MRX 501: a novel ultrasound contrast agent with therapeutic properties.
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Animals↗
Biomedical subjects
G Wu
Publications and source records attributed to G Wu.
Protocol for microneurography with concentric needle electrodes.
In 1968, the method of human percutaneous microneurography with solid tungsten electrodes was introduced. Since then many investigators used this technique to study peripheral mechanisms in the somatosensory, motor and autonomic systems of conscious humans. Although some modifications of the method were described, the basic construction of the recording electrode has remained the same over the years. In the present protocol we describe in detail the procedures of microneurography using a thin diameter concentric needle electrode. There are some advantages with the concentric electrodes in comparison with the tungsten needles: (1) the electrical and mechanical properties of the electrode are stable which allows repeated use, (2) its restricted and one-dimensionally directed recording area provides the possibility to study topographical aspects within even a part of a peripheral nerve fascicle, and (3) multi-channel recordings can be achieved by adding more recording surfaces to the electrode. Based on recent investigations evaluating the recording properties of concentric electrodes we propose a novel procedure for signal analysis where template matching is incorporated. The analyses described in this protocol might also be applicable for extracellular recordings from muscle or elsewhere within the nervous system.
Arm↗
Induction of axon-like and dendrite-like processes in neuroblastoma cells.
Neuroblastoma cells are widely utilized models for the study of the neuritic outgrowth phase of neuronal differentiation, but relatively few such studies have attempted to identify the nature of the process outgrowths. This identification will be necessary in developing strategies for utilizing these models to distinguish the underlying mechanisms involved in axonogenesis vs dendritogenesis. In an effort to identify procedures for inducing specific types of neurite outgrowth, and for distinguishing axon- from dendrite-like processes, we have subjected two neuroblastoma cell lines to a variety of stimuli previously shown to induce neurite outgrowth in these cells. These include neuraminidase, ionomycin, KCl+dibutyryl cAMP, cholera toxin B subunit, retinoic acid, dibutyryl cAMP (alone), GM1 ganglioside, and low serum. The first four of these (group 1) gave rise to neurites with axon-like characteristics, including immunostaining that was positive for phosphorylated high molecular weight neurofilament protein (NF-H) and synaptic vesicle protein-2 (SV2), but negative for microtubule-associated protein-2 (MAP2). The next three treatments (group 2) resulted in dendrite-like processes, as evidenced in immunostaining that was positive for MAP2 and negative for NF-H and SV2. Neurites produced by low serum had mixed properties. These cytoskeletal differences were supported by immunoblot analysis with antisera to the above cytoskeletal proteins. Striking morphological differences were also noted, group 2-induced neurites being significantly shorter with more branch points than those generated by group 1 stimulants. Time of exposure to stimulatory agent was crucial in determining expression of the neuritic phenotype. Correlation with previous studies suggests that axon-like neurites result from stimulants which elevate intracellular Ca2+, a dependence not previously reported to our knowledge. Dendrite-like process outgrowth, on the other hand, does not appear to depend on altered intracellular Ca2+.
Animals↗
Clustering of slowly adapting type II mechanoreceptors in human peripheral nerve and skin.
The intrafascicular organization of human peripheral nerves was studied with percutaneous microneurography in the median, radial and peroneal nerves with one-surface or two-surface thin diameter concentric needle electrodes. Data from 33 recording sites containing 47 slowly adapting type II (SAII) units were analysed. At many sites two, sometimes even three, neighbouring SAII units were recorded from the explored nerve fascicle and they had adjacent or even overlapping cutaneous receptive fields. Among pairs of SAII units found at the same site, one unit often had ongoing discharge, whereas the other was silent under resting conditions. The neighbouring SAII units were optimally activated by stretching the skin in different directions. Stretching the same skin area in different directions produced different unit recruitment. Clustered SAII units were often found in sites where Pacinian afferents and skin sympathetic activity were also recorded. No significant difference was observed in the degree of grouping of SAII units either between recordings obtained with one-surface versus two-surface electrodes or between glabrous and hairy skin. The data do not support the notion that myelinated fibres are randomly organized in peripheral nerve fascicles. Instead, the findings suggest that SAII units tend to be clustered in human peripheral nerves. Furthermore, the response of groups of SAII units to skin stretch suggests that they play a role in proprioception. Dual channel recordings with two-surface concentric needle electrodes may provide a novel approach to study fibre organization in human peripheral nerves and the behaviour of groups of nerve fibres.
