RESUMEN
Prymnesium parvum causes harmful algal blooms worldwide that are often associated with massive fish-kills and subsequent economic losses. Most of our knowledge of the toxicity of P. parvum derives from bioassays since methods for the identification and quantification of their toxins have been lacking. Recently, a quantitation method was developed for the causative lytic toxins, the prymnesins. Here, we for the first time present data on the influence of irradiance on cellular content and production of prymnesins under nutrient replete conditions in two P. parvum strains, which both produce B-type prymnesins. Large differences were observed between the two strains with regard to the influence of irradiance on prymnesin cell quota and production rates. At the highest irradiance level (550 µmol photons m-2 s-1), the cellular prymnesin quota was thirty times higher in strain K-0081 strain than in K-0374. The cellular prymnesin quota and production rates were closely linked to rates of growth and photosynthesis in strain K-0081, while this was not the case for K-0374. Yet, growth rate did explain the differences in prymnesin quota in the two strains. Consequently, the maximum prymnesin production rate (414 attomol cell-1 d-1) was only about three times higher in strain K-0081 than in K-0374, and revealed an optimum at the same irradiance of 200 µmol photons m-2 s-1 in both strains. At low irradiance levels, the difference in production rates between both strains became smaller, with 41 and 49 attomol cell-1 d-1 for K-0081 and K-0374, respectively. The carbon content of prymnesins made up for â¼3% and <1% of the total cellular carbon content in strains K-0081 and K-0374, respectively. The fraction of extracellular dissolved prymnesins was measured for strain K-0081, where it accounted for 14-30% of total prymnesin concentration in the cultures, irrespective of irradiance. The concentrations of prymnesins released to the water by the K-0081 strain were not significantly influenced by irradiance. Overall, we observed comparable responses in growth and photosynthesis of both tested strains toward changes in irradiance. However, the effects of irradiance on prymnesin quota and production rates were remarkably different between the two strains.
Asunto(s)
Haptophyta , Animales , Floraciones de Algas Nocivas , Lipoproteínas/metabolismo , Lipoproteínas/toxicidad , FotosíntesisRESUMEN
BACKGROUND: Helicobacter pylori is a bacterium that affects about 50% of the world population and, despite being often asymptomatic, it is responsible of several gastric diseases, from gastritis to gastric cancer. The protein Lpp20 (HP1456) plays an important role in bacterium survival and host colonization, but the possibility that it might be involved in the etiology of H. pylori-related disorders is an unexplored issue. Lpp20 is a lipoprotein bound to the external membrane of the bacterium, but it is also secreted inside vesicles along with other two proteins of the same operon, i.e. HP1454 and HP1457. RESULTS: In this study we determined the crystal structure of Lpp20 and we found that it has a fold similar to a carcinogenic factor released by H. pylori, namely Tipα. We demonstrate that Lpp20 promotes cell migration and E-cadherin down-regulation in gastric cancer cells, two events recalling the epithelial-mesenchymal transition (EMT) process. Differently from Tipα, Lpp20 also stimulates cell proliferation. CONCLUSIONS: This identifies Lpp20 as a new pathogenic factor produced by H. pylori that promotes EMT and thereby the progression of cancer to the metastatic state.
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Antígenos Bacterianos/química , Proteínas Bacterianas/química , Transición Epitelial-Mesenquimal/efectos de los fármacos , Helicobacter pylori/patogenicidad , Lipoproteínas/química , Antígenos Bacterianos/inmunología , Antígenos Bacterianos/toxicidad , Cadherinas/análisis , Línea Celular Tumoral , Movimiento Celular/efectos de los fármacos , Proliferación Celular/efectos de los fármacos , Humanos , Lipoproteínas/inmunología , Lipoproteínas/toxicidad , Pliegue de Proteína , Estructura Secundaria de Proteína , Neoplasias Gástricas/etiología , Neoplasias Gástricas/patologíaRESUMEN
OBJECTIVES: The underlying mechanism of atherosclerosis and visceral obesity remains unknown.The purpose of this study was to test the hypothesis that atherosclerosis and visceral obesity are caused by an immune response to native plasma lipoproteins, and the atherogenic and adipogenic effects of the antibodies to native lipoproteins stem from the androgen deficiency that is created. METHODS: Wistar rats were immunized with native human (nh) low-density (LDL) or high-density lipoproteins (HDL). Visceral fat, aortic wall structure, and testosterone levels were studied. RESULTS: Immunization with nhLDL or nhHDL induced in rats increased visceral abdominal fat and perivascular adipose tissue volume, the appearance of epicardial fat, and atherosclerosis-like changes in the aortic wall: accumulation of leucocytes, destruction of the intima, and disruption of the media structure. Immunized rats produced antibodies to native plasma lipoproteins, while there was no difference between immunized and adjuvant-injected rats with regard to the level of antibodies to oxidized LDL. The immune response to nhHDL caused testosterone disturbances, but it is not associated with visceral obesity and atherosclerosis. CONCLUSION: The immune response to native lipoproteins is atherogenic and adipogenic and testosterone is not involved in the atherogenic and adipogenic effects of antibodies to lipoproteins.
