RESUMEN

Oregano essential oil has long been known for its health-promoting benefits. Here, we report its activity against viral replication. Oregano oil was found to specifically inhibit lentiviruses, such as human and simian immunodeficiency viruses (HIV and SIV), irrespective of virus tropism, but not hepatitis C virus, adenovirus 5 (ADV5), Zika virus, and influenza (H1N1) virus. Oregano oil's most abundant components, carvacrol and its isomer, thymol, were shown to block virus-target cell fusion while not perturbing other stages of the virus life cycle. We detected changes in virus particle density, suggesting that cholesterol depletion from the HIV-1 envelope membrane reduces virus entry. Furthermore, infection was rescued by adding exogenous cholesterol. The evolution of viral resistance to carvacrol supported this mechanism of action with the identification of mutations in the viral gp41 fusion protein that counteracted cholesterol depletion. In addition, resistance to carvacrol emerged later than typically observed for other clinically used drugs, strengthening its antiviral potential. Structure-activity relationship studies revealed key motifs of carvacrol and thymol required for HIV neutralization and identified previously unknown active analogs. Carvacrol was also shown to additively cooperate with antiretroviral therapy. In sum, oregano oil and improved carvacrol and thymol analogs could be considered to supplement current HIV therapeutics.IMPORTANCE Oregano essential oil has multiple benefits in traditional medicine, cosmetics, and food industries. Carvacrol and its analog, thymol, are well-described components of oregano oil. Here, we show that these compounds inhibit HIV-target cell fusion independently of viral tropism. Our results suggest that carvacrol and thymol alter the cholesterol content of the viral membrane, blocking HIV-1 entry into the target cell. Resistance to carvacrol has selected for viruses with mutations in the viral envelope glycoprotein, gp41. This protein is known for its interaction with cholesterol present in membrane lipid rafts. Together, these results demonstrate the potential of therapies targeting the viral envelope membrane, and oregano oil is a safe supplement to antiretrovirals, potentially delaying disease progression and resistance development.

Asunto(s)

Cimenos/farmacología , Proteína gp41 de Envoltorio del VIH/metabolismo , VIH-1/metabolismo , Origanum/química , Aceites de Plantas/farmacología , Internalización del Virus/efectos de los fármacos , Animales , Colesterol/genética , Colesterol/metabolismo , Cimenos/química , Farmacorresistencia Viral , Proteína gp41 de Envoltorio del VIH/genética , Infecciones por VIH/tratamiento farmacológico , Infecciones por VIH/genética , Infecciones por VIH/metabolismo , VIH-1/genética , Células HeLa , Humanos , Macaca mulatta , Mutación , Aceites de Plantas/química

RESUMEN

There is a constant need to improve antiretrovirals against HIV since therapy is limited by cost, side effects and the emergence of drug resistance. Kudzu is a climbing vine from which the root extract (Pueraria lobata), rich in isoflavones and saponins, has long been used in traditional Chinese medicine for a variety of purposes, from weight loss to alcoholism prevention. Here we show that Kudzu root extract significantly inhibits HIV-1 entry into cell lines, primary human CD4+T lymphocytes and macrophages, without cell-associated toxicity. Specifically, Kudzu inhibits the initial attachment of the viral particle to the cell surface, a mechanism that depends on the envelope glycoprotein gp120 but is independent from the HIV-1 cell receptor CD4 and co-receptors CXCR4/CCR5. This activity seems selective to lentiviruses since Kudzu inhibits HIV-2 and simian immunodeficiency virus, but does not interfere with Hepatitis C, Influenza, Zika Brazil and adenovirus infection. Importantly, depending on the dose, Kudzu can act synergistically or additively with the current antiretroviral cocktails against HIV-1 and can block   viruses resistant to the fusion inhibitor Enfuvirtide. Together our results highlight Kudzu's root extract value as a supplement to current antiretroviral therapy against HIV.

