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The modification of behavior in response to experience is crucial for animals to adapt to environmental changes. Although factors such as neuropeptides and hormones are known to function in the switch between alternative behavioral states, the mechanisms by which these factors transduce, store, retrieve, and integrate environmental signals to regulate behavior are poorly understood. The rate of locomotion of the nematode Caenorhabditis elegans depends on both current and past food availability. Specifically, C. elegans slows its locomotion when it encounters food, and animals in a food-deprived state slow even more than animals in a well-fed state. The slowing responses of well-fed and food-deprived animals in the presence of food represent distinct behavioral states, as they are controlled by different sets of genes, neurotransmitters, and neurons. Here we describe an evolutionarily conserved C. elegans protein, VPS-50, that is required for animals to assume the well-fed behavioral state. Both VPS-50 and its murine homolog mVPS50 are expressed in neurons, are associated with synaptic and dense-core vesicles, and control vesicle acidification and hence synaptic function, likely through regulation of the assembly of the V-ATPase complex. We propose that dense-core vesicle acidification controlled by the evolutionarily conserved protein VPS-50/mVPS50 affects behavioral state by modulating neuropeptide levels and presynaptic neuronal function in both C. elegans and mammals., National Institutes of Health (U.S.) (Grant GM024663)

Quels sont les enjeux politiques, culturels et économiques auxquels fait face l'industrie du doublage au Canada? Peut-on parler d'acculturation du Canada par les États-Unis sans parler d'acculturation du Canada francophone par la France? Quel contrôle le Canada exerce-t-il?, What are the political, cultural and economic challenges facing the Canadian dubbing industry? Is it possible to discuss cultural assimilation of Canada by the USA without discussing that of French Canada by France? How much control does Canada have?

Human neutrophils have been shown recently to express both the beta and the gamma chains of the interleukin-2 receptor (IL-2R). IL-15, a cytokine that has recently been cloned and characterized, was found to share many of the biological functions of IL-2 and is known to mediate signals through IL-2R beta and IL-2R gamma. In recent studies, we observed that IL-2 exerts few effects on various neutrophil functions, but information on IL-15-neutrophil interactions is lacking. In this study, we observed that IL-15, in contrast to IL-2, induces important morphological cell shape changes that are typical of activated neutrophils. Furthermore, phagocytosis of opsonized sheep red blood cells was significantly increased by IL-15 but not by IL-2. However, similar to IL-2, IL-15 did not modulate the oxidative burst response. Furthermore, we observed that de novo RNA synthesis is increased in neutrophils by IL-15 along with de novo protein synthesis, whereas no significant effect of IL-2 was noted. Among the different proteins that were found to be upregulated by IL-15, one was identified by microsequencing as the cytoskeletal protein actin. Finally, we found that IL-15 delays apoptosis of neutrophils more efficiently than IL-2 when evaluated by both microscopic observations and flow cytometry procedures. Furthermore, this phenomenon was dose-dependent (10 to 500 ng/mL), and, at 500 ng/mL, IL-15 delayed apoptosis as strongly as granulocyte-macrophage colony-stimulating factor. This study is the first to show that IL-15 is a significant neutrophil agonist. Moreover, in view of the differential effects of IL-15 and IL-2 on this cell type, our results support the existence of a specific IL-15R component(s) on human neutrophils.

Neutrophils and macrophages represent the first line of defense against microbial invaders. However, the role of phagocytes in host response to viral infection is poorly understood. We have previously shown that Epstein-Barr virus (EBV) interacts with human monocytes and modulates cytokine production in this cell type, but its effects on neutrophils are still unknown. In the present study, we investigated the presence of EBV receptor (CR2 or CD21) on neutrophils by cytofluorometry using five different anti-CD21 monoclonal antibodies (MoAbs), as well as fluoroscein isothiocyanate-EBV (FITC-EBV). Whereas no significant amount of neutrophils reacted with anti-CD21 MoAbs, studies with FITC-EBV indicated that viral particles bind to 30% of cells (in some individuals, EBV binds to more than 50% of neutrophils). This interaction is specific as it was completely inhibited by nonconjugated virus or with labeled virus preincubated with neutralizing MoAbs. After EBV treatment, cellular aggregation was observed in neutrophil cultures, an indication that neutrophils were activated. Although EBV did not induce respiratory burst activity in neutrophils, pretreatment with infectious particles enhanced (priming effect) the fMLP-induced O2-release in neutrophils. Instead of restricting our analysis to specific cytokine genes, we investigated the effects of EBV on neutrophil transcriptional events in general. The effect of this virus on de novo synthesis of total cellular RNA was first investigated by measuring the incorporation of [5–3H] uridine into total RNA. The results showed that RNA synthesis in neutrophils was significantly increased (2.3- to 21.3-fold) by EBV compared with the unstimulated controls. Live and UV-inactivated virus markedly induced RNA synthesis, whereas heat-inactivated virus lost this ability. Induction of RNA transcription was EBV specific, as an EBV-neutralizing antiserum abolished this effect. Induction of protein synthesis was also studied by measuring the incorporation of [35S] methionine and [35S] cysteine into secreted and intracellular proteins in neutrophils incubated with EBV. The synthesis of both secreted and cytoplasmic proteins was induced by EBV. One- and two-dimensional gel electrophoresis analysis showed that EBV modulates protein synthesis, because activation of the synthesis of certain proteins was accompanied by the inhibition of others. Interleukin-1 beta (IL-1 beta) and IL-1 receptor antagonist (IL-1Ra) synthesis was found to be induced by EBV. Therefore, modulation of host-response proteins such as IL-1Ra could be one of the many mechanisms by which this virus avoids rejection.(ABSTRACT TRUNCATED AT 400 WORDS)