Asunto(s)
Aorta/inmunología , Aterosclerosis/inmunología , Inmunidad Celular/inmunología , Lipoproteínas/toxicidad , Obesidad Abdominal/inmunología , Testosterona/inmunología , Animales , Aorta/efectos de los fármacos , Aorta/metabolismo , Aterosclerosis/sangre , Aterosclerosis/inducido químicamente , Humanos , Inmunidad Celular/efectos de los fármacos , Masculino , Obesidad Abdominal/sangre , Obesidad Abdominal/inducido químicamente , Ratas , Ratas Wistar , Testosterona/sangreRESUMEN
Inflammation has long been implicated as a contributor to pathogenesis in neurobrucellosis. Many of the associated neurocognitive symptoms of neurobrucellosis may be the result of neuronal dysfunction resulting from the inflammatory response induced by Brucella abortus infection in the central nervous system. In this manuscript, we describe an immune mechanism for inflammatory activation of microglia that leads to neuronal death upon B. abortus infection. B. abortus was unable to infect or harm primary cultures of mouse neurons. However, when neurons were co-cultured with microglia and infected with B. abortus significant neuronal loss occurred. This phenomenon was dependent on TLR2 activation by Brucella lipoproteins. Neuronal death was not due to apoptosis, but it was dependent on the microglial release of nitric oxide (NO). B. abortus infection stimulated microglial proliferation, phagocytic activity and engulfment of neurons. NO secreted by B. abortus-activated microglia induced neuronal exposure of the "eat-me" signal phosphatidylserine (PS). Blocking of PS-binding to protein milk fat globule epidermal growth factor-8 (MFG-E8) or microglial vitronectin receptor-MFG-E8 interaction was sufficient to prevent neuronal loss by inhibiting microglial phagocytosis without affecting their activation. Taken together, our results indicate that B. abortus is not directly toxic to neurons; rather, these cells become distressed and are killed by phagocytosis in the inflammatory surroundings generated by infected microglia. Neuronal loss induced by B. abortus-activated microglia may explain, in part, the neurological deficits observed during neurobrucellosis.
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Brucella abortus/patogenicidad , Muerte Celular/fisiología , Inflamación/metabolismo , Microglía/microbiología , Microglía/fisiología , Neuronas/patología , Fagocitosis/fisiología , Animales , Antígenos Bacterianos/toxicidad , Proteínas de la Membrana Bacteriana Externa/toxicidad , Muerte Celular/genética , Células Cultivadas , Embrión de Mamíferos , Regulación Bacteriana de la Expresión Génica/fisiología , Inflamación/inducido químicamente , Inflamación/patología , Lipopolisacáridos/farmacología , Lipoproteínas/metabolismo , Lipoproteínas/toxicidad , Ratones , Ratones Endogámicos BALB C , Ratones Transgénicos , Factor 88 de Diferenciación Mieloide/genética , Factor 88 de Diferenciación Mieloide/metabolismo , Neuronas/citología , Neuronas/efectos de los fármacos , Óxido Nítrico/metabolismo , Prosencéfalo/citología , Receptor Toll-Like 2/genética , Receptor Toll-Like 2/metabolismo , Receptor Toll-Like 4/deficiencia , Receptor Toll-Like 4/genéticaRESUMEN
Many positive signalling pathways of osteoclastogenesis have been characterized, but negative signalling pathways are less well studied. Here we show by microarray and RNAi that guanine nucleotide-binding protein subunit α13 (Gα13) is a negative regulator of osteoclastogenesis. Osteoclast-lineage-specific Gna13 conditional knockout mice have a severe osteoporosis phenotype. Gna13-deficiency triggers a drastic increase in both osteoclast number and activity (hyper-activation), mechanistically through decreased RhoA activity and enhanced Akt/GSK3ß/NFATc1 signalling. Consistently, Akt inhibition or RhoA activation rescues hyper-activation of Gna13-deficient osteoclasts, and RhoA inhibition mimics the osteoclast hyperactivation resulting from Gna13-deficiency. Notably, Gα13 gain-of-function inhibits Akt activation and osteoclastogenesis, and protects mice from pathological bone loss in disease models. Collectively, we reveal that Gα13 is a master endogenous negative switch for osteoclastogenesis through regulation of the RhoA/Akt/GSK3ß/NFATc1 signalling pathway, and that manipulating Gα13 activity might be a therapeutic strategy for bone diseases.
