Miracle-dz/UniFAR-FTUs huggingface dataset:
Characterization of synaptic vesicles and related neuronal features in nerve growth factor and ras oncogene differentiated PC12 cells

PC12 cells can differentiate into neuron‐like cells after treatment with either nerve growth factor (NGF) or transduction with a retrovirus which expresses the K‐ras oncogene. The concomitant treatment of NGF plus ras differentiates PC12 cells further than either agent alone with respect to neurite outgrowth, acetylcholinesterase levels, and most strikingly, the number of synaptic vesicle (SV) clusters. These SV clusters in PC12 cell neurites closely resemble those in the presynaptic terminals of neurons. Such SV clusters have not been described in cell lines previously. The SV clusters from all three differentiated groups (NGF, ras, and NGF plus ras) were similar in size, shape, and configuration, except that the ones in the doubly treated group occur in higher frequency and have more vesicles. The synaptic nature of these vesicle clusters was demonstrated by their regulated depletion after potassium stimulation. Furthermore, these vesicle clusters stained positively for two SV‐associated proteins, synapsin I and synaptophysin, by EM immunocytochemistry (ICC). Such SV clusters in a cell line are very useful for characterizing the regulated release of SVs and the distribution of SV‐related antigens in intact cells. Analysis by SDS‐gel electrophoresis and immunoblotting indicated that synapsin I levels are higher in all three differentiated groups compared to untreated cells; whereas synaptophysin levels are lower in cells exposed to NGF alone or with NGF and ras double treatment. Possible convergence and/or divergence on the mechanisms of NGF and ras differentiation in PC12 cells are discussed. © 1995 Wiley‐Liss, Inc. This article is a US Government work and, as such, is in the public domain in the United States of America.
Effects of Lidocaine on the Anesthetic Requirements for Nitrous Oxide and Halothane

The effects of various plasma concentrations of lidocaine on nitrous oxide anesthesia in man and halothane requirements in the dog were studied. The response to incision of the skin was observed in 20 patients who were anesthetized with nitrous oxide, 70 per cent inspired, and oxygen, 30 per cent, plus various plasma levels of lidocaine. In addition, changes in the MAC of halothane in dogs were observed at various levels of lidocaine. In both circumstances lidocaine concentrations of 3 to 6 Mg/ml decreased anesthetic requirements approximately 10 to 28 per cent. At clinically common concentrations of lidocaine, significant decreases in anesthetic requirements should be anticipated
Fluoroquinolone ( cipro ̄ oxacin ) secretion by human intestinal epithelial ( Caco-2 ) cells

1 Human intestinal epithelial Caco-2 cells were used to investigate the mechanistic basis of transepithelial secretion of the ̄uoroquinolone antibiotic cipro ̄oxacin. 2 Net secretion and cellular uptake of cipro ̄oxacin (at 0.1 mM) were not subject to competitive inhibition by sulphate, thiosulphate, oxalate, succinate and para-amino hippurate, probenecid (10 mM), taurocholate (100 mM) or bromosulphophthalein (100 mM). Similarly tetraethylammonium and N'methylnicotinamide (10 mM) were without eect. 3 Net secretion of cipro ̄oxacin was inhibited by the organic exchange inhibitor 4,4'-diisothiocyanostilbene-2-2'-disulphonic acid (DIDS, 400 mM). 4 Net secretion of cipro ̄oxacin was partially inhibited by 100 mM verapamil, whilst net secretion of the P-glycoprotein substrate vinblastine was totally abolished under these conditions. Cipro ̄oxacin secretion was unaltered after preincubation of cells with two anti-P-glycoprotein antibodies (UIC2 and MRK16), which both signi®cantly reduced secretory vinblastine ̄ux (measured in the same cell batch). Cipro ̄oxacin (3 mM) failed to inhibit vinblastine net secretion in Caco-2 epithelia, and was not itself secreted by the P-glycoprotein expressing and vinblastine secreting dog kidney cell line, MDCK. 5 Net secretion and cellular uptake of cipro ̄oxacin (at 0.1 mM) were not subject to alterations of either cytosolic or medium pH, or dependent on the presence of medium Na, Cl or K in the bathing media. 6 The substrate speci®city of the cipro ̄oxacin secretory transport in Caco-2 epithelia is distinct from both the renal organic anion and cation transport. A role for P-glycoprotein in cipro ̄oxacin secretion may also be excluded. A novel transport mechanism, sensitive to both DIDS and verapamil mediates secretion of cipro ̄oxacin by human intestinal Caco-2 epithelia.
Role of the nanos homolog during sea urchin development