Asunto(s)

Fármacos Anti-VIH/farmacología , VIH-1/efectos de los fármacos , Extractos Vegetales/farmacología , Raíces de Plantas/química , Pueraria , Acoplamiento Viral/efectos de los fármacos , Animales , Células Cultivadas , Sinergismo Farmacológico , Enfuvirtida , Proteína gp120 de Envoltorio del VIH/metabolismo , VIH-1/fisiología , Humanos , Extractos Vegetales/química , Replicación Viral/efectos de los fármacos

RESUMEN

Hypersynchronous neuronal excitation manifests clinically as seizure (ictogenesis), and may recur spontaneously and repetitively after a variable latency period (epileptogenesis). Despite tremendous research efforts to describe molecular pathways and signatures of epileptogenesis, molecular pathomechanisms leading to chronic epilepsy remain to be clarified. We hypothesized that epigenetic modifications may form the basis for a cellular memory of epileptogenesis, and used a primary neuronal cell culture model of the rat hippocampus to study the translation of massive neuronal excitation into persisting changes of epigenetic signatures and pro-epileptogenic target gene expression. Increased spontaneous activation of cultured neurons was detected 3 and 7 days after stimulation with 10 µM glutamate when compared to sham-treated time-matched controls using calcium-imaging in vitro. Chromatin-immunoprecipitation experiments revealed short-term (3 h, 7 h, and 24 h) and long-term (3 d and 2 weeks) changes in histone modifications, which were directly linked to decreased expression of two selected epilepsy target genes, e.g. excitatory glutamate receptor genes Gria2 and Grin2a. Increased promoter methylation observed 4 weeks after glutamate stimulation at respective genes suggested long-term repression of Gria2 and Grin2a genes. Inhibition of glutamatergic activation or blocking the propagation of action potentials in cultured neurons rescued altered gene expression and regulatory epigenetic modifications. Our data support the concept of a cellular memory of epileptogenesis and persisting epigenetic modifications of epilepsy target genes, which are able to turn normal into pro-epileptic neurons and circuits.

Asunto(s)

Epigénesis Genética , Epilepsia/genética , Expresión Génica/fisiología , Hipocampo/citología , Neuronas/fisiología , Potenciales de Acción/efectos de los fármacos , Animales , Animales Recién Nacidos , Células Cultivadas , Metilación de ADN/efectos de los fármacos , Metilación de ADN/genética , Epigénesis Genética/efectos de los fármacos , Epigénesis Genética/fisiología , Fármacos actuantes sobre Aminoácidos Excitadores/farmacología , Expresión Génica/efectos de los fármacos , Ácido Glutámico/farmacología , Proteínas Asociadas a Microtúbulos/metabolismo , Modelos Biológicos , Neuronas/efectos de los fármacos , Quinoxalinas/farmacología , Ratas , Ratas Wistar , Receptores AMPA/genética , Receptores AMPA/metabolismo , Receptores de N-Metil-D-Aspartato/genética , Receptores de N-Metil-D-Aspartato/metabolismo , Bloqueadores de los Canales de Sodio/farmacología , Tetrodotoxina/farmacología , Valina/análogos & derivados , Valina/farmacología

RESUMEN

N-nitrosodimethylamine (NDMA) is a xenobiotic widespread in human environment capable of regulating the lifespan of immune cells. In this study, we examined the roles of the tumor necrosis factor-related apoptosis-inducing ligand (TRAIL)/death receptor 5 (DR5) complex and the Fas molecule in the induction of the extrinsic apoptosis pathway in human neutrophils (polymorphonuclear neutrophils (PMNs)) and peripheral blood mononuclear cells (PBMCs) exposed to NDMA. Also we assessed these proteins ability to trigger the intrinsic apoptosis pathway in those cells. For this purpose, we examined the expression of Fas-associated protein with death domain, truncated Bid (tBid) proteins, and apoptogenic factors such as apoptosis-inducing factor, Smac/Diablo, Omi/HtrA2, and caspase-3 as an indication of accomplished apoptosis phenomenon. PMNs and PBMCs were isolated from whole blood by density gradient centrifugation using Polymorphrep. Apoptotic cells were assessed with flow cytometry using a ready-made kit. The expression of proapoptotic molecules was investigated by Western blot analysis of PMNs and PBMCs treated with NDMA and/or rhTRAIL. The obtained results confirm the proapoptotic effects of NDMA on the examined human leukocytes and indicate an active participation of the TRAIL/DR5 complex and Fas protein in the process of apoptosis. Moreover, the research revealed distinct mechanisms of intrinsic apoptosis pathway activation between PMNs and PBMCs exposed to NDMA, as confirmed by the different levels of tBid, Smac/Diablo, Omi/HtrA2, and caspase-3 expression in those cells.