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Subunidades alfa de la Proteína de Unión al GTP G12-G13/metabolismo , Glucógeno Sintasa Quinasa 3 beta/metabolismo , Factores de Transcripción NFATC/metabolismo , Osteogénesis/fisiología , Proteínas Proto-Oncogénicas c-akt/metabolismo , ADP Ribosa Transferasas , Animales , Densidad Ósea , Toxinas Botulínicas , Femenino , Subunidades alfa de la Proteína de Unión al GTP G12-G13/genética , Regulación de la Expresión Génica/efectos de los fármacos , Regulación de la Expresión Génica/fisiología , Silenciador del Gen , Glucógeno Sintasa Quinasa 3 beta/genética , Compuestos Heterocíclicos con 3 Anillos/farmacología , Humanos , Leucocitos Mononucleares/fisiología , Lipoproteínas/toxicidad , Factor Estimulante de Colonias de Macrófagos , Masculino , Ratones , Factores de Transcripción NFATC/genética , Ovariectomía , Proteínas Proto-Oncogénicas c-akt/genética , Ligando RANK , Transducción de Señal/fisiologíaRESUMEN
Chronic kidney disease (CKD) is associated with an enhanced oxidative stress and deep modifications in lipid and lipoprotein metabolism. First, many oxidized lipids accumulate in CKD and were shown to exert toxic effects on cells and tissues. These lipids are known to interfere with many cell functions and to be pro-apoptotic and pro-inflammatory, especially in the cardiovascular system. Some, like F2-isoprostanes, are directly correlated with CKD progression. Their accumulation, added to their noxious effects, rendered their nomination as uremic toxins credible. Similarly, lipoproteins are deeply altered by CKD modifications, either in their metabolism or composition. These impairments lead to impaired effects of HDL on their normal effectors and may strongly participate in accelerated atherosclerosis and failure of statins in end-stage renal disease patients. This review describes the impact of oxidized lipids and other modifications in the natural history of CKD and its complications. Moreover, this review focuses on the modifications of lipoproteins and their impact on the emergence of cardiovascular diseases in CKD as well as the appropriateness of considering them as actual mediators of uremic toxicity.
Asunto(s)
Lípidos/toxicidad , Lipoproteínas/toxicidad , Insuficiencia Renal Crónica/metabolismo , Toxinas Biológicas/toxicidad , Uremia/etiología , Animales , Humanos , Estrés OxidativoRESUMEN
The endotoxin lipopolysaccharide (LPS) promotes sepsis, but bacterial peptides also promote inflammation leading to sepsis. We found, intraperitoneal administration of live or heat inactivated E. coli JE5505 lacking the abundant outer membrane protein, Braun lipoprotein (BLP), was less toxic than E. coli DH5α possessing BLP in Swiss albino mice. Injection of BLP free of LPS purified from E. coli DH5α induced massive infiltration of leukocytes in lungs and liver. BLP activated human polymorphonuclear cells (PMNs) ex vivo to adhere to denatured collagen in serum and polymyxin B independent fashion, a property distinct from LPS. Both LPS and BLP stimulated the synthesis of platelet activating factor (PAF), a potent lipid mediator, in human PMNs. In mouse macrophage cell line, RAW264.7, while both BLP and LPS similarly upregulated TNF-α and IL-1ß mRNA; BLP was more potent in inducing cyclooxygenase-2 (COX-2) mRNA and protein expression. Peritoneal macrophages from TLR2-/- mice significantly reduced the production of TNF-α in response to BLP in contrast to macrophages from wild type mice. We conclude, BLP acting through TLR2, is a potent inducer of inflammation with a response profile both common and distinct from LPS. Hence, BLP mediated pathway may also be considered as an effective target against sepsis.
Asunto(s)
Proteínas de la Membrana Bacteriana Externa/toxicidad , Endotoxemia/genética , Proteínas de Escherichia coli/toxicidad , Lipopolisacáridos/toxicidad , Lipoproteínas/toxicidad , Animales , Adhesión Celular/efectos de los fármacos , Ciclooxigenasa 2/genética , Ciclooxigenasa 2/inmunología , Endotoxemia/inducido químicamente , Endotoxemia/inmunología , Endotoxemia/mortalidad , Regulación de la Expresión Génica , Humanos , Interleucina-1beta/genética , Interleucina-1beta/inmunología , Hígado/efectos de los fármacos , Hígado/inmunología , Hígado/patología , Pulmón/efectos de los fármacos , Pulmón/inmunología , Pulmón/patología , Macrófagos Peritoneales/efectos de los fármacos , Macrófagos Peritoneales/inmunología , Macrófagos Peritoneales/patología , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Neutrófilos/efectos de los fármacos , Neutrófilos/inmunología , Neutrófilos/patología , Peroxidasa/genética , Peroxidasa/inmunología , Factor de Activación Plaquetaria/genética , Factor de Activación Plaquetaria/inmunología , Cultivo Primario de Células , Células RAW 264.7 , Análisis de Supervivencia , Factor de Necrosis Tumoral alfa/genética , Factor de Necrosis Tumoral alfa/inmunologíaRESUMEN
RATIONALE: To prevent or combat infection, increasing the effectiveness of the immune response is highly desirable, especially in case of compromised immune system function. However, immunostimulatory therapies are scarce, expensive, and often have unwanted side-effects. ß-glucans have been shown to exert immunostimulatory effects in vitro and in vivo in experimental animal models. Oral ß-glucan is inexpensive and well-tolerated, and therefore may represent a promising immunostimulatory compound for human use. METHODS: We performed a randomized open-label intervention pilot-study in 15 healthy male volunteers. Subjects were randomized to either the ß -glucan (nâ=â10) or the control group (nâ=â5). Subjects in the ß-glucan group ingested ß-glucan 1000 mg once daily for 7 days. Blood was sampled at various time-points to determine ß-glucan serum levels, perform ex vivo stimulation of leukocytes, and analyze microbicidal activity. RESULTS: ß-glucan was barely detectable in serum of volunteers at all time-points. Furthermore, neither cytokine production nor microbicidal activity of leukocytes were affected by orally administered ß-glucan. CONCLUSION: The present study does not support the use of oral ß-glucan to enhance innate immune responses in humans. TRIAL REGISTRATION: ClinicalTrials.gov NCT01727895.
