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Cannabidiol (CBD) is a substance chemically derived from Cannabis sativa and discussed to be non-psychoactive. According to the FDA, marijuana is classified as a schedule I substance; however, hemp which is defined as extracts from marijuana including cannabinoids containing less than 0.3% tetrahydrocannabinol (THC), is excluded from that controlled substance act and available at local convenience stores in the US as it is seen as an herbal supplement. CBD is purported to be used for various medical and psychiatric conditions: depression, anxiety, post-traumatic stress disorder, Alzheimer's or other cognitive illnesses as well as pain. There is also a new trend to use CBD for the treatment of opioid use disorder. The one CBD product on the market that is FDA approved for the treatment of childhood epilepsy forms Dravet and Lennox-Gastaut syndromes is available under the name Epidiolex. There is a significant difference between this medication and the over-the-counter CBD products that contain very inconsistent strengths of CBD, if they contain it at all, and vary in percentage even from sample to sample. Frequently the so-called CBD products are not containing any CBD at all, but mostly containing THC. This article is a systematic review of literature reviewing the available clinical data on CBD, for use in various medical and psychiatric conditions with focus on a review of the pharmacology and toxicity. Resources used were ORVID, PubMed, MEDLINE, PsychINFO, EMBASE with keywords CBD, cannabidiol, hemp and cannabinoids.
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Marijuana is the most consumed illicit drug in the world, with over 192 million users. Due to the current legalization push of marijuana in the United States, there has been a lack of oversight regarding its public health policies, as marijuana advocates downplay the drug's negative effects. This paper's approach is from a public health perspective, focusing specifically on the cases of violence amongst some marijuana users. Here, we present 14 cases of violence with chronic marijuana users that highlight reoccurring consequences of: marijuana induced paranoia (exaggerated, unfounded distrust) and marijuana induced psychosis (radical personality change, loss of contact with reality). When individuals suffering from pre-existing medical conditions use marijuana in an attempt to alleviate their symptoms, ultimately this worsens their conditions over time. Although marijuana effects depend on the individual's endocannabinoid receptors (which control behavioral functions, like aggression) and the potency level of tetrahydrocannabinol (THC) in the drug, scientifically documented links between certain marijuana users and violence do exist. Wider public awareness of the risks and side effects of marijuana, as well as a more prudent health policy, and government agency monitoring of the drug's composition, creation, and distribution, are needed and recommended.
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Cannabis , Fumar Marihuana , Violencia , Política de Salud , Humanos , Política Pública , Estados UnidosRESUMEN
Understanding how metapopulations persist in dynamic working landscapes requires assessing the behaviors of key actors that change patches as well as intrinsic factors driving turnover. Coupled human and natural systems (CHANS) research uses a multidisciplinary approach to identify the key actors, processes, and feedbacks that drive metapopulation and landscape dynamics. We describe a framework for modeling metapopulations in CHANS that integrates ecological and social data by coupling stochastic patch occupancy models of metapopulation dynamics with agent-based models of land-use change. We then apply this framework to metapopulations of the threatened black rail (Laterallus jamaicensis) and widespread Virginia rail (Rallus limicola) that inhabit patchy, irrigation-fed wetlands in the rangelands of the California Sierra Nevada foothills. We collected data from five diverse sources (rail occupancy surveys, land-use change mapping, a survey of landowner decision making, climate and reservoir databases, and mosquito trapping and West Nile virus testing) and integrated them into an agent-based stochastic patch occupancy model. We used the model to (1) quantify the drivers of metapopulation dynamics, and the potential interactions and feedbacks among them; (2) test predictions of the behavior of metapopulations in dynamic working landscapes; and (3) evaluate the impact of three policy options on metapopulation persistence (irrigation district water cutbacks during drought, incentives for landowners to create wetlands, and incentives for landowners to protect wetlands). Complex metapopulation dynamics emerged when landscapes functioned as CHANS, highlighting the importance of integrating human activities and other ecological processes into metapopulation models. Rail metapopulations were strongly top-down regulated by precipitation, and the black rail's decade-long decline was caused by the combination of West Nile virus and drought. Theoretical predictions of the two metapopulations' responses to dynamic landscapes and incentive programs were complicated by heterogeneity in patch quality and CHANS couplings, respectively. Irrigation cutbacks during drought posed a serious extinction risk that neither incentive policy effectively ameliorated.