Asunto(s)

Dimetilnitrosamina/toxicidad , Leucocitos Mononucleares/efectos de los fármacos , Neutrófilos/efectos de los fármacos , Adulto , Apoptosis/fisiología , Células Cultivadas , Femenino , Humanos , Leucocitos Mononucleares/metabolismo , Masculino , Neutrófilos/metabolismo , Receptores del Ligando Inductor de Apoptosis Relacionado con TNF/metabolismo , Ligando Inductor de Apoptosis Relacionado con TNF/metabolismo , Adulto Joven , Receptor fas/metabolismo

RESUMEN

The aim of this work was to assess the role of ethanol-derived acetate and acetate-mediated histone acetylation in arachidonic acid-induced stress in HepG2 cells and cells overexpressing CYP2E1. Cells were grown for 7 days with 1 mM sodium acetate or 100 mM ethanol; their acetylated histone proteins and histone deacetylase 2 expression was quantified using Western blot. Ethanol- or acetate-pretreated cells were also treated for 24 h with 60 µM arachidonic acid to induce oxidative stress. Cytotoxicity was estimated by lactate dehydrogenase release, 3-[4,5-dimethylthiazolyl-2] 2,5-diphenyltetrazolium bromide test, and by DNA damage, while oxidative stress was quantified using dichlorofluorescein diacetate. Cells grown with ethanol or acetate had increased acetylated histone H3 levels in both cell types and elevated acetylated histone H4 levels in cells overexpressing CYP2E1 but not in naïve cells. In cells overexpressing CYP2E1 grown with ethanol, expression of histone deacetylase 2 was reduced by about 40 %. Arachidonic acid altered cell proliferation and was cytotoxic mostly to cells engineered to overexpress CYP2E1 but both effects were significantly lower in cells pretreated with ethanol or acetate. Cytotoxicity was also significantly decreased by 4-methylpyrazole--a CYP2E1 inhibitor and by trichostatin--an inhibitor of histone deacetylases. In cells pretreated with acetate or ethanol, the oxidative stress induced by arachidonic acid was also significantly lower. Our data indicate that histone hyperacetylation may in some extent protect the cells against oxidative stress. It is possible that acetate may act as an antioxidant at histone level. This mechanism may be relevant to alcohol-induced liver injury.

Asunto(s)

Ácido Araquidónico/metabolismo , Enfermedad Hepática Inducida por Sustancias y Drogas/etiología , Citocromo P-450 CYP2E1/metabolismo , Etanol/toxicidad , Acetatos/administración & dosificación , Acetatos/metabolismo , Acetilación , Antioxidantes/metabolismo , Etanol/metabolismo , Fomepizol , Células Hep G2 , Histonas/metabolismo , Humanos , Ácidos Hidroxámicos/farmacología , Estrés Oxidativo/efectos de los fármacos , Pirazoles/farmacología

RESUMEN

PURPOSE: The role of MAP kinases in the activation of AP-1 (c-Jun, c-Fos) and NF-κB p65 engaged in the regulation of iNOS expression in human neutrophils (PMNs) exposed to N-nitrosodimethylamine (NDMA) was analyzed in the study. MATERIAL AND METHODS: The study included a group of 20 healthy individuals. Isolated human PMN were incubated in the presence of NDMA. Selective MAP kinases inhibitors were used. The expression of proteins in the cytoplasmic and nuclear fractions was assessed using Western blot method. RESULTS: The results show that NDMA intensifies iNOS, c-Jun, NF-κB p65 and IκB-α expression in the analyzed PMNs. The blocking of the p38 pathway led to lower iNOS expression, and higher expression of c-Jun and c-Fos in the cytoplasmic fraction, and also lower c-Jun expression in the nuclear fraction of PMNs exposed to NDMA. A decrease in iNOS expression in the cytoplasmic fraction, and also c-Jun in both fractions of the examined cells, was observed as a result of JNK pathway inhibition. The blocking of the ERK5 pathway led to higher iNOS, c-Jun and c-Fos expression in the cytoplasmic fraction, and higher c-Jun expression in the nuclear fraction of PMNs exposed to NDMA. The study also demonstrated that blocking of the p38 and JNK pathways resulted in higher expression of NF-κB p65 and IκB-α in the cytoplasmic fraction and their lower expression in the nuclear fraction of these cells. CONCLUSION: Our data indicate the role of MAP kinases p38 and JNK in the activation of c-Jun and NF-κB p65 transcription factors engaged in the regulation of iNOS expression in human neutrophils exposed to NDMA. However ERK5 kinase is not involved in the regulation of iNOS and NO production by those cells.

