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Indirect evidence for two distinct mechanisms of DNA cleavage at the oriT of RP4 during conjugation.

Mobilization of a potential Campylobacter shuttle vector from Escherichia coli K-12 into C. hyointestinalis generated a small plasmid harboring a hybrid oriT nick region. This is presumed to have resulted from the cleavage and religation of the oriT of RP4 and the putative nick region of the shuttle vector. These data imply that two distinct DNA cleavage reactions occur at the oriT site of RP4 during conjugal transfer.

Base Sequence↗

From rolling to arrest on blood vessels: leukocyte tap dancing on endothelial integrin ligands and chemokines at sub-second contacts.

In order to extravasate the bloodstream at specific sites of inflammation or antigen presentation, circulating leukocytes must rapidly translate specific adhesive and stimulatory signals into firm adhesion. Leukocyte arrest is nearly exclusively mediated by integrin receptors. Recent in vitro and in vivo evidence suggests that specialized integrins support reversible tethers that slow down selectin-initiated rolling of leukocytes prior to their arrest. In situ activation of integrin avidity by ligand and chemokine signaling can take place within fractions of seconds, resulting either in augmented reversible adhesions or immediate arrest on the vascular endothelium. The ability of leukocyte integrins to rapidly respond to these in situ avidity modulators appears to depend on preformed affinity and clustering states, which are internally regulated by cytoskeletal constraints on integrin conformation and mobility. We discuss potential regulatory mechanisms by which a given set of chemokine receptors and integrins may interact to rapidly generate high avidity, shear-resistant integrin-mediated leukocyte arrest on vascular endothelium.

Animals↗

Discrete intracellular Ca(2+) pools coupled to two distinct Ca(2+) influx pathways in human platelets.

Ca(2+) influx is an important event associated with platelet activation and regulated by the content of intracellular Ca(2+). Previous studies have suggested two different Ca(2+) pools and two Ca(2+) influx pathways exist in platelets. In the present study, we have investigated the regulation of thrombin- and thapsigargin-induced Ca(2+) entry into human platelets, using fluorescent indicators to monitor Ca(2+) mobilization and membrane potential. It was found that depletion of thapsigargin-sensitive Ca(2+) stores was coupled to Ca(2+) influx through a Ca(2+)-selective pathway. Additional release of Ca(2+) from the thapsigargin-insensitive pool by thrombin caused the opening of a nonselective cation channel.

Blood Platelets↗

How do grazers affect periphyton heterogeneity in streams?

The effects of grazing by stream invertebrates on algal biomass and spatial heterogeneity were tested experimentally in flow-through microcosms with natural substrates (rocks). One experiment tested the effects of fixed densities of three species of grazers (the caddisfly Allomyia sp. and two mayflies, Epeorus deceptivus and Baetis bicaudatus) on periphyton. Baetis was tested with and without chemical cues from fish predators, which reduced grazer foraging activity to levels similar to the less mobile mayfly (Epeorus). Mean algal biomass (chlorophyll a; chl a) was reduced in grazer treatments compared to ungrazed controls, but there were no differences among grazer treatments. Algal heterogeneity (Morisita index) increased with grazer mobility, with the highest heterogeneity occurring in the Baetis-no fish treatment (most mobile grazer) and the lowest in the caddisfly treatment (most sedentary grazer). A second experiment used a three factorial design, and tested whether initial resource distribution (homogeneous vs. heterogeneous), Baetis density (high vs. low) and fish odor (present vs. absent) affected grazer impact on algal resources. Abundances of Baetis and chl a on individual rocks were recorded to explore the mechanisms responsible for the observed distributions of algae. Initial resource heterogeneity was maintained despite being subjected to grazing. Mean chl a was highest in controls, as in experiment I, and effects of Baetis on algal biomass increased with grazer density. There were no fish effects on algal biomass and no effects of grazer density or fish on algal heterogeneity. At the scale of individual rocks Baetis was unselective when food was homogeneously distributed, but chose high-food rocks when it was heterogeneously distributed. Results of these mechanistic experiments showed that Baetis can track resources at the scale of single rocks; and at moderate densities mobile grazers could potentially maintain periphyton distributions observed in natural streams.

