Rapid Methods for Determining Decarboxylase Activity: Arginine Decarboxylase

A rapid biochemical method for the determination of arginine decarboxylase (EC 4.1.1.19) activity has been developed for use in the routine clinical microbiology laboratory and correlated with similar procedures for ornithine and lysine decarboxylase (EC 4.1.1.18) systems. It is based on the detection of agmatine, the amine end product formed during growth on a synthetic medium containing arginine as the key amino acid. A modified diacetyl reagent is used to detect this amine after a differential butanol extraction of the cultures. This procedure can be used to detect this amine after a 1- to 4-hr incubation period (with the use of an initial concentrated inoculum) or with an overnight culture. Thus, both an indirect measurement based on the alkalinization of the medium and a lengthy incubation period were avoided. Parameters for optimal enzyme activity and the pertinent enzyme systems involved in arginine and agmatine catabolism are discussed in detail.     

Interactions of GTP with the ATP-grasp domain of GTP-specific succinyl-CoA synthetase

Two isoforms of succinyl-CoA synthetase exist in mammals, one specific for ATP and the other for GTP. The GTP-specific form of pig succinyl-CoA synthetase has been crystallized in the presence of GTP and the structure determined to 2.1 A resolution. GTP is bound in the ATP-grasp domain, where interactions of the guanine base with a glutamine residue (Gln-20beta) and with backbone atoms provide the specificity. The gamma-phosphate interacts with the side chain of an arginine residue (Arg-54beta) and with backbone amide nitrogen atoms, leading to tight interactions between the gamma-phosphate and the protein. This contrasts with the structures of ATP bound to other members of the family of ATP-grasp proteins where the gamma-phosphate is exposed, free to react with the other substrate. To test if GDP would interact with GTP-specific succinyl-CoA synthetase in the same way that ADP interacts with other members of the family of ATP-grasp proteins, the structure of GDP bound to GTP-specific succinyl-CoA synthetase was also determined. A comparison of the conformations of GTP and GDP shows that the bases adopt the same position but that changes in conformation of the ribose moieties and the alpha- and beta-phosphates allow the gamma-phosphate to interact with the arginine residue and amide nitrogen atoms in GTP, while the beta-phosphate interacts with these residues in GDP. The complex of GTP with succinyl-CoA synthetase shows that the enzyme is able to protect GTP from hydrolysis when the active-site histidine residue is not in position to be phosphorylated.     

Differential effects of weather,plant phenology and predators on the seasonal variation of aphids on cabbage

The aphids Lipaphis erysimi pseudobrassicae (Davis) and Myzus persicae (Sulzer) pose serious threats to the production of cruciferous crops in the tropics. Understanding their population dynamics is important for developing integrated pest management programmes to minimize their damage to crops. This study investigated the effects of climatic factors, natural enemies and plant age on the population dynamics of these pests. The population density of aphids and their natural enemies in 20 cabbage plants, and weather conditions were monitored for five cropping seasons from 2019 to 2021 in two agroecological zones of Ghana (Coastal Savannah and Deciduous Forest zones). The highest population density of L. e pseudobrassicae was recorded in January (dry season) in both agroecological zones, while the highest population density for M. persicae occurred in September (minor rainy season) and August (dry spell) in the Coastal Savannah and Deciduous Forest zones, respectively. The highest aphid densities were noted to occur during periods with low relative humidity and low rainfall. The population density of L. e. pseudobrassicae was significantly negatively related to plant age, air temperature and relative humidity, and positively related to syrphids (Paragus borbonicus) and spiders in the Coastal Savannah zone, while in the Deciduous Forest zone, it was significantly positively related to coccinellids. On the other hand, M. persicae population density was significantly positively related to syrphids and coccinellids in the Deciduous Forest zone. Rainfall negatively affected syrphids in the Coastal Savannah zone, while air temperature positively affected syrphids and negatively affected spiders in the Deciduous Forest zone. Coccinellids had a significant positive relationship with relative humidity in the Deciduous Forest zone. This study provides important insights into the key factors that regulate aphid population densities on cabbage and will support development of timely interventions to manage these pests.     

Hevin/SC1, a matricellular glycoprotein and potential tumor-suppressor of the SPARC/BM-40/Osteonectin family

Hevin is an extracellular matrix-associated, secreted glycoprotein belonging to the secreted protein acidic and rich in cysteine (SPARC) family of matricellular proteins. It contains three conserved structural domains that are implicated in the regulation of cell adhesion, migration, and proliferation. Hevin is expressed during embryogenesis and tissue remodeling and is especially prominent in brain and vasculature. Its down-regulation in a number of cancers and the possibility of its functional compensation by SPARC has led to recent interest in hevin as a tumor suppressor and regulator of angiogenesis.     