Asunto(s)

Dimetilnitrosamina/farmacología , Sistema de Señalización de MAP Quinasas/efectos de los fármacos , Sistema de Señalización de MAP Quinasas/fisiología , Neutrófilos/enzimología , Óxido Nítrico Sintasa de Tipo II/metabolismo , Adulto , Regulación Enzimológica de la Expresión Génica/efectos de los fármacos , Humanos , Proteínas Quinasas JNK Activadas por Mitógenos/metabolismo , Persona de Mediana Edad , Proteína Quinasa 7 Activada por Mitógenos/antagonistas & inhibidores , Proteína Quinasa 7 Activada por Mitógenos/metabolismo , Neutrófilos/efectos de los fármacos , Óxido Nítrico/metabolismo , Factor de Transcripción AP-1/metabolismo , Factor de Transcripción ReIA/metabolismo , Xenobióticos/farmacología , Proteínas Quinasas p38 Activadas por Mitógenos/metabolismo

RESUMEN

Picrotoxin is a neurotoxin found in the berries of Anamirta cocculus, a plant native to Southeast Asia. Picrotoxin has potential for being used as a biological weapon since the toxin is relatively easy to isolate and purify. Limited information exists on the stability and detection of picrotoxin added to foods before or after processing. The objective of this study was to determine the stability of picrotoxin during yogurt manufacture and storage. Direct, cup-set yogurt was produced by using methods that mimic the conditions used in full-scale production of yogurt. Milk (full-fat or low-fat) was pasteurized at 85 degrees C for 30 min, and then cooled to 43 degrees C. Yogurt starter culture (thermophilic culture or thermophilic + probiotic culture) and picrotoxin (200 mug/mL milk) were added. Samples of yogurt during fermentation (5 to 6 h, 43 degrees C) and during 30 d refrigerated (4 to 6 degrees C) storage were analyzed for pH, titratable acidity, and picrototoxin levels. Regardless of starter culture used or fat content of milk, there were no significant differences in the pH and titratable acidities of the picrotoxin-spiked yogurt and the control yogurt (no added picrotoxin) during fermentation and up to 4 wk of refrigerated storage. The color or texture of the yogurt was not affected by addition of picrotoxin. Levels of picrotoxinin and picrotin (components of picrotoxin) in yogurt, as measured by LC/MS (APCI(+)/SIR) did not change significantly during fermentation and storage. A separate experiment determined that addition of picrotoxin to milk before pasteurization (85 degrees C, 30 min) did not affect picrotoxin stability. These results indicate that picrotoxin is stable in yogurt during manufacture and storage.

Asunto(s)

Manipulación de Alimentos/métodos , Conservación de Alimentos/métodos , Picrotoxina/análisis , Yogur/análisis , Frío , Estabilidad de Medicamentos , Fermentación , Calor , Concentración de Iones de Hidrógeno , Picrotoxina/química , Factores de Tiempo

RESUMEN

GaN and InN nanocrystals in silica glasses prepared by the sol-gel method were studied by transmission electron microscopy techniques. Morphology, structure and phase composition of silica gel containing Ga or In as function of the calcination and nitridation temperature were investigated.

RESUMEN

The spectrometric analysis of extracts from tobacco and tobacco smoke revealed the presence of pentobarbital in the analyzed substances. Tobacco samples and tobacco smoke were extracted with chloroform, determinations were performed with the Perkin-Elmer Autosystem XL system, on a Turbo Mass spectrometer. Subject to analysis were 4 cigarette brands manufactured in Poland and raw, unprocessed tobacco. The presence of pentobarbital in the analyzed samples was confirmed by the analysis of the mass spectrum of the substance, as well as by comparison of retention time with standard of pentobarbital. The determined pentobarbital concentrations in tobacco amounted to 3-6 microg/cigarette, and in tobacco smoke they were approximately 45% lower. In case of tobacco extracts it can with high probability be excluded that pentobarbital is synthesized during chromatographical analysis. The presence of pentobarbital in tobacco is thus beyond question.

