Synthesis of the ABC framework of tamulamides A and B

Synthesis of the fused tetrahydrofuran motif comprising the ABC rings of the marine ladder polyethers tamulamides A and B has been achieved via two different polyepoxide cascade strategies. Investigations into a triepoxide cascade under aqueous conditions revealed the importance of the electronic nature of the cascade end-group with this initial approach. Ultimately, a diepoxide cascade under basic conditions proved most successful, providing the ABC tetrahydropyran triad in 41% yield.     

Principles of optimization and scaling of certain processes in the vaccine production technology. 3. Method of determination of the optimal method of thermal sterilization of sugar solutions

One of the most widespread methods of thermal sugar sterilization is the use of saturated water vapour under pressure in the capacity of heat source. Guaranteed sterility of solutions should be reached under conditions providing the least sugar splitting. However, in practice the integral effect of the thermal field on the solution is as a rule not taken into consideration. A new calculation method permitting to determine the amount of sugar split in the varying thermal field.     

TFLUX: a general purpose program for the interpretation of radioactive tracer experiments.

TFLUX is a computer program which simulates radioactive tracer experiments using intact cells under steady-state conditions. It is designed for the naive computer user to use, providing English-like input. An example is described in which compartmentation and flux through the citric acid cycle in Dictyostelium discoideum are simulated.     

Label-free protein and pathogen detection using the atomic force microscope

The atomic force microscope (AFM) uses a sharp micron-scale tip to scan and amplify surface features, providing exceptionally detailed topographical information with magnification on the order of x10(6). This instrument is used extensively for quality control in the computer and semiconductor industries and is becoming a progressively more important tool in the biological sciences. Advantages of the AFM for biological application include the ability to obtain information in a direct, label-free manner and the ability to image in solution, providing real-time data acquisition under physiologically relevant conditions. A novel application of the AFM currently under development combines its surface profiling capabilities with fixed immuno-capture using antibodies immobilized in a nanoarray format. This provides a distinctive platform for direct, label-free detection and characterization of viral particles and other pathogens.     

Effects of mine acid on the longevity and reproductive rate of the gastrotricha Lepidodermella squammata (Dujardin)

Effects of mine acid on longevity and reproductive rate of the parthenogenic gastrotrich Lepidodermella squammata were studied under laboratory culture conditions. Water from unpolluted and polluted streams was used, directly or mixed, to establish a series of test conditions at pH 8.1, 7.1, 6.4, 5.2, 4.6 and 3.3. Eggs, cultured individually under each test condition, were observed at 12 h intervals for hatching, daughter egg laying and death. Data representing 50 animals under each test condition were used in the construction of a series of life tables. From these were calculated maximal life expectancy (e_x), net reproductive rate per individual lifetime (R_o) and intrinsic rate of natural increase (r_max).Values of e_x, R_o and r_max were maximal at pH 7.1 and were reduced slightly at the higher pH; e_x was greatly reduced and R_o zero at pH 6.4 and 5.2; and e_x was zero at pH 4.6 and 3.3. Analysis of variance tests indicate significant differences between e_x values of L. squammata cultured at pH 8.1 and those cultured at pH 7.1, but no differences between R_o or r_max.Associated with the decrease in pH was an increase in total conductivity and a decrease in carbonate alkalinity and hence in carbonate conductivity. It appears that L. squammata is capable of living and reproducing at pH 6.0 to 6.5 under field conditions low in carbonates, providing non-carbonate ions are not abundant, or under field conditions high in non-carbonate ions, providing sufficient carbonates are present.     

A Possible Mechanism of Blocking of Limbic Motor Seizure Reactions Induced by Activation of the Thalamic Reticular Nucleus

Experiments were directed toward elucidation of the role of the thalamic reticular nucleus (R) in the modulation of generalized seizure reactions under kindling conditions and of the mechanisms mediating the effects of stimulation of the above nucleus on seizure activity. It was shown that activation of the thalamic R in rats limits generalization of the seizure reactions both in the course of development of seizures of limbic genesis (evoked by stimulation of the hippocampus) and under conditions of the existence of a pre-formed epileptic nidus. Tetanic stimulation of the R in cats under conditions of acute experiments induced significant facilitation of IPSPs in thalamo-cortical neurons of the ventrolateral thalamic nucleus. This effect is rather long-lasting and may be considered a mechanism providing blocking of generalized seizures under kindling conditions. Neirofiziologiya/Neurophysiology, Vol. 37, No. 4, pp. 352–361, July–August, 2005.     

