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Isocitrate lyase of conifers (Pinus pinea).

1. Isocitrate lyase has been purified about 60 times from the conifer Pinus pinea. A first characterization was made. 2. The high instability is an important feature of this enzyme from higher plants, this causes serious problems in the purification and characterization. 3. A substantial agreement with the data from the literature was found for what concerns pH dependence of Vmax and pKm, the effect of bivalent cations and the requirement of Mg2+. 4. Kinetic studies gave evidence for a mechanism ordered uni-bi with glyoxylate being the last product released, kinetic constants were calculated, no evidence for cooperative effects was found. 5. Equilibrium constant by Haldane method calculation agrees with value calculated with isocitrate lyase from the bacterium Pseudomonas indigofera.

Cations, Divalent↗

Free-radical scavenging action of medicinal herbs from Ghana: Thonningia sanguinea on experimentally-induced liver injuries.

The antioxidant action of medicinal herbs used in Ghana for treating various ailments was evaluated in vitro and in vivo. Five plants, Desmodium adscendens, Indigofera arrecta, Trema occidentalis, Caparis erythrocarpus, and Thonningia sanguinea were tested for their free radical scavenging action by their interaction with 1,1-diphenyl-2-picrylhydrazyl (DPPH). Of these five plants, only Thonningia sanguinea was found to scavenge the DPPH radical. Lipid peroxidation in liver microsomes induced by H2O2 was also inhibited by T. sanguinea. The hepatoprotective effect of T. sanguinea was studied on acute hepatitis induced in rats by a single dose of galactosamine (GalN, 400 mg/kg, IP) and in mice by carbon tetrachloride (CCl4, 25 microl/kg, IP). GalN induced hepatotoxicity in rats as evidenced by an increase in alanine aminotransferase (ALT) and glutathione (GSH) S-transferase activities in serum was significantly inhibited when T. sanguinea extract (5 ml/kg, IP) was given to rats 12 hr and 1 hr before GalN treatment. The activity of liver microsomal GSH S-transferase, which is known to be activated by oxidative stress, was increased by the GaIN treatment and this increase was blocked by T. sanguinea pretreatment. Similarly, T. sanguinea pretreatment also inhibited CCl4-induced hepatotoxicity in mice. These data indicate that T. sanguinea is a potent antioxidant and can offer protection against GalN- or CCl4-induced hepatotoxicity.

Alanine Transaminase↗

Screening of Yemeni medicinal plants for antibacterial and cytotoxic activities.

Ethanolic extracts of 20 selected plant species used by Yemeni traditional healers to treat infectious diseases were screened for their antibacterial activity against both Gram-positive and Gram-negative bacteria, as well as for cytotoxic activity. Fourteen of the ethanolic extracts showed variable degrees of antibacterial activity. The active ethanolic extracts were partitioned between ethyl acetate and water for a first separation. The ethyl acetate extract of Lawsonia inermis was found to be the most active one against all bacteria in the test system. Other promising results could be obtained from extracts of Aloe perryi, Indigofera oblongifolia, Meriandra benghalensis and Ziziphus spina christi. Additionally, the ethanolic extracts of the 20 plants under investigation were tested for their cytotoxic effects on FL-cells using the neutral red assay. Extracts of Calotropis procera, Chenopodium murale, Pulicaria orientalis, Tribulus terrestris and Withania somniferum displayed a remarkable activity.

Anti-Bacterial Agents↗

Diversity of bradyrhizobia from 27 tropical Leguminosae species native of Senegal

We isolated 71 slow-growing bacterial strains from nodules of 27 native leguminous plants species in Senegal (West-Africa) belonging to the genera Abrus, Alysicarpus, Bryaspis, Chamaecrista, Cassia, Crotalaria, Desmodium, Eriosema, Indigofera, Moghania, Rhynchosia, Sesbania, Tephrosia, and Zornia playing an ecological role and having agronomic potential in arid regions. The isolates were characterised by restriction fragment length polymorphism (RFLP) analysis of PCR-amplified 16S rDNA and comparative SDS-PAGE of whole-cell proteins; reference strains of the different known rhizobial species and groups were included as references. We conclude that these nodule isolates are diverse, and form several phylogenetic subgroups inside Bradyrhizobium. Nodulation tests performed on 5 plant species demonstrated host specificity among the strains studied.