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Ecología , Modelos Biológicos , Animales , Aves , California , Ecosistema , Humanos , Dinámica PoblacionalRESUMEN
Cholesterol esterification in high density lipoproteins (HDLs) by lecithin:cholesterol acyltransferase (LCAT) promotes unesterified cholesterol (UC) transfer from red cell membranes to plasma in vitro. However, it does not explain the transfer of UC from most peripheral cells to interstitial fluid in vivo, as HDLs in afferent peripheral lymph are enriched in UC. Having already reported that the endogenous cholesterol esterification rate (ECER) in lymph is only 5% of that in plasma, we have now explored the underlying mechanism. In peripheral lymph from 20 healthy men, LCAT concentration, LCAT activity (assayed using an optimized substrate), and LCAT specific activity averaged, respectively, 11.8, 10.3, and 84.9% of plasma values. When recombinant human LCAT was added to lymph, the increments in enzyme activity were similar to those when LCAT was added to plasma. Addition of apolipoprotein AI (apo AI), fatty acid-free albumin, Intralipid, or the d < 1.006 g/ml plasma fraction had no effect on ECER. During incubation of lymph plus plasma, the ECER was similar to that observed with buffer plus plasma. When lymph was added to heat-inactivated plasma, the ECER was 11-fold greater than with lymph plus buffer. Addition of discoidal proteoliposomes of apo AI and phosphatidycholine (PC) to lymph increased ECER 10-fold, while addition of apo AI/PC/UC disks did so by only six-fold. We conclude that the low ECER in lymph is due to a property of the HDLs, seemingly substrate inhibition of LCAT by excess cell-derived UC. This is reversed when lymph enters plasma, consequent upon redistribution of UC from lymph HDLs to plasma lipoproteins.
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The mechanisms by which LDLs and HDLs cross the vascular endothelium from plasma into interstitial fluid are not understood, and have never been studied in humans in vivo. We determined whether the plasma-to-lymph clearance rates of LDL and HDL conform with those predicted by passive ultrafiltration through intercellular pores, or if it is necessary to invoke an active process such as receptor-mediated transcytosis. Plasma and afferent peripheral lymph were collected under steady-state conditions from 30 healthy men, and assayed for seven globular proteins of molecular radii 2.89-8.95 nm, complement C3, and apo AI, apo AII, and apo B. Plasma-to-lymph clearance rates of the seven proteins fitted the relation expected for molecules of their size when transported through two populations of pores of radius 4.95 and 20.1 nm. The same model parameters were then found to accurately predict the clearance rates of both HDL and LDL. The apparent clearance of complement C3, previously shown to be secreted by cultured endothelium, exceeded that predicted by the model. We conclude that the transport of HDL and LDL from plasma into interstitial fluid across the peripheral vascular endothelium in healthy humans can be explained by ultrafiltration without invoking an additional active process such as transcytosis.
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Endotelio Vascular/metabolismo , Lipoproteínas HDL/metabolismo , Lipoproteínas LDL/metabolismo , Microvasos/metabolismo , Adulto , Anciano , Apolipoproteínas/sangre , Apolipoproteínas/metabolismo , Transporte Biológico , Difusión , Humanos , Lipoproteínas HDL/sangre , Lipoproteínas LDL/sangre , Linfa/metabolismo , Masculino , Persona de Mediana Edad , Ultrafiltración , Adulto JovenRESUMEN
Inhibition of cholesteryl ester transfer protein (CETP) lowers plasma low-density lipoprotein cholesterol concentration and raises high-density lipoprotein (HDL) cholesterol, suggesting it might prevent cardiovascular disease (CVD). From the outset, however, the concept has been controversial owing to uncertainty about its effects on HDL function and reverse cholesterol transport (RCT). Although there has long been good evidence that CETP inhibition reduces atherosclerosis in rabbits, the first information on CETP as a CVD risk factor in a prospectively followed cohort was not published until after the first Phase 3 trial of a CETP inhibitor had begun. The worrying finding that CVD incidence was related inversely to plasma CETP has since been reproduced in each of five further prospective cohort studies. Similar results were obtained in subjects on or off statin therapy, for first and second CVD events, and for mortality as well as CVD morbidity. Additionally, two recent studies have found alleles of the CETP gene that lower hepatic CETP secretion to be associated with an increased risk of myocardial infarction. Meanwhile, CETP gene transfer in mice was found to increase RCT from peripheral macrophages in vivo, and human plasma with high CETP activity was shown to have a greater capacity to remove cholesterol from cultured cells than plasma with low activity. This mounting evidence for a protective function of CETP has been given remarkably little attention, and indeed was not mentioned in several recent reviews. It appears to show that CETP inhibition does not test the HDL hypothesis as originally hoped, and raises a pressing ethical issue regarding two Phase 3 trials of inhibitors, involving more than forty thousand subjects, which are currently in progress. As the weight of evidence now clearly supports an adverse effect of CETP inhibition on CVD, an urgent review is needed to determine if these trials should be discontinued.
