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Statistical mechanics of charged polymers in electrolyte solutions: a lattice field theory approach.

The lattice field theory approach to the statistical mechanics of a classical Coulomb gas [R.D. Coalson and A. Duncan, J. Chem. Phys. 97, 5653 (1992)] is generalized to include charged polymer chains. Saddle-point analysis is done on the functional integral representing the partition function of the full system. Mean-field level analysis requires extremization of a real-valued functional which possesses a single minimum, thus guaranteeing a unique solution. The full mean-field equations for such a coupled system are derived, as well as the leading (one-loop) fluctuation corrections. Two different numerical real-space lattice procedures are developed to implement the generalized theory; these are applied to the problem of a charged polymer confined to a spherical cavity in an electrolyte solution. The results provide insight into the physics of confined polyelectrolytes.

Journal Article↗

Bacterial binding protein-dependent permeases: characterization of distinctive signatures for functionally related integral cytoplasmic membrane proteins.

Bacterial binding protein-dependent transport systems belong to the superfamily of ABC transporters, which is widely distributed among living organisms. Their hydrophobic membrane proteins are the least characterized components. The primary structures of 61 integral membrane proteins from 35 uptake systems were compared in order to characterize a short conserved hydrophilic segment, with a consensus EAA---G---------I-LP, located approximately 100 residues from the C-terminus. Secondary structure predictions indicated that this conserved region might be formed by two amphipathic alpha-helices connected by a loop containing the invariant G residue. We classified the conserved motifs and found that membrane proteins from systems transporting structurally related substrates specifically display a greater number of identical residues in the conserved region. We determined a consensus for each class of membrane protein and showed that these can be considered as signatures.

ATP-Binding Cassette Transporters↗

Post-natal stem cells as participants in complex systems and the emergence of tissue integrity and function.

This issue of Pediatric Diabetes contains articles with scientific data, their interpretation, and a discussion of the respective field of stem cells and diabetes - its achievements and controversies. Through our daily laboratory activities, we employ a reductionist approach and often a deterministic interpretation of data. We would like to make this contribution a means of adding a somewhat different perspective to this collection of essays. This conceptual piece is merely intended to be an invitation to further thoughts and discussions.

Animals↗

Variations in the proportion of glycolytic/non-glycolytic energy substrates modulate sperm membrane integrity and function in diluted boar samples stored at 15-17 degrees C.

In this work the role of energy substrates in the maintenance of boar-sperm survival during storage at 15-17 degrees C was tested. For this purpose, boar spermatozoa were stored at 15-17 degrees C in several defined media with separate combinations of a monosaccharide, glucose and a non-monosaccharide, either citrate or lactate, energy substrates. Our results indicate that the medium containing the highest concentration of glucose together with low lactate levels was the most suitable to maintain sperm quality for 168 h at 15-17 degrees C. This was confirmed after observation of the results of the percentages of viability and altered acrosomes, the osmotic resistance test, the hyperosmotic resistance test and the rhythm of L-lactate production. The survival ability of boar sperm was greater in this experimental medium than in the standard Beltsville Thawing Solution extender, which contains only glucose as an energy substrate, although at a concentration far higher than that of all the tested experimental media. Our results indicate that the exact composition, more than the pure quantity of energy substrates, is a very important modulatory factor which affects survival ability of boar sperm in refrigeration. Thus, the exact combination of several energy substrates would have to be taken into account when optimizing the design of commercial extenders to store boar spermatozoa at 15-17 degrees C.

Acrosome↗

Signal attenuation as a function of integrator time constant and signal duration.

In some laboratories, eyeblink electromyogram is rectified and smoothed before the analog signal is digitized. The relationship among the integrator time constant, the duration of the response, and the attenuation of the data was investigated by passing a number of signals at durations of 5, 10, 20, 50, and 100 ms into a contour-following integrator (Coulbourn model S76-01), which first rectifies and then smooths (integrates) the signal. Integrator time constants of 10, 42, 70, 144, and 300 ms were used. The output of the integrator was sampled (1,000 Hz) and stored on a Macintosh SE computer. The integrator attenuated the signal, and the amount of attenuation increased as the duration of the signal decreased and as the time constant of the integrator increased. For brief inputs, the integrator does not charge fully, so only the early portion of the charge limb is present, and this early portion is the steepest part of the charge limb. Therefore, the longer the time constant, the less likely that small or brief responses will be detected.