Animals↗

Endometrial phospholipase A2 enzymes and their regulation by steroid hormones.

The presence of two phospholipase A2 (PLA2) enzymes, designated PLA2(i) and PLA2(ii), has been demonstrated in human endometrium. These enzymes differ with respect to pH and calcium requirements, location within the tissue and regulation by steroid hormones. Phospholipase A2(i) is calcium dependent, optimally active at pH 7.5-9.0 and present mainly in the glandular component of the endometrium. Changes in activity occur during the menstrual cycle which are indicative of regulation by ovarian steroids. Conversely, PLA2(ii) is calcium independent, optimally active at pH 7.0 and located predominantly in the stromal layer. Wide variation in PLA2(ii) activity was found between individual subjects and there was no relationship with the stage of the menstrual cycle. Activity was, however, much higher in pathological endometrium and in endometrium from subjects with severe dysmenorrhoea. Triton X-100 activated PLA2(i) but not PLA2(ii). In cultured explants of endometrium, both enzymes were inhibited by progesterone whereas oestradiol and dexamethasone had no effect. However, progesterone priming followed by treatment with oestradiol caused a 2-fold stimulation of PLA2(i) but not PLA2(ii). Phospholipase A2 is favoured as the rate-limiting step in the generation of arachidonic acid for prostaglandin synthesis. However, our studies so far do not support a direct relationship between PLA2 and endometrial concentrations of prostaglandins, which implies that other important regulatory steps are involved. Other enzymes which are potentially capable of mobilizing arachidonic acid should also be investigated.

Calcium↗

Determination of a novel xanthine derivative, MKS 213-492, in plasma by high-performance liquid chromatography with electrochemical detection.

A reversed-phase liquid chromatographic method with coulometric detection has been developed for the determination of free MKS 213-492 (a novel xanthine derivative) in plasma. This method is based on a simple and rapid plasma extraction procedure using precolumn-switching. The oxidation of this pharmacologically active xanthine derivative was optimized with respect to applied potential, pH of mobile and rinsing phase and rinsing time. The detection limit for MKS 213-492 was found to be 54 pg/injection.

Chromatography, High Pressure Liquid↗

High-performance liquid chromatography with mercury cathode electrochemical detection: application to lipid hydroperoxide analysis.

Lipid hydroperoxide species can be analyzed with high sensitivity and specificity, using reversed-phase high-performance liquid chromatography with reductive mode electrochemical detection on a mercury drop cathode [HPLC-ED(Hg)]. The purpose of this study was to examine different variables in the operation of HPLC-ED(Hg) and to select optimal conditions for the analysis of several biologically relevant peroxides, including species derived from cholesterol, cholesteryl linoleate, oleate, linoleate, and two synthetic phosphatidylcholines. Parameters such as operating potential and mobile-phase solvent proportions, electrolyte composition, and ionic strength were evaluated for each peroxide class. Under optimal conditions, we have achieved baseline separation of four cholesterol hydroperoxide species, not only from one another, but also from phospholipid hydroperoxides; detection limits were < 0.3 pmol and < 30 pmol for the cholesterol and phospholipid hydroperoxides, respectively.

Animals↗

Crosstalk between calcium- and cyclic AMP-mediated signalling systems and the short-term modulation of bile flow in normal and cholestatic rat liver.

The flow of bile is subject to short-term modulation by glucagon and calcium-mobilizing hormones. Of potential relevance is the crosstalk between the second messenger-mediated signal transducing systems of these agonists. This latter point has revealed an area of investigation that should enable further insights to be made into a physiological network that interrelates bile flow, hepatocellular calcium movements and hormone action. This information in turn may provide insights into the etiology and treatment of human and animal diseases in which cholestasis is an underlying feature.

Animals↗

Isolation and characterization of the bacteriophage WO from Wolbachia, an arthropod endosymbiont.

