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And tnfalpha inhibit cortical neuron dendrite beats by dre sale online development Prenatal infection and risk for schizophrenia:Il1, il6, and tnf inhibit cortical neuron dendrite development John h gilmore1, 2, lars fredrik jarskog1, 2, swarooparani vadlamudi2 and jean m lauder3 1UNC Schizophrenia Research Center, Chapel Hill, NC, USA2Department of Psychiatry, Chapel Hill, NC, USA3Department of Cell and Developmental Biology University of North Carolina at Chapel Hill, Chapel Hill, NC, USACorrespondence:Jh gilmore, department of psychiatry, cb 7160, university of north carolina at chapel hill, chapel hill, nc 275997160, usa. Top of pageabstractprenatal exposUre to infection increases risk for schizophrenia, and we have hypothesized that inflammatory cytokines, generated in response to maternal infection, alter neUron development and increase risk for schizophrenia.Primary mixed neUronal cUltUres were obtained from e18 rats and exposed to 0, 100, or 1000 Units(U)/ml of IL1, TNF, IL6, or IL1+TNF for 44 h.Map2positive neurons were randomly identified for each condition and the number of primary dendrites, nodes, and total dendrite length was determined.We found that 100 u of tnf significantly reduced the number of nodes(27%, p=0.02)And total dendritic length(14%, p=0.04), but did not affect overall neuron survival.A measure of 100 u il1+tnf significantly reduced the number of primary dendrites(17%, p=0.006), nodes(32%, p=0.001),And total dendritic length(30%, pTop of pageINTRODUCTIONMaternal infection during pregnancy increases the risk of the offspring developing schizophrenia and other neurodevelopmental disorders.The weight of the evidence indicates that maternal influenza infection during pregnancy is associated with a higher incidence of schizophrenia in offspring(Reviewed in mcgrath and murray, 2003;Bagalkote et al, 2001).Other types of maternal infections have also been implicated, including pneumonia and diphtheria(Watson et al, 1984;O'callaghan et al, 1994), rubella(Brown et al, 2000b), measles, varicellazoster, and polio(Torrey et al, 1988;Suvisaari et al, 1999).Recent studies have moved beyond epidemiologic association with studies that link infections in individual mothers with schizophrenia in their adult children.Respiratory infections in the second trimester increase risk for schizophrenia(Brown et al, 2000a).In this same cohort, serologic evidence of maternal exposure to influenza also increased the risk of schizophrenia in offspring(Brown et al, 2004a).Finally, the offspring of mothers with elevated igg and igm levels, and antibodies to herpes simplex virus type 2 during pregnancy, have an increased risk for schizophrenia(Buka et al, 2001a). The pathological mechanisms responsible for increased risk of schizophrenia in offspring after maternal infection remain largely unstudied.Hypotheses about the role of infection in the etiology of schizophrenia have focused on direct infection of the developing fetus(Yolken and torrey, 1995)Or the generation of antibodies that crossreact with neuronal antigens(Wright et al, 1993).As a variety of infections are associated with increased risk of schizophrenia, a feature common to all infections would be a likely candidate mechanism.We have hypothesized that cytokines generated in response to maternal infection alter early brain development and increase risk for schizophrenia(Gilmore and jarskog, 1997).Cytokines, including interleukin1(Il1), interleukin6(Il6), and tumor necrosis factor(Tnf), have a variety of effects on the central nervous system(Cns)And are expressed by glial and neuronal elements within the cns(Bartfai and schultzberg, 1993;Hopkins and rothwell, 1995;Rothwell and hopkins, 1995).Cytokines, including il1, il6, and tnf, regulate normal brain development and have been implicated in abnormal brain development(Merrill, 1992;Mehler and kessler, 1994;Mehler and kessler, 1997).Expression of cytokine mrna in the cns is developmentally regulated in mouse, rat, sheep, and human brain(Burns et al, 1993;Gadient and otten, 1994;Pousset, 1994;Mousa et al, 1999;Dziegielewska et al, 2000), an indication of the important role that cytokines play in neurodevelopment. Il1, il6, and tnf are elevated in the maternal unit after maternal infection in human pregnancies(Hillier et al, 1993;Fortunato et al, 1996;Yoon et al, 2003)And in animal models(Fidel et al, 1994;Urakubo et al, 2001).Maternally generated cytokines cross the placenta and regulate cell growth and development in the fetus(Medlock et al, 1993;Letterio et al, 1994;Stallmach and karolyi, 1994;Li et al, 1995;Mcduffie et al, 2001).An additional source of cytokines in prenatal infection may be the placenta, as the human placenta synthesizes il1, il6, and tnf in response to infection(Fortunato et al, 1996;Taniguchi et al, 1991;Menon et al, 1995).Finally, the fetus itself can mount an inflammatory response, especially of il6, in the face of maternal infection(Gomez et al, 1998;Yoon et al, 2003).Il1, il6, and tnf cross the blood barrier in mature rodents(Banks et al, 1991;Guiterrez et al, 1993;Banks et al, 1994).Finally, the blood barrier is incomplete