Arousal↗

Ca2+ imaging of mouse neocortical interneurone dendrites: Ia-type K+ channels control action potential backpropagation.

GABAergic interneurones are essential in cortical processing, yet the functional properties of their dendrites are still poorly understood. In this first study, we combined two-photon calcium imaging with whole-cell recording and anatomical reconstructions to examine the calcium dynamics during action potential (AP) backpropagation in three types of V1 supragranular interneurones: parvalbumin-positive fast spikers (FS), calretinin-positive irregular spikers (IS), and adapting cells (AD). Somatically generated APs actively backpropagated into the dendritic tree and evoked instantaneous calcium accumulations. Although voltage-gated calcium channels were expressed throughout the dendritic arbor, calcium signals during backpropagation of both single APs and AP trains were restricted to proximal dendrites. This spatial control of AP backpropagation was mediated by Ia-type potassium currents and could be mitigated by by previous synaptic activity. Further, we observed supralinear summation of calcium signals in synaptically activated dendritic compartments. Together, these findings indicate that in interneurons, dendritic AP propagation is synaptically regulated. We propose that interneurones have a perisomatic and a distal dendritic functional compartment, with different integrative functions.

Action Potentials↗

Incorporation of protease K into larval insect membrane vesicles does not result in disruption of integrity or function of the pore-forming Bacillus thuringiensis delta-endotoxin.

Bacillus thuringiensis delta-endotoxins insert into the brush border membranes of insect larval cells to form ion channels. A possible interaction of these toxins with a cytoplasmic component was examined by preloading vesicles from insect larval cells with protease K followed by incubation with toxin. There was no evidence for toxin antigens smaller than the intact toxin in extracts of solubilized vesicles, nor was there an effect of the inclusion of protease K on either of two functional properties, the formation of toxin aggregates or of ion pores. These toxins, physically and functionally, appear to be confined to the membrane.

Animals↗

Intermediate filaments: molecular structure, assembly mechanism, and integration into functionally distinct intracellular Scaffolds.

The superfamily of intermediate filament (IF) proteins contains at least 65 distinct proteins in man, which all assemble into approximately 10 nm wide filaments and are principal structural elements both in the nucleus and the cytoplasm with essential scaffolding functions in metazoan cells. At present, we have only circumstantial evidence of how the highly divergent primary sequences of IF proteins lead to the formation of seemingly similar polymers and how this correlates with their function in individual cells and tissues. Point mutations in IF proteins, particularly in lamins, have been demonstrated to lead to severe, inheritable multi-systemic diseases, thus underlining their importance at several functional levels. Recent structural work has now begun to shed some light onto the complex fine tuning of structure and function in these fibrous, coiled coil forming multidomain proteins and their contribution to cellular physiology and gene regulation.

Amino Acid Sequence↗

Contractile function, sarcolemma integrity, and the loss of dystrophin after skeletal muscle eccentric contraction-induced injury.

The purpose of this study was to evaluate the integrity of the muscle membrane and its associated cytoskeleton after a contraction-induced injury. A single eccentric contraction was performed in vivo on the tibialis anterior (TA) of male Sprague-Dawley rats at 900 degrees /s throughout a 90 degrees -arc of motion. Maximal tetanic tension (Po) of the TAs was assessed immediately and at 3, 7, and 21 days after the injury. To evaluate sarcolemmal integrity, we used an Evans blue dye (EBD) assay, and to assess structural changes, we used immunofluorescent labeling with antibodies against contractile (myosin, actin), cytoskeletal (alpha-actinin, desmin, dystrophin, beta-spectrin), integral membrane (alpha- and beta-dystroglycan, sarcoglycan), and extracellular (laminin, fibronectin) proteins. Immediately after injury, P0 was significantly reduced to 4.23 +/- 0.22 N, compared with 8.24 +/- 1.34 N in noninjured controls, and EBD was detected intracellularly in 54 +/- 22% of fibers from the injured TA, compared with 0% in noninjured controls. We found a significant association between EBD-positive fibers and the loss of complete dystrophin labeling. The loss of dystrophin was notable because organization of other components of the subsarcolemmal cytoskeleton was affected minimally (beta-spectrin) or not at all (alpha- and beta-dystroglycan). Labeling with specific antibodies indicated that dystrophin's COOH terminus was selectively more affected than its rod domain. Twenty-one days after injury, contractile properties were normal, fibers did not contain EBD, and dystrophin organization and protein level returned to normal. These data indicate the selective vulnerability of dystrophin after a single eccentric contraction-induced injury and suggest a critical role of dystrophin in force transduction.