Journal Article↗

Micro-Raman spectroscopy of natural and synthetic indigo samples.

In this work indigo samples from three different sources are studied by using Raman spectroscopy: the synthetic pigment and pigments from the woad (Isatis tinctoria) and the indigo plant (Indigofera tinctoria). 21 samples were obtained from 8 suppliers; for each sample 5 Raman spectra were recorded and used for further chemometrical analysis. Principal components analysis (PCA) was performed as data reduction method before applying hierarchical cluster analysis. Linear discriminant analysis (LDA) was implemented as a non-hierarchical supervised pattern recognition method to build a classification model. In order to avoid broad-shaped interferences from the fluorescence background, the influence of 1st and 2nd derivatives on the classification was studied by using cross-validation. Although chemically identical, it is shown that Raman spectroscopy in combination with suitable chemometric methods has the potential to discriminate between synthetic and natural indigo samples.

Journal Article↗

Isolation and characterization of isocitrate lyase from a thermophilic Bacillus sp.

Isocitrate lyase was isolated in homogeneous state from a thermophilic Bacillus. The enzyme has a mol.wt. of 180000 and a pI of 4.5 and contains threonine as the N-terminal residue. It resembles in size the cognate enzyme from the mesophilic bacterium Pseudomonas indigofera, but is smaller than the enzyme from the eukaryotic fungus Neurospora crassa. All three lyases are tetramers and similar in amino acid composition, but the thermophile enzyme is distinctive from its mesophilic coutnerparts in possessing a lower catalytic-centre activity, greater resistance to chemical and thermal denaturation and fewer thiol groups and in being strongly activated by salts. Salt activation, by 0.4M-KCl, is about 3-fold at 30 degrees C and pH 6.8 and weakens progressively as the temperature or pH is raised. The activation is probably due to a change in the enzyme conformation caused by the electrolyte modifying the interaction between charged groups or between hydrophobic groups in protein. The possible significance of the salt activation, of the relative paucity of thiol groups and of the greater resistance to chemical denaturants is discussed. Besides its effect on the Vmax., KCl produces large increases in the magnitude of several kinetic parameters. A rise in reaction temperature from 30 to 55 degrees C produces a somewhat similar result. In view of these peculiar features, the patterns of inhibition of enzyme activity by compounds such as succinate and phosphoenolpyruvate were examined at 30 and 55 degrees C in the presence and absence of KCl.

Amino Acids↗

Catabolism of arginine, citrulline and ornithine by Pseudomonas and related bacteria.

The distribution of the arginine succinyltransferase pathway was examined in representative strains of Pseudomonas and related bacteria able to use arginine as the sole carbon and nitrogen source for growth. The arginine succinyltransferase pathway was induced in arginine-grown cells. The accumulation of succinylornithine following in vivo inhibition of succinylornithine transaminase activity by aminooxyacetic acid showed that this pathway is responsible for the dissimilation of the carbon skeleton of arginine. Catabolism of citrulline as a carbon source was restricted to relatively few of the organisms tested. In P. putida, P. cepacia and P. indigofera, ornithine was the main product of citrulline degradation. In most strains which possessed the arginine succinyltransferase pathway, the first step of ornithine utilization as a carbon source was the conversion of ornithine into succinylornithine through an ornithine succinyltransferase. However P. cepacia and P. putida used ornithine by a pathway which proceeded via proline as an intermediate and involved an ornithine cyclase activity.

Acyltransferases↗

Comparative studies on the degradation of guanidino and ureido compounds by Pseudomonas.

The utilization of guanidino and ureido compounds was studied in several Pseudomonas species. Multiple routes of agmatine catabolism were found. All members of the homology group I of Pseudomonas use the initial deamination of agmatine to carbamoylputrescine which is subsequently converted to putrescine. In Pseudomonas indigofera, the catabolism of agmatine can also occur via an initial hydrolysis of the amidino group to putrescine catalyzed by an agmatine amidinohydrolase. A third pathway was found in Pseudomonas cepacia, namely oxidative deamination producing guanidinobutyraldehyde catalyzed by agmatine dehydrogenase, followed by formation of guanidinobutyrate and removal of urea by guanidinobutyrate amidinohydrolase to produce 4-aminobutyrate. Novel amidino-hydrolases were characterized in P. putida for the utilization of arcaine and audouine, and in P. cepacia for arcaine, homoarginine and guanidinovalerate. Guanidinovalerate amidinohydrolase was also detected in P. doudoroffii. Some of these amidinohydrolases accept more than one substrate, e.g., guanidinobutyrate and guanidinovalerate utilization by P. doudoroffii and P. cepacia, the catabolism of arcaine and audouine by P. putida, and the degradation of arcaine and homoarginine by P. cepacia.