On estimating the role of diffraction in electron cyclotron absorption at the periphery of the plasma column

The propagation and absorption of microwave radiation at the periphery of a tokamak plasma under ECR conditions are considered. For microwaves propagating in a quasi-transverse direction, the range of plasma parameters is determined in which the effective plasma permittivity can be approximated by a piecewise linear function. With this approximation, it is possible to obtain analytic solutions to the wave equation and to use them to estimate the width of the power deposition region for different modes of microwave launching. A detailed analysis is given of tangential launching—the propagation of microwaves along a tangent to the resonance magnetic surface at which they begin to be absorbed under ECR conditions. Using the ITER tokamak as an example, it is shown that this launching method is most efficient in providing the narrowest power deposition profile at the plasma periphery. The results obtained are of interest for the problems of suppressing tearing-mode instabilities by localized ECR heating.     

Photo-induced unfolding and inactivation of bovine carbonic anhydrase in the presence of a photoresponsive surfactant

Photoreversible changes in the conformation and enzymatic activity of bovine carbonic anhydrase have been investigated as a function of photoresponsive surfactant concentration and light conditions. The light-responsive surfactant undergoes a photoisomerization from the relatively hydrophobic trans isomer under visible light to the relatively hydrophilic cis isomer upon UV illumination, providing a means to photoreversibly control enzyme–surfactant interactions. Small-angle neutron scattering and dynamic light scattering measurements, along with fluorescence spectroscopy, indicate that carbonic anhydrase unfolds upon addition of the surfactant under visible light, while only a small degree of unfolding is observed under UV light. Therefore, the enzyme is completely inactivated in the presence of the trans surfactant, while 40% of the native activity is preserved under UV light, providing a photoreversible “on/off switch” of enzyme activity. Small-angle neutron scattering data provide details of the in vitro conformational changes of the enzyme in response to the photosurfactant and light, with the enzyme found to aggregate as a result of photosurfactant-induced unfolding. Fourier transform infrared (FT-IR) spectroscopy further provides information on the secondary structure changes of the protein in the presence of photosurfactant.     

Nectar‐providing plants enhance the energetic state of herbivores as well as their parasitoids under field conditions

Abstract 1. The use of flowering vegetation has been widely advocated as a strategy for providing parasitoids and predators with nectar and pollen. However, their herbivorous hosts and prey may exploit floral food sources as well. 2. Previous laboratory studies have shown that not all flower species are equally suitable in providing accessible nectar. Relatively little is known about actual nectar exploitation under field conditions. 3. The present study investigates nectar exploitation by the pest, Plutella xylostella, and its parasitoid, Diadegma semiclausum, under field conditions and examines whether floral nectar exploitation in the field can be predicted based on controlled laboratory studies. 4. Insects were collected from fields bordered by flowering margins containing Fagopyrum esculentum, Lobularia maritima, Anethum graveolens, Centaurea jacea or the grass Lolium perenne (control). Whole insect bodies were individually assayed by HPLC to establish their sugar profile as a measure of the level of energy reserves and the degree of food source use. 5. The average overall sugar content of P. xylostella and D. semiclausum collected in fields bordered by flowering margins was significantly higher than those of individuals collected from grass‐bordered control plots. To the authors’ knowledge, this represents the first demonstration that nectar‐providing plants enhance the energetic state of herbivores under field conditions. 6. In contrast to earlier laboratory studies, the present study did not find elevated sugar contents in P. xylostella and D. semiclausum individuals collected from fields bordered by buckwheat (F. esculentum). 7. The present study shows widespread sugar feeding by both the herbivore and its parasitoid. It also shows that laboratory studies establishing nectar exploitation under controlled conditions can not always be extrapolated to actual exploitation under field conditions. This emphasises the importance of studying field‐collected insects with regard to food source use and nutritional status.     

Neuromediator provision of the organs of immune system in benzpyrene intoxication]

The density of adrenergic innervation and relative content of catecholamines were investigated in rat lymphoid organs under different type of benzpyrene treatment. It was found that both ante- and postnatal influence of toxicant leads to a decrease of neurotransmitter providing of the thymus, spleen, mesenterial, iliac and popliteal lymph nodes. On the contrary, when mixed ante- and postnatal benzpyrene influence has place, the adrenergic innervation density and relative content of catecholamines are increased. We suppose that benzpyrene treatment of pregnant animals has specific "training" effect for monoaminergic systems of foetus and causes the increase of neurotransmitter providing of immunocompetent organs in conditions of repeat body and toxicant meeting.     