Animals↗

Removal of sarcolemmal sialic acid residues results in a loss of sarcolemmal functioning and integrity.

Treatment of neonatal rat heart cells with neuraminidase results in a large increase of cellular-associated Ca2+. The study described below was designed to test the hypothesis that neuraminidase produces its effects by increasing the transient Ca2+ channel current, as proposed by Yee et al. (24). This ICa,T can be inactivated by dodecylsulfate (DDS) (17). The experimental data show that 1) the increase in cellular Ca2+ during neuraminidase treatment cannot be explained by an increased ICa,T; 2) neuraminidase treatment has a much more profound effect on sarcolemmal permeability than has been recognized previously; and 3) the effect of neuraminidase treatment can be prevented by 50 microM DDS. The study indicates that glycocalyx-lipid bilayer interactions are important in maintenance of selective permeability of the sarcolemma. The protective effect of 50 microM DDS is probably mediated by insertion of the negatively charged amphiphilic molecule in the sarcolemma, although the exact mechanism remains to be elucidated.

Animals↗

Endogenous nitric oxide required for an integrative respiratory function in the cat brain.

1. The involvement of nitric oxide (NO) in the respiratory function of the pons was examined by microinjecting NO synthase-related drugs into discrete regions of the pontine respiratory group (PRG) in decerebrate and decerebellate cats. 2. Microinjection of N omega-nitro-L-arginine (L-NNA, inhibitor of NO synthase), but not D-NNA (the inactive enantiomer), significantly prolonged the duration of inspiration in all 10 cats when lung inflation was withheld. 3. The prolongation of inspiration produced by L-NNA was partially reversed in three cats by microinjections of L-arginine (NO synthase substrate) at the same sites. 4. We conclude that endogenous production of NO from L-arginine in the PRG region is involved in the normal function of the pontine pneumotaxic mechanism. These findings provide the first conclusive evidence that endogenous NO formation is involved in the mechanisms associated with respiratory rhythm generation.

Animals↗

Integrated cardiovascular function in the conscious streptozotocin-diabetic deoxycorticosterone-acetate-hypertensive rats.

Blood pressure, heart rate, and left ventricular function were measured in conscious diabetic Sprague-Dawley rats subjected to 5 weeks of deoxycorticosterone acetate (DOCA) treatment which was started 1 week following intravenous injection of streptozotocin (STZ) (60 mg/kg) to induce diabetes mellitus. It was found that chronic administration of DOCA in nondiabetic animals caused an increase in blood pressure and functional parameters of left ventricle, and a decrease in heart rate and plasma insulin levels. Normotensive diabetic rats exhibited hyperglycemia, hypoinsulinemia, and a lower body weight as compared with control animals but did not show significant abnormalities in cardiovascular function. DOCA-hypertensive STZ-diabetic rats had similar hyperglycemia, milder hypoinsulinemia, and a significantly lower rate of left ventricular relaxation and systolic blood pressure compared with the nondiabetic DOCA-hypertensive animals. It is concluded that the addition of DOCA hypertension to intact 6-week STZ-diabetic Sprague-Dawley rats results in the occurrence of cardiac dysfunction.

Animals↗

Mitochondrial integrity and function in atherogenesis.

BACKGROUND: Coronary atherosclerotic disease remains the leading cause of death in the Western world. Although the exact sequence of events in this process is controversial, reactive oxygen and nitrogen species (RS) likely play an important role in vascular cell dysfunction and atherogenesis. Oxidative damage to the mitochondrial genome with resultant mitochondrial dysfunction is an important consequence of increased intracellular RS. METHODS AND RESULTS: We examined the contribution of mitochondrial oxidant generation and DNA damage to the progression of atherosclerotic lesions in human arterial specimens and atherosclerosis-prone mice. Mitochondrial DNA damage not only correlated with the extent of atherosclerosis in human specimens and aortas from apolipoprotein E(-/-) mice but also preceded atherogenesis in young apolipoprotein E(-/-) mice. Apolipoprotein E(-/-) mice deficient in manganese superoxide dismutase, a mitochondrial antioxidant enzyme, exhibited early increases in mitochondrial DNA damage and a phenotype of accelerated atherogenesis at arterial branch points. CONCLUSIONS: Mitochondrial DNA damage may result from RS production in vascular tissues and may in turn be an early event in the initiation of atherosclerotic lesions.