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Estrogens are recognized as modulators of immune responses in mammals and teleosts. While it is known that the effects of estrogens are mediated via leukocyte-specific estrogen receptors (ERs) in humans and mice, leucocyte-specific estrogen receptor expression and the effects of estrogens on this cell population is less explored and poorly understood in teleosts. Here in, we verify that channel catfish (Ictalurus punctaus) leukocytes express ERα and ERß2. Transcripts of these isoforms were detected in tissue-associated leukocyte populations by PCR, but ERß2 was rarely detected in PBLs. Expression of these receptors was temporally regulated in PBLs following polyclonal activation by concanavalin A, lipopolysaccharide or alloantigen based on evaluation by quantitative and end-point PCR. Examination of long-term leukocyte cell lines demonstrated that these receptors are differentially expressed depending on leukocyte lineage and phenotype. Expression of ERs was also temporally dynamic in some leukocyte lineages and may reflect stage of cell maturity. Estrogens affect the responsiveness of channel catfish peripheral blood leukocytes (PBLs) to mitogens in vitro. Similarly, bactericidal activity and phorbol 12-myristate 13-acetate induced respiratory burst was modulated by 17ß-estradiol. These actions were blocked by the pure ER antagonist ICI 182780 indicating that response is, in part, mediated via ERα. In summary, estrogen receptors are expressed in channel catfish leukocytes and participate in the regulation of the immune response. This is the first time leukocyte lineage expression has been reported in teleost cell lines.
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Receptor alfa de Estrógeno/genética , Receptor beta de Estrógeno/genética , Estrógenos/metabolismo , Proteínas de Peces/genética , Regulación de la Expresión Génica , Ictaluridae/genética , Leucocitos/inmunología , Animales , Línea Celular , Proliferación Celular , Concanavalina A/farmacología , Receptor alfa de Estrógeno/metabolismo , Receptor beta de Estrógeno/metabolismo , Proteínas de Peces/metabolismo , Ictaluridae/inmunología , Isoantígenos/farmacología , Leucocitos/metabolismo , Lipopolisacáridos/farmacología , Mitógenos/farmacología , Reacción en Cadena en Tiempo Real de la Polimerasa/veterinariaRESUMEN
Since the discovery in the 1970s that plasma levels of high-density lipoprotein cholesterol (HDL-C) are inversely associated with cardiovascular outcome, it has been postulated that HDL is anti-atherogenic and that increasing HDL-C levels is a promising therapeutic strategy. However, the recent failure of three orally active, HDL-C-raising agents has introduced considerable controversy, prompting the question of whether increasing the cholesterol cargo of HDL in a non-selective manner is an effective pharmacological approach for the translation of its atheroprotective and vasculoprotective activities. The interrelationships between HDL-C concentration, HDL particle number and levels of diverse HDL particle subpopulations of defined composition are complex, as are their relationships with reverse cholesterol transport and other anti-atherogenic functions. Such complexity highlights the incompleteness of our understanding of the biology of HDL particles. This article examines the HDL hypothesis in molecular and mechanistic terms, focusing on features that have been addressed, those that remain to be tested, and potential new targets for future pharmacological interventions.