Adaptation, Physiological↗
Maternal dietary protein deficiency decreases nitric oxide synthase and ornithine decarboxylase activities in placenta and endometrium of pigs during early gestation.
Little is known about the mechanism responsible for retarded placental and fetal growth induced by maternal dietary protein malnutrition. On the basis of the recent finding that nitric oxide (NO) and polyamines (products of L-arginine) play an important role in embryonic and placental development, the present study was designed to determine whether protein deficiency decreases placental and endometrial activities of NO synthase (NOS) and ornithine decarboxylase (ODC) (the first and key regulatory enzyme in polyamine synthesis). Primiparous gilts selected genetically for low or high plasma total cholesterol concentrations (low line and high line, respectively) were mated and then fed 1.8 kg/d of isocaloric diets containing 13% or 0.5% crude protein. At d 40 or 60 of gestation, they were hysterectomized, and placenta and endometrium were obtained for incubations, NOS and ODC assays, and measurements of free amino acids and polyamines. Maternal dietary protein restriction decreased arginine and ornithine concentrations, constitutive and inducible NOS activities and NO production, as well as ODC activity and polyamine concentrations in placenta and endometrium of both lines of gilts. Placental NO synthase activity and NO generation were lower in high line gilts than in low line gilts. ODC activities and polyamine concentrations in placenta and endometrium were decreased at d 60 compared with d 40 of gestation. These changes in placental and endometrial synthesis of NO and polyamines during early gestation may be a mechanism responsible for reduced placental and fetal growth in protein-deficient gilts and for altered conceptus development in high line gilts.
Amino Acids↗
Maternal dietary protein deficiency decreases amino acid concentrations in fetal plasma and allantoic fluid of pigs.
This study was conducted to test the hypothesis that maternal dietary protein deficiency decreases amino acid availability to the fetus, thereby contributing to retarded fetal growth. Primiparous gilts selected genetically for low or high plasma total cholesterol concentrations (low line and high line, respectively) were mated, and then fed 1.8 kg/d of isocaloric diets containing 13% or 0.5% crude protein. At d 40 or 60 of gestation, they were hysterectomized, and maternal and fetal blood samples as well as amniotic and allantoic fluids were obtained for analyses of amino acids, ammonia and urea. Dietary protein restriction decreased (P < 0.05) the following: 1) maternal plasma concentrations of urea at d 40 and 60 of gestation; 2) fetal plasma concentrations of alanine, arginine, branched-chain amino acids (BCAA), glutamine, glycine, lysine, ornithine, proline, taurine, threonine and urea at d 60 of gestation; 3) amniotic and allantoic fluid concentrations of urea at d 40 and 60 of gestation; and 4) allantoic fluid concentrations of alanine, arginine, BCAA, citrulline, cystine, glycine, histidine, methionine, proline, serine, taurine, threonine and tyrosine at d 40 of gestation, in gilts of both genetic lines. At d 60 of gestation, protein deficiency decreased (P < 0.05) allantoic fluid concentrations of arginine, cystine, glycine, taurine and tyrosine in low line gilts and of cystine, glutamine, ornithine, serine, taurine and tyrosine in high line gilts. Low line and high line gilts also differed remarkably in allantoic fluid concentrations of arginine, glutamine, ornithine and ammonia at d 40 and 60 of gestation. Our results suggest the following: 1) protein-deficient gilts maintain maternal plasma concentrations of amino acids by mobilizing maternal protein stores and decreasing oxidation of amino acids during the first half of gestation; 2) protein deficiency may impair placental transport of amino acids from the maternal to the fetal blood; and 3) low line and high line gilts differ in fetal amino acid metabolism. Decreases in concentrations of the essential and nonessential amino acids in the fetus may be a mechanism whereby maternal dietary protein restriction results in fetal growth retardation.
