Productivity improvement in xanthan gum fermentation using multiple substrate optimization

A novel and more comprehensive formulation of the optimal control problem that reflects the operational requirements of a typical industrial fermentation has been proposed in this work. This formulation has been applied to a fed-batch bioreactor with three control variables, i.e., feed rates of carbon source, nitrogen source, and an oxygen source, to result in a 148.7% increase in product formation. Xanthan gum production using Xanthomonas campestris has been used as the model system for this optimization study, and the liquid-phase oxygen supply strategy has been used to supply oxygen to the fermentation. The formulated optimization problem has several constraints associated with it due to the nature of the system. A robust stochastic technique, differential evolution, has been used to solve this challenging optimization problem. The infinite dimensional optimization problem has been approximated to a finite dimensional one by control vector parametrization. The state constraints that are path constraints have been addressed by using penalty functions and by integrating them over the total duration to ensure a feasible solution. End point constraints on final working volume of the reactor and on the final residual concentrations of carbon and nitrogen sources have been included in the problem formulation. Further, the toxicity of the oxygen source, H(2)O(2), has been addressed by imposing a constraint on its maximum usable concentration. In addition, the initial volume of the bioreactor contents and feed concentrations have been handled as decision variables, which has enabled a well-grounded choice for their values from the optimization procedure; adhoc values are normally used in the industry. All results obtained by simulation have been validated experimentally with good agreements between experimental and simulated values.     

On-line monitoring and control of substrate concentrations in biological processes by flow injection analysis systems

Concentrations of substrates, glucose, and ammionia in biological processes have been on-line monitored by using glucose-flow injection (FIA) and ammonia-FIA systems. Based on the on-line monitored data the concentrations of substrates have been controlled by an on-off controller, a PID controller, and a neural network (NN) based controller. A simulation program has been developed to test the control quality of each controller and to estimate the control parameters. The on-off controller often produced high oscillations at the set point due to its low robustness. The control quality of a PID controller could have been improved by a high analysis frequency and by a short residence time of sample in a FIA system. A NN-based controller with 3 layers has been developed, and a 3(input)-2(hidden)-1(output) network structure has been found to be optimal for the NN-based controller. The performance of the three controllers has been tested in a simulated process as well as in a cultivation process ofSaccharomyces cerevisiae, and the performance has also been compared to simulation results. The NN-based controller with the 3-2-1 network structure was robust and stable against some disturbances, such as a sudden injection of distilled water into a biological process.     

Diagnosis of Helicobacter pylori

The different invasive and noninvasive diagnostic tests for Helicobacter pylori have been applied mainly in emerging countries. Molecular methods have been developed, especially a test for detection of H. pylori and its clarithromycin resistance directly from stools. The long-term effects of eradication on histologic lesions have been studied in a meta-analysis and the prognostic value of post-treatment in gastric mucosa-associated lymphoid tissue lymphoma has been assessed. An operating link for gastritis assessment (the OLGA staging) has also been published. Attempts to simplify the urea breath test protocol have been made, and new stool antigen tests have been proposed and compared to those previously available.     

Adenylate cyclase of the causative agent of plague: its purification and properties

Three forms of adenylate cyclase have been detected in Y. pestis: membrane-bound, cytoplasmic and extracellular. Extracellular adenylate cyclase has been purified so as to achieve a homogeneous state, and some of its physicochemical parameters have been investigated. In the process of purification the initial preparation of this enzyme has been subjected to heating at 100 degrees C for 15 minutes, fractionation with ammonium sulfate, and gel filtration on Sephadex G-100. The homogeneity of adenylate cyclase has been confirmed by electrophoresis in 7.5% polyacrylamide gel and precipitation by the plague agglutinating serum. The enzyme has been found to have a molecular weight of 30,000 daltons and to show the optimum activity at pH 7.0-7.2 and at a temperature between 37 and 40 degrees C. Monospecific rabbit serum to the homogeneous preparation of adenylate cyclase has been obtained.     