Amidohydrolases↗

Experimental poisoning of horses and cattle with Swainsona canescens var horniana.

Horses and cattle fed swainsona (Swainsona canescens var horniana) over a period of 8 to 10 weeks lost condition and became incoordinated and hypersensitive. Histological examination of tissues from affected animals revealed the characteristic changes of widespread cellular vacuolation and axonal spheroids in the CNS. Cattle withdrawn from the toxic plant after being fed for varying periods up to 8 weeks returned to normal. Serum alpha-mannosidase activity declined significantly in cattle during the test period, whereas in horses that activity rose. Serum alkaline phosphatase levels increased significantly in cattle but not in horses. The similarity of the clinical signs of disease in horses was noted to those seen in Indigofera linnaei poisoning (Birdsville disease). Differential diagnosis can be made by botanical observations and by an increased frequency of vacuolated lymphocytes in the blood in swainsona poisoning.

Animals↗

Genotypic characterization of Bradyrhizobium strains nodulating small Senegalese legumes by 16S-23S rRNA intergenic gene spacers and amplified fragment length polymorphism fingerprint analyses.

We examined the genotypic diversity of 64 Bradyrhizobium strains isolated from nodules from 27 native leguminous plant species in Senegal (West Africa) belonging to the genera Abrus, Alysicarpus, Bryaspis, Chamaecrista, Cassia, Crotalaria, Desmodium, Eriosema, Indigofera, Moghania, Rhynchosia, Sesbania, Tephrosia, and Zornia, which play an ecological role and have agronomic potential in arid regions. The strains were characterized by intergenic spacer (between 16S and 23S rRNA genes) PCR and restriction fragment length polymorphism (IGS PCR-RFLP) and amplified fragment length polymorphism (AFLP) fingerprinting analyses. Fifty-three reference strains of the different Bradyrhizobium species and described groups were included for comparison. The strains were diverse and formed 27 groups by AFLP and 16 groups by IGS PCR-RFLP. The sizes of the IGS PCR products from the Bradyrhizobium strains that were studied varied from 780 to 1,038 bp and were correlated with the IGS PCR-RFLP results. The grouping of strains was consistent by the three methods AFLP, IGS PCR-RFLP, and previously reported 16S amplified ribosomal DNA restriction analysis. For investigating the whole genome, AFLP was the most discriminative technique, thus being of particular interest for future taxonomic studies in Bradyrhizobium, for which DNA is difficult to obtain in quantity and quality to perform extensive DNA:DNA hybridizations.

Bacterial Typing Techniques↗

Structural basis of the thermostability of monomeric malate synthase from a thermophilic Bacillus.

Malate synthases from a thermophilic Bacillus and Escherichia coli have been isolated in a high state of purity. Molecular weights of these two proteins determined in the native state and after denaturation in sodium dodecyl sulfate-mercaptoethanol show that the enzymes are monomeric. This conclusion is supported, for the thermophile enzyme, by the result of an electrophoretic analysis of that protein after treatment with dimethylsuberimidate and denaturation. The thermophilic Bacillus malate synthase is considerably more thermostable than its mesophilic counterparts from E. coli, Bacillus licheniformis, and Pseudomonas indigofera. It is, however, markedly labilized by an increase in the ionic strength of the medium brought about by the addition of 0.2 M potassium chloride or in pH above 9. Increased ionic strength has little effect on the thermostability of the mesophilic bacterial malate synthases. These observations provide strong support for the idea that monomeric proteins in thermophiles owe their unusual heat stability to the presence of salt bridges in their tertiary structure.

Bacillus↗

Occurrence of isocitrate lyase in a thermophilic bacillus species.