Allantois↗
Intestinal mucosal amino acid catabolism.
The small intestine is not only responsible for terminal digestion and absorption of nutrients, but it also plays an important role in catabolism of arterial glutamine and dietary amino acids. Most of glutamine and almost all of glutamate and aspartate in the diet are catabolized by small intestinal mucosa, and CO2 accounts for 56-64% of their metabolized carbons. The small intestinal mucosa also plays an important role in degrading arginine, proline and branched-chain amino acids, and perhaps methionine, lysine, phenylalanine, threonine, glycine and serine in the diet, such that 30-50% of these dietary amino acids are not available to extraintestinal tissues. Dietary amino acids are major fuels for the small intestinal mucosa and are essential precursors for intestinal synthesis of glutathione, nitric oxide, polyamines, purine and pyrimidine nucleotides, and amino acids (alanine, citrulline and proline), and are obligatory for maintaining intestinal mucosal mass and integrity. Because intestinal amino acid catabolism plays an important role in modulating dietary amino acid availability to extraintestinal tissues, it has important implications for the utilization efficiency of dietary protein and amino acids in animals and humans.
Amino Acids↗
Energy metabolism of rat colonocytes changes during the tumorigenic process and is dependent on diet and carcinogen.
Alterations in ATP production, intracellular energy levels and mitochondrial function have been shown to trigger cytokinetic events in vitro, including inhibition of cell division, abnormal or blocked differentiation and inhibition of apoptosis. Changes in colonic cytokinetics are directly related to colon tumorigenesis but alterations in energy metabolism during the tumorigenic process have never been reported. We conducted a 2 x 2 x 3 factorial design study in 120 male Sprague-Dawley rats with two diets (pectin or cellulose-supplemented), two injected subgroups (with or without the carcinogen azoxymethane, AOM) and three termination time points (6, 16 and 36 wk post-second injection). Colonocytes were isolated and incubated with their primary energy substrates (radiolabeled butyrate, glucose, glutamine and beta-hydroxybutyrate) for 60 min. Production of lactate, ketone bodies and CO2 were determined. At 6 wk, there were no significant differences in metabolism among treatments. In contrast, at 16 wk, AOM-injected rats had dramatically lower rates of CO2 production (P < 0.001) from both glucose and butyrate and lower rates of lactate and ketone body production than their saline counterparts. At 36 wk, when tumors developed, the depressed production of lactate and ketone bodies seen in AOM-injected rats at 16 wk returned to control values. However, in AOM-injected rats, CO2 production from glucose and butyrate remained depressed. Cellulose feeding resulted in decreased oxidation of glucose, butyrate and glutamine and an increased production of ketone bodies from butyrate by colonocytes compared with pectin feeding at 36 wk. We conclude that colonocyte energy metabolism differs in AOM-injected rats vs. saline controls and changes during tumorigenesis, and suggest a relationship between intracellular energy status and changes in cell kinetics. This is the first report that such a relationship may exist in vivo.
3-Hydroxybutyric Acid↗
Binding and lysing of blood clots using MRX-408.
