride (68%) and potassium dichromate (67%). The most common allergens for each of the following relevance categories were as follows - present: p-phenylenediamine, thiuram mix, potassium dichromate; past: colophony, nickel sulfate, cobalt chloride; exposed: neomycin, gold sodium thiosulfate, fragrance mix and unknown: gold sodium thiosulfate, potassium dichromate, cobalt chloride. Evaluating relevance is difficult, and the results of relevance scoring vary widely between different groups. A consensus on the definition and standardization of the concept of relevance is needed.

Allergens↗

Survey of antibiotic resistance in an integrated marine aquaculture system under oxolinic acid treatment.

The consequences of antibiotic use in aquatic integrated systems, which are based on trophic interactions between different cultured organisms and physical continuity through water, need to be examined. In this study, fish reared in a prototype marine integrated system were given an oxolinic acid treatment, during and after which the level of resistance to this quinolone antibiotic was monitored among vibrio populations from the digestive tracts of treated fish, co-cultured bivalves and sediments that were isolated on thiosulfate-citrate-bile-sucrose. Oxolinic acid minimum inhibitory concentration distributions obtained from replica plating of thiosulfate-citrate-bile-sucrose plates indicated that a selection towards oxolinic acid resistance had occurred in the intestines of fish under treatment. In contrast, and despite oxolinic acid concentrations higher than minimum inhibitory concentrations of susceptible bacteria, no clear evolution of resistance levels was detected either in bivalves or in sediments.

Animals↗

Spirochaeta smaragdinae sp. nov., a new mesophilic strictly anaerobic spirochete from an oil field.

An obligately anaerobic spirochete designated strain SEBR 4228T (T = type strain) was isolated from an oil field of Congo, Central Africa. The strain grew optimally with a sodium chloride concentration of 5% (sodium chloride concentration) growth range 1.0-10%) at 37 degrees C (growth temperature range 20-40 degrees C) and pH of 7.0-7.2 (pH growth range pH 5.5-8.0). Strain SEBR 4228T grew on carbohydrates (glucose, fructose, ribose, D-xylose, galactose, mannitol and mannose), glycerol, fumarate, peptides and yeast extract. Yeast extract was required for growth and could not be replaced by vitamins. It reduced thiosulfate and sulfur, to H2S. Glucose was oxidised to lactate, acetate, CO2 and H2S in the presence of thiosulfate but in its absence lactate, ethanol, CO2 and H2 were produced. Fumarate was fermented to acetate and succinate. The G + C content of strain SEBR 4228T was 50%. Strain SEBR 4228T was spiral shaped measuring 5-30 by 0.3-0.5 micron and was motile with a corkscrew-like motion. Electron microscopy revealed the presence of periplasmic flagella in a 1-2-1 arrangement. Strain SEBR 4228T possessed features typical of the members of the genus Spirochaeta. 16S rRNA sequence analysis revealed that it was closely related to Spirochaeta bajacaliforniensis (similarity 98.6%). The lack of DNA homology with S. bajacaliforniensis (38%), together with other phenotypic differences, indicated that strain SEBR 4228T is a new species, which we have designated Spirochaeta smaragdinae. The type strain is SEBR 4228T (= DSM 11293).

DNA, Ribosomal↗

Gold--a controversial sensitizer. European Environmental and Contact Dermatitis Research Group.

Until recently, gold allergy was considered to be extremely rare. Gold has been used and worshipped for thousands of years without any obvious complaints of skin problems, either in those participating in mining and other ways of prospecting, or in those wearing jewellery. When studies on contact allergy to gold sodium thiosulfate were published at the beginning of the 1990s, the allergic nature of the reported positive patch test reactions to gold was questioned. The major argument for such questioning was the lack of demonstrable clinical relevance in most positive reactors. A major reason for the questioning may have been confusion in differentiating between contact allergy and allergic contact dermatitis. To arrive at a diagnosis of allergic contact dermatitis, 3 steps have, in principle, to be fulfilled: (i) establishment of contact allergy; (ii) demonstration of present exposure; (iii) assessment of clinical relevance, i.e., causing or aggravating a contact dermatitis. In this paper, these steps are discussed with regard to gold. With our present knowledge of contact allergy-allergic contact dermatitis, we do not recommend including gold sodium thiosulfate in the standard series. It should be applied for scientific purposes and when allergic contact dermatitis from gold is suspected.

