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Excitation-contraction coupling in amphioxus muscle cells.

1. Excitation-contraction coupling was studied in myotomal muscles of amphioxus, Branchiostoma californiense.2. The action potential of a muscle cell produces a twitch with a rise time of 30-40 msec at 11 degrees C and its Q(10) is about 2.2.3. The twitch increases in amplitude with increasing external Ca concentration and is abolished in Ca-free saline (1 mM-EGTA and 55.7 mM-MgCl(2)); the twitch amplitude is suppressed by Co or La ions.4. Caffeine at concentrations above 1 mM in the external saline causes a prolongation of the action potential and a contracture which lasts several minutes.5. After exposure to caffeine the responsiveness of the muscle to subsequent applications of caffeine recovers in normal saline in 20-30 minutes but not in Ca-free saline.6. The amplitude of the caffeine contracture is independent of the external Ca concentration and is unaltered after the twitch is eliminated in Ca-free saline.7. After exposure to caffeine a full-sized twitch can be obtained before the responsiveness to caffeine shows any significant recovery.8. It is concluded that the twitch is produced by the Ca influx resulting from the increased permeability of the muscle cell membrane to Ca during the action potential and that the Ca mobilized by caffeine is not necessary to the initiation of the twitch.9. Electronmicroscopy shows the existence of sarcoplasmic reticulum.

Action Potentials↗

Identification of human endogenous retroviruses with complex mRNA expression and particle formation.

Retroviruses comprise strains with considerable disease potential in animals and humans. In addition to exogenous strains transmitted horizontally, endogenous proviruses are transmitted through the germ line. Some of these endogenous retroviruses can be pathogenic in mice and possibly in other animal species. They may also be considered as mobile genetic elements with the potential to produce mutations. In humans, genomic DNA contains numerous endogenous retroviral sequences detected by their partial relatedness to animal retroviruses. However, all proviruses sequenced so far have been found to be defective. In this communication, we describe the expression of a family of human endogenous retrovirus sequences (HERV-K) in GH cells, a teratocarcinoma cell line producing the human teratocarcinoma-derived retrovirus (HTDV) particles previously described by us. Four viral mRNA species could be identified, including a full-length mRNA. The other three subgenomic mRNAs are generated by single or double splicing events. This expression pattern is reminiscent of the more complex control of virus gene regulation observed, for example, with lenti- or spumavirus strains, although HERV-K shows no sequence homology to human T-lymphotropic virus or human immunodeficiency virus. Sequence analysis of expressed HERV-K genomes revealed non-defective gag genes, a prerequisite for particle formation. Open reading frames were also observed in pol and env. Antisera raised against recombinant gag proteins of HERV-K stained HTDV particles in immunoelectron microscopy, linking them to the HERV-K family.

Base Sequence↗

The relationship between functional mobility and the intensity of physical therapy intervention in children with traumatic brain injury.

PURPOSE: The purposes of this study were to describe the types of physical therapy (PT) procedural interventions used during inpatient rehabilitation for children and adolescents with traumatic brain injury (TBI) and to examine the relationship between intensity of intervention and mobility changes. METHODS: We examined a consecutive series of 80 records of children and adolescents (mean age = 10.3 years) with TBI. Intensity of PT was calculated as the number of 15-minute PT units of service per day/length of stay. We used the Pediatric Evaluation of Disability Inventory (PEDI) to calculate mobility change scores between admission and discharge and the achievement of a minimal clinically important difference (MCID) consisting of an 11-point change. We used a series of hierarchical and logistic regression analyses to identify the relationship of PT intensity to mobility changes, while controlling for potential confounding variables. RESULTS: Therapeutic exercise was the most frequently recorded intervention. On average, PT intensity was provided at a level of 3.1 units (46.5 minutes) per day. After controlling for severity and age, intensity was significantly related to change in PEDI mobility scores; this effect was removed when controlling for admission scores. The MCID was related to PT intensity, even when controlling for initial status. CONCLUSIONS: Although the magnitude of the correspondence is small, the intensity of PT intervention in an inpatient rehabilitation hospital is related to positive changes in functional mobility scores and the achievement of an MCID. However, initial mobility status is an essential factor in the interpretation of change scores. Future studies are needed to determine whether controlled variations of PT service intensity have a differential effect on mobility recovery during inpatient rehabilitation.