in the fetus(Adinolfi, 1985), making it very likely that systemically generated cytokines gain entry into the fetal brain. Several recent studies support the hypothesis that cytokines play a key role in the association between maternal infection, altered brain development, and risk for schizophrenia.Maternal blood levels of tnf(Buka et al, 2001b)And il8(Brown et al, 2004b)Are elevated in pregnancies in which the offspring goes on to develop schizophrenia.Three animal models of maternal infection have recently been advanced as models of schizophrenia that also support our hypothesis.Maternal infection with human influenza virus in mice causes abnormalities in prepulse inhibition(Shi et al, 2003), and maternal exposure to Escherichia coli cell wall endotoxin, lipopolysaccharide(Lps), disrupts sensorimotor gating in the offspring(Borrell et al, 2002).Finally, maternal exposure to polyi:C, a synthetic doublestranded rna that stimulates a cytokine response, causes prepulse inhibition abnormalities(Shi et al, 2003)And disrupted latent inhibition(Zuckerman and weiner, 2003;Zuckerman et al, 2003).In the influenza model, no virus is detected in the fetal brain(Shi et al, 2003), suggesting that the immune responses, especially cytokines, are the likely mediators of the abnormal brain development that leads to longterm behavioral changes.In the polyi:C and lps models, it is likely that the immune response to the challenge plays a major role in the mechanism of action, as no infectious agent is present.We have shown that maternal lps exposure increases cytokine expression in the placenta and amniotic fluid of rats(Urakubo et al, 2001). Cytokines can be neurotoxic to developing neurons, as they decrease survival of serotonergic and dopaminergic neurons(Jarskog et al, 1997), hippocampal neurons(Araujo and cotman, 1995), and cortical neurons(Gelbard et al, 1993;Jeohn et al, 1998;Marx et al, 2001).The neuropathology of cerebral cortex in schizophrenia is subtle and does not appear to involve neuron loss, but rather loss of neuropil(Selemon et al, 2003), dendrites and spines(Glantz and lewis, 2000;Broadbelt et al, 2002), and synaptic markers(Glantz and lewis, 1997).The actions of cytokines on developmental processes that may give rise to schizophrenia, such as dendrite development, are less well known.This study was conducted to determine the effect of cytokines on cortical neuron dendrite development.We hypothesized that the inflammatory cytokines il1, il6, and tnf, which are generated in response to maternal infection, would decrease the development of dendrites on embryonic cortical neurons in vitro. Top of pagematerials and methodscell cUltUreprimary cortical neUronal cells were isolated as described previoUsly(Marx et al, 2001).Briefly, frontal cerebral cortex was dissected from whole brains of embryonic day 18 spragUe rats(Charles river, raleigh, nc)And the meninges removed.The trimmed tissUes were dissociated in 0.125% trypsin and 0.5 mM EDTA for 20 min at 37 The reaction was terminated by adding an eqUal volUme of DUlbecco's modified Eagle's mediUm+10% fetal bovine serUm+5 Units(U)/ml penicillin/streptomycin.The cells were further dissociated by mechanical trituration, followed by filtering through the nylon mesh and centrifugation.The cells(5 104/ml/well)Were plated on 18mm glass coverslips coated with polydlysine(100 g/ml)In 12well culture plates.Cells were cultured in neurobasal medium supplemented with b27(Invitrogen, carlsbad, ca), 0.5 mM Lglutamine, 25 M Lglutamate and 5 U/ml penicillin/streptomycin.After 4 h, media was exchanged and cultures were treated with tnf, il6(R systems, minneapolis, mn), IL1(Boehringer mannheim, ridgefield, ct), or TNF+IL1 for 44 h at concentrations of 0, 100, or 1000 U/ml, three wells per condition.Specific activity per manufacturer:Tnf, 60 u/ng;Il6, 12.5 beats by dre online sale U/ng;And il1, 50 u/ng.Three cultures from three different litters were studied for a total n=9 per treatment condition. Immunohistochemistryafter 44 h of cytokine exposure, cultures were rinsed in hank's balanced salt solution, fixed in 4% paraformaldehyde, and permeabilized with 0.2%.Triton x100 in phosphatebuffered saline(Pbs)And incubated overnight with goat polyclonal antibody map2(1:100).To confirm that neurons in culture expressed the respective cytokine receptors, three cover glasses each were incubated overnight with goat polyclonal antibodies for tnf receptor1(1:100)And tnf receptor2(1:100), or rabbit polyclonal IL1 receptor1(1:500)Or il6 receptor(1:200).Map2 and cytokine receptor antibodies were purchased from santa cruz(Santa cruz, ca).To determine if glial cells were present, cultures were incubated overnight with monoclonal mouse antigfap(Boehringer mannheim, 1:500).Cultures were then processed using the avidin peroxidase complex(Abc)Method(Vector).Following immunochemistry and osmium tetraoxide intensification, cultures were rinsed in pbs and counterstained with toluidine blue.A negative control experiment was also performed to rule out nonspecific staining in the absence of primary antibodies.In this experiment, abc immunochemistry procedures were followed, but no primary antibody was