RATIONALE AND OBJECTIVES: A thrombus-specific ultrasound contrast agent, MRX-408, has been developed recently. This agent consists of phospholipid-coated microbubbles with a ligand capable of targeting the GPIIb/IIIa receptor, thereby allowing the microbubbles to bind with thrombi rich in activated platelets. In vitro and in vivo animal experiments have been conducted to examine imaging enhancement and sonothrombolysis using this agent compared with a nontargeted agent. METHODS: For clot binding, blood-smeared slides were incubated with microbubbles and examined under a light microscope. Change in backscatter signals from the blood clots after binding was examined by both an ultrasound scanner and two single-element transducers arranged in a transmitter-receiver pair. For clot lysis, either 1-MHz or 20-KHz ultrasound was used to enhance the lysing effects of MRX-408 with or without urokinase. RESULTS: Evidence of binding was demonstrated under a microscope. In vitro experiments showed that the "acoustic signature", or properties, of blood clots changed after binding. Clots became more echogenic and nonlinear. In vivo fundamental ultrasound imaging confirmed that as a result of binding, blood clots were more visible, the area of detection was improved, and shadowing behind clots was more noticeable. Under 1-MHz ultrasound and 30 minutes of treatment, lysis efficiency reached 34% with MRX-408, whereas there was no visible clot lysis with saline. CONCLUSION: The results of these preliminary studies show that as a contrast agent, MRX-408 enhanced clots under ultrasound imaging and facilitated sonothrombolysis with or without thrombolytic drugs.
Animals↗
Arabidopsis Rho-related GTPases: differential gene expression in pollen and polar localization in fission yeast.
The Rho small GTP-binding proteins are versatile, conserved molecular switches in eukaryotic signal transduction. Plants contain a unique subfamily of Rho-GTPases called Rop (Rho-related GTPases from plants). Our previous studies involving injection of antibodies indicated that the pea Rop GTPase Rop1Ps is critical for pollen tube growth. In this study we show that overexpression of an apparent Arabidopsis ortholog of Rop1Ps, Rop1At, induces isotropic cell growth in fission yeast (Schizosaccharomyces pombe) and that green fluorescence protein-tagged Rop1At displays polar localization to the site of growth in yeast. We found that Rop1At and two other Arabidopsis Rops, Rop3At and Rop5At, are all expressed in mature pollen. All three pollen Rops fall into the same subgroup as Rop1Ps and diverge from those Rops that are not expressed in mature pollen, suggesting a coupling of the structural conservation of Rop GTPases to their gene expression in pollen. However, pollen-specific transcript accumulation for Rop1At is much higher than that for Rop3At and Rop5At. Furthermore, Rop1At is specifically expressed in anthers, whereas Rop3At and Rop5At are also expressed in vegetative tissues. In transgenic plants containing the Rop1At promoter:GUS fusion gene, GUS is specifically expressed in mature pollen and pollen tubes. We propose that Rop1At may play a predominant role in the regulation of polarized cell growth in pollen, whereas its close relatives Rop3At and Rop5At may be functionally redundant to Rop1At in pollen.
Amino Acid Sequence↗
Prediction of serum vancomycin concentrations using one-, two- and three-compartment models with implemented population pharmacokinetic parameters and with the Bayesian method.
Although previous studies have shown that vancomycin has a complicated pharmacokinetic profile requiring description using a two- or, better, three-compartment model, until recently predictions of serum vancomycin concentrations have been mainly based on one- or two-compartment models using computer software packages. In this study, we have predicted serum vancomycin concentrations in 59 patients using one-, two- and three-compartment models with implemented population pharmacokinetic parameters in the Abbott PKS program and by use of the Bayesian method. The percentage errors of predictions made using the one-compartment model were smaller when either the Bayesian method or implemented population pharmacokinetic parameters were used (medians of -8.61% and -9.49%, respectively). Predictions using the one-compartment model with the Bayesian method were less biased (median of -1.52 microgmL(-1). The best predictions were those made using the three-compartment model with the Bayesian method-they were most accurate (median of 3.40 microgmL(-1) and highly precise (median of 11.53 microg(2)mL(-1)). The results suggest that predictions made using the one-compartment model with implemented population pharmacokinetic parameters are preferable if no samples are available, otherwise predictions made using the three-compartment model with the Bayesian method are preferable. The results also supported our previous argument that the greater the number of compartments involved in individualization, the better the predictions obtained using the Bayesian method.