pH-sensitive hydrogel based on a novel photocross-linkable copolymer

A pH sensitive hydrogel has been prepared by a UV irradiation technique. Starting polymer was the PHM (poly hydroxyethylaspartamide methacrylated) obtained from polyaspartamide (PHEA) partially derivatized with methacrylic anhydride (MA). This new copolymer has been further derivatized with succinic anhydride (SA) to obtain PHM-SA that has been cross-linked by UV irradiation to form a pH sensitive hydrogel. The network, recovered after washing as a powder, has been been characterized by FT-IR spectrophotometry and particle size distribution analysis. Moreover, to have information about water affinity of the prepared sample, swelling measurements have been carried out in aqueous media mimicking biological fluids. The possibility to employ the prepared hydrogel as a pH-sensitive drug delivery system (DDS) has been investigated. In particular, ibuprofen ((S)(+)4-isobutyl-alpha-methylphenyl-acetic acid), chosen as a model drug, has been entrapped into the PHM-SA hydrogel, and in vitro release studies have showed that its release rate depends on different swelling of the network as a function of the environmental pH.     

Simulation of the bis-(penicillamine) enkephalin in ammonium chloride solution: a comparison with sodium chloride

In order to quantify specific ion effects, a simulation study of bis(penicllamine) enkephalin, also known as DPDPE, has been performed in aqueous ammonium chloride solution and has been compared to a previous simulation of DPDPE in aqueous sodium chloride solution. Global thermodynamics have been calculated for a model system and the solution environment around DPDPE has been characterized. Associations of ions with DPDPE have been investigated. The observed differences between sodium chloride solution and ammonium chloride solution suggest that individual cations affect the solvation and peptide binding properties of a given anion.     

Imaging erythrocytes under physiological conditions by atomic force microscopy

Since its invention in the mid 1980s atomic force microscopy has revolutionised the way in which surfaces can be imaged. Close to atomic resolution has been achieved for some materials and numerous images of molecules on surfaces have been recorded. Atomic force microscopy has also been of benefit to biology where protein molecules on surfaces have been studied and even whole cells have been investigated. Here we report a study of red blood cells which have been imaged in a physiological medium. At high resolution, the underlying cytoskeleton of the blood cell has been resolved and flaws in the cytoskeleton structure may be observed. Comparison of the normal 'doughnut' shaped cells with swollen cells has been undertaken. Differences in both the global properties of the cells and in the local features in cytoskeleton structure have been observed.     

Marine mammal chemoreception

The presence or absence of a chemoreceptive capacity in marine mammals has drawn relatively little attention from the research community outside the Soviet Union. Toothed whales are typically labelled anosmic (lacking a sense of smell) since they do not have the peripheral olfactory structures typically associated with terrestrial mammals. Baleen whales are known to possess reduced olfactory tracts; their olfactory bulbs also may be reduced or absent. Although the neural structures that mediate taste in terrestrial mammals have been reported to be present in both groups of whales, cetaceans have been considered to have a poor sense of taste because typical mammalian taste receptors have been thought to be absent. Soviet researchers, however, recently have reported that gustatory receptors are present on some cetacean tongues and that the tongue of Tursiops truncatus appear to be well innervated. These workers also have been conducting investigations which seem to be aimed at describing a specialized gustatory capability in cetaceans. No experimental work has been reported by Western scientists. Little work has been done by either Western or Soviet researchers with regard to chemoreception among the other orders of marine mammals (Pinnipedia, Carnivora and Sirenia). Pinnipedia are typically labelled microsmatic (having a poor sense of smell); research has been restricted to histological examination of the nasal pathways, and neural anatomy. Sea otters are credited with a keen sense of smell, but no quantitative work has been reported. The chemosensory abilities of Sirenia remain unknown. The tongues of non-cetacean marine mammals have been histologically examined and found to resemble those of terrestrial mammals. No other investigations of gustation in non-cetacean marine mammals have been reported.     

Interferons: Purification and Physicochemical Aspects

Antiviral interferon activity in any one species can be exhibited by a variety of substances that differ in their physical and chemical properties, but the nature of these differences is not understood. Conditions that can lead to the formation of diverse types of interferons have been outlined. Reasons have been adduced why, for certain purposes, purification of interferons is desirable or even necessary, and examples have been presented to show how and to what extent this has been achieved. In spite of some very high purification factors, not a single interferon has been obtained as a pure substance. Therefore, all available knowledge of physical and chemical properties has been obtained by indirect means.     