Journal Article↗

Hypoxia increases group IIA phospholipase A(2) expression under inflammatory conditions in rat renal mesangial cells.

Hypoxia evokes a common mechanism of oxygen sensing mediated by hypoxia-inducible transcription factors (HIF) in many mammalian cells. This study investigated the effect of hypoxia on group-IIA secretory phospholipase A(2) (sPLA(2)-IIA) expression in renal mesangial cells. Stimulation of cells with IL-1beta under normoxic conditions (21% O(2)) is known to induce expression and secretion of the group sPLA(2)-IIA. This induction is further enhanced by constantly reducing the O(2) concentration to 1% O(2), and is accompanied by increased sPLA(2) activity. To see whether hypoxia potentiates IL-1beta-induced sPLA(2)-IIA gene expression, a 2.67-kb fragment of the rat sPLA(2)-IIA promoter was fused to a luciferase reporter construct and used to transfect mesangial cells. Hypoxia alone is not able to activate the sPLA(2) promoter, whereas it significantly enhances IL-1beta-stimulated promoter activity. A deletion mutant of the promoter that lacks the two putative hypoxia responsive elements (HRE) is devoid of the potentiating effect of hypoxia. Moreover, site-directed mutagenesis of either of the two HRE is sufficient to abolish the potentiating effect of hypoxia. Electrophoretic mobility shift assays show that HIF-2alpha, which is the only HIF subtype expressed in mesangial cells, binds to both HRE in the sPLA(2)-IIA promoter. In summary, the data show that in an inflammatory setting hypoxia is able to potentiate sPLA(2)-IIA expression and activity in renal mesangial cells, and thereby may critically contribute to enhanced formation of inflammatory lipid mediators seen in a diverse range of kidney diseases.

Animals↗

Dietary intakes of nitrate, nitrite and NDMA in the Finnish Mobile Clinic Health Examination Survey.

Concern about potential health hazards of nitrate, nitrite and N-nitroso compounds necessitates calculations of exposures to these compounds and their distribution in normal populations. This study describes dietary intake of nitrate (NO3-), nitrite (NO2-) and N-nitrosodimethylamine (NDMA) among 5304 adult men and 4750 women, who participated in the Finnish Mobile Clinic Health Examination Survey in 1967-72. Food consumption data for each individual over the preceding year were collected by a dietary history interview. Intakes of nitrate, nitrite and NDMA from vegetables, fruits, cheese, meat and fish products were calculated using available values mainly derived from Finland and other countries in northern Europe. Nitrate and nitrite from drinking water were not included in the study. Mean daily dietary intake of nitrate was 77 mg, of nitrite 5.3 mg, and of NDMA 0.05 microgram respectively. Intake of NDMA from beer, estimated in a part of the study population, was 0.07 microgram per day. More than 90% of dietary nitrate was derived from vegetables, including potatoes. Nitrite was mainly provided by cured meat products. Cured meat products and smoked and salted fish were important food sources of NDMA. The total daily intake of nitrate was similar in men and women, whereas intakes of nitrite and NDMA were higher in men than in women. The diet of farmers was characterized by lower amounts of nitrate, nitrite and NDMA, whereas white collar workers and those employed in industry had higher intakes. Current smokers were exposed to higher dietary intakes of nitrate, nitrite and NDMA than non-smokers. Intakes of dietary nitrate, nitrite and NDMA estimated on an individual level are suggested to be useful in evaluating the health effects of these compounds in epidemiological studies.

Adolescent↗

Estimation and comparison of zeta-potentials of silica-based anion-exchange type porous particles for capillary electrochromatography from electrophoretic and electroosmotic mobility.