Asunto(s)
Inmunidad Innata/efectos de los fármacos , beta-Glucanos/administración & dosificación , Administración Oral , Candida albicans/crecimiento & desarrollo , Células Cultivadas , Citocinas/metabolismo , Humanos , Leucocitos/citología , Leucocitos/efectos de los fármacos , Leucocitos/metabolismo , Lipopolisacáridos/toxicidad , Lipoproteínas/toxicidad , Masculino , Proyectos Piloto , Polidesoxirribonucleótidos/farmacología , Factor de Necrosis Tumoral alfa/metabolismo , Adulto Joven , beta-Glucanos/sangreRESUMEN
BACKGROUND: Leptospiral glycolipoprotein (GLP) is a potent and specific Na/K-ATPase inhibitor. Severe pulmonary form of leptospirosis is characterized by edema, inflammation and intra-alveolar hemorrhage having a dismal prognosis. Resolution of edema and inflammation determines the outcome of lung injury. Na/K-ATPase activity is responsible for edema clearance. This enzyme works as a cell receptor that triggers activation of mitogen-activated protein kinase (MAPK) intracellular signaling pathway. Therefore, injection of GLP into lungs induces injury by triggering inflammation. METHODS: We injected GLP and ouabain, into mice lungs and compared their effects. Bronchoalveolar lavage fluid (BALF) was collected for cell and lipid body counting and measurement of protein and lipid mediators (PGE2 and LTB4). The levels of the IL-6, TNFα, IL-1B and MIP-1α were also quantified. Lung images illustrate the injury and whole-body plethysmography was performed to assay lung function. We used Toll-like receptor 4 (TLR4) knockout mice to evaluate leptospiral GLP-induced lung injury. Na/K-ATPase activity was determined in lung cells by nonradioactive rubidium incorporation. We analyzed MAPK p38 activation in lung and in epithelial and endothelial cells. RESULTS: Leptospiral GLP and ouabain induced lung edema, cell migration and activation, production of lipid mediators and cytokines and hemorrhage. They induced lung function alterations and inhibited rubidium incorporation. Using TLR4 knockout mice, we showed that the GLP action was not dependent on TLR4 activation. GLP activated of p38 and enhanced cytokine production in cell cultures which was reversed by a selective p38 inhibitor. CONCLUSIONS: GLP and ouabain induced lung injury, as evidenced by increased lung inflammation and hemorrhage. To our knowledge, this is the first report showing GLP induces lung injury. GLP and ouabain are Na/K-ATPase targets, triggering intracellular signaling pathways. We showed p38 activation by GLP-induced lung injury, which was may be linked to Na/K-ATPase inhibition. Lung inflammation induced by GLP was not dependent on TLR4 activation.
Asunto(s)
Leptospira interrogans , Lipopolisacáridos/toxicidad , Lipoproteínas/toxicidad , Lesión Pulmonar/inducido químicamente , Lesión Pulmonar/enzimología , ATPasa Intercambiadora de Sodio-Potasio/antagonistas & inhibidores , Animales , Línea Celular Tumoral , Inhibidores Enzimáticos/toxicidad , Células Endoteliales de la Vena Umbilical Humana/efectos de los fármacos , Células Endoteliales de la Vena Umbilical Humana/enzimología , Humanos , Lesión Pulmonar/patología , Masculino , Ratones , Ratones Endogámicos C57BL , ATPasa Intercambiadora de Sodio-Potasio/metabolismoRESUMEN
The use of surfactin and mycosubtilin as an eco-friendly alternative to control lettuce downy mildew caused by the obligate pathogen Bremia lactucae was investigated. Preliminary ecotoxicity evaluations obtained from three different tests revealed the rather low toxicity of these lipopeptides separately or in combination. The EC50 (concentration estimated to cause a 50 % response by the exposed test organisms) was about 100 mg L(-1) in Microtox assays and 6 mg L(-1) in Daphnia magna immobilization tests for mycosubtilin and 125 mg L(-1) and 25 mg L(-1) for surfactin, respectively. The toxicity of the mixture mycosubtilin/surfactin (1:1, w/w) was close to that obtained with mycosubtilin alone. In addition, the very low phytotoxic effect of these lipopeptides has been observed on germination and root growth of garden cress Lepidium sativum L. While a surfactin treatment did not influence the development of B. lactucae on lettuce plantlets, treatment with 100 mg L(-1) of mycosubtilin produced about seven times more healthy plantlets than the control samples, indicating that mycosubtilin strongly reduced the development of B. lactucae. The mixture mycosubtilin/surfactin (50:50 mg L(-1)) gave the same result on B. lactucae development as 100 mg L(-1) of mycosubtilin. The results of ecotoxicity as well as those obtained in biocontrol experiments indicated that the presence of surfactin enhances the biological activities of mycosubtilin. Mycosubtilin and surfactin were thus found to be efficient compounds against lettuce downy mildew, with low toxicity compared to the toxicity values of chemical pesticides. This is the first time that Bacillus lipopeptides have been tested in vivo against an obligate pathogen and that ecotoxic values have been given for surfactin and mycosubtilin.