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Enfermedades Cardiovasculares/prevención & control , Ensayos Clínicos como Asunto , Drogas en Investigación/uso terapéutico , Hipolipemiantes/uso terapéutico , Lipoproteínas HDL/agonistas , Modelos Biológicos , Terapia Molecular Dirigida , Animales , Apolipoproteína A-I/agonistas , Apolipoproteína A-I/genética , Apolipoproteína A-I/metabolismo , Enfermedades Cardiovasculares/sangre , Enfermedades Cardiovasculares/metabolismo , Proteínas de Transferencia de Ésteres de Colesterol/antagonistas & inhibidores , Proteínas de Transferencia de Ésteres de Colesterol/metabolismo , Drogas en Investigación/efectos adversos , Drogas en Investigación/metabolismo , Humanos , Hipolipemiantes/efectos adversos , Hipolipemiantes/metabolismo , Lipoproteínas HDL/sangre , Lipoproteínas HDL/metabolismo , Lipoproteínas HDL/uso terapéutico , Terapia Molecular Dirigida/efectos adversos , Niacina/efectos adversos , Niacina/metabolismo , Niacina/uso terapéutico , Fosfatidilcolina-Esterol O-Aciltransferasa/efectos adversos , Fosfatidilcolina-Esterol O-Aciltransferasa/genética , Fosfatidilcolina-Esterol O-Aciltransferasa/metabolismo , Fosfatidilcolina-Esterol O-Aciltransferasa/uso terapéutico , Proteínas Recombinantes/efectos adversos , Proteínas Recombinantes/metabolismo , Proteínas Recombinantes/uso terapéutico , Regulación hacia Arriba/efectos de los fármacosRESUMEN
The life cycles of VLDLs and most LDLs occur within plasma. By contrast, the role of HDLs in cholesterol transport from cells requires that they readily gain access to and function within interstitial fluid. Studies of lymph derived from skin, connective tissue, and adipose tissue have demonstrated that particles as large as HDLs require transport through lymphatics to return to the bloodstream during reverse cholesterol transport. Targeting HDL for therapeutic purposes will require understanding its biology in the extravascular compartment, within the interstitium and lymph, in health and disease, and we herein review the processes that mediate the transport of HDLs and chylomicrons through the lymphatic vasculature.
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Quilomicrones/metabolismo , Lipoproteínas HDL/metabolismo , Vasos Linfáticos/metabolismo , Animales , Aterosclerosis/metabolismo , Transporte Biológico , Colesterol/metabolismo , Líquido Extracelular/metabolismo , HumanosRESUMEN
BACKGROUND: The mobile intercellular fluid flowing to and in the lymphatics contains filtered plasma products and substances synthesized and excreted by tissue cells. Among them are signaling proteins such as cytokines, chemokines, enzymes, and growth factors. They act locally in autocrine and paracrine systems regulating cell metabolism, proliferation, and formation of the ground matrix. They play an immunoregulatory role in infections, wound healing, and tumor cell growth. METHODS AND RESULTS: In this study we measured the concentration of selected cytokines, chemokines, tissue enzymes, and growth factors in tissue fluid/lymph drained from normal human leg soft tissues. Legs exposed to infections and trauma often result in development of lymphedema. Lymph was drained from superficial calf lymphatics using microsurgical techniques. Our studies showed generally higher concentrations of cytokines, chemokines, enzymes, and growth factors in lymph than in serum. The total protein L/S ratio was 0.22, whereas that of various lymph signaling proteins ranged between 1 and 10. CONCLUSIONS: This indicates that in addition to proteins filtered from blood, local cells contribute to lymph concentration by own production, depending on the actual cell requirement. Moreover, there were major individual differences of lymph levels with simultaneous stable serum levels. This suggests existence of a local autonomous regulatory humoral mechanism in tissues, not reflected in serum.
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Citocinas/metabolismo , Péptidos y Proteínas de Señalización Intercelular/metabolismo , Linfa/metabolismo , Metaloproteinasa 9 de la Matriz/metabolismo , Inhibidor Tisular de Metaloproteinasa-1/metabolismo , Inhibidor Tisular de Metaloproteinasa-2/metabolismo , Voluntarios Sanos , Humanos , Pierna/irrigación sanguínea , Linfa/química , Vasos Linfáticos/irrigación sanguínea , Vasos Linfáticos/metabolismo , Masculino , Suero , Transducción de Señal/fisiologíaRESUMEN
Four studies present the first evidence showing that public (vs. private) provocation augments triggered displaced aggression by increasing the perceived intensity of the provocation. This effect is shown to be independent of face-saving motivation. Following a public or private provocation, Study 1 participants were induced to ruminate or were distracted for 20 min. They then had an opportunity to aggress against another person who either acted in a neutral or mildly annoying fashion (viz. triggering event). As expected, the magnitude of the greater displaced aggression of those who ruminated before the triggering event compared with those distracted was greater under public than private provocation. Study 2 replicated the findings of Study 1 and confirmed that public provocations are experienced as more intense. Studies 3 and 4 both manipulated provocation intensity directly to show that it mediated the moderating effect of public/private provocation found in Study 1. The greater intensity of a public provocation increases reactivity to a subsequent trigger, which in turn, augments triggered displaced aggression.