Mobilities of different chromatographic particles obtained from two electrokinetic methods were determined and compared. The particles were all based on porous silica, between 3 and 15 microm diameter, and were either native, or derivatized. As intermediate of chemical modification 3-mercaptopropyl-modified silica particles (TP-silica) are obtained. These particles were finally transformed into weakly basic anion exchangers with O-9-(tert-butylcarbamoyl)quinine (tBuCQN) as chiral selector. The electrophoretic mobility of the particles was determined from their migration velocity in an electric field using microelectrophoresis. Electrokinetic chromatography with a capillary column packed with the same particles was used to measure the electroosmotic flow generated. All measurements were carried out in background electrolytes of equal ionic strength (10(-2) mol/L), at pH varying between 3.5 and 9.5. From these data a rough estimation of the zeta-potential was made, taking Helmholtz-Smoluchowski conditions into consideration. With both methods the zeta-potential of the native silica particles is negative throughout, and its value increases with pH. The weakly basic tBuCQN particles have positive zeta-potentials at pH lower than about 7.5, but exhibit a negative zeta-potential above this pH, indicating the dominating effect of residual silanol groups at the silica surface. The zeta-potential for these anion-exchange particles ranged between +30 and -40 mV. The zeta-potentials derived with electrophoresis and electroosmosis agree, showing the adequacy of the approach, although many limitations must be taken into account in the treatment of the electrokinetic phenomena in such porous systems. These restrictions in interpreting mobility and zeta-potential were discussed.

Anion Exchange Resins↗

Platelets in patients with homozygous familial hypercholesterolemia are sensitive to Ca(2+)-mobilizing activity of low density lipoproteins.

While some data suggest that Ca(2+)-mobilizing effects of low density lipoprotein (LDL) in platelets are mediated by a specific membrane receptor, the data about the nature of this lipoprotein-binding site are contradictory. This work was performed in order to assess possible involvement of apolipoprotein (apo) B,E receptor, present in most cell types. To answer the question we compared effects of LDL in normal platelets and those obtained from patients with homozygous familial hypercholesterolemia (HFH), characterized by absence of functional apo B,E receptors. We have found that in accordance with previous results LDL induced instant reversible elevation of free cytoplasmic calcium concentration ([Ca2+]i) in fura-2-loaded platelets. The effect was observed both in healthy and HFH groups. Neither half-maximal effective concentrations nor maximal effects of LDL differed significantly between two groups. Ca(2+)-mobilizing effects of lipoproteins were potentiated about 4-fold by epinephrine and completely blocked by prostaglandin E1 both in platelets of healthy and HFH subjects. The similarity of lipoprotein effects in control and HFH platelets is evidence that apo B,E receptor does not mediate the Ca(2+)-mobilizing activity of LDL in this cell type.

Adenosine Diphosphate↗

Leaching behaviour of pharmaceuticals in soil-testing-systems: a part of an environmental risk assessment for groundwater protection.

The leaching behaviour of six selected pharmaceuticals was tested in different soils. Leaching experiments are a part of environmental risk assessment to estimate the distribution and fate of these pharmaceuticals in the environment. Based on the results of this assessment their mobility in soil and their potential to contaminate groundwater was evaluated. When assessing the leaching behaviour of these compounds, the influence of the properties (e.g. grain size distribution, pH, Corg) of different soils has to be taken into account. The test results indicated that the leaching potential found could be rated as low for diazepam, ibuprofen, ivermectin and carbamazepine. Therefore, contamination of the groundwater with these substances seems to be unlikely if the groundwater level is covered with sufficient layers of the soils investigated. Clofibric acid and iopromide were very mobile under the experimental conditions and thus, groundwater contamination would be possible if the soil is exposed to these pharmaceuticals, i.e. wastewater irrigation. These results are more or less in agreement with groundwater monitoring data found in the literature for ibuprofen and diazepam which were in general not present in groundwater, while clofibric acid and iopromide were frequently detected. However, a discrepancy was found for carbamazepine, since it occurs very often in groundwater. This discrepancy might be explained by the fact that the leaching tests were performed with soil, whereas in reality the groundwater contamination occurs mainly over river sediments and sub soil from receiving waters.

Carbon↗

Leukocyte mobilization from the guinea pig spleen by muscarinic cholinergic stimulation.