At inflammatory sites, neutrophils are stimulated by a range of proinflammatory molecules which elicit a number of cellular responses. Considerable information on the cytoplasmic events that occur following activation of neutrophils at the cell membrane level already exists. In this study, we have focused on the ability of neutrophil agonists to initiate nuclear signaling events by investigating the induction of de novo RNA synthesis. Of a total of 14 different known potent leukocyte agonists, only three had a significant effect on the induction of RNA synthesis in neutrophils; the formylated oligopeptide formyl-methionyl-leucylphenylalanine (fMet-Leu-Phe), granulocyte-macrophage colony-stimulating factor, and tumor necrosis factor alpha. All three agonists induced de novo RNA synthesis in neutrophils at concentrations known to be optimal for the activation of a number of other cellular responses occurring in inflammation. Of significance was the observation that activation of RNA synthesis in neutrophils is a G-protein-mediated event, is also dependent on tyrosine phosphorylation, but is not influenced by cAMP. Finally, we have demonstrated that all three agonists also induce de novo synthesis of a limited number of proteins, with granulocyte-macrophage colony-stimulating factor and fMet-Leu-Phe having the most potent effect. These studies define the effects of neutrophil agonists on de novo RNA and protein synthesis in a proinflammatory context and suggest that these events in neutrophils occur in a restricted fashion, highly dependent on the stimuli present at sites of inflammation.

We tested a wide range of pro-inflammatory cytokines for their capacity to activate protein synthesis in neutrophils as analyzed b y [35S] methionine metabolic labelling experiments. Of all the cytokines tested, only GM-CSF and TNF alpha stimulated significant synthesis and secretion of a 23 kD protein which resolved into two bands on two dimensional gels. Under non-reducing conditions on one dimensional gels, its migration pattern remained the same indicating that the two bands most likely represent isoforms of the same protein. Immunoisolation studies using antibodies directed against size-relevant molecules did not lead to the identification of this molecule. The fact that this 23 kD molecule is induced in a highly specific and selective manner by GM-CSF and TNF alpha indicates that it may play a key role in some of the responses of neutrophils to these two cytokines. Therefore, full characterization of this 23 kD protein could provide important new knowledge on the mechanisms by which these two cytokines exert their biological effects on neutrophils.

The primary objective of this study was to yield normal range values, with a current perception threshold technique, that could be used to assess the functionality of the third division of the trigeminal nerve on a healthy population. Moreover, we wanted to evaluate the impact of gender and training on these values.Standardized current perception threshold (CPT) measures using constant alternating current sinusoid waveform stimulus at 5, 250, and 2,000 HZ were obtained from 50 healthy patients at the mental foramen area bilaterally using a Neurometer current perception threshold device (Neurotron Inc, Baltimore, MD) (2,000 Hz specifically stimulates Abeta fibers, 250 Hz Adelta fibers, and 5 Hz stimulates C fibers).The mean CPT values for the 2,000, 250, and 5 Hz groups were respectively, 157.6 +/- 54.67, 53.10 +/- 27.64, and 33.44 +/- 23.17 mAmp. These values were used to construct a CPT scale to classify patients in the hyperesthetic, normative, or hypoesthetic range. There were no significant differences when comparing CPT values in men and women except in the 2,000 Hz group (P.02; n = 23). In addition, the test was carried out a first time on the right side and a second time on the left. This training procedure showed a significant decrease in the CPT values in men at 2,000 Hz (P.01; n = 23) for the second measure.The Neurometer can be beneficial for the evaluation and intraneural localization of sensory dysfunctions associated with the third division of the trigeminal nerve. It also could be used for initial diagnosis and subsequent evaluation of the patient's neurologic status through the course of their condition.