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Agresión/psicología , Ira , Motivación , Pensamiento , Femenino , Humanos , Masculino , Factores de TiempoRESUMEN
Although much is known about the remodeling of high density lipoproteins (HDLs) in blood, there is no information on that in interstitial fluid, where it might have a major impact on the transport of cholesterol from cells. We incubated plasma and afferent (prenodal) peripheral lymph from 10 healthy men at 37°C in vitro and followed the changes in HDL subclasses by nondenaturing two-dimensional crossed immunoelectrophoresis and size-exclusion chromatography. In plasma, there was always initially a net conversion of small pre-ß-HDLs to cholesteryl ester (CE)-rich α-HDLs. By contrast, in lymph, there was only net production of pre-ß-HDLs from α-HDLs. Endogenous cholesterol esterification rate, cholesteryl ester transfer protein (CETP) concentration, CE transfer activity, phospholipid transfer protein (PLTP) concentration, and phospholipid transfer activity in lymph averaged 5.0, 10.4, 8.2, 25.0, and 82.0% of those in plasma, respectively (all P < 0.02). Lymph PLTP concentration, but not phospholipid transfer activity, was positively correlated with that in plasma (r = +0.63, P = 0.05). Mean PLTP-specific activity was 3.5-fold greater in lymph, reflecting a greater proportion of the high-activity form of PLTP. These findings suggest that cholesterol esterification rate and PLTP specific activity are differentially regulated in the two matrices in accordance with the requirements of reverse cholesterol transport, generating lipid-poor pre-ß-HDLs in the extracellular matrix for cholesterol uptake from neighboring cells and converting pre-ß-HDLs to α-HDLs in plasma for the delivery of cell-derived CEs to the liver.
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Apolipoproteína A-I/metabolismo , Líquido Extracelular/metabolismo , Lipoproteínas/metabolismo , Adulto , Humanos , Masculino , Adulto JovenRESUMEN
United States (U.S.) energy policy includes an expectation that bioenergy will be a substantial future energy source. In particular, the Energy Independence and Security Act of 2007 (EISA) aims to increase annual U.S. biofuel (secondary bioenergy) production by more than 3-fold, from 40 to 136 billion liters ethanol, which implies an even larger increase in biomass demand (primary energy), from roughly 2.9 to 7.4 EJ yr(-1). However, our understanding of many of the factors used to establish such energy targets is far from complete, introducing significgant uncertainty into the feasibility of current estimates of bioenergy potential. Here, we utilized satellite-derived net primary productivity (NPP) data-measured for every 1 km(2) of the 7.2 million km(2) of vegetated land in the conterminous U.S.-to estimate primary bioenergy potential (PBP). Our results indicate that PBP of the conterminous U.S. ranges from roughly 5.9 to 22.2 EJ yr(-1), depending on land use. The low end of this range represents the potential when harvesting residues only, while the high end would require an annual biomass harvest over an area more than three times current U.S. agricultural extent. While EISA energy targets are theoretically achievable, we show that meeting these targets utilizing current technology would require either an 80% displacement of current crop harvest or the conversion of 60% of rangeland productivity. Accordingly, realistically constrained estimates of bioenergy potential are critical for effective incorporation of bioenergy into the national energy portfolio.