Asunto(s)
Antifúngicos/farmacología , Lactuca/microbiología , Lipopéptidos/farmacología , Oomicetos/efectos de los fármacos , Péptidos Cíclicos/farmacología , Control Biológico de Vectores/métodos , Enfermedades de las Plantas/microbiología , Antifúngicos/toxicidad , Sinergismo Farmacológico , Lipopéptidos/toxicidad , Lipoproteínas/farmacología , Lipoproteínas/toxicidad , Oomicetos/crecimiento & desarrollo , Péptidos Cíclicos/toxicidadRESUMEN
Phagocytes release inflammatory mediators to defense harmful stimuli upon bacterial invasion, however, excessive inflammatory reaction leads to tissue damage and manifestation of pathological states. Therefore, targeting on uncontrolled inflammation seems feasible to control numerous inflammation-associated diseases. Under the drug screening process of synthetic diphenylpyrazole derivatives, we discovered compound yuwen02f1 possesses anti-inflammatory effects in decreasing the release of pro-inflammatory cytokines including TNFα and IL-6, nitric oxide, reactive oxygen species (ROS) as well as inhibiting migration of LPS-stimulated phagocytes. In addition, we observed that the molecular mechanism of yuwen02f1-mediated anti-inflammation is associated with decreasing phosphorylation of MAPK molecules including ERK1/2, JNK and p38, and attenuating translocation of p47(phox) and p67(phox) to the cell membrane. Yuwen02f1 also reverses IκBα degradation and attenuates the expression of NFκB-related downstream inducible enzymes like iNOS and COX-2. Furthermore, we found that yuwen02f1 attenuates some pathological syndromes of LPS-induced sepsis and adjuvant-induced arthritis in mice, as evidenced by decreasing the cytokine production, reversing thrombocytopenic syndrome, protecting the mice from tissue injury in septic mice, and attenuating paw edema in arthritic mice as well. These results suggest that yuwen02f1 is a potential anti-inflammatory agent for alleviating syndromes of acute and chronic inflammatory diseases as evidenced by attenuating the generation of cytokines and down-regulating the expression of iNOS and COX-2 through the blockade of ROS generation and NADPH oxidase, NFκB and MAPK activation pathways in LPS-stimulated phagocytes.
Asunto(s)
Artritis Experimental/inducido químicamente , Endotoxemia/inducido químicamente , Furanos/farmacología , Lipoproteínas/toxicidad , Proteínas Quinasas Activadas por Mitógenos/metabolismo , FN-kappa B/metabolismo , Pirazoles/farmacología , Animales , Antiinflamatorios/farmacología , Artritis Experimental/tratamiento farmacológico , Línea Celular , Movimiento Celular/efectos de los fármacos , Supervivencia Celular/efectos de los fármacos , Ciclooxigenasa 2/genética , Ciclooxigenasa 2/metabolismo , Endotoxemia/tratamiento farmacológico , Furanos/química , Regulación de la Expresión Génica/efectos de los fármacos , Humanos , L-Lactato Deshidrogenasa/genética , L-Lactato Deshidrogenasa/metabolismo , Macrófagos , Masculino , Ratones , Ratones Endogámicos ICR , Proteínas Quinasas Activadas por Mitógenos/genética , Estructura Molecular , Monocitos , Óxido Nítrico/metabolismo , Óxido Nítrico Sintasa de Tipo II/genética , Óxido Nítrico Sintasa de Tipo II/metabolismo , Pirazoles/química , Especies Reactivas de OxígenoRESUMEN
Leptospira cause disease through a toxin-mediated process by inducing vascular injury, particularly a small-vessel vasculitis. Breakdown of vessel endothelial cell integrity may increase vessel permeability which is correlated with the changes of tight junction and/or apoptosis in vessel endothelial cells. The specific toxin responsible remains unidentified. In this study, we amplified outer membrane protein LipL32 from the genome of Leptospira interrogans serovar Lai, and it was subcloned in pET32a(+) vector to express thioredoxin(Trx)-LipL32 fusion protein in Escherichia coli BL21(DE3). The protein was expressed and purified, and Trx-LipL32 was administered to culture with human umbilical vein endothelial cells (HUVEC) to elucidate the role of leptospiral outer membrane proteins in vessel endothelial cell. The purified recombinant protein was capable to increase the permeability of HUVECs. And the protein was able to decrease the expression of ZO-1 and induce F-actin in HUVECs display thickening and clustering. Moreover, apoptosis of HUVEC was significantly accelerated. But the fusion partner had no effect in these regards. It is possible that LipL32 is involved in the vessel lesions.