Acetylsalicylic acid has long been the only nonsteroidal anti-inflammatory drug recommended for the treatment and prevention of thromboembolic diseases. More recently, new compounds have been used in patients with vascular diseases. However, these drugs are often associated with longer bleeding times and greater operative risk. In most surgical specialties, the question always arises as to whether antiplatelet therapy should be stopped before elective surgery. If so, for how long? If not, what are the risks? This article reviews the various antiplatelet drugs in use today, focusing on their mode of action, their effects on platelet function and the associated operative risks. It also proposes an algorithm for decision making in this setting, based on the literature and an understanding of the mechanisms of action of this class of drugs.

In our study of the modulation of the expression of inflammation-related genes in neutrophils, we have found a gene called CLECSF6 (C-type lectin superfamily 6). CLECSF6 expresses two mRNA species at low levels in resting neutrophils. Here, we describe for the first time the sequence of the short mRNA version. It lacks amino acids that are likely to affect the functionality of its protein product. GM-CSF, IL-3, IL-4, and IL-13 caused an accumulation of the short CLECSF6 mRNA in neutrophils. The surface expression of the CLECSF6 protein was reduced by TNF-α, IL-1α, LPS, and Matrigel®. CLECSF6 bears the immunoreceptor tyrosine-based inhibition motif (ITIM) involved in signal transduction resulting in the inhibition of leukocyte activation. We propose that some neutrophil activators modulate the expression of CLECSF6 at the mRNA (GM-CSF, IL-3, IL-4, and IL-13) or protein (TNF-α, IL-1α, LPS, and Matrigel®) levels in ways that block ITIM-based transduction of anti-inflammatory signals and therefore promote inflammation.

Background The submental route for endotracheal intubation has been proposed as an alternative to tracheotomy in the surgical management of patients with maxillofacial trauma. The purpose of this study was to review our experience with this procedure. Methods Medical records of 25 patients who had surgical reduction of midfacial or panfacial fractures while securing their airway with submental intubation were reviewed. After standard orotracheal intubation, a passage was created by blunt dissection with a hemostat clamp through the floor of the mouth in the submental area. The proximal end of the orotracheal tube was pulled through the submental incision. Surgery was completed with minimal interference from the endotracheal tube. At the end of surgery, the tube was pulled back to the usual oral route. Results Mean duration of surgery was 7.9 hours (range, 2-16 hours). Mean duration of postoperative mechanical ventilation was 5.2 days (range, 1-24 days). Fourteen of these patients required prolonged (>24 hours) postoperative mechanical ventilation because of associated injuries. Two patients later required a tracheotomy because of prolonged respiratory failure. One patient died of multiple organ failure. One complication of the submental intubation was observed: a superficial infection of the submental wound. Conclusion Submental intubation is a simple technique associated with a low morbidity. It is an attractive alternative to tracheotomy in the surgical management of selected cases of maxillofacial trauma.

Differential display-polymerase chain reaction (DD-PCR) was used to evaluate changes in mRNA expression of granulocyte-macrophage colony-stimulating factor (GM-CSF) treated human neutrophils to better understand how this cytokine affects the functions of neutrophils at the molecular level. Although a variety of cDNA fragments were identified as modulated by GM-CSF with the use of DD-PCR, one fragment in particular, NGS-17 (neutrophil GM-CSF-stimulated fragment #17), was characterized. The NGS-17 fragment hybridized to a 3.8-kb mRNA that encodes for a protein of a predicted molecular mass of 47.6 kDa. After cloning and sequencing, this gene was found to code for the recently sequenced tapasin or TAP-A protein. Immunoprecipitation and immunoblotting studies using anti-tapasin antibodies showed that tapasin is expressed in neutrophils and is associated with the MHC class I-TAP complex. Moreover, tapasin expression was found to be induced by dimethyl sulfoxide and by retinoic acid in HL-60 cells. This is the first report on the expression of tapasin in human neutrophils. It provides novel information, at the molecular level, on how GM-CSF enhances the functions of these cells. J. Leukoc. Biol. 65: 205–210; 1999.