Adult↗
Effect of the number of samples on Bayesian and non-linear least-squares individualization: a study of cyclosporin treatment of haematological patients with multidrug resistance.
We have studied whether the prediction of drug concentrations improves as the number of samples used for individualization is increased, and whether the Bayesian method of individualization is superior to the non-linear least-squares method. Data were obtained from ten adult haematological patients with multidrug resistance who were treated with cyclosporin. The predictions of blood-cyclosporin concentrations were made using the Abbott PKS program. The number of samples used for individualization was increased from 1 to 30 for the Bayesian method and from 4 to 30 for the non-linear least-squares method. Linear regression, percentage prediction error, and absolute and relative predictive performance were used to evaluate the predictions. The results show that the Bayesian method affords greater precision than the non-linear least-squares method, but that the non-linear least-squares method is more accurate and results in less bias. Whereas for linear regression predictions improve as the number of samples is increased, other evaluations show improvement in the range from 5 to 11 samples; linear regression, percentage prediction errors and prediction bias support the opinion that the Bayesian method progressively becomes the non-linear least-squares method as the number of samples used for individualization is increased, but the accuracy and precision of prediction do not support this opinion. The study supports the statement that Bayes' law requires parameters from an infinite population, otherwise the advantage of the Bayesian method might be marginal.
Adult↗
Recent developments in solid-state nuclear magnetic resonance of quadrupolar nuclei and applications to biological systems.
Recent advances in nuclear magnetic resonance (NMR) methodology and improvements in high-field NMR instrumentation have generated a new wave of research interests in the application of solid-state NMR to the study of quadrupolar nuclei. These developments now permit increasingly complex biological systems to be probed by quadrupolar NMR. In this review I describe a few recent developments in NMR studies of quadrupolar nuclei and demonstrate the potential of solid-state quadrupolar NMR in the study of biological systems. In particular, I discuss the application of solid-state NMR of (17)O, 67Zn, 59Co, 23Na, and 39K nuclei with a prognosis for future work.
Anisotropy↗
Adipose tissue partitioning of limit-fed beef cattle and beef cattle with ad libitum access to feed differing in adaptation to heat.
We compared fat distribution and lipoprotein lipase (LPL) activity in steers differing in adaptability to the subtropics. Steers were fed a grain diet (3.13 Mcal ME/kg DM) at limited (150 kcal ME x kg[-.75] x d[-1]; .23 kg ADG) or ad libitum levels for 140 d, then slaughtered. Sixteen British- (8 Angus, 8 Hereford; S), 16 Boran- (R), 16 Brahman- (B), and 16 Tuli- (T) cross steers from MARC III composite cows were used. Adipose tissue samples from perirenal, omental, and subcutaneous depots were analyzed for LPL activity. Carcass measurements including omental, external, and seam fat trim from 1/ 2 of the carcass were measured. Subcutaneous fat had greater (P < .05) LPL activity than fat from the other depots. Generally, there were no differences (P > .05) in fat distribution for steers fed at limited levels. Means for ADG, slaughter weights, carcass weights, yield grades, and carcass lipid weights for S and B fed for ad libitum intake were greater (P < .05) than those for T and R. Marbling was greatest (P < .05) for S and did not differ (P > .05) for the other breeds with ad libitum intake. Factor analysis of fat depots for animals with ad libitum intake indicated that Bos taurus cattle differing in adaptation to heat deposited fat differently; S deposited greater (P < .05) proportions of carcass fat and T deposited greater (P < .05) proportions of internal fat. It seems that accumulation of internal fat is detrimental for ADG for Bos taurus cattle.
Acclimatization↗
Physiological adaptations in adipose tissue of Brahman vs Angus heifers.