Asunto(s)
Proteínas de la Membrana Bacteriana Externa/toxicidad , Células Endoteliales/microbiología , Leptospira interrogans/patogenicidad , Lipoproteínas/toxicidad , Actinas/metabolismo , Apoptosis , Proteínas de la Membrana Bacteriana Externa/genética , Proteínas de la Membrana Bacteriana Externa/aislamiento & purificación , Permeabilidad Capilar/efectos de los fármacos , Células Cultivadas , Clonación Molecular , Escherichia coli/genética , Expresión Génica , Vectores Genéticos , Humanos , Lipoproteínas/genética , Lipoproteínas/aislamiento & purificación , Proteínas de la Membrana/antagonistas & inhibidores , Fosfoproteínas/antagonistas & inhibidores , Proteínas Recombinantes/genética , Proteínas Recombinantes/aislamiento & purificación , Proteínas Recombinantes/toxicidad , Proteína de la Zonula Occludens-1RESUMEN
Increasingly over the past century, seasonal fish kills associated with toxic blooms of Prymnesium parvum have devastated aquaculture and native fish, shellfish, and mollusk populations worldwide. Protracted blooms of P. parvum can result in major disturbances to the local ecology and extensive monetary losses. Toxicity of this alga is attributed to a collection of compounds known as prymnesins, which exhibit potent cytotoxic, hemolytic, neurotoxic and ichthyotoxic effects. These secondary metabolites are especially damaging to gill-breathing organisms and they are believed to interact directly with plasma membranes, compromising integrity by permitting ion leakage. Several factors appear to function in the activation and potency of prymnesins including salinity, pH, ion availability, and growth phase. Prymnesins may function as defense compounds to prevent herbivory and some investigations suggest that they have allelopathic roles. Since the last extensive review was published, two prymnesins have been chemically characterized and ongoing investigations are aimed at the purification and analysis of numerous other toxic metabolites from this alga. More information is needed to unravel the mechanisms of prymnesin synthesis and the significance of these metabolites. Such work should greatly improve our limited understanding of the physiology and biochemistry of P. parvum and how to mitigate its blooms.
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Chrysophyta/química , Peces/fisiología , Invertebrados/efectos de los fármacos , Lipoproteínas/toxicidad , Sistema Nervioso/efectos de los fármacos , Animales , Chrysophyta/fisiología , Lipoproteínas/química , Lipoproteínas/aislamiento & purificación , Lipoproteínas/metabolismo , Venenos/química , Venenos/aislamiento & purificación , Venenos/metabolismo , Venenos/toxicidadAsunto(s)
Eucariontes/fisiología , Floraciones de Algas Nocivas/fisiología , Lipoproteínas/toxicidad , Toxinas Marinas/toxicidad , Venenos/toxicidad , Contaminantes Químicos del Agua/toxicidad , Fenómenos Químicos , Concentración de Iones de Hidrógeno , Lipoproteínas/análisis , Lipoproteínas/química , Toxinas Marinas/análisis , Toxinas Marinas/química , Modelos Químicos , Venenos/análisis , Venenos/química , Contaminantes Químicos del Agua/análisis , Contaminantes Químicos del Agua/químicaRESUMEN
The harmful algal bloom species Prymnesium parvum has caused millions of dollars in damage to fisheries around the world. These fish kills have been attributed to P. parvum releasing a mixture of toxins in the water. The characterized toxins, reported as prymnesin-1 and -2, have structural similarities consistent with other known ionizable compounds (e.g., ammonia). We investigated whether pH affects the toxicity of P. parvum under conditions representative of inland Texas reservoirs experiencing ambient toxicity from bloom formation. We evaluated pH influences on toxicity in laboratory and field samples, and modeled the physicochemical properties of prymnesins. Aquatic toxicity to a model fish and cladoceran was reduced by lowering pH in samples obtained from reservoirs experiencing P. parvum blooms; similar observations were confirmed for experiments with laboratory cultures. A pKa value of 8.9 was predicted for the prymnesins, which suggests that ionization states of these toxins may change appreciably over surface water pH of inland waters. These findings indicate that ionization states of toxins released by P. parvum may strongly influence site-specific toxicity and subsequent impacts to fisheries. Consequently, these results emphasize the importance of understanding processes that affect pH during P. parvum blooms, which may improve predictions of ambient toxicity.