included. Cell counting and dendrite morphologyprimary cortical neurons were cultured at low density(5 104 cells/ml/well)To allow morphological analysis of single neurons.Neuronal survival was analyzed by counting neuritebearing map2immunostained cells with clear neuronal morphology at 200 magnification using an ocular grid.In all, 12 grid areas were examined per coverslip using a blinded, randomized procedure(0.25 mm2 per grid area 12 per coverslip=total sample area of 3.00 mm2 per coverslip).Cell counts were converted to percentages of intraexperimental controls. For morphometry, the first 10 single neurons that had two or more dendrites encountered from the center of the coverslip were digitally captured at 60.Dendrites of chpbts scanned cells were traced using neurolucida and neuroexplorer software(Micro bright field, williston, vt).Morphometry data from individual neurons were collected for a number of primary dendrites per neuron, branch points per neuron, and total dendrite length per neuron(Figure 1).Primary dendrites were defined as neurites originating at the neuronal somata.A total of 90 neurons were studied for each treatment condition. Figure 1. ImmunoDeteCtion of reCeptors for il1, il6, AnD tnf on emBryoniC CortiCAl neurons in vitro using AntiBoDies to(A)Il1r,(B)Il6r,(C)Tnfr1, anD(D)Tnfr2(60 magnification). Full figure and legend(161K) Data analysisttests and oneway analysis of variance(Anova)With post hoc dunnett's multiple comparison tests were performed.Significance was set at p Top of pageresultscortical neurons in culture expressed receptors for il1, il6, and tnf(Types 1 and 2);Figure 1).There were no gfappositive cells present in the cultures.Representative map2positive neurons and their dendrite tracings are presented in figure 2.At a concentration of 100 U, tnf significantly reduced the number of nodes(27%, p=0.02)And total dendrite length(14%, p=0.04;Table 1)Of map2labeled cortical neurons in culture.Il1+tnf(100 U)Significantly reduced the number of primary dendrites(17%, p=0.006), nodes(32%, p=0.001),And total dendrite length(30%, pTable 1).There were no significant effects of il1 or il6 on the number of primary dendrites, nodes, or total dendrite length at this concentration. At 100 U, there were no significant effects of IL1, IL6, TNF, orIl1+tnfon neuron survival (P>0.05 for each cytokine). Figure 2. Representative map2labeled neurons and their dendrite tracings from neurolucida: (A and b)Control; (C and d)100 U of IL1+TNF(60 magnification).Tnf also significantly reduced the number of primary dendrites(24%, pppppp+TNF significantly reduced the number of primary dendrites(17%, ppp(14%, pp(17%;P+tnf(22%, pFigure 4). Figure 3. Morphometric analysis of cortical neurons in primary culture treated with 1000 u of il1, il6, tnf, or il1+tnf for 44 h.Each cytokine significantly decreased primary dendrite number, number of nodes, and total dendrite length.Results of anova are presented with each experiment.Post hoc dunnett's test:Pp


Posted: 06:06, 28/3/2014
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Cryptococcus louboutin pas cher breaks ristianlbtinaussurespaser out of the opportunistic box James w.Kronstad, won hee jung, nature reviews microbiology 9, 193203(March 2011)The ability of cryptococcus neoformans and cryptococcus gattii to influence the intracellular environment of macrophages during fungal proliferation, expulsion and transfer between cells is probably dependent on exported fungal factors.Secretion is clearly necessary for cell surface delivery of known virulence christian louboutin paris soldes factors such as the polysaccharide capsule, the enzyme laccase(Which synthesizes melanin)And the enzymes phospholipase b and urease.Many of these factors are delivered, at least in part, by membranebound, extracellular vesicles that have been observed to traverse the cell wall80, 81, 82(See the figure;Dotted arrows indicate potential, but unproven, steps in the pathway).These socalled 'virulence factor delivery bags' may be released by the fusion of multivesicular bodies with the plasma membrane thus suggesting a role for golgi trafficking and/or by blebbing from the plasma membrane(Capturing cytosolic material to form vesicles)11, 15, 81, 82.Although little is known about the intracellular trafficking functions that package virulence factors into vesicles for export, there is some evidence both for a role for exocytosis in the export of internallysynthesized capsule polysaccharide, and for regulation of secretion by the cyclic amp signalling pathway15, 83, 84, 85, 86.The extracellular vesicles are produced by c.Neoformans during infection of the mouse macrophagelike cell line j774.16, and similar vesicles that contain capsule polysaccharide accumulate in the cytoplasm of infected macrophages in vivo and in vitro43, 81.These phagocytic cells show permeabilization of the phagosomal membrane, and it is tempting to speculate that cryptococcal vesicles and/or exported enzymes(For example, phospholipase b)Play a role in this modification29, 43, 44.The extracellular vesicles from the supernatants of fungal cultures are internalized by macrophagelike cells(The mouse cell line raw 264.7), and these vesicles also induce a dosedependent production of nitric oxide, tumour necrosis factor, interleukin10 and transforming growth factor by the mouse cells87.