Wolbachia is a group of obligate symbiotic bacteria found in many insects and other arthropods. The presence of Wolbachia alters reproduction in the host, but the mechanisms are unknown. Molecular biological studies of Wolbachia have delayed significantly, and one of the reasons is the lack of transformation techniques of this bacterium. In the present study, bacteriophage particles were isolated from Wolbachia for the first time. The purified phage had an isometric head that was approximately 40 nm in diameter and contained linear double-stranded DNA of approximately 20 kbp. Partial sequence information (total of 20,484 bp) revealed that there were 24 open reading frames including a structural gene module, and genes for replication and lysogenic conversion. This bacteriophage is the only known mobile genetic element potentially used for transformation of Wolbachia.

Animals↗

Large thoracolumbar meningomyelocele defects: incidence and clinical experiences with different modalities of latissimus dorsi musculocutaneus flap.

In this study, we report a series of large thoracolumbar meningomyelocele (MMS) defects. The total incidence per 1000 live birth is 0.6, the male incidence per 1000 live birth is 0.56, and the female incidence per 1000 live birth is 0.47. The study was conducted on 23 infants, all suffered from large thoracolumbar MMC defects, and our treatment techniques are modifications of procedures previously described. The timing of surgery ranged between 2 and 5 days after birth, and the follow-up was ranged between 3 and 7 years. Different modalities were used for reconstruction: bilateral proximally based skin island, muscle pedicle latissimus dorsi (LD), myocutaneous flap (seven patients), bilateral bipedicled LD, myocutaneous flaps (nine patients), and distally based skin island, muscle pedicle LD, myocutaneous flaps (seven patients). All the patients tolerated procedures smoothly, and because LD muscle may be important for mobility in a potentially paraplegic wheelchair-bound patient, the author presents some modifications in order to preserve the muscle function. Criteria for flap selection will be discussed.

Female↗

Mobility of heavy metals from tailings to stream waters in a mining activity contaminated site.

In this paper the results of a recent characterization of Rio Piscinas (SW of Sardinia, Italy) hydrological basin are reported. In such area (about 50 km2), previous mining activities caused a serious heavy metal contamination of surface waters, groundwater, soils and biota. Acid mine drainage phenomena were observed in the area. The main sources of contamination are the tailings stored in mine tunnels and abandoned along fluvial banks. A methodological approach was adopted in order to identify relations between tailings and water contamination. Representative samples of tailings and stream sediments samples were collected. XRD analyses were performed for mineralogical characterization, while acid digestion was carried out for determining metal contents. Batch sequential leaching tests were performed in order to assess metal mobility. Also groundwater and stream water were sampled in specific locations and suitably characterized. All information collected allowed the understanding of the effect of tailings on water contamination, thus contributing to the qualitative prediction of pollution evolution on the basis of metal mobility. Finally, a potential remediation strategy of stream water is proposed.

Geologic Sediments↗

Characterization of latex particles for aqueous polymeric coating by electroacoustic method.

Concentrated dispersions are used as coating dispersions for aqueous polymeric coatings, and characterization of the actual concentrated dispersions is an important in the pharmaceutical industry. The commonly used aqueous coating polymers are acrylic polymers and cellulose derivatives. We conducted a characterization study of polymethacrylate-based aqueous polymeric latex for aqueous coating, Eudragit L30D-55 (A-latex) and Eudragit RL30D (C-latex), by electroacoustic method. Colloidal Vibration Current (CVI) is one of the most important parameters relating to dynamic electrophoretic mobility and zeta potential, so we evaluated this parameter first. Volume fractions var of the latex in concentrated dispersions affect the CVIs according to the theory of Dukhin et al. The A-latex and C-latex CVI(*)s which were corrected with regard to the effect of volume fraction by the theoretical equation were nearly constant independent of phi in the ranges >0.04 and >0.03, respectively. The zeta potentials and colloidal stabilities of the concentrated dispersions were evaluated using an electroacoustic method by altering the pH and salt concentration. A-latex strongly aggregated at and below pH 2.5 and at and beyond 0.06 mol/L of electrolyte concentration. Regarding C-latex, instable dispersion was observed at 0.3 and 1 mol/L of NaCl concentration. The total potential energy of interaction between pairs of latex particles was changed by altering the salt concentration in this dispersion based on DLVO theory. The experimental results of stability in the concentrated latex dispersions can be explained by the total interaction energies.

Acoustics↗

Those interfering little RNAs! Silencing and eliminating chromatin.