Acetylcholine receptor-controlled ion flux in electroplax membrane vesicles: Identification and characterization of membrane properties that affect ion flux measurements

Summary Several intrinsic properties of acetylcholine receptor-rich membrane vesicles prepared fromElectrophorus electricus, which need to be considered in measurements of receptor-mediated ion flux, have been identified. One of these properties is a slow exchange of inorganic ions in the vesicles. The slow exchange of ions is not related to the receptor-mediated flux of ions and accounts for 30–35% of the efflux observed. A method to separate this process from the receptor-controlled flux has been developed. It has also been shown, using a light-scattering method, that aggregation-disaggregation of the vesicles can interfere with the efflux measurements, and a method to overcome this problem has been developed. The difference in the amplitude of effluxes induced by saturating amounts of carbamylcholine and gramicidin has been investigated and has been shown not to be due to a receptor-controlled process; therefore, the amplitude difference does not need to be considered in understanding the receptor-controlled process.     

The Organization for Disaster in British Columbia

Plans are being formulated for organizing an Emergency Health Service in British Columbia. The same organization that is being created to deal with civilian casualties after an enemy attack should, it is felt, be able to assist in major natural disasters. Such an organization will have to remain ready but inactive for long periods; therefore the responsibility for planning, organizing and directing this service has been given to the Health Branch, Provincial Department of Health Services and Hospital Insurance.A committee has been struck to advise the Health Branch, and a Director and a Health Supplies Officer have been appointed. Agreement has been reached as to who is to be assigned to work in the Emergency Health Service in the event of a nuclear attack, and sites have been selected for the stockpiling of medical equipment.The Province has been divided into zones, and Emergency Health Service Directors have been appointed for each zone. Their duties include, among many others, organizing Advanced Treatment Centres and Emergency Hospitals, and developing an Emergency Public Health Service, an Emergency Blood Transfusion Laboratory and a Mortuary Service.     

Application of a chemoenzymatic glycosylation method to α-chymotrypsin and Candida rugosa lipase surface modifications

A recently developed chemoenzymatic glycosylation procedure has been successfully applied on two hydrolytic enzymes, α-chymotrypsin and Candida rugosa lipase. First, a number of sucrose molecules have been bound to the surface lysine residues and then, lengthening of the glycosidic chains has been carried out by the action of a levansucrase from Bacillus subtilis. For both steps, reaction conditions have been studied in order to obtain a range of glycosylation degrees. The influence of glycoside binding on biocatalyst surface characteristics has been assessed and a progressive increase in global enzyme hydrophilic character with glycosylation has been observed. Besides, the study of hydrolytic activity and kinetic constants showed that the performed modifications brought about a certain decrease in enzyme hydrolytic activity and very slight variations in enzyme-substrate affinity.     

New antibiotic resistance Loci in the ribosomal region of yeast mitochondrial DNA

A large number of mitochondrial antibiotic-resistant mutants have been isolated following mutagenesis with manganese. These include several different phenotypic classes of mutants, as distinguished by cross-resistance patterns, that have been found to be allelic at cap1 or ery1; some have been found to be heteroallelic.--Seven chloramphenicol-resistant mutants have been identified that are nonallelic by recombination tests with the three loci (cap1, spi1 and ery1) previously identified in the ribosomal region. Four of these are allelic with each other and define a new locus, cap3; two others are allelic and define another new locus, cap2; the seventh maps at yet a different locus, cap4. One new spiramycin-resistant mutant has been identified that defines still another new locus, spi2. A variety of genetic techniques have been used to map these loci within the ribosomal region of the mitochondrial genome.-Manganese has been shown to be effective in inducing the mutation from omega(-) to omega(n) in many mutants that experience a simultaneous mutation at the closely linked cap1 locus. The omega(n) mutation has also been described in the cap4 mutant, and this locus has been shown to be more closely linked to omega than cap1 is to omega.     

Circadian rhythmicity of prolactin secretion in Huntington's chorea

Prolactin circadian secretion has been studied in 16 males and in 8 females with Huntington's Chorea, in order to evaluate changes of the hormone's titres and of their periodicity in a condition where alterations of neurotransmission have been documented. Prolactin levels have been found normal, enhanced and low levels have been observed in one male case. Sleep connected surge has been found to be present in all cases. A harmonic analysis of periodical changes has been performed by a Fourier's cosines series. The changes in the occurence of different harmonic frequencies in respect to normal controls are not significant. No correlation could be drawn among clinical indices, hormone levels and the feature of the occurring harmonics.     