Posted: 06:02, 28/3/2014
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The creativity of john updike The last time i saw john updike, he was seated at charlie rose massive round table;Impeccably dressed in a tan suit, starched white shirt and a speckled salmoncolored tie.The last epistle would be his last. Updike died on january 27th from the ravages of lung cancer.Updike and andrew iaelorstasen wyeth, my two most consistent muses, passed away 11 days apart and i am orphaned again.There is something about the possibility of the next that has been a bond to my literary and artistic heroes.I still tenaciously cling to the belief mark twain alive and well. This past october, updike talked with sam tanenhaus of the new york times about the craft of his writing.Don think of myself as writing stylishly, i think of myself as trying to write with precision, what my mind eye conjures up.So if out of this the sentences become shapely and vivid, that great, but i mostly concerned with trying to deliver to the reader, my images, and my sense of human behavior and landscape.Find that the main charge, let call it, that i get out of writing, is when i feel that i gotten something down accurately.The main bliss, whether i read henry green or nabokov or proust or tolstoy, is the sense that they described precisely a certain moment of experience, whether it a dress, a chair or how a person face looks. Much like wyeth, updike had the ability to breath life into the seemingly mundane.A trip to the blood bank.An underused swimming pool.A man walking his wife friend home while snow michael kors taschen online begins to fall. There is no clearer example, than his autobiographical musings selfconsciousness.You are there and not there. Even mark twain best friend, william dean howell said, were always there for him, but he wasn always there for you.It was less a chide than an observation that twain was somewhere else in his mind.This is about living a dual life, liver and observer.Updike describes those minute and intimate details that make up the fabric of the extraordinary that lies hidden in the ordinary. Since i am an uncle many times over, i will share one of my favorite updike essays, my uncle death. Died while shaving;When i was told of this, i pictured him staggering back heavily, stricken, his own amazed face in the mirror the last thing he ever saw.His face flashed there for him, hung there, slipped backward;And then the mirror was full of the blank bathroom wall.I pictured this so sharply i seemed to have been there. At his funeral, i felt, for the first time my adult height.The manatees are not a family of breeders, and the number of relatives was small;Walking up the aisle to the front pew with my parents, my aunt and my two cousins, i felt tall and prominent.Walking back down the aisle after the service, i caught from the faces of those still seated, an odd motionless, intent look, almost an odor of sympathy and curiosity and reverence for grief. Cannot reach him.I can remember nothing about him that is quite real except his death;He is like a celestial body which only an eclipse renders measurable.He was six feet, four inches tall, but his immensity was narrowshouldered, small boned and unmuscular.He was vain of having, for so outsized a man, rather small feet.He usually wore neat black loafers, virtually slippers, of english leather, and sprawling, soddenly in a chair, he generally contrived to thrust his feet forward on the floor, or up on a stool, so they were noticed.I can remember my mother i must have been ten or eleven teasing him about his dainty feet.I cannot recapture her words, but she was still slim then, and her pose as she spoke head tilted back, hands half lifted stuck in my my mind;She so seldom struck an unmotherly attitude that it was as if a strange spirit had come for a moment and possessed her body.My uncle, presumably, responded with a dry flutter of the sheepish gallantry that he seemed to reserve for my mother and waitresses in restaurants.My mother seemed exempt from the rather lazy distaste with which uncle viewed the rest of the world, and perhaps as her son, i was included in this exemption, for he was kind to me.Felt tall and prominent.Walking back down the aisle after the service, i caught from the faces of those still seated, an odd motionless, intent look, almost an odor of sympathy and curiosity and reverence for grief.While it is viewed from the prism of an older version of himself, he selfconsciously remembers that was included in this exemption, for he was kind to me.It is as much about updike as it is his uncle. My uncle carl funeral was the first funeral i ever experienced.I was about the same age as updike when his uncle died.I remember my uncle widow wailing.It is impossible to know another grief, but i do remember my mother whispering to my grandmother that it was bit much.I remember the chairs arranged at the grave site and how there was a kind of hierarchy of who sat where.And i remember