Effect of Melatonin on the Intensity of Adenosine Production in the Rat Forebrain under Conditions of Acute Hypoxia and Varied Photoperiodicity

We studied the effect of injections of melatonin and modifications of the duration of illumination on the activity of 5-nucleotidase, an enzyme providing synthesis of adenosine, in the forebrain of juvenile male albino rats. The measurements were performed under conditions of acute hypobaric hypoxia. We found that, under conditions of natural illumination, neither isolated injections of melatonin nor acute hypoxia noticeably changed the activity of 5-nucleotidase. At the same time, acute hypoxia combined with melatonin injections increased the activity of this enzyme. A similar noticeable rise in the activity of 5-nucleotidase was observed after melatonin injections in normoxic animals kept in constant darkness, and in rats subjected to hypoxia without the above injections but under conditions of constant illumination. These data allow us to suppose that melatonin (whose level in the extracellular medium is a factor providing synchronization of endogenous temporal rhythms) stimulates 5-nucleotidase-mediated production of adenosine in brain neurons. Acute hypoxia promotes such an effect of melatonin.     

Development of high-cell-density fermentation for heterologous interleukin 1\ production in Kluyveromyces lactis controlled by the PH05 promoter

The use of a phosphate organic form taken up by Kluyveromyces lactis removes repression of the PHO5 promoter and releases heterologous interleukin 1\ synthesis while providing sufficient phosphate for growth. The oxidative metabolism of high-cell-density fed-batch and chemostat cultures was thus maintained under derepressed protein synthesis conditions. Interleukin 1\ production was then growth-associated, an unusual mode of protein synthesis regulation under the control of the PHO5 promoter.     

The role of antioxidant enzymes in photoprotection

The enzymatic component of the antioxidant system is discussed as one of the defensive mechanisms providing protection against excessive light absorption in plants. We present an analysis of attempts to improve stress tolerance by means of the creation of transgenic plants with elevated antioxidant enzyme activities and conclude that the effect of such transgenic manipulation strongly depends on the manner in which the stress is imposed. The following factors may diminish the differences in photosynthetic performance between transgenic plants and wild type under field conditions: effective functioning of the thermal dissipation mechanisms providing a primary line of defense against excessive light, long-term adjustments of the antioxidant system and other photoprotective mechanisms, the relatively low level of control over electron transport exerted by the Water–Water cycle, especially under warm conditions, and a decrease in the content of the transgenic product during leaf aging.     

Differential usage of storage carbohydrates in the CAM bromeliad Aechmea 'Maya' during acclimation to drought and recovery from dehydration

CAM requires a substantial investment of resources into storage carbohydrates to account for nocturnal CO₂ uptake, thereby restricting carbohydrate partitioning to other metabolic activities, including dark respiration, growth and acclimation to abiotic stress. Flexible modulation of carbon flow to the different competing sinks under changing environmental conditions is considered a key determinant for the growth, productivity and ecological success of the CAM pathway. The aim of the present study was to examine how shifts in carbohydrate partitioning could assure maintenance of photosynthetic integrity and a positive carbon balance under conditions of increasing water deprivation in CAM species. Measurements of gas exchange, leaf water relations, malate, starch and soluble sugar (glucose, fructose and sucrose) contents were made in leaves of the CAM bromeliad Aechmea ‘Maya’ over a 6‐month period of drought and subsequently over a 2‐month period of recovery from drought. Results indicated that short‐term influences of water stress were minimized by elevating the level of respiratory recycling, and carbohydrate pools were maintained at the expense of export for growth while providing a comparable nocturnal carbon gain to that in well‐watered control plants. Longer term drought resulted in a disproportionate depletion of key carbohydrate reserves. Sucrose, which was of minor importance for providing substrate for the dark reactions under well‐watered conditions, became the major source of carbohydrate for nocturnal carboxylation as drought progressed. Flexibility in terms of the major carbohydrate source used to sustain dark CO₂ uptake is therefore considered a crucial factor in meeting the carbon and energy demands under limiting environmental conditions. Recovery from CAM‐idling was found to be dependent on the restoration of the starch pool, which was used predominantly for provision of substrate for nocturnal carboxylation, while net carbon export was limited. The conservation of starch for the nocturnal reactions might be adaptive with regard to responding efficiently to a return of water stress.     