A thermophilic, sporeforming bacterium has been isolated from soil on a medium containing acetate as a carbon source. This organism is similar to Bacillus stearothermophilus in most respects but differs in its inability to hydrolyze starch. Isocitrate lyase is present in cell-free extracts of organisms grown in a medium with acetate as a carbon source. The specific activity was 400 times lower in extracts of organisms utilizing glucose as a carbon source. With crude extracts, enzyme activity was strongly stimulated by Mg(++), but cysteine and ethylenediaminetetraacetate had little effect. It appeared to be more heat-stable than the pure isocitrate lyase from Pseudomonas indigofera.

Bacillus↗

Laribacter hongkongensis gen. nov., sp. nov., a novel gram-negative bacterium isolated from a cirrhotic patient with bacteremia and empyema.

A bacterium was isolated from the blood and empyema of a cirrhotic patient. The cells were facultatively anaerobic, nonsporulating, gram-negative, seagull shaped or spiral rods. The bacterium grows on sheep blood agar as nonhemolytic, gray colonies 1 mm in diameter after 24 h of incubation at 37 degrees C in ambient air. Growth also occurs on MacConkey agar and at 25 and 42 degrees C but not at 4, 44, and 50 degrees C. The bacterium can grow in 1 or 2% but not 3, 4, or 5% NaCl. No enhancement of growth is observed with 5% CO(2). The organism is aflagellated and nonmotile at both 25 and 37 degrees C. It is oxidase, catalase, urease, and arginine dihydrolase positive, and it reduces nitrate. It does not ferment, oxidize, or assimilate any sugar tested. 16S rRNA gene sequencing showed that there are 91 base differences (6.2%), 112 base differences (7.7%), and 116 base differences (8.2%) between the bacterium and Microvirgula aerodenitrificans, Vogesella indigofera, and Chromobacterium species, respectively. The G+C content (mean and standard deviation) is 68.0% +/- 2.43%, and the genomic size is about 3 Mb. Based on phylogenetic affiliation, the bacterium belongs to the Neisseriaceae family of the beta-subclass of Proteobacteria. For these reasons, a new genus and species, Laribacter hongkongensis gen. nov., sp. nov., is proposed, for which HKU1 is the type strain. Further studies should be performed to ascertain the potential of this bacterium to become an emerging pathogen.

Bacteremia↗

Toxicological and toxicogenetic effects of plants used in popular medicine and in cattle food.

Toxicological and toxicogenetic effects of aqueous (tea) and hexanic fruit extract of Indigofera suffruticosa Mill. and hydroalcoholic root extract of Solanum agrarium Stendt. were evaluated in Balb C male mice intraperitoneally exposed. A hepatotoxic effect was observed just for animals treated with aqueous fruit extract of I. suffruticosa. In relation to the toxicogenetic effect, just the group treated with 12.5% of toxic dose of aqueous fruit extract of I. suffruticosa showed a statistically significant increase in the frequency of cells with chromosome aberrations (cytogenetic effect), although a slight increase was also observed for the highest dose (25% of LD50) of hydroalcoholic root extract of S. agrarium. The results obtained show that before S. agrarium is used as medicine and before the wide use of I. suffruticosa in cattle food, careful evaluation must be done.

Animal Feed↗

Nitrate pollution in groundwater and strategies to reduce pollution.

The input-intensive rainfed tropical ecosystem, where wet season (WS) rice (Oriza sativa L.)-dry season (DS) diversified high-value upland crops like vegetables predominate, has resulted in a problem of a large leakage of N into the environment, thereby polluting the water. Excessive use of N fertilizer in high-value crops grown in DS is economically motivated. Out of twenty water sources evaluated in a watershed with a total area of 265 ha located in Magnuang, Ilocos Norte, Philippines, twelve had near or above the World Health Organization's (WHO) NO3-N limit for drinking water of 10 ppm. Soil mineral N (upper 100 cm) observed in seven rice-sweet pepper (Capsicum annuum L.) farmers' fields ranged from 111 to 694 kg ha(-1) which decreased by 10 to 68% in plots with dry-to-wet (DTW) crops like indigo, indigo+mungo and corn. In fallow plots where mineral N was either maintained or increased, it showed movement to lower soil profiles demonstrating NO3 leaching without a crop. On average, maize (Zea mays L.) captured 176 kg N ha(-1) and indigo (Indigofera tinctoria L.) 194 kg N ha(-1). In both fallow and planted plots, mineral N declined to low levels at 100% water-filled pore spaces (WFPS) before rice transplanting. A strategy for including indigo plus maize as a N-catch crop is proposed to decrease NO3 leaching and maximize N use efficiency in a rice-sweet pepper cropping system.