The nanos genes play important roles in the development of primordial germ cells in animal species. In the sea urchin, Hemicentrotus pulcherrimus, small micromere descendants specifically express HpNanos mRNA and this expression continues in the left coelomic pouch, which produces the major component of the adult rudiment. In this study, we showed that morpholino knockdown of HpNanos resulted in a delay of primary mesenchyme cell ingression and a decrease in the number of cells comprising the left coelomic pouch. Knockdown analysis in chimeras and whole embryos revealed the disappearance of small micromere descendants from the archenteron tip. Furthermore, the expression of HpNanos mRNA was induced in other cell lineages in the HpNanos‐knockdown and micromere‐deleted embryos. Taken together, our results suggest that HpNanos is involved in the inductive interaction of small micromere descendants with other cell lineages, and that HpNanos is required for the survival of small micromere descendants. Developmental Dynamics 238:2511–2521, 2009. © 2009 Wiley‐Liss, Inc.
Inhibition of the Mitochondrial KATP Channel by Long-chain Acyl-CoA Esters and Activation by Guanine Nucleotides*

The mitochondrial KATP channel (mitoKATP) is highly sensitive to ATP, which inhibits K+ flux with K1/2 values of 20-40 μM. This raises the question, how can mitoKATP be opened in the presence of physiological concentrations of ATP? We measured K+ flux in liposomes reconstituted with purified mitoKATP and found that guanine nucleotides are potent activators of this channel. ATP-inhibited K+ flux was completely reactivated by both GTP (K1/2 = 7 μM) and GDP (K1/2 = 140 μM). These ligands had no effect in the absence of ATP. The K1/2 for ATP inhibition exhibited quadratic dependence on [GTP] and [GDP], consistent with two binding sites for guanine nucleotides. We also found that palmitoyl-CoA and oleoyl-CoA inhibited K+ flux through reconstituted mitoKATP with K1/2 values of 260 nM and 80 nM, respectively. This inhibition was reversed by GTP (K1/2 = 232 μM) as well as by the K+ channel openers cromakalim (20 μM) and diazoxide (10 μM). Inhibition of mitoKATP by long-chain acyl-CoA esters, like that of ATP, exhibited an absolute requirement for Mg2+ ions. We propose that the open-closed state of the mitochondrial KATP channel is determined by the relative cytosolic concentrations of GTP and long-chain acyl-CoA esters.
The Role of Adding Hyaluronidase to Fluoroscopically Guided Caudal Steroid and Hypertonic Saline Injection in Patients with Failed Back Surgery Syndrome: A Prospective, Double‐Blinded, Randomized Study

Background: Failed back surgery syndrome (FBSS) has been reported to account for up to 40% of patients with chronic low back pain. Epidural fibrosis may be responsible for up to 30% of all cases of FBSS. Perineural fibrosis can interfere with cerebrospinal fluid‐mediated nutrition, rendering the nerve root hyperesthetic and hypersensitive to compression. Traditionally, steroid injection has been used to treat low back pain and radiculopathy. The addition of hyaluronidase to the injectate has been reported to reduce the degree of fibrosis.
A novel missense mutation in the GTPase activating protein homology region of TSC2 in two large families with tuberous sclerosis complex