the american flag folded tightly and given gently to my aunt. But i remember nearly every detail about the service and the postfuneral gathering at my uncle home.I recall the sublime taste of ginger ale that flowed freely.I saw walls filled with porcelain figurines he collected while he was stationed as a colonel in germany.But mostly, i remember that it was the first sibling death for my mother.Much like updike uncle, my mother and carl had an especially close relationship and even at 16, i understood this and it was difficult to gauge her own grief. Updike acute observation and his mingling of memory and detail continue to move me.He is often the catalyst to discovering what i missed in life by being equally self conscious.The creativity of john updike is about honoring his gift of observation and dancing with the alphabet. In updike honor and with much feeling, i am opening up a bottle of ginger ale and it will flow freely.I will miss him. 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Posted: 06:00, 28/3/2014
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Anchorage climber completes longtime alaska range goal He took care of a tooth that's been nagging him since 1993. Rowe, a 50yearold anchorage schoolteacher, scaled the alaska range's 9, 050foot broken tooth on the last day in may, becoming what is believed to be the first person to climb routes on all five of the major peaks in the mooses tooth group moose's tooth(10, 335 feet), Bear Tooth(10, 070), Eye Tooth(9, 000 feet), Sugar Tooth(8, 000)And broken tooth. Broken tooth was the first of the five nearly vertical peaks to draw rowe's attention, and for 20 years it teased, tantalized and taunted him. He tried climbing it for the first time in 1993.He failed.He tried four or five more times before putting broken tooth on hold to climb the other four peaks.Then he returned to broken tooth, again and again and again. The 12th time was the charm. Rowe and climbing partner peter haeussler recorded the fifth ascent of broken tooth last week, climbing on ice rapidly turning into waterfalls and surviving an avalanche that rowe called"The scariest thing that ever happened to me. " Each man took his turn on the corniced summit, with rowe relishing his quick moment in a place he worked so hard to reach. "I think broken tooth is the most strikingly beautiful mountain i've ever seen in my life,"He said. "I think it's the beauty of the peak that keeps bringing me back. "I've attempted it from every direction east, south, west, and i even went to buckskin glacier last year and attempted it from the north.I've climbed every side of that mountain, and i just feel sort of a kinship with it.I've certainly never tried to climb another mountain 12 times. " The rotten tooth Broken tooth's steepness makes it an elusive goal, as does the condition of its summit. "It's called broken tooth because the whole top has been eroded away, so there's a lot of rotten rock,"Rowe said. "That's the thing that has turned us back multiple times.The rock is so deteriorated you can't put any anchors into it and you can't climb it safely.It also doesn't have an easy route up it. " That's why broken tooth, which rises from coffee glacier in the central alaska range, is seldom climbed.And that's why rowe is believed to be the first person to scale broken, sugar, eye, bear and moose's tooth, said spokeswoman maureen mclaughlin of denali national park.Park rangers are unaware of any other climbers making the same claim. "(S)Ince the broken tooth has seen very few ascents, the chance that the full set has been climbed is very slim,"She said. Summits of sugar beats by dre sale cheap tooth are also rare when rowe and hauessler climbed it in 2007, theirs was just the second ascent of the peak.Rowe said he made four attempts in three years before he could cross off sugar tooth from his todo list. Erik rieger of the american alpine journal said in an email that he couldn't confirm if rowe's fivetooth climbing achievement is a first or not.Last year, climbers freddie wilkinson and renan ozturk became the first to traverse the mooses tooth massif's skyline in a single push, he said, but that traverse doesn't include broken tooth. Also, he wrote, there is the question of what exactly makes up the mooses tooth massif: "The broken tooth isn't exactly part of the main mooses tooth massif;If that's your criteria you'd have to take into account the wisdom tooth, the incisor, etc., which are more or less surrounding peaks. " Partner drdrestudio in climb Rowe might still have a tooth problem if not for haeussler, a climbing partner willing to rappel where he'd been repelled before. "This was the third attempt for me, and i woulda been perfectly happy to go somewhere else,"Haeussler said,"But he was so into it, so i said, ok, jay, we either need to get up this thing or we need to have an intervention or something. " Haeussler, an anchorage research geologist, has climbed moose's tooth, eye tooth, sugar tooth and now broken tooth.He did all but moose's tooth with rowe. "I haven't done bear tooth,"He said, and he has no plans to.He loves to climb, but there's no moby dick of mountains calling to him.