Agriculture↗

Nutritional support for chronic myelogenous and other leukemias: a review of the scientific literature.

Chronic myelogenous leukemia (CML) is a slowly progressive disease characterized by the overproduction of granulocytes (neutrophils, eosinophils, and basophils). A blood smear shows moderate elevations in white blood cell counts that may persist for years and be benign. Platelets are increased in number, although their function is impaired, resulting in symptoms of easy bleeding (purpura, swollen gums). Conventional medical treatment is a marrow transplant and alkylating agents, which are usually prescribed only during crisis. Several nutrients and botanicals have been studied for use in CML, including vitamin A and all-trans retinoic acid (Retin-A), vitamin D3, vitamin E, vitamin B12, indirubin (found in herbs including Indigofera tinctoria and Isatis tinctoria), and Curcuma longa. This article briefly reviews the scientific literature on the therapeutic use of these nutrients for CML.

Antineoplastic Agents↗

[Morphological and histological studies on the Chinese drug shan-dou-gen].

The Chinese drug "Shan-Dou-Gen" has been used for removing toxic heat, promoting the subsidence of swelling and soothing the sore throat since the ancient time. The authors made a survey of the drug "Shan-Dou-Gen" available in drug markets as well as the plant origin from the drug producing districts. The results showed that the drug "Shan-Dou-Gen" used in different regions in China at present are the roots or rhizomes derived from 9 species: Sophora tonkinensis Gagnep. (Leguminosae), Menispermum dauricum DC. (Menispermaceae), Indigofera amblyantha Craib (Leguminosae), I. carlesii Carib, I., fortunei Craib, I. decoa Lindl. var. ichangensis Y. Y. Fang et C. Z. Zheng, I. kirilowii Maxim. et Palibin, I. potaninii Craib, and Beesia calthaefolia (Maxim.) Ulbr. (Ranuculaceae). In this paper, the morphological characters of the crude drugs are described, compared and illustrated with photographs. The histological structures of the used parts are described, compared and illustrated with line drawings. The morphological and histological similarities and differences found among the above 9 species are summarized, and the key for the identification of the crude drugs is provided. As the drug "Shan-Dou-Gen" derived from different species has different actions and dosages, it is necessary to give different names to different species and use them correctly.

Medicine, Chinese Traditional↗

[Population dynamics of endangered plant species Abies chensiensis].

In order to know the endangered status and causes of Abies chensiensis in Qinlin Mountains, a field investigation on 18 plots was conducted on its age structure, life table and fecundity, and its population dynamics were predicted by time sequence model. The analysis on the age structure of Abies chensiensis populations showed that there were fewer young individuals, but middle-aged and old individuals were relatively rich. The population D in Abies chensiensis-Indigofera amblyantha-Carex lanceolata association showed a relatively stable development tendency, while other four populations (A, B, C and E) in Abies chensiensis-Pinus tabulaeformis-Sinarundinaria nitida-Carex lanceolata association, Abies chensiensis-Quercus aliena var. acutserrata-Litsea pungens-Carex lanceolata association, Abies chensiensis-Betula albo-sinensis-Sinarundinaria nitida-Duchesnea indica association, and Abies chensiensis-Pinus tabulaeformis-Smilax stans-Carex lanceolata association all showed an obviously declining tendency. The analysis on the life tables and survival curves showed that the survival curve of Abies chensiensis populations belonged to Deevey III, and the death peak of different populations was in the period of 60--100 years old. The number difference among populations reflected the population habitat. Time sequence prediction indicated the numbers of old individuals would be increased at the beginning, and decreased finally in 20, 40, and 80 years. It was difficult to maintain the population stability. Analysis on 10 ecological factors showed that tree coverage, soil organisms and air humidity influenced population positively, and human disturbance and sunlight influenced population negatively. In situ conservation should be taken as the most important management countermeasure, and natural regeneration should be promoted. At the same time, artificial population should be expanded.

Abies↗