Vascular functioning and the water balance of ripening kiwifruit (Actinidia chinensis) berries

Indirect evidence suggests that water supply to fleshy fruits during the final stages of development occurs through the phloem, with the xylem providing little water, or acting as a pathway for water loss back to the plant. This inference was tested by examining the water balance and vascular functioning of ripening kiwifruit berries (Actinidia chinensis var. chinensis 'Hort16A') exhibiting a pre-harvest 'shrivel' disorder in California, and normal development in New Zealand. Dye labelling and mass balance experiments indicated that the xylem and phloem were both functional and contributed approximately equally to the fruit water supply during this stage of development. The modelled fruit water balance was dominated by transpiration, with net water loss under high vapour pressure deficit (D(a)) conditions in California, but a net gain under cooler New Zealand conditions. Direct measurement of pedicel sap flow under controlled conditions confirmed inward flows in both the phloem and xylem under conditions of both low and high D(a). Phloem flows were required for growth, with gradual recovery after a step increase in D(a). Xylem flows alone were unable to support growth, but did supply transpiration and were responsive to D(a)-induced pressure fluctuations. The results suggest that the shrivel disorder was a consequence of a high fruit transpiration rate, and that the perception of complete loss or reversal of inward xylem flows in ripening fruits should be re-examined.     

The 5' untranslated region of the maize alcohol dehydrogenase gene provides efficient translation of mRNA in plants under stress conditions

Reduced level of expression of most cell proteins under stress conditions is determined by low efficiency of cap-dependent translation of corresponding mRNAs. The maize gene encoding alcohol dehydrogenase, adh1, is an example of a gene which mRNA is efficiently translated under hypoxia. Using reporter gene assay we showed that the leader sequence of adh1 mRNA, provides efficient translation of reporter gene gfp in Nicotiana benthamiana cells under hypoxia and heat shock. The presence of this leader sequence in 5' UTR of mRNA does not change the level of expression in aerobic conditions, but under hypoxia and heat shock the levels of reporter gfp expression were reduced about 5-10 fold in the absence of leader and remained unaffected in its presence in 5'UTR. We found that this leader sequence does not change the level of mRNA stability and does not exhibit promoter activity. Consequently, leader sequence acts as translational enhancer providing efficient translation of mRNA in plant cells under stress conditions. Introduction of this sequence into standard expression cassettes may be used for development of new systems of expression of target proteins in plants, efficient under stress conditions.     

Mutational analysis of the aggregation-prone and disaggregation-prone regions of acylphosphatase

We have performed an extensive mutational analysis of aggregation and disaggregation of amyloid-like protofibrils of human muscle acylphosphatase. Our findings indicate that the regions that promote aggregation in 25% (v/v) 2,2,2 trifluoroethanol (TFE) are different from those that promote disaggregation under milder conditions (5% TFE). Significant changes in the rate of disaggregation of protofibrils in 5% TFE result not only from mutations situated in the regions of the sequence that play a key role in the mechanism of aggregation in 25% TFE, but also from mutations located in other regions. In order to rationalise these results, we have used a modified version of the Zyggregator aggregation propensity prediction algorithm to take into account structural rearrangements of the protofibrils that may be induced by changes in solution conditions. Our results suggest that a wider range of residues contributes to the stability of the aggregates in addition to those that play an important kinetic role in the aggregation process. The mutational approach described here is capable of providing residue-specific information on the structure and dynamics of amyloid protofibrils under conditions close to physiological and should be widely applicable to other systems.     

31P-NMR study of kinetics of 2,3-diphosphoglycerate degradation in human erythrocytes during their depletion

2.3-Diphosphoglycerate degradation kinetics was studied under conditions of human erythrocyte depletion, using the method of 31P-NMR of high performance. The kinetic curve was shown to have a plateau during the first 1.5-2 hours after the beginning of depletion. Possible mechanisms providing for the appearance of such a plateau are discussed.     

Real time monitoring of biofilm development under flow conditions in porous media

Biofilm growth can impact the effectiveness of industrial processes that involve porous media. To better understand and characterize how biofilms develop and affect hydraulic properties in porous media, both spatial and temporal development of biofilms under flow conditions was investigated in a translucent porous medium by using Pseudomonas fluorescens HK44, a bacterial strain genetically engineered to luminesce in the presence of an induction agent. Real-time visualization of luminescent biofilm growth patterns under constant pressure conditions was captured using a CCD camera. Images obtained over 8 days revealed that variations in bioluminescence intensity could be correlated to biofilm cell density and hydraulic conductivity. These results were used to develop a real-time imaging method to study the dynamic behavior of biofilm evolution in a porous medium, thereby providing a new tool to investigate the impact of biological fouling in porous media under flow conditions.