Tuberous sclerosis complex (TSC) is an autosomal dominant disorder (OMIM 191092) characterised by autism, seizures, mental retardation, benign tumours of the brain, heart, kidney, lung, and skin, and malignant tumours of the kidney.1 TSC has a wide range of phenotypic variability, with some subjects severely affected and others only mildly affected. There are two TSC genes, TSC1 on chromosome 9q342 and TSC2 on chromosome 16p13.3Approximately two thirds of cases of TSC appear to result from de novo germline mutations. In 1994, an extended four generation family with 19 affected members was reported in which 34 members (17 affected with TSC and 17 unaffected) underwent both physical and psychiatric assessments.4 The majority of the affected subjects had mild physical expression of TSC, but there was significant clustering of neuropsychiatric disorders among affected subjects compared with their unaffected relatives. The disorders that were over-represented included mood disorder, anxiety disorder, and autism. The largest difference was observed in anxiety disorder, which was seen in 10 of the affected subjects and in two of the unaffected subjects (p=0.016). One affected child had pervasive developmental disorder and one had autism. Analysis of this family suggested that TSC could present phenotypically with mild physical signs and symptoms, but with significant neuropsychiatric disease. Linkage to the TSC2 gene locus on chromosome 16p13.3 was shown with a lod score of over 3.4 We report here the identification of a missense mutation in exon 34 of the TSC2 gene in affected members of this family. We also examined a second four generation family5from the same geographical area as the …
Optimizing Coding and Reimbursement to Improve Management of Alzheimer's Disease and Related Dementias

The objectives of this study were to review the diagnostic, International Classification of Disease, 9th Revision, Clinical Modification (ICD‐9‐CM), diagnosis related groups (DRGs), and common procedural terminology (CPT) coding and reimbursement issues (including Medicare Part B reimbursement for physicians) encountered in caring for patients with Alzheimer's disease and related dementias (ADRD); to review the implications of these policies for the long‐term clinical management of the patient with ADRD; and to provide recommendations for promoting appropriate recognition and reimbursement for clinical services provided to ADRD patients.
Evidence-based practice: a critical appraisal.

Welcome to this special issue on evidence-based practice and occupational therapy. It has been a privilege to be the guest editor and to learn about evidence-based practice in the Netherlands (Ammeraal and Coppers, 2012; Döpp et al., 2012), Norway (Aas et al., 2012), the Republic of Ireland (Stronge and Cahill, 2012) and the United States (Winkle et al., 2012). In this editorial, I will draw out some themes from the five papers and offer a personal perspective on evidence-based occupational therapy. Evidence-based practice is framed as an evolving, contested concept with many achievements and some notable challenges ahead. Some theoretical frameworks, references and online resources are signposted to help the reader explore the nuances of evidence-based practice. In 2007, evidence-based medicine (EBM) was voted as one of the 15 greatest medical advances since 1840 (BMJ 2007). The success is unsurprising given that evidence-based practice is about clinical efficacy. This focus is stated in two simple questions: Does it work? And is it cost effective? These questions are pertinent to everyone with a stake in health care throughout the world. This includes policy makers responsible for funding healthcare systems; organisations that provide health care; and multi-disciplinary teams and individual practitioners incorporating research results into clinical practice and, most importantly, to patients who want ethical, efficacious treatment. The term EBM first appeared in the medical literature in the early 1990s when it was presented as a fundamentally new approach to the teaching and practice of medicine (Dawes et al., 2005). This was because EBM promotes the examination of evidence from clinical research and requires new skills, especially critical appraisal. Such critique challenges custom and practice, making it legitimate to question authority or eminence-based practice (Isaacs and Fitzgerald, 1999). A critical attitude towards one’s own practice and to the evidence is, in my opinion, one of the greatest achievements of the evidence-based movement. Critical
Consumption of Microwaved Food induces Oxidative Stress in the Liver and Brain of Sprague-Dawley Rats

Introduction: Microwaves violently vibrate the water molecules in food, thus creating internal friction. The radiation, created by microwaves results in the destruction and deformation of food molecules, plus the formation of new radiolytic compounds. Methods: In our present study, the susceptibility of microwave radiations on food fed to rats is studied by measuring antioxidant enzymes catalase, reduced glutathione and superoxide dismutase as well as measuring the level of TBARS (Thiobarbituric Acid Reacting Substances). Results: It was observed from the result that the activities of antioxidant enzymes decreased drastically (p<0.05) in rats fed the microwaved food as compared to their controls. Conclusion: Microwave radiation affects the oxidative state of the liver and brain macromolecules. Damage caused by free radicals is aggravated by a reduction in protective antioxidant enzymes caused by the MW interactions in biological tissues leads to production of ROS that eventually affect the status of the measured physiological parameters.
Quantitative dual‐probe microdialysis: mathematical model and analysis