Important interactions between the immune system and the nervous and endocrine systems have become increasingly accepted. The present results demonstrate that the cholinergic agonist carbacholine greatly increased the number of granulocytes and lymphocytes in the splenic venous blood, but not arterial blood, shortly after administration to guinea pigs. The effect was largely blocked by pretreatment with atropine. In contrast, animals treated with indomethacin had a decreased number of leukocytes in both splenic venous and arterial blood. A decrease in relative splenic weight due to carbacholine treatment was also blocked by atropine. However, cholinergic leukocyte mobilization, or that previously observed after adrenergic stimulation, may not be caused by capsule contraction since it is not accompanied by mobilization of erythrocytes. Furthermore, indomethacin, which potentiates the response of splenic smooth muscle to adrenergic stimuli, blocked the effect of noradrenaline (NA) on leukocyte mobilization.

Animals↗

An electroacoustic and high-frequency dielectric response study of stagnant layer conduction in emulsion systems.

Stagnant layer conduction (or anomalous surface conduction) in perfluoromethyldecalin (PFMD) and n-hexadecane emulsions has been measured by electroacoustics and verified by high-frequency dielectric response experiments. The electroacoustic technique can detect the presence of stagnant layer conduction from the salt dependence of the dynamic mobility. As the indifferent electrolyte concentration is increased from low values (<5 mM), the zeta-potential and droplet size, estimated from the dynamic mobility by the normal procedures, gradually increase in magnitude until the size plateaus and the zeta-potential begins to decrease with added salt in the usual fashion. When stagnant layer conduction is taken into account, the dynamic mobility can be fitted to a constant size distribution and more realistic zeta-potential values with varying electrolyte concentration. High-frequency dielectric response has been used to measure the total conduction in a PFMD emulsion system. Very good agreement between these two independent techniques verifies the existence of conduction behind the shear plane and demonstrates that electroacoustics alone can detect and quantify its extent. This is possible because of the unique character of the AcoustoSizer procedure, which estimates both particle size and zeta-potential from the same signal.

Journal Article↗

Some problems of zeta potential determination in electrophoretic measurements on lipid membranes.

Electrophoretic mobilities were measured for cardiolipin, phosphatidylinositol and phosphatidylserine liposomes in solutions of potassium chloride. The zeta potential as a function of ionic strength deviates significantly from the predictions of the double-layer theory in the 10(-3)-5.10(-2) M range. This might be due to changes in pH in the course of the experiment and/or to the inapplicability of the Smoluchowski equation at low ionic strengths. Taking into account the relaxation effects results in theoretical curves, calculated for various particle sizes at low ionic strength, which are in better agreement with the experiment. However, for quantitative comparison with experimental data, further investigations are necessary.

Cardiolipins↗

[Pacemaker dysfunction during use of a mobile telephone].

Ubiquitious electronic equipment increases the number of potential sources of electromagnetic interference with sufficient energy to influence a pacemaker function. Besides others the mobile phone play important role. A 76 year-old female patient with pacemaker was hospitalized because of bilateral congestive heart failure. During ECG telemetry a short episode of pacemaker dysfunction was recorded. In a period of 15 seconds at first a pulse output inhibition occurred, then an undersensing was present. In all the 24-hour recording no other similar episode was observed. Such an utterly atypical picture lead to a direct question on mobile telephone calling. The patient confirmed that during a visit of her relatives she was really using a mobile phone. The GSM (Global System for Mobile Communication) phones operate on a carrier frequency of 900 MHz which is modulated generally in pulses about 8 Hz and 2 Hz. These signals can be falsely detected as an intrinsic heart activity by a pacemaker or they are analysed to be an electromagnetic interference. As a result a pacemaker inhibition, an asynchronous pacing and in dual chamber systems also a ventricular triggering can be present. These reactions usually do not occur if the distance between phone and pacemaker is higher than 10 cm. With some measures patients with pacemakers can safely use mobile phones. In pacemaker dependent patients a careful approach is necessary.

Aged↗

Effects of dissolved organic matter from animal waste effluent on chlorpyrifos sorption by soils.