Asunto(s)
Eucariontes/fisiología , Floraciones de Algas Nocivas/fisiología , Lipoproteínas/toxicidad , Toxinas Marinas/toxicidad , Venenos/toxicidad , Contaminantes Químicos del Agua/toxicidad , Animales , Bioensayo , Fenómenos Químicos , Cyprinidae/fisiología , Daphnia/fisiología , Concentración de Iones de Hidrógeno , Larva/efectos de los fármacos , Dosificación Letal Mediana , Lipoproteínas/química , Longevidad/efectos de los fármacos , Toxinas Marinas/química , Modelos Químicos , Venenos/química , Pruebas de Toxicidad Aguda , Contaminantes Químicos del Agua/químicaRESUMEN
The pathogenesis of Mycoplasma pneumoniae infection is considered to be in part attributable to excessive immune responses. In this study, we investigated whether synthetic lipopeptides of subunit b of F0F1-type ATPase (F0F1-ATPase), NF-kappaB-activating lipoprotein 1 (N-ALP1), and N-ALP2 (named FAM20, sN-ALP1, and sN-ALP2, respectively) derived from M. pneumoniae induce cytokine and chemokine production and leukocyte infiltration in vivo. Intranasal administration of FAM20 and sN-ALP2 induced infiltration of leukocyte cells and production of chemokines and cytokines in bronchoalveolar lavage fluid, but sN-ALP1 failed to do so. The activity of FAM20 was notably higher than that of sN-ALP2. FAM20 and sN-ALP2 induced tumor necrosis factor alpha (TNF-alpha) through Toll-like receptor 2 in mouse peritoneal macrophages. Moreover, in the range of low concentrations of lipopeptides, FAM20 showed relatively high activity of inducing TNF-alpha in mouse peritoneal macrophages compared to synthetic lipopeptides such as MALP-2 and FSL-1, derived from Mycoplasma fermentans and Mycoplasma salivarium, respectively. These findings indicate that the F0F1-ATPase might be a key molecule in inducing cytokines and chemokines contributing to inflammatory responses during M. pneumoniae infection in vivo.
Asunto(s)
Inflamación/inducido químicamente , Lipoproteínas/toxicidad , Pulmón/efectos de los fármacos , Mycoplasma pneumoniae/metabolismo , Administración Intranasal , Animales , Citocinas , Regulación de la Expresión Génica/efectos de los fármacos , Leucocitos , Lipoproteínas/administración & dosificación , Lipoproteínas/síntesis química , Lipoproteínas/metabolismo , Pulmón/citología , Pulmón/patología , Ratones , Ratones Endogámicos C57BL , Conformación ProteicaRESUMEN
Tolerization with bacterial lipoprotein (BLP) affords a significant survival benefit in sepsis. Given that high mobility group box protein-1 (HMGB1) is a recognized mediator of sepsis-related lethality, we determined if tolerization with BLP leads to alterations in HMGB1. In vitro, BLP tolerization led to a reduction in HMGB1 gene transcription. This was mirrored at the protein level, as HMGB1 protein expression and release were reduced significantly in BLP-tolerized human THP-1 monocytic cells. BLP tolerance in vivo led to a highly significant, long-term survival benefit following challenge with lethal dose BLP in C57BL/6 mice. This was associated with an attenuation of HMGB1 release into the circulation, as evidenced by negligible serum HMGB1 levels in BLP-tolerized mice. Moreover, HMGB1 levels in peritoneal macrophages from BLP-tolerized mice were reduced significantly. Hence, tolerization with BLP leads to a down-regulation of HMGB1 protein synthesis and release. The improved survival associated with BLP tolerance could thus be explained by a reduction in HMGB1, were the latter associated with lethality in BLP-related sepsis. In testing this hypothesis, it was noted that neutralization of HMGB1, using anti-HMGB1 antibodies, abrogated BLP-associated lethality almost completely. To conclude, tolerization with BLP leads to a down-regulation of HMGB1, thus offering a novel means of targeting the latter. HMGB1 is also a mediator of lethality in BLP-related sepsis.