Asunto(s)

Hipnóticos y Sedantes/análisis , Nicotiana/química , Pentobarbital/análisis , Cromatografía de Gases y Espectrometría de Masas , Polonia

RESUMEN

BACKGROUND: In our previous study we found that rhsIL-6R, along with recombinant human interleukin-6, plays a regulatory role in the immune response by modulating the tumour necrosis factor-alpha(TNF-alpha) expression and its production by peripheral blood mononuclear cells (PBMC). We also suggested that sIL-6R with IL-6 secreted by human PMN (neutrophils) influenced the TNF-alpha expression and its production by autologous PBMC. AIMS: Since soluble gp130 (sgp130) is a natural inhibitor for sIL-6R/interleukin-6 responses, in the present study we estimated an effect of exogenous recombinant human sgp130 and sgp130 secreted by PMN on the TNF-alpha expression and its production by PBMC. METHODS: Cells were isolated from whole blood of healthy persons. The PMN were cultured in 96-well plates for 1 h at 37 degrees C in a humidified incubator with 5% CO2. After the incubation, the culture supernatant of PMN was removed and added to the PBMC. PBMC were incubated for 1 h at 37 degrees C in the same conditions. Cytoplasmic protein fractions of PMN and, for comparative purpose of PBMC, were analysed for presence of sgp130 by western blotting with the use of monoclonal antibody capable of detecting this protein. In the culture supernatants of PMN we examined the concentrations of sgp130 by human enzyme-linked immunosorbent assay. TNF-alpha was measured at the protein levels as well as the mRNA levels. RESULTS AND CONCLUSIONS: The present results revealed that exogenous recombinant human sgp130modulates the TNF-alpha expression and production by PBMC. In contrast, we did not find any effect of sgp130 secreted by PMN on the TNF-alpha expression and its production by autologous PBMC.

Asunto(s)

Antígenos CD/fisiología , Leucocitos Mononucleares/metabolismo , Glicoproteínas de Membrana/fisiología , Factor de Necrosis Tumoral alfa/metabolismo , Antígenos CD/análisis , Antígenos CD/farmacología , Western Blotting , Células Cultivadas , Medios de Cultivo/química , Receptor gp130 de Citocinas , Ensayo de Inmunoadsorción Enzimática , Leucocitos Mononucleares/efectos de los fármacos , Glicoproteínas de Membrana/análisis , Glicoproteínas de Membrana/farmacología , ARN Mensajero/metabolismo , Proteínas Recombinantes/farmacología , Factor de Necrosis Tumoral alfa/análisis , Factor de Necrosis Tumoral alfa/genética

RESUMEN

PURPOSE: The aim of this study was to estimate the production of interleukin-15 (IL-15) by neutrophils (PMNs) and peripheral blood mononuclear cells (PBMC) confronted with the serum levels of IL-15 in patients with Lyme disease. MATERIAL AND METHODS: PMN and PBMC were isolated from heparinized whole blood of patients. The cells were incubated for 18 hs at 37 degrees C in a humidified incubator with 5% CO2. After 18 hs incubation, supernatant was removed and assessed for IL-15 using ELISA kits. RESULTS: The results obtained showed significant increase in the ability of patient's PMNs and peripheral blood mononuclear cells (PBMC) to release IL-15. Although PBMC produced higher concentrations of IL-15 than PMN, the quantitative dominance of PMN in the peripheral blood suggest a significant role for these cells in the defense reactions controlled by this cytokine. Similar changes in the secretion of IL-15 by PMN and PBMC in patient group may be caused by the same regulatory mechanisms which influence the functional abnormalities of the cells examined. CONCLUSIONS: A change in the ability of PMN and PBMC to release IL-15 may have various implications for the cellular and humoral response to the Borrelia burgdorferi (B.b.) infection in patients with Lyme disease.

Asunto(s)

Interleucina-15/biosíntesis , Leucocitos Mononucleares/metabolismo , Enfermedad de Lyme/inmunología , Neutrófilos/metabolismo , Adulto , Anciano , Femenino , Humanos , Enfermedad de Lyme/metabolismo , Masculino , Persona de Mediana Edad

RESUMEN

The biological activities of IL-18 include its ability to induce the production of inflammatory cytokines such as IL-1 or IL-8 by immunocompetent cells. Our previous study demonstrated that rhIL-18 induces IL-1beta and, to a lesser exted, the secretion of IL-1beta regulatory proteins involving interleukin-1 receptor antagonist (IL-IRa) and soluble interleukin-1 receptor II (sIL-1RII) by neutrophils (PMN), suggesting a significant role of IL-18 in the reactions mediated by IL-1beta. In this study, we estimated the effect of rhIL-18 on the induction of IL-6 and its soluble receptors - sIL-6Ralpha and sgp130 by these cells. Results obtained indicate that IL-18 is a promising candidate for the enhanced secretion of IL-6 by human neutrophils. In contrast, we have not found a significant effect of IL-18 on the release of both soluble receptors of IL-6. The influence of IL-18 on the IL-6 production by PMN appears to indicate a potential role of IL-18 in the early steps of the inflammatory cascade and other immune reactions mediated by IL-6.