Interleukin-4 (IL-4) and IL-13 are cytokines that share many biological activities. We have previously demonstrated that IL-13 affects a number of neutrophil responses, and here we extend our observations to IL-4. We present, for the first time, direct evidence for the presence of functional IL-4 receptors on human neutrophils. We report that IL-4 induces RNA synthesis in a concentration-dependent manner and, based on observations of the induction of morphological cell shape changes and spreading onto glass, we demonstrate that IL-4 activates neutrophil cytoskeletal rearrangements. We further show that IL-4 is a potent activator of de novo protein synthesis in neutrophils, and we identify by microsequencing one of these proteins as the cytoskeletal protein actin. We were also able to demonstrate for the first time that actin is cleaved into at least two fragments of approximately 30 kDa (pI 5.4) and approximately 25 kDa (pI 5.0) in neutrophils. Finally, we report that IL-4 delays neutrophil apoptosis, as assessed by morphological observations from cytocentrifuge preparations, as well as by measurement of differences in staining by flow cytometry with both propidium iodide and Hoechst reagent. Taken together, we conclude that IL-4 is a more potent neutrophil agonist than previously believed. We discuss the possibility that the induction of the de novo synthesis of actin by IL-4 is related to the mechanism by which this cytokine delays apoptosis; in addition, the cleavage of this protein is likely to contribute to the apoptotic process.

It was recently shown that interleukin-13 (IL-13) induces the expression and release of the IL-1 decoy receptor (type II) in human neutrophils along with the production of the IL-1 receptor antagonist. In the present study we focused on further studying the modulatory effects of IL-13 on these cells. We found that recombinant human IL-13 (rhIL-13) induces cellular morphological changes in neutrophils that are typical of activated cells. Furthermore, rhIL-13 was shown to increase tyrosine phosphorylation of several neutrophil proteins. We also demonstrate that this cytokine stimulates de novo RNA synthesis in a concentration-dependent fashion as measured by [5-3H]uridine incorporation and that rhIL-13 induces the synthesis of several neutrophil proteins according to high-resolution two-dimensional gel electrophoresis of metabolically [35S]-labeled cells. We observed that the IL-8 levels in the external milieu of IL-13-stimulated cells was almost fivefold increased when compared with control cells. Finally, we observed that rhIL-13 has no significant effect on phagocytosis and apoptosis. Taken together, our results demonstrate that IL-13 is a modulator of several human neutrophil functions. This leads us to conclude that the modulatory role of IL-13 on human neutrophils, a potentially important anti-inflammatory cytokine, is more complex than previously believed. Furthermore, because we show that the synthesis of several as yet unidentified proteins was up-regulated by IL-13, our findings open new avenues of research on the effects of this cytokine on human neutrophils.

1. 1. Ram seminal plasma α-glucosidase has been purified in order to increase our knowledge of this enzyme and of its role in epididymal physiology. 2. 2. Since the enzyme behaved differently from other known acid α-glucosidases and was not affinity-adsorbed on dextran gels, another approach had to be used. 3. 3. The final procedure included an ethanol precipitation step, sequential chromatography on hydroxylapatite and DEAE Sepharose CL-6B, isoelectric focusing in polyacrylamide gels and ultrafiltration. 4. 4. The resulting purification factor of α-glucosidase was 9822 with an overall yield of 5%. 5. 5. The purified material consisted of several isoforms with a mol. wt of 105,000 and isoelectric points varying between 4 and 5.

Canine prostate fluids and seminal plasma contain a major androgen-dependent protein which was identified as a proteolytic enzyme exhibiting an Arg-esterase activity. This protease, as characterized, is shown to be present as a two-chain structure held together by at least one disulfide bridge and composed of approximately 220 amino acids. Amino acid sequence determination of both chains has revealed a clear homology to other known amino acid sequences of serine proteases. Furthermore, the comparison of the presented 58 amino acids of the Arg-esterase with the other sequences revealed a very strong homology (larger than 50%) to members of the kallikrein family. The two chain structure could thus result from autolysis of a single chain enzyme in the ‘kallikrein autolysis loop’. Amino acid composition of the canine prostatic enzyme suggests that it is related, but not identical, to pancreatic canine kallikrein.