Leaf morphology,rather than plant water status,underlies genetic variation of rice leaf rolling under drought

Soil drying causes leaf rolling in rice, but the relationship between leaf rolling and drought tolerance has historically confounded selection of drought‐tolerant genotypes. In this study on tropical japonica and aus diversity panels (170–220 genotypes), the degree of leaf rolling under drought was more affected by leaf morphology than by stomatal conductance, leaf water status, or maintenance of shoot biomass and grain yield. A range of canopy temperature and leaf rolling (measured as change in normalized difference vegetation index [ΔNDVI]) combinations were observed among aus genotypes, indicating that some genotypes continued transpiration while rolled. Association mapping indicated colocation of genomic regions for leaf rolling score and ΔNDVI under drought with previously reported leaf rolling genes and gene networks related to leaf anatomy. The relatively subtle variation across these large diversity panels may explain the lack of agreement of this study with earlier reports that used small numbers of genotypes that were highly divergent in hydraulic traits driving leaf rolling differences. This study highlights the large range of physiological responses to drought among rice genotypes and emphasizes that drought response processes should be understood in detail before incorporating them into a varietal selection programme.     

Packaging of transducing DNA by bacteriophage P1

Summary P1 transduces bacterial chromosomal markers with widely differing frequencies. We use quantitative Southern hybridisations here to show that, despite this, most markers are packaged at similar levels. Exceptions are a group of markers near 2 min and another at 90 min which seem to be packaged at levels two-to threefold higher. We thus conclude that certain marker frequency variations in transduction can be explained by differences in packaging level, but that most cannot. The limited range in packaging levels suggests that P1 can initiate the packaging of chromosomal DNA from many sites. This idea is supported by our failure to find any chromosomal sequences with homology to the phage pac site and by the occurrence of hybridising bands which seem to suggest sequential packaging from a large number of specific sites. We eliminate the possibility that chromosomal DNA packaging is the result of endonucleolytic cutting by the P1 res enzyme.     

Rapid Methods for Determining Decarboxylase Activity: Ornithine and Lysine Decarboxylases

A rapid biochemical method for the determination of ornithine and lysine decarboxylase (EC 4.1.1.18) activity has been developed for use in the routine clinical laboratory. It is based on the detection of the amine end product produced in response to the single key amino acid added to a synthetic medium. A modified ninhydrin reagent is used to detect the amine after a chloroform extraction. This procedure can be used with a 1- to 4-hr incubation period (utilizing an initial concentrated inoculum) or with an overnight culture. Thus, measurements based on the alkalinization of the medium after a lengthy incubation period are avoided. Optimal parameters for enzyme activity are discussed.     

Expression of the Escherichia coli pcnB gene is translationally limited using an inefficient start codon: a second chromosomal example of translation initiated at AUU

Expression of the gene pcnB, encoding the dispensable Escherichia coli poly(A) polymerase (PAPI), which is toxic when overproduced, was investigated. Its promoter was identified and found to be moderately strong when used to express a beta-galactosidase reporter. Expression levels were not affected by increasing or decreasing PcnB concentration. Translation of pcnB was found to initiate from the non-canonical initiation codon AUU. The only other coli gene reported to use AUU as initiation codon is infC, which encodes the initiation factor IF-3. AUU, in common with other rarely used initiation codons, is discriminated against by IF-3, resulting in the aborting of most AUU-promoted initiation events. This enables AUU to form part of an autoregulatory circuit controlling IF-3 production. We show that InfC discrimination reduces PcnB production fivefold. This is the first instance of this mechanism being used to limit severely the production of a potentially toxic product.     

Insects of the Muni-Pomadze Ramsar site

A variety of collection methods were used to inventory the insect diversity of the Muni-Pomadze Ramsar site along the Ghana coastline. A total of 75 butterfly species in five families were collected and identified. Twenty-six percent of the butterfly species were open country species. The butterflies were all typical of the coastal zone and no endangered or narrowly endemic species were recorded. The Muni-Pomadze site was also rich in other insect species (67 insect species belonging to 15 orders) as a result of the diverse terrestrial habitats surrounding the lagoon. Butterfly species composition changed with habitat indicating a fine-grained response by the butterfly communities to habitat changes. Data from long-term monitoring of butterfly communities at Muni-Pomadze could prove useful as indicators of habitat quality.