Posted: 07:58, 25/3/2014
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Council issues fines warning ristianlbtinpaserfemme as seagull complaints soar Business owners and staff said they were faced with running the gauntlet just to attend work.Ian lawson, of ian lawson kitchens in park hill road, said the birds were bombing people as they got in and out of cars. Elsie martlew, council member responsible for the environment, said they were seeing an problem with the christian louboutin pas cher gulls in carlisle. Determined to tackle it, but we need the public to support our efforts, she said. Realise that it will take a number of years to make a significant impact and property owners can help by acting on the advice given to their premises.This month, the council unveiled a new toughened sack to protect purple rubbish bags from the birds ripping them open in the search for food. The sack which would affect the majority of the remaining 5, 538 properties still on a weekly waste collection in carlisle, is undergoing public christian louboutin escarpin noir consultation with a final decision expected by the council in july. Whitehaven town centre is also overrun by seagulls flying vermin and no, they were not here before us they followed the easy food trail and leave excrement all over, with the attendent diseases that kids do not appreciate.


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Amazon drought and forest fire prediction system devised Researchers have devised a model to anticipate drought and forest fires in the amazon rainforest. The research, which used precipitation records dating back to 1970 and hotspots tracked by the moderate resolution imaging spectroradiometer(Modis)Aboard nasa satellites, finds a strong correlation between sea surface temperatures in the tropical north atlantic and subsequent drought in the western amazon.Drought in the amazon is increasingly associated with forest fires due to landclearing fires set by agricultural developers and cattle ranchers.Coupled with its existing satellitebased deforestation monitoring capabilities, the warning system could prove useful to brazil as it aims to reduce deforestation rates under its national climate action plan. The study, which is published in geophysical research letters, comes on the heels of another paper, published in the same issue, that reports an increase in droughts in the beats by dre uk amazon. That research led by jose marengo of brazil's national institute for space research(Inpe)Identifies the 2005 chpbtsbydreuk and 2010 droughts in the amazon as the worst on record.It finds that since the mid1970s droughts have increased in the amazon region, while the length of the typical dry season has expanded.The study says droughts are"Aggravated"When there was low rainfall in the previous wet season. The 2010 drought affected some 3 million hectares of forest, nearly 60 percent more than the 1.9 million square kilometers affected during the 2005 drought.Both events triggered severe drops in water levels for amazon tributaries, stranding communities and stalling river transportation.Fires and associated 'haze' increased markedly during both droughts. Scientists have warned for years that climate change could turn much of the amazon into a tinderbox.Other research suggests that by midcentury, much of the amazon could be at risk of dieoff due to climate changeinduced drought, the effects of which are worsened by deforestation, forest degradation, and fragmentation.