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Biocombustibles/provisión & distribución , Agricultura , Comunicaciones por Satélite , Árboles , Estados UnidosRESUMEN
Peptides secreted by adipose tissue (adipokines) may enter blood via capillaries or lymph. The relative importance of these pathways for a given adipokine might influence its biological effects. Because this has not been studied in any species, we measured the concentrations of seven adipokines and eight nonsecreted proteins in afferent peripheral lymph and venous plasma from 12 healthy men. Data for nonsecreted proteins were used to derive indices of microvascular permeability, which in conjunction with the molecular radii of the adipokines were used to estimate the amounts leaving the tissue via capillaries. Transport rates via lymph were estimated from the lymph adipokine concentrations and lymph flow rates and total transport (secretion) as the sum of this and capillary transport. Concentrations of nonsecreted proteins were always lower in lymph than in plasma. With the exception of adiponectin, adipokine concentrations were always higher in lymph (P < 0.01). Leptin and MCP-1 were secreted at the highest rates (means: 43 µg/h or 2.7 nmol/h and 32 µg/h or 2.4 nmol/h, respectively). IL-6 and MCP-1 secretion rates varied greatly between subjects. The proportion of an adipokine transported via lymph was directly related to its molecular radius (r(s) = +0.94, P = 0.025, n = 6), increasing from 14 to 100% as the radius increased from 1.18 (IL-8) to 3.24 nm (TNFα). We conclude that the lymph/capillary partitioning of adipokines is a function of molecular size, which may affect both their regional and systemic effects in vivo. This finding may have implications for the physiology of peptides secreted by other tissues.
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Adipocitos/metabolismo , Adipoquinas/metabolismo , Tejido Adiposo/metabolismo , Permeabilidad Capilar/fisiología , Sistema Linfático/fisiología , Adipoquinas/sangre , Adiponectina/sangre , Adiponectina/metabolismo , Adulto , Transporte Biológico , Humanos , Leptina/sangre , Leptina/metabolismo , Vasos Linfáticos/metabolismo , MasculinoRESUMEN
Although rumination following a provocation can increase aggression, no research has examined the processes responsible for this phenomenon. With predictions derived from the General Aggression Model, three experiments explored the impact of two types of post-provocation rumination on the processes whereby rumination augments aggression. In Experiment 1, relative to distraction, self-focused rumination uniquely increased the accessibility of arousal cognition, whereas provocation-focused rumination uniquely amplified the accessibility of aggressive action cognition. In Experiment 2, provocation-focused rumination uniquely increased systolic blood pressure. In Experiment 3, both types of rumination increased aggressive behaviour relative to a distraction condition. Angry affect partially mediated the effects of both provocation- and self-focused rumination on aggression. Self-critical negative affect partially mediated the effect of self-focused rumination but not provocation-focused rumination. These findings suggest that provocation-focused rumination influences angry affect, aggressive action cognition, and cardiovascular arousal, whereas self-focused rumination increases self-critical negative affect, angry affect, and arousal cognition. These studies enhance our understanding of why two types of post-provocation rumination increase aggressive behaviour.
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Agresión/psicología , Emociones , Pensamiento , Femenino , Humanos , Masculino , Modelos PsicológicosRESUMEN
Channel catfish Ictalurus punctatus express two Ig isotypes: IgM and IgD. Although catfish IgM has been extensively studied at the functional and structural levels, much less is known about IgD. In this study, IgM(+)/IgD(+) and IgM(-)/IgD(+) catfish B cell populations were identified through the use of anti-IgM and anti-IgD mAbs. Catfish IgM(+)/IgD(+) B cells are small and agranular. In contrast, IgM(-)/IgD(+) B cells are larger and exhibit a plasmablast morphology. The use of cell sorting, flow cytometry, and RT-PCR demonstrated that IgD(+) B cell expression varies among individuals. For example, some catfish have <5% IgM(-)/IgD(+) B cells in their PBLs, whereas in others the IgM(-)/IgD(+) B cell population can represent as much as 72%. Furthermore, IgD expressed by IgM(-)/IgD(+) B cells preferentially associates with IgL σ. Comparatively, IgM(+)/IgD(+) B cells can express any of the four catfish IgL isotypes. Also, transfection studies show that IgD functions as a typical BCR, because Igδ-chains associate with CD79a and CD79b molecules, and all membrane IgD transcripts from sorted IgM(-)/IgD(+) B cells contain viable VDJ rearrangements, with no bias in family member usage. Interestingly, all secreted IgD transcripts from IgM(+)/IgD(+) and IgM(-)/IgD(+) B cells were V-less and began with a leader spliced to Cδ1. Importantly, transfection of catfish clonal B cells demonstrated that this leader mediated IgD secretion. Together, these findings imply that catfish IgM(-)/IgD(+) B cells likely expand in response to certain pathogens and that the catfish IgD Fc-region, as has been suggested for human IgD, may function as a pattern recognition molecule.