Dermatitis, Allergic Contact↗

Dosimetry of a thyroid uptake detected in seed migration survey following a patient's iodine-125 prostate implant and in vitro measurements of intentional seed leakages.

As a quality control procedure, a post-implant seed migration survey has been accomplished on 340 prostate cancer patients since November 2001. Pulmonary seed embolization and intracardiac seed embolization have been detected. A case of thyroid uptake due to leaking iodine-125 (I-125) sources was also seized. In order to determine the dose to the thyroid, a dosimetry method was developed to link in vivo measurements and the cumulated dose to the thyroid. The calculated source leakage half-life in the case was approximately 15 days based on the measurements and the estimated cumulated dose to thyroid was 204 cGy. It is concluded that one seed was leaking. In order to verify the in vivo measurements, intentional in vitro seed leakage tests were performed. A seed was cut open and placed in a sealed glass container filled with a given volume of saline. The I-125 concentration in the saline was subsequently measured over a period of six months. Consistent in vivo and in vitro results were obtained. Recent incidents of seed leaks reported from other centers have drawn practitioners' attention to this problem. In order to make the measurements more useful, the seed leakage tests were expanded to include I-125 seeds from six other vendors. The results show that the leakage half-lives of those seeds varied from nine days to a half-year. Two seed models demonstrated least leakage. Since the measurements lasted for six months, the escape of iodine resulted from oxidation of iodide in the saline was a concern for the measurement accuracy. As a reference, another set of leakage tests were performed by adding sodium thiosulfate salt (Na2S2O3 x 5 H2O) to the saline. Sodium thiosulfate is a reducing agent that prevents the conversion of iodide to iodate so as to minimize I-125 evaporation. As a result, significantly shortened leakage half-lives were observed in this group. Seed agitation was also performed and no significant deviations of the leakage rates were observed. Considering the body fluid is more complex than saline, the in vivo leakage half-life, in case a source leak is encountered, may vary significantly from what is presented in this paper due to chemical reactions. In vivo measurements thus may produce a more accurate estimation of leakage half-life and thyroid uptake dose.

Brachytherapy↗

Development and validation of a neutralizer system for in vitro evaluation of some antiseptics.

A neutralizer system was developed and validated for use in the in vitro bactericidal evaluation of three commonly used antiseptics, namely, Hibiclens (4% [wt/vol] chlorhexidine gluconate), Betadine (7.5% [wt/vol] povidone-iodine), and pHisoHex (3% [wt/vol] hexachlorophene). The neutralizer finally selected after a screening of 12 potential candidates consisted of 3% Asolectin, 10% Tween 80, and 0.3% sodium thiosulfate in the recovery agar. This neutralizer system was tested and validated for its neutralizing capacity for the three antiseptics, as well as for its lack of inherent bactericidal action against Staphylococcus aureus and a number of gram-negative bacteria of clinical significance, With no more than a 10-fold dilution of the antiseptic, the selected neutralizer system was 100% effective in neutralizing all the bacteriostatic carry-over of the three antiseptics and was also completely without any inherent bactericidal action against all the test organisms used. Sodium sulfite (considered to be a potential inactivator for iodophores such as Betadine), even in concentrations as low as 0.1%, was found to be ineffective or inherently bactericidal, whereas 0.3% sodium thiosulfate, in combination with Asolectin and Tween 80, was adequate (effective as well as non-bactericidal) and was considered to be essential for the neutralization of the three test antiseptics, namely, Hibiclens, Betadine, and pHisoHex.

Anti-Infective Agents, Local↗

Chemolithotrophic sulfur-oxidizing bacteria from the galapagos rift hydrothermal vents.