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Posted: 02:59, 19/3/2014
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Alpha production and inflammatory activity in crohn's patients with fistulizing disease Tnf 308a promoter polymorphism is associated with enhanced tnf production and inflammatory activity in drdrebtshdphones crohn's patients with fistulizing disease Segundo gonz md, phd1, luis rodrigo md2, jes mart bs3, antonio l md3, dolores fuentes md2, pilar ni md2, valle cadah md2, cristina saro md4, m angeles dieguez md3 and carlos l phd3received 12 june 2002;Accepted 30 october 2002. Top of pageabstractobjective:Tumor necrosis factor(Tnf)Plays a key role in the inflammatory response and pathogenesis of crohn's disease(Cd).Tnf 308a polymorphism within the tnf gene promoter has been associated with enhanced tnf production in vitro.The aim of this study was to investigate the effect of tnf promoter polymorphism at 308 on the susceptibility and phenotypic expression of fistulizing cd. Methods:The distribution of 308 tnf genotypes was analyzed in 50 patients with fistulizing cd and 100 healthy matched controls.Tnf, interleukin1, and interleukin6 serum levels were measured by elisa.Serum amyloida, creactive protein, 1antitrypsin, 1acid glycoprotein, and haptoglobin were beats by dre sale uk measured by nephelometry. Results:No significant differences were found in the allele frequencies of beats by dre outlet the polymorphism between patients and controls.However, compared with 308gg patients, those carrying 308ag had a significant increase of serum levels of tnf(58 79 vs 8 19 pg/ml, p(3645vs 16 20 pg/ml, p = 0.048), and acute phase proteins(Apps).308A carriers had also a higher frequency of arthritis(66% vs 26%, p = 0.039).The logistic regression model showed that the patients carrying 308a polymorphism had a relative risk for developing arthritis of 5.45(95% CI = 1.1 No other clinical or analytical findings were predictive for the risk of development of arthritis.


Posted: 01:46, 17/3/2014
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Posted: 01:45, 17/3/2014
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Alpha causes apoptosis in epidermal keratinocytes after subcutaneous injection of tnf Correspondence:Dr xianmin meng, department of dermatology and cutaneous biology, jefferson medical college, thomas jefferson university, 450 blsb, 233 south 10th street, philadelphia, pa 19107 Received 19 march 1999;Revised 18 june 1999;Accepted 12 http://www.teamskyuser.co.uk/beats-by-dr-dre-pro.html july 1999. To the drdre editor: Apoptosis is known as programmed cell death and has been recognized as an active regulator in the process of cell proliferation and differentiation.Some activators such as hormones p53 and uvb, can induce apoptosisIn vitro and in vivo(Mccall cohen 1991;Gniadecki et al.1997).Tumor necrosis factor(Tnf)Is an inflammatory cytokine, which is generally thought to have an antitumor effect.It has been demonstrated that tnf can also induce apoptosis in hepatocytes(Senaldi et al.1998), human keratinocytes(Reinartz et al.1996), and normal cytotrophoblasts(Garcia et al.1996)In vitro;However, there is a contrary report in which tnf is shown not to induce apoptosis in cultured human keratinocytes(Benassi et al.1997), and no report has ever demonstrated induction of apoptosis in TNF transgenic mice(Probert et al.1995;Crew et al.1998;Nakama et al.1998).We have previously reported that the human interleukin10(Meng et al.1998)And the human interleukin6(Sawamura et al.1998)Gene can be introduced into epidermal keratinocytes in vivo using the plasmid dna injection method.Here, we have examined the biologic effect of transgenic tnf by introducing the beats by dre outlet human tnf gene into epidermal keratinocytes using this method. First, we constructed a human tnF expression plasmid(PhtnF).The coding region oF human tnF cdna was ampliFied using polymerase chain reaction(Pcr)Based on its cdna sequence(Wang et al.1985).For subcloning, primers contained restriction enzyme sites at the 5'ends(F:5'AACTCGAGATGAGCACTGAAAGCATGATCCGG3', R:5'AAGCGGCCGCTCACAGGGCAATGATCCCAAAG3').Subcloning was performed using a pcy4b expression vector(Meng et al.1998).Plasmid containing the tnf cdna without any eukaryotic promoter, named phtnf(Was used as a negative control.Plasmid phtnf was intradermally injected into the dorsalSkin of hirosaki hairless rat(Hhr)At a concentration of 0.2 g per l(Meng et al.1998).To determine whether keratinocytes could express human tnf mrna after introduction of phtnf, reversetranscriptase(Rt)Pcr was performed using rna samples derived from