Asunto(s)
Proteínas Bacterianas/toxicidad , Proteína HMGB1/inmunología , Tolerancia Inmunológica , Lipoproteínas/toxicidad , Macrófagos Peritoneales/inmunología , Sepsis/inmunología , Animales , Anticuerpos/farmacología , Línea Celular , Regulación hacia Abajo/efectos de los fármacos , Regulación hacia Abajo/inmunología , Proteína HMGB1/biosíntesis , Humanos , Tolerancia Inmunológica/efectos de los fármacos , Macrófagos Peritoneales/metabolismo , Macrófagos Peritoneales/patología , Masculino , Ratones , Sepsis/inducido químicamente , Sepsis/metabolismo , Sepsis/prevención & control , Transcripción Genética/efectos de los fármacos , Transcripción Genética/inmunologíaRESUMEN
The importance of the biological function and activity of lipoproteins from the outer or cytoplasmic membranes of Gram-positive and Gram-negative bacteria is being increasingly recognized. It is well established that they are like the endotoxins (lipopolysaccharide (LPS)), which are the main amphiphilic components of the outer membrane of Gram-negative bacteria, potent stimulants of the human innate immune system, and elicit a variety of proinflammatory immune responses. Investigations of synthetic lipopeptides corresponding to N-terminal partial structures of bacterial lipoproteins defined the chemical prerequisites for their biological activity and in particular the number and length of acyl chains and sequence of the peptide part. Here we present experimental data on the biophysical mechanisms underlying lipopeptide bioactivity. Investigation of selected synthetic diacylated and triacylated lipopeptides revealed that the geometry of these molecules (i.e. the molecular conformations and supramolecular aggregate structures) and the preference for membrane intercalation provide an explanation for the biological activities of the different lipopeptides. This refers in particular to the agonistic or antagonistic activity (i.e. their ability to induce cytokines in mononuclear cells or to block this activity, respectively). Biological activity of lipopeptides was hardly affected by the LPS-neutralizing antibiotic polymyxin B, and the biophysical interaction characteristics were found to be in sharp contrast to that of LPS with polymyxin B. The analytical data show that our concept of "endotoxic conformation," originally developed for LPS, can be applied also to the investigated lipopeptide and suggest that the molecular mechanisms of cell activation by amphiphilic molecules are governed by a general principle.
Asunto(s)
Lipoproteínas/metabolismo , Lipoproteínas/toxicidad , Fragmentos de Péptidos/metabolismo , Fragmentos de Péptidos/toxicidad , Membrana Celular/efectos de los fármacos , Células Cultivadas , Fenómenos Químicos , Química Física , Humanos , Lipopolisacáridos/farmacología , Lipoproteínas/síntesis química , Lipoproteínas/química , Activación de Macrófagos/efectos de los fármacos , Macrófagos/efectos de los fármacos , Macrófagos/enzimología , Macrófagos/metabolismo , Estructura Molecular , Fragmentos de Péptidos/síntesis química , Fragmentos de Péptidos/química , Polimixina B/farmacología , Unión Proteica , Conformación Proteica , Receptor Toll-Like 2/antagonistas & inhibidores , Receptor Toll-Like 2/metabolismo , Factor de Necrosis Tumoral alfa/biosíntesisRESUMEN
The two Bacillus subtilis strains (DM-03 and DM-04) were isolated from two extremely different habitats; one from the traditional fermented food and another one from a petroleum contaminated soil sample. These strains produced quantitatively and qualitatively different cyclic lipopeptides isoforms under laboratory culture conditions. MALDI-TOF mass spectral analysis revealed that lipopeptide profile varied according to the producing B. subtilis strains; iturins and surfactins isoforms were pre-dominant cyclic lipopeptides produced by B. subtilis DM-03 and DM-04 strains, respectively. A comparative study showed that these strains possessed distinct preferences for the carbon and nitrogen substrates, temperature and pH for optimal growth and biosurfactant production. Our study documented that the cyclic lipopeptide isoforms produced by the respective strains played an important role in the utilization of available hydrophobic substrate(s) from their natural habitats and conferred some kind of competitive advantage to the producing B. subtilis strains in their parent ecological niche.
Asunto(s)
Bacillus subtilis/crecimiento & desarrollo , Bacillus subtilis/metabolismo , Ambiente , Lipoproteínas/metabolismo , Bacillus subtilis/genética , Biomasa , Medios de Cultivo , Cartilla de ADN , Escherichia coli/efectos de los fármacos , Microbiología de Alimentos , Concentración de Iones de Hidrógeno , Kluyvera/efectos de los fármacos , Lipoproteínas/toxicidad , Pruebas de Sensibilidad Microbiana , Polimorfismo de Longitud del Fragmento de Restricción , Microbiología del Suelo , Especificidad de la Especie , Espectrometría de Masa por Láser de Matriz Asistida de Ionización Desorción , Tensoactivos/metabolismo , Temperatura , Levaduras/efectos de los fármacosRESUMEN
There have been no reports of the topical application of micafungin to the eye. The aim of this study was to evaluate the safety of topical instillation of 0.1% micafungin ophthalmic solution in rabbit eyes. In New Zealand white rabbits (n = 6), 50 microL of 0.1% micafungin solution was topically instilled to 1 eye, and 50 microL of sterile saline was applied to the other eye. Both eyedrops were administered hourly from 7 A.M. for 7 days. Measurements were conducted on corneal thickness, intraocular pressure, endothelial cell density, and lactate dehydrogenase (LDH) activity of tear samples. The eyes were examined slit-lamp biomicroscopically and histopathologically. Topical micafungin application for 1 week did not induce any changes in intraocular pressure, endothelial cell density, and tear LDH. Corneal thickness after instillation was slightly, but significantly, smaller in the micafungin group than in the control group (P = 0.0156, paired t test), but this difference disappeared within 24 hours after the final instillation. Biomicroscopy and histopathology revealed no significant toxic influence of micafungin application on the cornea. Topical instillation of micafungin solution had no apparent toxicity to the cornea. These results warrant future studies on the efficacy of micafungin ophthalmic solution against corneal fungal infection.