The Identity of Hexokinase Activities from Mitochondrial and Cytoplasmic Fractions of Rat Brain Homogenates

Cytoplasmic hexokinase (ATP: D-hexose 6-phosphotransferase, EC 2.7.1.1) was purified from the soluble fraction of a rat brain homogenate by a procedure that included a unique affinity elution of the enzyme from Blue Dextran-Sepharose. The purified enzyme was examined with respect to properties in which the impure cytoplasmic enzyme has been reported to differ from the solubilized mitochondrial enzyme. These included the ability to bind to mitochondria, inhibition by quercetin, effect of pH on activity, and kinetics. In all regards the purified mitochondrial and cytoplasmic enzymes appeared identical. In addition, comparative peptide maps after partial proteolysis showed no detectable differences. These results do not support the view that there exist distinct mitochondrial and cytoplasmic forms of hexokinase, the latter being permanently relegated to a cytoplasmic location and unable to participate in a dynamic equilibrium with the mitochondrially-bound enzyme. Alternatives are proposed to explain previous results that had been interpreted as indirect evidence for the existence of a distinct cytoplasmic hexokinase.     

Fluence response relationship of carotenogenesis inNeurospora crassa

The fluence response of the blue light induced carotenoid synthesis inNeurospora is biphasic. Using fluence rates between 0.3 and 40 Wm^-2, increasing illumination times beyond 16 min (at 20°C) result in a second rise of the amount of carotenoids synthesized in the subsequent dark period. On altering the temperature, the transition point to the second phase of the response is shifted to shorter/longer illumination periods with increasing/decreasing temperature, respectively. The transition point can also be shifted by administering high fluence rates of near UV light: The start of the second phase is already triggered after an irradiation time of 2 min. The findings suggest that elements of the transduction sequence become depleted and senstivity recovers in a temperature-dependent process. The biphasic response and the effects of UV light are discussed in relation to the transduction mechanism and to the ecological significance.Presented in part at the meeting of the Deutsche Botanische Gesellschaft, September 1978, Marburg, Federal Republic of Germany     

Stomatal responses to light in the facultative Crassulacean acid metabolism species, Portulacaria afra

Guard cell responses to light are mediated by guard cell chlorophyll and by a specific blue light photoreceptor. Gas exchange and epidermal peel techniques were employed to investigate these responses in the facultative Crassulacean acid metabolism (CAM) species, Portulacaria afra (L.) Jacq. In P. afra individuals performing C₃ metabolism, red light stimulated an increase in leaf conductance in intact leaves and stomatal opening in isolated epidermal peels, indicating the presence in guard cells of the chlorophyll-mediated response to light. Under a background of continuous red illumination, conductance exhibited transient increases following pulses of blue but not red light, indicating that the specific stomatal response to blue light was also operative. In contrast, in CAM individuals, conductance in gas exchange experiments and stomatal opening in epidermal peel experiments were not stimulated by red light. In CAM plants, conductance did not increase following blue light pulses administered over a range of temperatures, vapor pressure differences (VPD), ambient CO₂ concentrations and background red light intensities. These results indicate that P. afra does possess typical guard cell responses to light when performing C₃ metabolism. The metabolic pathways mediating these responses are either lost or inhibited when CAM is induced.     

Antibiotic-resistantArthrobacter sp. andPseudomonas sp. responses in the rhizosphere of blue grama after herbage removal

Summary Streptomycin-resistant Pseudomonas and Arthrobacter were isolated from semi-arid grassland soil and their relative responses in the rhizosphere of blue grama (Bouteloua gracilis) subjected to herbage removal were evaluated. Using plants grown in normal soil, the two bacteria showed differential responses to herbage removal, which were most marked in the rhizoplane, where the Pseudomonas showed a two-log unit increase over a 60 hour period, while Arthrobacter, in contrast, exhibited a one-log unit decrease in viable counts for at least 48 hours after defoliation, responses which are similar to those observed in root exudate medium experiments by earlier workers. These results suggest that the rhizoplane may be a critical environment for interaction of these two types of microorganisms, and that sequential responses of the root-associated soil microorganisms may occur after herbage removal from this important rangeland plant. These responses are most likely associated with increased exudate release following herbage removal, which has been best documented using blue grama grown under sterile conditions.     