Acid alpha-glucosidase and L-carnitine (a well-known epididymal marker) were measured in rete testis and epididymal fluids of adult male rams. These fluids were collected by selective catheterization or by a micropuncture technique, respectively. Both parameters remained at a low and constant level in rete testis and all along caput and corpus epididymidis. Then they increased at equivalent rates in cauda epididymidis to much higher levels than those in seminal plasma (5 mU/mg protein and 10 mM, respectively). An optimum pH study of alpha-glucosidase activity in these fluids showed two well-separated peaks in rete testis and caput epididymal fluids around pH 4 and 7, respectively, but only a single peak at pH 4 in cauda epididymidis that was comparable to the one in seminal plasma. Sucrose density gradient fractions analyzed for their enzyme content in the absence or presence of sodium dodecyl sulfate (1% w/v), a selective inhibitor of acid alpha-glucosidase activity, allowed the demonstration of two enzyme forms at pH 6.8 in rete testis fluid sedimenting in the 7S and 4S regions of the gradient, while a unique 4S form was encountered in cauda epididymidis and in seminal plasma. Although the fate of the minor 7S component of the rete testis fluid in its epididymal transit is presently unknown, similarities between the enzyme in cauda epididymidis and seminal plasma are strong enough to support the hypothesis that epididymis contributes primarily to the acid alpha-glucosidase content of ram seminal plasma.

FRANCIS James Child mentions a Veronese broadside of 1629 as textual evidence of the existence of 'Lord Randal'" (No.12) in Italy as early as the 17th century.2 The ballad, he says, is also found in several other European countries, among which France is not mentioned. And yet, on the eastern coast of Canada, formerly called Acadie, a French 'Lord Randal', classed as 'Le testament du garqon empoisonnd' in Conrad Laforte's Catalogue3 and at the National Museum of Man in Ottawa, has been repeatedly collected. A comparative study of versions of those two ballads will shed light on the nature and extent of contacts between the francophone and the anglophone communities in Eastern Canada, as well as on the role of translation in the formation of the traditional repertoire. Child publishes fifteen versions of 'Lord Randal,' which he identifies with letters A to O. These versions come from manuscript or printed transcriptions, and were collected in Scotland, in Ireland, in Suffolk, England, and in New England in the United States. Because of variations in the narrative structure, only four of Child's versions concern us here: versions A, B, C, and D, all from Scotland. According to Child, they are 19th century texts, transcribed either at the beginning or in the middle of the last century. Child selects 'Lord Randal' as a critical title even though the hero bears various other names, because, he says, 'that name occurs in one of the better versions, and because it has become famous through Scott's Minstrelsy.'4 'Lord Randal' is widely known in North America, where many versions were collected, in particular in the Maritime Provinces of Canada and in New England.5 Helen Creighton publishes two versions and a fragmentary text from Nova Scotia.6 Phillips Barry prints fifteen versions in his British Ballads from Maine. One of his texts, version H entitled 'Lord Ronald my Son,' was sent to him from New Brunswick by a lady who had learnt this song from her mother in Scotland. The informant of Barry's L version quitted New Brunswick and settled in Maine before 1870. Version G was collected from an old lady who learned it in her youth in Ireland. Helen Hartness Flanders also publishes nineteen versions or fragments of 'Lord Randal' collected in New England.8 Some of her versions proceed from Ireland and Scotland, while one informant acknowledges having learnt her version (B) from her mother, born in Abercorn, Quebec, in 1853. French versions of 'Lord Randal' were collected on the Atlantic coast of Canada, despite the fact that this song is unknown in France. In French, the

In addition to other known markers of the human prostate, it was shown that the prostatic fraction of the split ejaculate was rich in a 16-kDa protein with properties not described previously. This protein was purified from human seminal plasma using ammonium sulfate precipitation, DEAE-Sepharose CL-6B ion exchange chromatography, and gel filtration on Sephadex G-100. The purified protein showed a single prominent spot on two-dimensional gel electrophoresis. The sequence of the first 40 amino acids that could be positively identified was identical to that of a prostatic secretory protein of 94 amino acids (PSP94) previously designated as β-inhibin. Antibodies produced in rabbits against the purified protein were used to develop a radioimmunoassay. These antibodies appeared to recognize only the NH2-terminal portion of the native molecule since they did not react with a synthetic peptide composed of the 28 C-terminal residues. The radioimmunoassay showed that the concentration of the protein was 1320 ± 183 μg/ml in the seminal plasma of adult fertile men and 1134 ± 136 μg/ml in vasectomized patients. In hypertrophic and adenocarcinomatous prostates, the concentrations were 326 ± 156 and 104 ± 23 μg/ml, respectively, while values were lower than 0.060 μg/ml in the testis, epididymis, vas deferens and liver. The blood plasma concentration was 0.019 ± 0.002 μg/ml in 23 asymptomatic men 45 to 65 years old and 0.115 ± 0.036 μg/ml in eight patients with prostate cancer. These results show that PSP94 is a major protein in prostatic secretions that warrants further study in the monitoring of prostate cancer patients.