Steady‐state microdialysis is a widely used technique to monitor the concentration changes and distributions of substances in tissues. To obtain more information about brain tissue properties from microdialysis, a dual‐probe approach was applied to infuse and sample the radiotracer, [3H]mannitol, simultaneously both in agar gel and in the rat striatum. Because the molecules released by one probe and collected by the other must diffuse through the interstitial space, the concentration profile exhibits dynamic behavior that permits the assessment of the diffusion characteristics in the brain extracellular space and the clearance characteristics. In this paper a mathematical model for dual‐probe microdialysis was developed to study brain interstitial diffusion and clearance processes. Theoretical expressions for the spatial distribution of the infused tracer in the brain extracellular space and the temporal concentration at the probe outlet were derived. A fitting program was developed using the simplex algorithm, which finds local minima of the standard deviations between experiments and theory by adjusting the relevant parameters. The theoretical curves accurately fitted the experimental data and generated realistic diffusion parameters, implying that the mathematical model is capable of predicting the interstitial diffusion behavior of [3H]mannitol and that it will be a valuable quantitative tool in dual‐probe microdialysis.
An evolutionary neuro-fuzzy approach to recognize on-line Arabic handwriting

The author describes a system that recognizes on-line Arabic cursive handwriting. In this system, a genetic algorithm is used to select the best combination of characters recognized by a fuzzy neural network. The handwritten words used in this system are modelled by a theory of movement generation. Based on this motor theory, the features extracted from each character are the neuro-physiological and biomechanical parameters of the equation describing the curvilinear velocity of the script. The evolutionary approach proposed permits the recognition of cursive handwriting with a segmentation procedure allowing overlapped strokes having neuro-physiological meaning.
The Structure of the Catalytic Domain of a Plant Cellulose Synthase and Its Assembly into Dimers[C][W][OPEN]

Small-angle x-ray scattering gives a glimpse at the solution structure of the catalytic domains of plant cellulose synthase and their dimerization. Dimerization through the plant-specific sequences of the catalytic domain provides important clues for how Zn-finger domains couple these fundamental scaffold units into large, multimeric synthase complexes. Cellulose microfibrils are para-crystalline arrays of several dozen linear (1→4)-β-d-glucan chains synthesized at the surface of the cell membrane by large, multimeric complexes of synthase proteins. Recombinant catalytic domains of rice (Oryza sativa) CesA8 cellulose synthase form dimers reversibly as the fundamental scaffold units of architecture in the synthase complex. Specificity of binding to UDP and UDP-Glc indicates a properly folded protein, and binding kinetics indicate that each monomer independently synthesizes single glucan chains of cellulose, i.e., two chains per dimer pair. In contrast to structure modeling predictions, solution x-ray scattering studies demonstrate that the monomer is a two-domain, elongated structure, with the smaller domain coupling two monomers into a dimer. The catalytic core of the monomer is accommodated only near its center, with the plant-specific sequences occupying the small domain and an extension distal to the catalytic domain. This configuration is in stark contrast to the domain organization obtained in predicted structures of plant CesA. The arrangement of the catalytic domain within the CesA monomer and dimer provides a foundation for constructing structural models of the synthase complex and defining the relationship between the rosette structure and the cellulose microfibrils they synthesize.
Quantitative determination of bacterial replication in vivo

A new methodology which permits the quantitative measurement of absolute bacterial replication in vivo is proposed. Mice were inoculated with mixtures of temperature-sensitive mutants and parental wild types, and the changes in the ratios of the two strains were measured. The number of wild-type generations was calculated from the declining ratios over time with the formula n = log (r0/rt)/log 2; n is the number of generations, and r0 and rt are the ratio of temperature-sensitive mutants to the parental wild type at time zero and at the times sampled throughout the experiment. The replication rate was determined by regression analysis. A mathematical argument for the formula is presented. Using this technique, we determined the mean generation times of Escherichia coli (33 min) and Pseudomonas aeruginosa (20 min) in the peritoneal cavities of mice, in the face of host clearance mechanisms during the first stages of infection.
Maternal mortality, United States and Canada, 1982-1997.