Asunto(s)

Interleucina-18/farmacología , Interleucina-6/metabolismo , Glicoproteínas de Membrana/metabolismo , Neutrófilos/efectos de los fármacos , Receptores de Interleucina-6/fisiología , Adulto , Antígenos CD , Células Cultivadas/efectos de los fármacos , Células Cultivadas/metabolismo , Medios de Cultivo Condicionados/química , Receptor gp130 de Citocinas , Humanos , Inflamación/fisiopatología , Neutrófilos/metabolismo , Proteínas Recombinantes/farmacología , Solubilidad

RESUMEN

Vascular endothelial growth factor (VEGF) is a multifunctional cytokine that plays a pivotal role in angiogenesis in vivo. In the present study we examined the ability of polymorphonuclear neutrophils (PMN) to secrete VEGF confronted with the serum levels in oral cavity cancer patients. To investigate whether VEGF may have a prognostic importance, its value in the serum and the culture supernatants was related to the clinical course of patients. The levels of VEGF in the culture supernatant of PMN from patients were significantly higher than those from control. Increased VEGF production by PMN according to clinical progression disease, observed in the present study, seems to suggest a stimulating role of tumour cells in VEGF production by PMN. Additionally, a decrease in the ability of PMN to VEGF release after surgery may be caused by a removal of the tumour mass and then the lack the effects of tumour cells on PMN function. Results obtained appear to suggest that PMN can contribute significantly to the initiation and amplification of tumour angiogenesis and metastasis in oral cavity cancer patients. Increased values of VEGF with progression of disease and decreased values after surgery treatment clearly suggest that VEGF can play a role as a tumour marker in oral cavity cancer patients.

Asunto(s)

Carcinoma de Células Escamosas/sangre , Factores de Crecimiento Endotelial/sangre , Péptidos y Proteínas de Señalización Intercelular/sangre , Linfocinas/sangre , Neoplasias de la Boca/sangre , Proteínas de Neoplasias/sangre , Neutrófilos/metabolismo , Adulto , Anciano , Carcinoma de Células Escamosas/patología , Carcinoma de Células Escamosas/cirugía , Técnicas de Cultivo de Célula , Femenino , Humanos , Masculino , Persona de Mediana Edad , Neoplasias de la Boca/patología , Neoplasias de la Boca/cirugía , Estadificación de Neoplasias , Pronóstico , Factor A de Crecimiento Endotelial Vascular , Factores de Crecimiento Endotelial Vascular

RESUMEN

Identifying and evaluating the priming agents for cytokine release by neutrophils might be helpful in controlling the innate immune response of the host. In the present study we examined the role of granulocyte/macrophage colony-stimulating factor (GM-CSF), interferon-gamma (IFN-gamma) and tumour necrosis factor-alpha (TNF-alpha) as priming agents for interleukin (IL)-1beta, IL-6 and TNF-alpha production by stimulated neutrophils from control subjects and malignant melanoma patients. When the cells from controls and patients were preincubated with primer agents, opsonized zymosan-stimulated inflammatory cytokine production was enhanced. The major neutrophil-priming factor for IL-6 secretion by polymorphonuclear leukocytes (PMNs) in the control and patient groups was TNF-alpha. However, GM-CSF and IFN-gamma are also significant primers. GM-CSF priming was critical for the release of TNF-alpha from PMNs in control and melanoma patients. The ability of GM-CSF, IFN-gamma and TNF-alpha to serve as effective priming agents for inflammatory mediator production by PMNs revealed a new role for these cytokines in the innate immune response of the melanoma-bearing host.