Blue adaptation: An experimental tool for the study of visual receptor mechanisms and behaviour of Drosophila

Physiological and behavioural studies with Drosophila to elucidate visual mechanisms have exploited the bi-stability of the visual pigment in the peripheral retinula cells R1–6, and the off-on switch action of blue and orange light. Measurements of flicker fusion and response waveform from both receptor and lamina regions prior and subsequent to blue adaptation, which induces a prolonged depolarising afterpotential and loss of visual function in R1–6, show these retinula cells to have a high fusion frequency and R7/8, the central retinula cells, a lower fusion frequency. Such measurements also allow analysis of the extracellular response in terms of contributing cells, and its potential for studying the fly's ability to respond to various potential visual cues such as a rotating plane of polarised light. Blue adapted flies fail to fixate normally a black stripe, confirming a role for R1–6 in orientation behaviour requiring a competent degree of acuity.Based on material presented at the European Neurosciences Meeting, Florence, September 1978     

A modified Coomassie Brilliant Blue staining method at nanogram sensitivity compatible with proteomic analysis

A more sensitive and convenient Coomassie Brilliant Blue (CBB) staining method for visualizing proteins was developed. Compared with the modifications include the supplement of 10% (v/v) methanol into the fixing solution, an increase of an additional sensitization step and CBB raised from 0.1 to 0.125%. The improved method can detect proteins at nanogram level. The improved method is more sensitive than Blue Silver and more convenient than the Silver protocol. Mass spectrometry results confirmed that it is suitable for subsequent proteomic research.     

Assessment of blue-stain resistance according to the EN 152 and a reverse test method using visual and computer-aided techniques

A computer technique for assessing blue-stained coated wood has been implemented for evaluating the discoloration of coatings and analysing the interior wood staining of samples subjected to testing according to European Standard EN 152. The comparison of visual assessment and computer-evaluated percentages of blue staining is based on a combination of correlation measures, principal components and cluster analysis. It appears difficult to imitate human evaluation with image processing, since computer ratings represent exact percentages, while subjective evaluations do not. Additionally, more specific techniques for exploring fungal growth in coated wood have been described. As EN 152 was specifically developed for testing efficacy of wood preservatives, a modified test methodology was elaborated for testing the efficacy of wood coatings, called here as the EN 152-reverse method. Furthermore three-dimensional (3D) reconstruction is validated as a tool for in-depth analysis of blue-stain disfigurement. This 3D visualisation indicates important differences in fungal infestation and proves its suitability for blue-stain resistance testing.     

Synthesis,characterization and in vitro biological evaluation of some novel 1,3,5-triazine–Schiff base conjugates as potential antimycobacterial agents

A series of some novel 1,3,5-triazine–Schiff base conjugates (1–32) have been synthesized and evaluated for their in vitro antimycobacterial activity against Mycobacterium tuberculosis H37Rv using Alamar Blue assay and the activity expressed as the minimum inhibitory concentration (MIC) in μg/mL. Compounds 4 (4-Methoxy-6-methyl-N-(3,4,5-trimethoxybenzylidene)-1,3,5-triazin-2-amine), 11 (4-Methoxy-6-methyl-N-(2-hydroxy-3-bromo-5-chloro-benzylidene)-1,3,5-triazin-2-amine) and 24 (4-Methoxy-6-methyl-N-(1-(2,5-dihydroxyphenyl)ethylidene)-1,3,5-triazin-2-amine) exhibited a significant activity at 3.125, 6.25 and 6.25 μg/mL, respectively, when compared with the antitubercular drugs such as ethambutol (3.125 μg/mL), pyrazinamide (6.25 μg/mL) and streptomycin (6.25 μg/mL) and it could be a potential starting point to develop new lead compounds in the fight against Mycobacterium tuberculosis H37Rv.