Nonpregnant yearling Brahman (n = 12) and Angus (n = 12) heifers were equally allocated to two dietary treatments in a replicated study to examine responses in lipid metabolism to nutritional treatments consisting of a moderate energy diet (2.0 Mcal ME/kg) fed at maintenance and a 2.5 x maintenance high-energy diet (2.4 Mcal ME/kg) fed for 30 d. In vitro lipogenesis and the activities of lipoprotein lipase (LPL) and hormone-sensitive lipase (HSL) were determined in perianal subcutaneous adipose tissue biopsies at the start and end of the trial. At the start of the trial, breeds had similar (P > .10) rates of lipogenesis and LPL activity. Brahman had greater (P < .05) HSL activity than Angus at the start of the trial and tended (P < .07) to have greater HSL activity at the end. Diet did not influence (P > .10) HSL activity. Heifers on the high-energy, higher-intake diet had greater lipogenesis (P < .001) and LPL activity (P < .01) than those on the moderate-energy diet. Inclusion of body condition score (BCS) nested within breed as a covariate explained breed differences for lipogenesis (P < .05). Thus, by including the covariate, the two breeds had similar (P > .10) rates of lipogenesis at the end of the trial. When adjusted for BCS nested within breed, Brahman had greater (P < .05) LPL activity than Angus.
Adaptation, Physiological↗
A K+ channel-blocking peptide from venom of Chinese scorpion Buthus martensii Karsch.
AIM: To purify and characterize a potassium channel blocker (BmP-3) from the venom of Chinese scorpion Buthus martensii Karsch. METHODS: 1. Purification was carried out by gel-filtration, cation-exchange, and reversed-phase chromatographies. N-terminal was directly sequenced by double-coupling manual method. Molecular weight was determined on an electrospray ionization mass spectrometer. Amino acid composition was analyzed after acidic hydrolysis for 20 h in HCl 6 mol.L-1 at 110 degrees C. 2. Toxicity tests were conducted in mice and cockroaches. 3. The inhibitory effects of BmP-3 on K+ channels were tested in acutely dissociated rat hippocampal pyramidal neurons using whole-cell patch-clamp configuration. RESULTS: 1. A pure peptide (BmP-3, 8.1 mg) was obtained, about 0.08% of total proteins of the venom. The N-terminal sequences were VGCEE and the molecular weight was 2938 in ESI-mass spectra. 2. No death occurred at the dosage of 200 micrograms in mice and 8 micrograms in cockroaches. 3. The peptide at 10 mumol.L-1 reduced the peak outward K+ currents by 63% +/- 4% in vitro. CONCLUSION: BmP-3 inhibited K+ channels.
Amino Acid Sequence↗
The effect of N-acetylglucosamine as a substrate for in vitro synthesis of glycosaminoglycans by human peritoneal mesothelial cells and fibroblasts.
The effect of N-acetylglucosamine (NAG) on in vitro synthesis of glycosaminoglycans by human peritoneal mesothelial cells and fibroblasts was studied. In contrast to isosmotic concentrations of glucose, NAG increases the synthesis of hyaluronan by mesothelial cells and fibroblasts in a dose-dependent manner. This effect of NAG can be demonstrated in the presence of increased glucose levels in a medium, or in a medium mixed with effluent dialysate obtained from continuous ambulatory peritoneal dialysis (CAPD) patients. Glucose inhibits synthesis of sulphated glycosaminoglycans by peritoneal mesothelial cells and fibroblasts, whereas NAG stimulates their production. Our results demonstrate that NAG is an effective stimulator of the in vitro glycosaminoglycans synthesis by human peritoneal mesothelial cells and fibroblasts.
Acetylglucosamine↗
[The application of low tidal volume pressure-controlled ventilation in patients with acute respiratory distress syndrome].
Low tidal volume pressure-controlled ventilation therapy was used in 32 patients with ARDS. There were resulted a lower peak inspiratory pressure (PIP) level than that of control group, thus a decline in incidence of overventilation and the overventilation induced mortality. The results suggest that low tidal volume pressure-controlled ventilation may consider as preferred ventilation mortality in management of ARDS.
Adolescent↗