Delivery of photoreactive psoralen derivatives to specific biological targets

Psoralens are bifunctional molecules which photoreact with the pyrimidine bases of nucleic acids to form monoadducts and diadducts, or interstrand cross-links. We have prepared psoralen derivatives with additional functional groups which can be specifically directed to chosen biological targets. A sulfhydryl-containing psoralen which can form site-specific cross-links in plasmid DNA has been used to study psoralen repair and mutagenesis. Cloned DNA containing psoralen monoadducts has been cross-linked to specific regions of viral RNA and used to probe virus assembly. A biotinylated psoralen derivative which binds specifically to avidin has been used to detect small amounts of DNA. Finally, a psoralen derivative of insulin has been used to deliver psoralen specifically to activated lymphocytes.     

Calculation of Vapour-Liquid Equilibria of Lennard-Jones Binary Systems by the Gibbs Ensemble Monte Carlo Simulation

Abstract

The Gibbs ensemble Monte Carlo simulation has been used to calculate vapour-liquid equilibria of a Lennard-Jones (LJ) binary mixture. The mixture studied is the LB-2-1 model which has been used in our previous calculations on PVT relation and density-dependent local composition. The P-x-y relation has been established at two different temperatures and used to determine vapour-liquid coexistence region in the PVTx space.     

Use of carboxypeptidase Y in studies of the mode of insertion of cytochrome b5 into lipid vesicles

Carboxypeptidase Y preparations from baker's yeast have been found to exhibit endopeptidase activity when cytochrome b5 was used as substrate. As the susceptibility of cytochrome b5 to attack by carboxypeptidase Y has been used to distinguish between two modes of insertion of cytochrome b5 into lipid bilayer, one which has the C terminal buried in the lipid bilayer and one which has a free C terminal, caution should be taken when employing carboxypeptidase Y preparations for this type of studies.     

巴戟天应用的名实沿革考

通过对古今文献的分析及实地考查认为:南北朝以前使用之主流巴戟天药材的原植物可能为五味子科植物铁箍散Schisandr a propinqua(Wall.)Baill.var.sinensis Oliv.,药材现称川巴戟或香巴戟;唐代至清末广为运用的品种为归州巴戟天,原植物为茜草科植物四川虎刺Dammacanthus officinarum Huang,药材现称鄂西巴戟天或恩施巴戟天;现今药典收载之巴戟天为清末发展的新品种,其原植物为茜草科植物巴戟天 Morinda officinalis How。     

A gamma-ray computed tomography scanner for the quantitative measurement of bone density

A special purpose gamma-ray computed tomography scanner has been developed for the precise measurement of bone density in the distal forearm. Details of the scanner hardware and computer analysis technique are given. Suitable phantoms have been used to test the operation of the scanner, which has been used to measure trabecular and cortical bone density with a precision better than 1%.     

In-Cell Solid-State NMR: An Emerging Technique for the Study of Biological Membranes

Biological molecular processes are often studied in model systems, which simplifies their inherent complexity but may cause investigators to lose sight of the effects of the molecular environment. Information obtained in this way must therefore be validated by experiments in the cell. NMR has been used to study biological cells since the early days of its development. The first NMR structural studies of a protein inside a cell (by solution-state NMR) and of a membrane protein (by solid-state NMR) were published in 2001 and 2011, respectively. More recently, dynamic nuclear polarization, which has been used to enhance the signal in solid-state NMR, has also been applied to the study of frozen cells. Much progress has been made in the past 5 years, and in this review we take stock of this new technique, which is particularly appropriate for the study of biological membranes.     

Pollen as food and medicine—A review

Pollen, the male gametophyte of flowering plants, is a high energy material, which is collected by insects and stored as food reserve. Pollen has been used traditionally by humans for religious purposes and as supplementary food. Pollen is a concentrated, energy and vitamin rich food that in contemporary times is not only consumed as a dietary component, but also is used in alternative medical treatments. Pollen has potential imporiance as a supplementary and survival food, and for conditioning of athletes. Pollen has been used medically in prostatitis, bleeding stomach ulcers and some infectious diseases, although such use has been questioned by the medical profession. Pollen may also be used for treatment and prevention of the high-altitude-sickness syndrome. Because some individuals are allergic to pollen, and various pollen species contain specific allergens, individual sensitivities must be tested before pollen is used as a treatment or as a supplementary food.     