RNA interference (RNAi) is widely used for knocking down expression of genes of interest and in systematic screens for desired phenotypes. In post-transcriptional gene silencing, double-stranded RNA triggers are processed to small interfering RNAs, which act to seek out and destroy homologous transcripts. A variety of organisms utilise the RNAi pathway to silence expression of potentially harmful endogenous mobile elements and to eliminate unnecessary sequences. In plants and fission yeast, RNAi can also mediate chromatin-based silencing resulting in transcriptional shutdown of homologous transcription units (transcriptional gene silencing) and the formation of centromeric heterochromatin. In metazoans, the expression of non-coding RNAs is often associated with the formation of silent chromatin domains but it remains to be determined if RNAi is involved.

Animals↗

Development of a fingerprint of Salvia miltiorrhiza Bunge by high-performance liquid chromatography with a coulometric electrode array system.

To standardize and control herbal medicines, a feasible approach and control system is necessary. In this paper, a high-performance liquid chromatography with a coulometric electrode array detector (HPLC-CEAD) system was applied to fingerprint Salvia miltiorrhiza Bunge (S. miltiorrhiza Bunge), a popular herbal medicine, for the first time. pH of mobile phase, working potentials and sample preparation were included in our research. Twenty-five common peaks were obtained from extracts of S. miltiorrhiza Bunge (Shandong province), more than that obtained in previous report. Fingerprints of S. miltiorrhiza Bunge from different locations were also studied. The content of main components varied in different samples. Overlapping ratio of peaks (ORP) in 10 batches of S. miltiorrhiza Bunge (Shandong province) was not less than 72.46%. In method validation, relative standard deviation (RSD) of relative retention times and relative peak areas were of not more than 3%. It was concluded that HPLC-CEAD system can be applied in fingerprinting herbal medicines.

Chromatography, High Pressure Liquid↗

Assistive devices for gait in Parkinson's disease.

While medications have improved the function of patients with Parkinson's disease (PD), over time most patients experience progressive problems with gait and balance. There have been important recent advances in the development of mechanical assistive devices that have the potential of improving mobility, but there are possible risks and the optimal role for such devices in PD has not been carefully studied. We review the available literature and provide practical information about available mobility assistive devices for patients with PD.

Gait↗

Sleep and intercontinental flights.

At present there are no 'high-tech' solutions to the problems that may beset intercontinental travellers. Indications for the use of drugs are limited, and their use must accord with Good Clinical Practice. Essentially, travellers must look after their sleep, as far as possible, during and after the flight. The most useful time for sleep during the flight must be anticipated, caution exercised in the use of hypnotics in-flight as reduced mobility is a potential risk factor for venous thrombosis, and a strategy adopted, whether flying east or west, to adapt as quickly as possible to the working hours of the new locality. After an eastward flight a hypnotic may be useful, but this strategy is seldom necessary after a westward flight unless the journey has involved crossing more than 5 or 6 time zones. The claim that melatonin accelerates the shift of the sleep-wakefulness cycle to a new time zone is controversial, and its recommended use may prejudice alertness during working hours. Exposure to artificial light and avoidance of ambient light at certain times of the day could prove to be of help-possibly in conjunction with drugs. However, effective and practical alterations in the light environment must be devised before such strategies can be considered with confidence.

Aircraft↗

23Na-nuclear magnetic resonance study of ionophore-mediated cation exchange between two populations of liposomes.

A model system to observe and investigate the transfer of Na+ ions between different internal compartments in suspension of liposomes was developed, and the exchange was followed by nuclear magnetic resonance spectroscopy. The experiments were performed under conditions of a Donnan equilibrium. Quantitative analysis of this three-site transmembrane exchange system allowed us to distinguish between direct and indirect exchange between liposomes. It also disclosed a "confining" effect on the exchange between the two populations of liposomes. This confining effect may have been due to an electrostatic field in the presence of a membrane potential. Donnan potentials and ionic compositions at equilibrium for the three-compartment system were calculated numerically. The model system may be used to explore further the effects of membrane potentials, surface potentials, and ionic mobilities on ion transport in biological (model) systems in general.

Biophysical Phenomena↗