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Subgrupos de Linfocitos B/inmunología , Linfocitos B/inmunología , Ictaluridae/inmunología , Inmunoglobulina D/inmunología , Animales , Western Blotting , Antígenos CD79/inmunología , Separación Celular , Citometría de Flujo , Expresión Génica , Genes de Inmunoglobulinas , Inmunoglobulina D/genética , Inmunoprecipitación , Reacción en Cadena de la Polimerasa de Transcriptasa InversaRESUMEN
In mammals, expression of the three alternatively spliced exons of the tyrosine phosphatase CD45 is regulated by the developmental and activation state of the cell. In comparison, the channel catfish, Ictalurus punctatus, CD45 homolog contains 18 functional alternatively spliced exons. Since very little is known about CD45 regulation in ectothermic vertebrates, this study examines the regulation of catfish CD45 mRNA isoform expression in clonal T and B cells in response to stimulation. Results show that mitogenic stimulation using catfish serum or concanavalin A induced expression of mRNAs for small CD45 isoforms, and isoform message expression was growth curve dependent, i.e. cells in logarithmic phase express message for smaller CD45 isoforms, whereas stationary phase cells express message for longer CD45 isoforms. In addition, cells treated with the protein synthesis inhibitor cycloheximide expressed message for longer CD45 isoforms, and treatment with lactacystin, which blocks protein degradation, rescued smaller isoform message expression. Collectively these data suggested that expression of CD45 isoforms, in catfish, at least at the mRNA level, is "constitutively dynamic" and highly dependent on extracellular stimuli.
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Linfocitos B/metabolismo , Ictaluridae , Antígenos Comunes de Leucocito/metabolismo , Isoformas de Proteínas/metabolismo , Linfocitos T/metabolismo , Acetilcisteína/análogos & derivados , Acetilcisteína/farmacología , Empalme Alternativo/efectos de los fármacos , Empalme Alternativo/inmunología , Animales , Linfocitos B/citología , Linfocitos B/efectos de los fármacos , Linfocitos B/inmunología , Células Clonales , Concanavalina A/inmunología , Concanavalina A/metabolismo , Cicloheximida/farmacología , Retroalimentación Fisiológica , Perfilación de la Expresión Génica , Inmunización , Antígenos Comunes de Leucocito/genética , Antígenos Comunes de Leucocito/inmunología , Activación de Linfocitos/efectos de los fármacos , Isoformas de Proteínas/genética , Isoformas de Proteínas/inmunología , Linfocitos T/citología , Linfocitos T/efectos de los fármacos , Linfocitos T/inmunologíaRESUMEN
BACKGROUND: The pre-nodal afferent lymph is the fluid which directly derives from the extracellular milieu from every parenchymal organ and, as it continues to circulate between the cells, it collects products deriving from the organ metabolism/catabolism. A comprehensive qualitative and quantitative investigation of the self-antigenic repertoire transported by the human lymph is still missing. METHODOLOGY/PRINCIPAL FINDINGS: A major difference between lymph and plasma could be visualized by FPLC and 2D gel in the amount of low molecular weight products corresponding to peptide fragments. Naturally processed peptides in normal pre-nodal human lymph were then fractionated by HPLC and characterized by multidimensional mass spectrometry. Analysis of more then 300 sequences identified self-peptides derived from both intracellular and extracellular proteins revealing the variety of catabolic products transported by human lymph. Quantitative analysis established that at least some of these peptides are present in the circulating lymph in nanomolar concentration. CONCLUSIONS/SIGNIFICANCE: The peptidome, generated by physiological tissue catabolism and transported by the pre-nodal lymph, is in addition to the self-peptidome generated in endosomal compartment. Unlike self antigen processed by local or nodal APC, which mostly produce epitopes constrained by the endosomal processing activity, self antigens present in the lymph could derived from a wider variety of processing pathways; including caspases, involved in cellular apoptosis, and ADAM and other metalloproteinases involved in surface receptor editing, cytokines processing and matrix remodeling. Altogether, expanding the tissue-specific self-repertoire available for the maintenance of immunological tolerance.