The decolorization of betacyanin by polyamines

The pigment betacyanin, the efflux of which has been used as a measure of membrane integrity in red beet discs, has now been shown to react with di- and polyamines with consequent decolorization.     

STUDIES IN RESPIROMETRY : I. A COMBINED GAS BURETTE-INTERFEROMETER RESPIROMETER

A machine has been described which combines volumetric and refractometric means of observing changes in carbon dioxide and oxygen concentrations and amounts in a closed system suitable for containing a respiring system. A means of thermoregulation has been described briefly. The theory upon which estimation is based has been outlined in considerable detail and generality; indicating how dependence upon absorption of gases by reagents can be eliminated completely, although they may be used in certain instances as a means of independent check. An experiment of this nature has been reported as an illustration of one use of the machine. Data of other experiments in which a moist environment was employed for the respiring system will be given in another communication.     

The synthesis of phenyl(2-3H)glyoxal.

A simple inexpensive method has been developed for the synthesis of [2-3H]acetophenone, which has been converted into phenyl[2-3H]glyoxal. The latter compound has been used to modify arginine residues in alkaline phosphatase from two sources, and also a sialidase.     

Elaboration of PCR MP diagnostic kit for genetic typing of bacterial strains

The rules of diagnostic kit elaboration for genetic typing of microorganisms, designed for epidemiological studies, have been shown in this paper. PCR MP method has been used for diagnostic kit elaboration. Well defined epidemiologically Enterococcus faecium strains have been applied as a research model. The optimisation of the method has been carried out using different amount of reagents and time of the particular stages. Critical parameters, which have significant influence on the quality of obtained results, have been assigned. Optimalised procedure, named PCR MP unique, has been validated for genetic typing of different species of microorganisms and its potential application for routine epidemiological studies. The PCR MP method has been successfully used for elaboration of diagnostic PCR MP unique-KIT, which allows intra-species differentiation of bacterial strains. The PCR MP unique-KIT enables fast, easy and cheap analysis of strains, using elementary laboratory equipment--gradient thermocycler.     

Simulation of gene pyramiding in Drosophila melanogaster

Gene pyramiding has been successfully practiced in plant breeding for developing new breeds or lines in which favorable genes from several different lines were integrated.But it has not been used in animal breeding,and some theoretical investigation and simulation analysis with respect to its strategies,feasibility and efficiency are needed before it can be implemented in animals.In this study,we used four different pure lines of Drosophila melanogaster,each of which is homozygous at a specific mutant gene ...     

Complementation analysis of Agrobacterium tumefaciens Ti plasmid mutations affecting oncogenicity

A wide host range cosmid vector has been constructed by insertion of the lambda cos site into the plasmid pRK2501. This cosmid, which is maintained in Agrobacterium tumefaciens and is compatible with the Ti plasmid, has been used to make a clone bank of the A. tumefaciens pTiA6 plasmid. Several pTiA6 cosmids have been used to complement Tn5-induced Ti plasmid mutations. Five avirulent mutations which map outside of the region of the plasmid maintained in plant tumours (T-DNA) could be complemented in a trans orientation. Two mutations which are located on a single HpaI restriction fragment outside of the T-DNA, as well as three mutations which map within the T-DNA region, could not be complemented in a trans orientation in a REC- host.     

Is there a future for therapeutic use of transcranial magnetic stimulation?

Repetitive transcranial magnetic stimulation (rTMS) has in recent years been used to explore therapeutic opportunities in a bewildering variety of conditions. Although there is good evidence that this technique can modify cortical activity, the rationale for its use in many of the conditions investigated so far is not clear. Here we discuss the effects of rTMS in healthy subjects and how it has been used in a number of neurological conditions. We argue that a better understanding of both the effects of rTMS and the pathological processes underlying the conditions for which it is used will reveal whether rTMS really does offer therapeutic potential and, if so, for which conditions.     