The increased use of animal waste-derived effluents for irrigation could result in the enhanced movement of pesticides through complexation with dissolved organic materials. Batch equilibrium studies were conducted to measure the interaction among soil, chlorpyrifos [O,O-diethyl O-(3,5,6-trichloro-2-pyridyl) phosphorothioate], and dissolved organic matter (DOM) from poultry, swine, and cow waste-derived lagoon effluents. All DOM was found to have a strong affinity for chlorpyrifos, resulting in reduced sorption of chlorpyrifos by soil, thus the potential for DOM-enhanced mobility. Effluent DOM was observed to sorb to soils. Thus, for increasingly higher soil mass to solution volume ratios, the effect of chlorpyrifos association with water-borne DOM on sorption decreases significantly. For high soil mass to solution volume ratios typical of soil profiles in the landscape, the potential for DOM-enhanced transport will be greatly attenuated. Dissolved organic matter concentration and the nonpolar nature of DOM in the lagoon effluent decreased with increasing residence time in the cells of the lagoon system, thus reducing the potential for DOM-enhanced transport.

Adsorption↗

Effect of dextran as a run buffer additive in drug-protein binding studies using capillary electrophoresis frontal analysis.

The study of drug-protein interactions by capillary electrophoresis frontal analysis requires establishment of a sufficient mobility difference between the mobility of the ligand and protein. The potential utility of dextran as a run buffer additive to manipulate the electrophoretic mobilities of low molecular weight ligands and protein in capillary electrophoresis frontal analysis binding studies was assessed. It was demonstrated that dextran was effective in improving the separation between the ligands warfarin and flurbiprofen and human serum albumin. Separation of ligand and protein increased with the concentration of added dextran (0-7.5% (w/w)), while molecular weight of the additive (70,000-2,000,000) only had a minor effect. The effect of dextran addition on viscosity and electrophoretic and electroosmotic mobilites was systematically studied. Optimal frontal analysis settings were a compromise between achieving satisfactory separation and acceptable analysis times without loss of plateau peak conditions. No effect of dextran upon the drug-human serum albumin interactions could be detected for the model ligands. Introduction of dextran into the electrophoresis buffer expands the applicability of capillary electrophoresis frontal analysis in drug research to binding interactions between proteins and low molecular weight ligands possessing similar electrophoretic mobilities.

Buffers↗

Particle-scale understanding of the bioavailability of PAHs in sediment.

This study reports results of sediment bioslurry treatment and earthworm bioaccumulation for polycyclic aromatic hydrocarbon (PAH) contaminants found in sediment dredged from Milwaukee Harbor. A significant finding was that bioslurry treatment reduced PAHs on the sediment clay/silt fraction but not on the sediment coal-derived fraction and that PAH reduction in the clay/silt fraction correlated with substantial reduction in earthworm PAH bioaccumulation. These findings are used to infer PAH bioavailability from characterization of particle-scale PAH distribution, association, and binding among the principal particle fractions in the sediment. The results are consistent with work showing that the sediment comprised two principal particle classes for PAHs, coal-derived and clay/silt, each having much different PAH levels, release rates, and desorption activation energies. PAH sorption on coal-derived particles is associated with minimal biodegradation, slow release rates, and high desorption activation energies, while PAH sorption on clay/silt particles is associated with significant potential biodegradability, relatively fast release rates, and lower desorption activation energies. These characteristics are attributed to fundamental differences in the organic matter to which the PAHs are sorbed. Although the majority of the PAHs are found preferentially on coal-derived particles, the PAHs on the clay/silt sediment fraction are more mobile and available, and thus potentially of greater concern. This study demonstrates that a suite of tests comprising both bioassays and particle-scale investigations provide a basis to assess larger-scale phenomena of biotreatment of PAH-impacted sediments and bioavailability and potential toxicity of PAH contaminants in sediments. Improved understanding of contaminant bioavailability aids decision-making on the effectiveness of biotreatment of PAH-impacted sediments and the likelihood for possible reuse of dredged sediments as reclaimed soil or fill.

Adsorption↗

Fate of antibiotic resistance genes during rural domestic wastewater treatment: Anaerobic unit as enrichment hotspot versus aerobic unit as attenuation zone.