keratinocytes in the treated area as previously described(Meng et al.1998).Pcr products were electrophoresed by a 2% agarose gel.Transgenic tnf in local keratinocytes was detected by elisa(Biosource international, camarillo, ca)And was expressed as nanograms per microgram of protein(Protein assay kit;Biorad, hercules, ca).Control samples were taken from the ptnf(Skin.Immunohistochemical staining for tnf was done using a polyclonal rabbit antihuman tnf antibody(Genzyme).Skin samples were taken 24 h aFter PhtnF injection and Frozen sections were used For the staining. To examine the biologic eFFect oF transgenic TNF in localSkin,Skin specimens were Fixed, paraFFinembedded, cut in 5 mm sections and stained with hematoxylin and eosin For routine histologic examination.To detect apoptotic cells in the epidermis, the terminal deoxynucleotidyl transferase(Tdt)Mediated dutpbiotin nick end labeling(Tunel)Technique was performed using an apoptosis detection kit(Apoptag, oncor, gaitheerburg, md). Our rEsults oF rtpCr showED thAt A 741 Bp BAnD wAs prEsEnt in thESAmplEs From phtnFtrAnsFErrED kErAtinoCytEs, whErEAs no BAnD wAs oBsErvED inSAmplEs From phtnF(KErAtinoCytEs(FigurE 1A).PCr with mrnA sAmplEs without A rt stEp wErE Also pErFormED AnD no tnF DnA BAnD wAs DEtECtED(DAtA not shown). REsults oF ELISA showED thAtInjECtion oF phTNF CAusED A signiFiCAnt inCrEAsE oF trAnsgEniC hTNF in loCAlKErAtinoCytEs(FigurE 1B). ImmunohistoChEmiCAl stAining gAvE A positivE stAining oFKErAtinoCytEsin phTNFInjECtED skin(FigurE 2A), whErEas a nEgativE rEsult was shown in phTNF(InjECtED skin(FigurE 2B). RoutinE stAining showED thAt somEKErAtinoCytEsAppEArED EosinophiliC in thE EpiDErmis (FigurE 2C). TUNELpositivE CElls wErE oBsErvED in thE EpiDErmis, inDiCAting thAt trAnsgEniC TNF inDuCED Apoptosis inKErAtinoCytEs(FigurE 2E).SAmplEs From phtnF(TrEAtED skin show nEgAtivE rEsults in Both thE routinE stAin AnD thE tunEl stAining(FigurE 2D, F). (A) TotAl RNA wAs ExtrACtED From rAtKErAtinoCytEsFrom thE gEnEtrAnsFErrED ArEA 24 h AFtEr phTNF or phTNF(InjECtion.RtpCr wAs pErFormED using primErs to AmpliFy tnF CDnA. A, phTNFInjECtion;B, phtnF(InjECtion;M, sizE mArkEr. (B)KErAtinoCytEswErE oBtAinED From thE gEnEtrAnsFErrED ArEA 24 h AFtEr phTNF or phTNF(InjECtion.TrAnsgEniC tnF lEvEls wErE AssAyED By ElisA.EACh vAluE shown rEprEsEnts thE mEAn sD oF six inDiviDuAl sAmplEs. TNF positivE stAining oFKErAtinoCytEswAs oBsErvED in phTNF trEAtED skin (A), whErEas a nEgativE rEsult was shown in phTNF(InjECtED skin(B). HEmAtoxylin AnD Eosin stAining showED thAt somEKErAtinoCytEsAppEArED EosinophiliC in thE EpiDErmis (C). TUNEL stAining showED TUNELpositivE CElls in thE EpiDErmis, suggEsting thAt trAnsgEniC TNF inDuCED Apoptosis inKErAtinoCytEs(E).SAmplEs From phtnF(TreateD skin showeD negative results both in the routine(D)And in the tunel staining(F).Scale bar:25 mm. Full figure and legend(170K) In this study, human tnf gene was introduced into rat skin in vivo by injection of its dna plasmid.The transferred htnf gene was expressed in local keratinocytes, and these keratinocytes produced transgenic htnf.Transgenic htnf was observed to have a local biologic effect in the epidermis where it induced apoptosis of keratinocytes.Our results demonstrated that apoptosis of epidermal keratinocytes could be induced by tnf plasmid in vivo. The molecular mechanisms of apoptosis are generally accepted to involve intracellular interactions induced by molecular signals.These signals, including fas ligand, tnf, and other antigens, can interact with receptors, including fas, tnf receptor, and t cell receptor, respectively.Messages from these receptors are transmitted to caspase.Some genes and proteins, such as cmyc, cfos, bcl2, and p53 are also involved.Finally, endonuclease is activated and this leads to dna fragmentation(Tartaglia et al.1993;Hale et al.1996;Tanuma1996).In this study, transgenic tnf was shown to bind tnf receptor in keratinocytes and induce apoptosis. As a reliable method of in vivo gene transfer, plasmid dna injection may be applied as keratinocyte gene therapy to treat certain skin diseases.In addition, this method can be useful for detection of biologic effects of certain genes in the skin in vivo.As for the possibility of clinical usefulness, keratinocyte gene therapy may be applied for some skin tumors by introducing tnf gene into tumor tissues by using the plasmid dna injection method.


Posted: 06:43, 13/3/2014
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Posted: 06:42, 13/3/2014
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Posted: 04:32, 6/3/2014
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