Asunto(s)

Factor Estimulante de Colonias de Granulocitos y Macrófagos/farmacología , Interferón gamma/farmacología , Interleucina-1/biosíntesis , Interleucina-6/biosíntesis , Neutrófilos/efectos de los fármacos , Factor de Necrosis Tumoral alfa/farmacología , Adulto , Anciano , Células Cultivadas , Relación Dosis-Respuesta a Droga , Ensayo de Inmunoadsorción Enzimática , Humanos , Melanoma/patología , Persona de Mediana Edad , Activación Neutrófila/efectos de los fármacos , Neutrófilos/metabolismo , Proteínas Recombinantes , Neoplasias Cutáneas/patología , Factor de Necrosis Tumoral alfa/biosíntesis

RESUMEN

N-nitrosodimethylamine (NDMA) causes the apoptosis of neutrophils in vitro experiments. This compound also has the ability to stimulate neutrophils for the production of reactive oxygen species. It has been decided to examine more closely whether the apoptosis of neutrophils by NDMA is caused by the influence of the radicals produced by these cells and whether the stimulation to undergo apoptosis of neutrophils is caused by NDMA in either the original form or by its metabolites. The experiment was conducted on rats. The animals were administered a one-time dose of NDMA intragastrically, 1.5 mg/kg. The research was conducted 1,2,4,12 h consecutively following NDMA administration. The concentration of NDMA in blood was evaluated by means of the gas chromatography method. The neutrophils were isolated from blood by means of differential centrifugation. Respiratory burst was assessed in cells, by means of the cytochrome c reduction method. The percentage of cells revealing morphological properties of apoptosis was determined under the fluorescent microscope. It has been observed that the activation of the respiratory burst is caused mainly by non-metabolised NDMA. Probably the non-metabolised molecules of this compound also have a decisive role in the initiation of apoptosis of neutrophils. It can be assumed that the main factor responsible for the apoptosis of neutrophil rats following a one-time NDMA administration is the induction of respiratory burst in neutrophils by this compound.

Asunto(s)

Apoptosis/efectos de los fármacos , Agonistas de Aminoácidos Excitadores/toxicidad , N-Metilaspartato/toxicidad , Neutrófilos/efectos de los fármacos , Especies Reactivas de Oxígeno/fisiología , Animales , Masculino , Ratas , Ratas Wistar , Estallido Respiratorio/efectos de los fármacos

RESUMEN

In this paper we have presented data on an environmental exposure to N-nitrosodimethylamine (NDMA) and factors which favour endogenous biosynthesis of this compound. The factors influencing metabolism and toxicity as well as health effect of exposure have been reported.

Asunto(s)

Dimetilnitrosamina/toxicidad , Contaminantes Ambientales/toxicidad , Animales , Dimetilnitrosamina/química , Dimetilnitrosamina/metabolismo , Contaminantes Ambientales/análisis , Contaminantes Ambientales/metabolismo , Humanos , Dosificación Letal Mediana

RESUMEN

Simultaneous evaluation of cytokines and their soluble receptor production and the serum levels can be helpful in understanding the local and systemic immune response of a tumor-bearing host. In the present study we examined serum levels of TNF-alpha, IL-6 and their soluble receptors: sTNFRp55, sTNFRp75 and sIL-6R confronted with their production by the polymorphonuclear neutrophils (PMN) from cancer patients. Examinations were carried out in patients with adenocarcinoma breast cancer and squamous cell carcinoma of the oral cavity and related to the clinical course and to different phases of therapy. Secretion of IL-6, sTNFRp55 and sTNFRp75 by PMN appeared to be dependent on tumor type, clinical progression of disease as well as on therapy, suggesting a significant role of these cells at different phases of the immune response to cancer associated with these mediators. Changes in values of TNF-alpha, IL-6 and their soluble receptors in sera of both cancer groups, dependent on tumor type, clinical progression and cancer therapy, could have a diagnostic and prognostic role in cancer disease.

Asunto(s)

Interleucina-6/sangre , Neoplasias/inmunología , Receptores de Interleucina-6/sangre , Receptores del Factor de Necrosis Tumoral/sangre , Factor de Necrosis Tumoral alfa/metabolismo , Adenocarcinoma/inmunología , Adenocarcinoma/terapia , Adulto , Anciano , Antígenos CD/sangre , Neoplasias de la Mama/inmunología , Neoplasias de la Mama/terapia , Carcinoma de Células Escamosas/inmunología , Carcinoma de Células Escamosas/terapia , Estudios de Casos y Controles , Femenino , Humanos , Interleucina-6/metabolismo , Masculino , Persona de Mediana Edad , Neoplasias de la Boca/inmunología , Neoplasias de la Boca/terapia , Neoplasias/terapia , Neutrófilos/inmunología , Neutrófilos/metabolismo , Receptores Tipo I de Factores de Necrosis Tumoral , Receptores Tipo II del Factor de Necrosis Tumoral , Solubilidad