BACKGROUND The 1998 public awareness campaign on Safe Motherhood called attention to the issue of maternal mortality worldwide. This paper focuses upon maternal mortality trends in the United States and Canada, and examines differentials in maternal mortality in the United States by maternal characteristics. METHODS Data from the vital statistics systems of the United States and Canada were used in the analysis. Both systems identify maternal deaths using the definition of the World Health Organization's International Classification of Diseases. Numbers of deaths, maternal mortality rates, and confidence intervals for the rates are shown in the paper. RESULTS Maternal mortality declined for much of the century in both countries, but the rates have not changed substantially between 1982 and 1997. In this period the maternal mortality levels were lower in Canada than in the United States. Maternal mortality rates vary by maternal characteristics, especially maternal age and race. CONCLUSIONS Maternal mortality continues to be an issue in developed countries, such as the United States and Canada. Maternal mortality rates have been stable recently, despite evidence that many maternal deaths continue to be preventable. Additional investment is needed to realize further improvements in maternal mortality.
An acute decrease in TCA cycle intermediates does not affect aerobic energy delivery in contracting rat skeletal muscle

We tested the hypothesis that an acute decrease in muscle TCA cycle intermediates during contraction would compromise aerobic energy delivery. Male Wistar rats were anaesthetized and the gastrocnemius–plantaris–soleus (GPS) muscle complex from one leg was isolated and perfused with a red cell medium containing either saline (Con) or cycloserine (Cyclo; 0.05 mg g−1), an inhibitor of alanine aminotransferase (AAT). After 1 h of perfusion, the GPS muscle was either snap frozen (Con‐Rest, n= 11; Cyclo‐Rest, n= 9) or stimulated to contract for 10 min (1 Hz, 0.3 ms, 2 V) with blood flow fixed at 30 ml min−1 (100 g)−1 and then snap frozen (Con‐Stim, n= 10; Cyclo‐Stim, n= 10). Maximal AAT activity was > 80% lower (P < 0.001) in both Cyclo‐treated groups (Rest: 0.61 ± 0.02; Stim: 0.63 ± 0.01 mmol (kg wet wt)−1 min−1; mean ±s.e. m. ) compared to Con (Rest: 3.56 ± 0.16; Stim: 3.92 ± 0.29). The sum of five measured TCAI (ΣTCAI) was reduced by 23% in Cyclo‐Rest versus Con‐Rest but this was not different (P= 0.08). However, after 10 min of contraction, the ΣTCAI was 25% lower (P= 0.006) in Cyclo‐Stim compared to Con‐Stim (1.88 ± 0.15 versus 2.48 ± 0.11 mmol (kg dry wt)−1). Despite the acute decrease in TCAI after Cyclo treatment, the contraction‐induced changes in markers of non‐oxidative energy provision (phosphocreatine, ATP and lactate) and the decline in tension after 10 min of stimulation were similar compared to Con. These data do not support the hypothesis that the total muscle concentration of TCAI is causally linked to the rate of mitochondrial respiration during contraction.
The role of iron in restless legs syndrome

The impressive relief from restless legs syndrome (RLS) symptoms provided by levodopa treatment indicates RLS is caused by a dopaminergic abnormality. But similar and more lasting relief also occurs for iron treatment in some patients. Thus there are two major putative causes for RLS: CNS dopaminergic abnormality and CNS iron insufficiency. This article presents the data documenting that both peripheral and CNS iron insufficiency occur with RLS symptoms. Brain iron insufficiency is supported by independently replicated cerebrospinal fluid and brain imaging studies for patients without iron deficiency (ID) anemia. Autopsy studies and intravenous iron treatment further link brain iron insufficiency to RLS. The brain iron insufficiency in patients with RLS is now well established. In this article the data are reviewed that support the following postulates combining dopaminergic and iron causes of RLS: (1) All conditions that compromise iron availability will increase the risk of RLS leading to a higher than expected prevalence of RLS in these conditions. (2) Some patients with RLS have marginal CNS iron status that can become insufficient when deprived of normal access to adequate peripheral iron or may be insufficient even with normal access to adequate peripheral iron. (3) The change or reduced CNS iron status produces RLS symptoms largely through its effects on the dopaminergic system and the corollary to 3. (4) Dopaminergic system abnormalities producing RLS symptoms will be included in those produced by brain ID. Study of the iron model of RLS offers hope for developing new treatment approaches and perhaps methods to prevent or cure the disorder. © 2007 Movement Disorder Society