Rural domestic wastewater treatment systems are important but understudied reservoirs for antibiotic resistance genes (ARGs), whose full-process migration mechanisms remain unclear. Herein, the contribution of each treatment unit of ARGs was investigated using metagenomic methods across two seasons in typical rural domestic wastewater treatment systems. Although a removal efficiency (69&#xa0;% in winter and 22&#xa0;% in summer) was observed for ARGs, higher antibiotic residues and temperature dramatically induced ARG occurrence in wastewater and horizontal gene transfer (HGT) risk during wastewater treatment. The ARG abundances in the anaerobic unit increased by 1.6-2.1 fold compared to the regulating pool, primarily driven by elevated mobile genetic element (MGE) activity. In sharp contrast, ARG reduction was achieved through ARG host removal and suppressed HGT potential in the aerobic unit. Notably, mobile ARGs were dominated by tetracycline resistance genes in winter and co-dominated by tetracycline and sulfonamide genes in summer, with most flanked by transposases. Key pathogenic hosts, including Klebsiella pneumoniae, Escherichia coli, and Pseudomonas aeruginosa carrying ARG-MGE complexes, were primarily concentrated in the regulating pool and the influent, forming high-risk upstream sources of dissemination. Partial least-squares path model highlighted MGEs as the primary drivers, and variance partitioning analysis indicated that MGEs account for 31&#xa0;% of the explained variation in ARGs during wastewater treatment. In summary, the anaerobic unit was an ARG enrichment hotspot, while the aerobic unit as ARG attenuation zone during wastewater treatment. These findings provide crucial evidence to optimize rural wastewater treatment processes and to target the control of antibiotic resistance.

Wastewater↗

Zeta Potential of Nanobubbles Generated by Ultrasonication in Aqueous Alkyl Polyglycoside Solutions.

A simple and convenient method to measure microelectrophoretic mobilities was proposed to determine the zeta potential of nanobubbles generated by ultrasonication. Bubbles in pure water solutions and in aqueous solutions of alkyl polyglycoside (AG) with different alkyl chain lengths and degrees of polymerization in the head group were sonicated with a palladium-coated electrode designed specially by the manufacturer. The zeta potentials of bubbles with ordinary cationic and ionic surfactants are consistent with others' previous results. The average size of the bubbles generated by sonication is in the range of 300 to 500 nm. The zeta potentials of bubbles in both pure water and AG solutions at all pH values are negative. As the chain length of AG increases, zeta potentials significantly decrease at high pH. For nonionic AG, a possible charging mechanism based on known mechanisms is suggested to explain the negative charge, known to be unusual. Even with a very high concentration of H(+) ions in solution the bubbles are charged negatively because the interface is covered with slightly acidic alcohol groups of AGs. At high pH, the less polar the surfactant, the more negative the charge, since nonpolar surfactant molecules induce the adsorption of OH(-) ions, rather than H(+) ions that prefer hydration by water molecules. Copyright 2000 Academic Press.

Journal Article↗

Degenerative changes in the function of neuromuscular junctions of Manduca sexta during metamorphosis.

In Manduca sexta the decline in neuromuscular function during metamorphic degeneration was compared in two muscles which differed characteristically with regard to pre- and postsynaptic physiological properties. In both muscles, morphological evidence indicated that a significant number of the active zones within the population of neuromuscular junctions on a given fiber were nonfunctional. Nevertheless, the degenerating nerve terminals were able to produce an above-threshold excitatory junction potential (EJP) which was facilitated in a manner characteristic of the muscle being observed. Abnormal findings during the early stages of degeneration included a larger than normal EJP, a decline in EJP amplitude over a 20 min period even with low frequencies of stimulation, an increase in EJP duration, a decline in muscle fiber resting potential amplitude with age, a decrease or disappearance of post-tetanic potentiation and long-term facilitation, and an increased likelihood that the motor nerve would fail to conduct a stimulus. The two muscles were qualitatively similar but quantitatively different with regard to these degenerative changes. It is suggested that this combination of relatively normal function with abnormal properties might be associated with the withdrawal of glial processes from the neuromuscular junctions, changes in the cable properties associated with shrivelling of the muscle fibers, and a decline in the metabolic functions supporting both muscle fiber resting potentials and those underlying transmitter synthesis, mobilization and release.

Animals↗