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.     

Spin-lattice relaxation of water protons in plant and animal cells

NMR-spin echo method has been used to study spin-lattice relaxation time of protons T1 in plant and animal cells ?? muscle tissue of fish, the cells of which unlike plant cells have no developed system of vacuoles, plastids and a solid cell wall. According to the values of T1 time a new NMR parameter K, a coefficient of relaxation effectiveness of a cell structure, has been calculated. This parameter can be used for quantitative characterization of the influence of different cell structures, the tissue water interact with, for a time of spin-lattice relaxation of water protons. It has been ascertained that the values of K coefficient in animal tissue and in storing tissues of some plants differ little; it may be stipulated by permanent transmembrane water exchange which occurs at high rate in the living cell. It has been concluded that there exists a certain similarity between water state in protoplast of plant and animal cells.     

Magic bullets for protein kinases

A chemical-genetic method for the generation of target-specific protein kinase inhibitors has been developed recently. This strategy utilizes a functionally silent active-site mutation to sensitize a target kinase to inhibition by a small molecule that does not inhibit wild-type kinases. Tyrosine and serine/threonine kinases are equally amenable to the drug-sensitization approach, which has been used to generate selective inhibitors of mutant Src-family kinases, Abl-family kinases, cyclin-dependent kinases, mitogen-activated kinases, p21-activated kinases and Ca(2+)/calmodulin-dependent kinases. The designed inhibitors are specific for the sensitized kinase in a cellular background where the wild-type kinase has been inactivated. By these means, kinase-sensitization has been used systematically to generate and analyze conditional alleles of several yeast protein kinases in vivo.     

High-performance liquid chromatographic determination of PEG 600 in human urine

Polyethylene glycols (PEGs) are non-ionic, water-soluble synthetic polymers which have been widely used for many applications. Since they are of very low toxicity and are readily excreted in urine, PEGs in the molecular weight range 400–6000 have been used extensively in the study of intestinal physiology in man. A high-performance liquid chromatographic (HPLC) method has been developed for the determination of PEG 600 in human urine, which includes a pre-column derivatisation step. The dibenzoate derivatives of PEG 600 can be quantitatively prepared, and this, coupled with ultraviolet detection at 230 nm, has greatly improved the limit of detection for the determination of PEGs by HPLC. A suitable extraction procedure has also been developed which enabled PEG levels in urine to be monitored with much greater sensitivity than any previously reported method.     

High-performance affinity chromatography: a powerful tool for studying serum protein binding

High-performance affinity chromatography (HPAC) is a method in which a biologically-related ligand is used as a stationary phase in an HPLC system. This approach is a powerful means for selectively isolating or quantitating agents in complex samples, but it can also be employed to study the interactions of biological systems. In recent years there have been numerous reports in which HPAC has been used to examine the interactions of drugs, hormones and other substances with serum proteins. This review discusses how HPAC has been used in such work. Particular attention is given to the techniques of zonal elution and frontal analysis. Various applications are provided for these techniques, along with a list of factors that need to be considered in their optimization and use. New approaches based on band-broadening studies and rapid immunoextraction are also discussed.     

Single-Molecule Microscopy Reveals Membrane Microdomain Organization of Cells in a Living Vertebrate

It has been possible for several years to study the dynamics of fluorescently labeled proteins by single-molecule microscopy, but until now this technology has been applied only to individual cells in culture. In this study, it was extended to stem cells and living vertebrate organisms. As a molecule of interest we used yellow fluorescent protein fused to the human H-Ras membrane anchor, which has been shown to serve as a model for proteins anchored in the plasma membrane. We used a wide-field fluorescence microscopy setup to visualize individual molecules in a zebrafish cell line (ZF4) and in primary embryonic stem cells. A total-internal-reflection microscopy setup was used for imaging in living organisms, in particular in epidermal cells in the skin of 2-day-old zebrafish embryos. Our results demonstrate the occurrence of membrane microdomains in which the diffusion of membrane proteins in a living organism is confined. This membrane organization differed significantly from that observed in cultured cells, illustrating the relevance of performing single-molecule microscopy in living organisms.