Animals↗

Ultrasonic energy. Effects on vascular function and integrity.

BACKGROUND: Ultrasonic energy transmitted via flexible wire probes provides a new means of ablating atherosclerotic plaque. We studied the effects of ultrasonic energy (20 kHz) delivered via a ball-tipped wire probe on arterial vasomotor behavior in rabbit thoracic aortas in a perfused whole-vessel model. METHODS AND RESULTS: After precontraction with phenylephrine (10(-5) M) or KCl (60 mM), the effects of ultrasonic energy (0.7-5.5 W x 60 seconds, 42-330 J) on arterial vasomotor behavior were measured using long-axis ultrasonic vessel imaging of the proximal (ultrasonic probe-treated) and distal (untreated) control segments. The efficacy of plaque ablation at these same probe-tip power outputs was evaluated in atherosclerotic, human cadaver iliofemoral arteries. Ultrasonic energy caused dose (energy)-dependent relaxation in rabbit aortas after precontraction with phenylephrine in arteries with endothelium (n = 8) and without endothelium (n = 8) (p less than 0.001 versus ultrasound treated at power outputs of 2.9 and 5.5 W). There was no difference in the relaxation dose responses between endothelialized and endothelially denuded segments (p = NS). Ultrasonic energy also caused significant relaxation (67 +/- 8%) after voltage-dependent precontraction with 60 mM KCl. Temperature measurements revealed less than 1 degrees C warming of the vessel wall during as long as 2 minutes of treatment at a power output of 5.5 W. Pathological examination showed no smooth muscle injury at (moderate) power outputs that caused arterial relaxation. At probe-tip power outputs of 2.9-5.5 W, ultrasonic energy recanalized two of two totally occluded cadaveric iliofemoral vessel segments. The ultrasonic ablation catheter was also demonstrated to cause arterial relaxation in a recanalized canine femoral artery in vivo. CONCLUSIONS: Ultrasonic energy delivered via a flexible-wire probe produces dose-dependent, endothelium-independent smooth muscle relaxation capable of reversing both receptor-mediated and voltage-dependent vasoconstriction in vitro. At moderate power outputs, this relaxation response does not appear to be due to thermal effects or irreversible smooth muscle cell injury. This vasorelaxant effect of ultrasonic energy is also apparent in vivo, at doses that effectively ablate atherosclerotic plaque, and may improve the safety of arterial recanalization using ultrasonic energy.

Animals↗

Glutamine-supplemented parenteral nutrition improves gut mucosa integrity and function in endotoxemic rats.

The effects of glutamine-supplemented parenteral nutrition on protein metabolism, small intestinal mucosal metabolism, morphology, and barrier function were studied in endotoxin-treated rats. Forty-six male Wistar rats were randomized to two groups of 23 animals each and received total parenteral nutrition solutions supplemented with either glutamine (GLN group) or glycine (GLY group) at 2% wt/vol. Endotoxemia was induced by continuous intravenous infusion of endotoxin at a dose of 2 mg/kg per day throughout the 4-day study period. The GLN group had a less-negative cumulative nitrogen balance (-14.0 +/- 132.8 mg of nitrogen in the GLN group and -86.8 +/- 161.7 mg of nitrogen in the GLY group, p < .05) and less cumulative excretion of urinary 3-methylhistidine (2910 +/- 593 nmol) than the GLY group (4447 +/- 933 nmol, p < .01). Jejunal mucosal glutaminase activity and the arterio-portal venous blood glutamine concentration differences were significantly higher in the GLN group compared with the GLY group (15.6 +/- 2.3 vs 11.1 +/- 1.9 mumol/g per minute, p < .05, and 181 +/- 52 vs 147 +/- 36 nmol/mL, p < .05, respectively). The morphology of the jejunal mucosa in the GLN group was significant for having greater mucosal weight (23.4 +/- 3.1 vs 17.6 +/- 2.5 mg/cm), villus height (445 +/- 75 vs 357 +/- 57 microns), crypt depth (197 +/- 34 vs 161 +/- 28 microns), and wall thickness (751 +/- 77 vs 648 +/- 102 microns) than the GLY group (p < .05).(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