Biochemistry and structural DNA nanotechnology: an evolving symbiotic relationship

Structural DNA nanotechnology is derived from naturally occurring structures and phenomena in cellular biochemistry. Motifs based on branched DNA molecules are linked together by sticky ends to produce objects, periodic arrays, and nanomechanical devices. The motifs include Holliday junction analogues, double and triple crossover molecules, knots, and parallelograms. Polyhedral catenanes, such as a cube or a truncated octahedron, have been assembled from branched junctions. Stiff motifs have been used to produce periodic arrays, containing topographic features visible in atomic force microscopy; these include deliberately striped patterns and cavities whose sizes can be tuned by design. Deliberately knotted molecules have been assembled. Aperiodic arrangements of DNA tiles can be used to produce assemblies corresponding to logical computation. Both DNA structural transitions and branch migration have been used as the basis for the operation of DNA nanomechanical devices. Structural DNA nanotechnology has been used in a number of applications in biochemistry. An RNA knot has been used to establish the existence of RNA topoisomerase activity. The sequence dependence of crossover isomerization and branch migration at symmetric sites has been established through the use of symmetric immobile junctions. DNA parallelogram arrays have been used to determine the interhelical angles for a variety of DNA branched junctions. The relationship between biochemistry and structural DNA nanotechnology continues to grow.     

Upper lobe lobectomy and concomitant thoracoplasty in pulmonary tuberculosis. A preliminary report

In this, a preliminary report, the advantages and disadvantages of upper lobe lobectomy and concomitant thoracoplasty in tuberculosis have been summarized. There has been no undue increase in the operative hazard, there have been no complications, and the immediate results have been entirely satisfactory.Twenty-six patients were subjected to a combined operation (17 upper lobectomy-thoracoplasty and nine pneumonectomy-thoracoplasty) with one postoperative death. Indications for primary lobectomy have been discussed. The technique of resection and concomitant thoracoplasty has been briefly described.     

Mechanism of sulfide-quinone reductase investigated using site-directed mutagenesis and sulfur analysis

Biological sulfide oxidation is a reaction occurring in all three domains of life. One enzyme responsible for this reaction in many bacteria has been identified as sulfide:quinone oxidoreductase (SQR). The enzyme from Rhodobacter capsulatus is a peripherally membrane-bound flavoprotein with a molecular mass of approximately 48 kDa, presumably acting as a homodimer. In this work, SQR from Rb. capsulatus has been modified with an N-terminal His tag and heterologously expressed in and purified from Escherichia coli. Three cysteine residues have been shown to be essential for the reductive half-reaction by site-directed mutagenesis. The catalytic activity has been nearly completely abolished after mutation of each of the cysteines to serine. A decrease in fluorescence on reduction by sulfide as observed for the wild-type enzyme has not been observed for any of the mutated enzymes. Mutation of a conserved valine residue to aspartate within the third flavin-binding domain led to a drastically reduced substrate affinity, for both sulfide and quinone. Two conserved histidine residues have been mutated individually to alanine. Both of the resulting enzymes exhibited a shift in the pH dependence of the SQR reaction. Polysulfide has been identified as a primary reaction product using spectroscopic and chromatographic methods. On the basis of these data, reaction mechanisms for sulfide-dependent reduction and quinone-dependent oxidation of the enzyme and for the formation of polysulfide are proposed.     

Cytoskeleton: actin and endocytosis--no longer the weakest link.

The actin cytoskeleton has long been believed to play a role in endocytosis, but its actual function in this process has been unclear. Now, three proteins that promote actin nucleation have been found to provide a link between the actin cytoskeleton and the endocytic machinery.     

The Effects of Rehydration and Oxygen on the Heat Resistance of High Vacuum Treated Spores

S ummary . Damage induced in bacterial spores by exposure to reduced pressures of the order of 10^-8 torr, has been assessed in terms of the differences in the heat resistance of the dried spores. The response of the spores has been assessed as a function of (a), the drying temperature from 0-65°; (b) the duration and level of rehydration; (c) the presence or absence of oxygen during heating. Comparison has also been made between spores dried to a given water level and spores rehydrated to the same water level after prolonged drying. Log survivors/heating time curves for treated spore samples have been constructed and have been shown to exhibit a shoulder at high survival levels and a linear portion below a surviving fraction of 0·1. These curves have been explained on the basis of the shoulder representing the time during which necessary structural changes occur in the spore, before the lethal mechanism responsible for the linear portion of the curve becomes operative. The heat response has been shown to be a function of the temperature of drying, and of the presence of oxygen during heating, the structural change itself being reversible by water.