RESUMEN

In the present study, we demonstrate an effect of rhIL-15 on the simultaneous secretion of IL-1beta and its natural inhibitors IL-1Ra and sIL-1RII by human neutrophils isolated from normal and tumour-bearing hosts (oral cavity cancer and melanoma patients) compared with serum IL-15 levels. We found an rhIL-15 influence on IL-beta and IL-1Ra secreted by PMN from healthy controls. In contrast, the PMNs from cancer patients were not sensitive to rhIL-15 stimulation. However, we found a priming effect of rhIL-15 on IL-1beta production by LPS-stimulated cells in oral cavity cancer. We also found no effect on sIL-1RII release by PMN from cancer patients.

Asunto(s)

Interleucina-15/inmunología , Interleucina-1/metabolismo , Neutrófilos/inmunología , Receptores de Interleucina-1/biosíntesis , Sialoglicoproteínas/metabolismo , Carcinoma de Células Escamosas/sangre , Carcinoma de Células Escamosas/inmunología , Humanos , Proteína Antagonista del Receptor de Interleucina 1 , Interleucina-15/farmacología , Melanoma/sangre , Melanoma/inmunología , Neoplasias de la Boca/sangre , Neoplasias de la Boca/inmunología , Neutrófilos/citología , Neutrófilos/efectos de los fármacos , Neutrófilos/metabolismo , Proteínas Recombinantes/farmacología , Células Tumorales Cultivadas

RESUMEN

An inflammation or an other malignant process may create a microenvironment that modulates the production and activity of cytokines and their regulators. In the present study we compared the secretion of IL-1beta and its regulatory proteins: IL-IRa and sIL-1RII by PMN and PBMC derived from patients with inflammation and patients with cancer disease of the same location. We also examined the serum levels of these mediators in groups of patients. The results obtained revealed changes in the secretion of IL-1beta and IL-1Ra which are more characteristic of PMN and PBMC from cancer patients than of the cells from patients with inflammation. In contrast, the secretion of sIL-1RII is more characteristic of PMN and PBMC derived from patients with inflammation. Furthermore, PMN appear to play more significant role in the secretion of IL-1Ra into the circulation of cancer patients than PBMC. In contrast, PBMC affect to a large extent the secretion of IL-1beta and sIL-1RII into the circulation of patients with inflammation than PMN. Concluding, the secretion of IL-1beta and its regulatory proteins may depend on the type of immune cells and the type of the disease.

Asunto(s)

Carcinoma de Células Escamosas/inmunología , Interleucina-1/biosíntesis , Leucocitos Mononucleares/inmunología , Neutrófilos/inmunología , Receptores de Interleucina-1/biosíntesis , Sialoglicoproteínas/biosíntesis , Adulto , Anciano , Carcinoma de Células Escamosas/sangre , Medios de Cultivo , Femenino , Humanos , Proteína Antagonista del Receptor de Interleucina 1 , Interleucina-1/sangre , Masculino , Persona de Mediana Edad , Receptores de Interleucina-1/sangre , Receptores Tipo II de Interleucina-1 , Sialoglicoproteínas/sangre

RESUMEN

Evaluation of the influence of N-nitrosodimethyloamine (NDMA) on the apoptosis of neutrophils of peripheral blood (PMN) and the expression of the IL-6R membrane receptor - in vitro research. The aim of the present work was the evaluation of N-nitrosodimethyloamine (NDMA) on the induction of apoptosis in the neutrophils of peripheral blood as well as the evaluation of the surface receptors for IL-6. The isolated neutrophils were incubated for 1 and 3 hours with NDMA of a concentration of 2.5, 5, 7.5, and 10 mg/ml. In the samples incubated for 1 hour a significant, dose- dependent increase of apoptosis in the examined cells was observed. In the cells incubated for 3 hours, the increase of apoptosis was observed only at concentration of NDMA of 2.5 and 5 mg/ml. In case of higher concentration used, probably necrotic processes dominated in the cells. No influence of NDMA on the expression of IL-6R was observed.

Asunto(s)

Apoptosis/efectos de los fármacos , Dimetilnitrosamina/farmacología , Neutrófilos/efectos de los fármacos , Receptores de Interleucina-6/biosíntesis , Células Cultivadas , Humanos , Técnicas In Vitro , Leucocitos Mononucleares/citología , Neutrófilos/citología , Neutrófilos/metabolismo