Three distinct physiological types of sulfur-oxidizing bacteria were enriched and isolated from samples collected at several deep-sea hydrothermal vents (2,550 m) of the Galapagos Rift ocean floor spreading center. Twelve strains of the obligately chemolithotrophic genus Thiomicrospira were obtained from venting water and from microbial mats covering surfaces in the immediate vicinity of the vents. From these and other sources two types of obligately heterotrophic sulfur oxidizers were repeatedly isolated that presumably oxidized thiosulfate either to sulfate (acid producing; 9 strains) or to polythionates (base producing; 74 strains). The former were thiobacilli-like, exhibiting a thiosulfate-stimulated increase in growth and CO(2) incorporation, whereas the latter were similar to previously encountered pseudomonad-like heterotrophs. The presence of chemolithotrophic sulfur-oxidizing bacteria in the sulfide-containing hydrothermal water supports the hypothesis that chemosynthesis provides a substantial primary food source for the rich populations of invertebrates found in the immediate vicinity of the vents.

Journal Article↗

Reduced sulfur in ashes and slags from the gasification of coals: availability for chemical and microbial oxidation.

This study was initiated to determine if reduced sulfur contained in coal gasifier ash and slag was available for microbial and chemical oxidation because eventual large-quantity landfill disposal of these solid wastes is expected. Continuous application of distilled water to a column containing a high-sulfur-content (4% [wt/wt]) gasifier slag yielded leachates with high sulfate levels (1,300 mg of sulfate liter) and low pH values (4.2). At the end of the experiment, a three-tube most-probable-number analysis indicated that the waste contained 1.3 x 10 thiosulfate-oxidizing bacteria per g. Slag samples obtained aseptically from the column produced sulfate under both abiotic and biotic conditions when incubated in a mineral nutrient solution. Both microbial and chemical sulfate syntheses were greatly stimulated by the addition of thiosulfate to the slag-mineral nutrient solution. These results led to a test of microbial versus chemical sulfur oxidation in ashes and slags from five gasification processes. Sulfate production was measured in sterile (autoclaved) and nonsterile suspensions of the solid wastes in a mineral nutrient solution. These ashes and slags varied in sulfur content from 0.3 to 4.0% (wt/wt). Four of these wastes demonstrated both chemical (2.0 to 27 mug of sulfate g day) and microbial (3.1 to 114 mug of sulfate g day) sulfur oxidation. Obvious relationships between sulfur oxidation rate and either sulfur content or particle size distribution of the wastes were not immediately evident. We conclude that the sulfur contained in all but one waste is available for oxidation to sulfuric acid and that microorganisms play a partial role in this process.

Journal Article↗

Improved Assay for Rhodanese in Thiobacillus spp.

Rhodanese (thiosulfate:cyanide sulfurtransferase; EC 2.8.1.1) catalyzes the conversion of thiosulfate and cyanide to thiocyanate and sulfite. Conventional rhodanese assays colorimetrically measure the formation of one or the other of the products. These assays suffer from the fact that there is significant nonbiological formation of these products in addition to the enzymatically catalyzed reaction. In the present report, we describe a modified procedure for assaying rhodanese in which a separate boiled control was prepared for each assay trial. The boiled control corrected for the nonbiological contributions to product formation.

Journal Article↗

Sulfur chemistry in bacterial leaching of pyrite.

In the case of pyrite bioleaching by Leptospirillum ferrooxidans, an organism without sulfur-oxidizing capacity, besides the production of tetra- and pentathionate, a considerable accumulation of elemental sulfur occurred. A similar result was obtained for chemical oxidation assays with acidic, sterile iron(III) ion-containing solutions. In the case of Thiobacillus ferrooxidans, only slight amounts of elemental sulfur were detectable because of the organism's capacity to oxidize sulfur compounds. In the course of oxidative, chemical pyrite degradation under alkaline conditions, the accumulation of tetrathionate, trithionate, and thiosulfate occurred. The data indicate that thiosulfate, trithionate, tetrathionate, and disulfane-monosulfonic acid are key intermediate sulfur compounds in oxidative pyrite degradation. A novel (cyclic) leaching mechanism is proposed which basically is indirect.

Journal Article↗