RESUMEN
The Barton-Zard reaction of ß-fluoro-ß-nitrostyrenes with ethyl α-isocyanoacetate was studied. The reaction was found to proceed in a highly chemoselective manner to form preferably 4-fluoropyrroles in up to 77% yield. The corresponding 4-nitrosubstituted pyrroles are formed as minor products of the reaction. The broad scope of ß-fluoro-ß-nitrostyrenes was demonstrated in the preparation of a variety of fluorinated pyrroles. The obtained experimental results are in perfect agreement with the data obtained by theoretical investigation of this reaction. The subsequent study of synthetic utility of monofluorinated pyrroles was performed to open a way for the development of a variety of functionalized pyrrole derivatives.
RESUMEN
The Diels-Alder reaction of ß-fluoro-ß-nitrostyrenes with cyclic 1,3-dienes was investigated. A series of novel monofluorinated norbornenes was prepared in up to 97% yield. The reaction with 1,3-cyclohexadiene permits the preparation of monofluorinated bicyclo[2.2.2]oct-2-enes. The kinetic data of the reactions with 1,3-cyclopentadiene and 1,3-cyclohexadiene were used to calculate activation parameters. Furthermore, the synthetic utility of the cycloadducts obtained was demonstrated.
RESUMEN
Autophagy is a highly conserved mechanism of eukaryotic cells implicated in cell homeostasis and elimination of intracellular pathogens. Functional polymorphisms in genes encoding for autophagy have been associated with susceptibility to inflammatory and infectious diseases, but data on severe infections are missing. The aim of the present study was to assess whether polymorphisms in genes encoding proteins involved in autophagy influence susceptibility to ventilator-associated pneumonia (VAP). Mechanically ventilated patients with VAP were studied. Genotyping for autophagy-related 16-like 1 (ATG16L1, rs2241880) functional polymorphism was performed using the TaqMan single-nucleotide assay. Monocytes were isolated from patients and stimulated with lipopolysaccharide (LPS). Tumor necrosis factor-α (TNF-α) was measured in the supernatants of monocytes using an enzyme-linked immunosorbent assay. Procalcitonin (PCT) was also measured in the serum of patients by an immuno-time-resolved amplified cryptate technology assay. A total of 155 patients with VAP were enrolled in the study. Carriage of the minor A allele of ATG16L1 was associated with septic shock with at least one organ failure (odds ratio (OR): 2.40, p: 0.036). TNF-α production was significantly greater among the carriers of the polymorphism presenting with at least one organ failure (p: 0.040). PCT was increased upon worsening to septic shock and organ failure only among carriers of the minor frequency A alleles. In a homogeneous cohort of septic patients with VAP, the carriage of autophagy polymorphisms predisposes to VAP severity and septic shock development. This may be related with predisposition to immunoparalysis.
Asunto(s)
Proteínas Portadoras/genética , Predisposición Genética a la Enfermedad , Neumonía Asociada al Ventilador/patología , Polimorfismo Genético , Sepsis/patología , Adolescente , Adulto , Anciano , Anciano de 80 o más Años , Proteínas Relacionadas con la Autofagia , Calcitonina/sangre , Péptido Relacionado con Gen de Calcitonina , Femenino , Genotipo , Técnicas de Genotipaje , Humanos , Masculino , Persona de Mediana Edad , Precursores de Proteínas/sangre , Factor de Necrosis Tumoral alfa/metabolismo , Adulto JovenRESUMEN
BACKGROUND: Recent evidence has suggested that deranged immune responses play a role in the pathogenesis of hidradenitis suppurativa (HS). OBJECTIVES: To investigate the role of single nucleotide polymorphisms (SNPs) of the tumour necrosis factor (TNF) and Toll-like receptor 4 (TLR4) genes in the physical course of HS; these genes encode for proteins implicated in the immune response of the host. METHODS: DNA was isolated from 190 patients with HS and 84 healthy controls. SNPs at the promoter regions -376G/A, -238G/A and -308G/A of the TNF gene and the Asp299Gly and Thr399Ile SNPs of the TLR4 gene were determined by polymerase chain reaction (PCR) and digestion of the PCR product by restriction enzymes; after electrophoresis on 2·0% agarose gel, products were visualized on under ultraviolet radiation. RESULTS: The presence of the -238 TNF gene polymorphism was associated with a predisposition to HS (P = 0·027). Susceptibility to the disease was strongly correlated with the presence of AGG/GGA/AGA/GAA TNF haplotypes in 32 (17%) patients compared with two (2%) controls (P < 0·001, odds ratio 8·30, 95% confidence interval 1·94-35·52). The frequency of HS exacerbations and disease severity were greater in patients carrying any of the GAG/AGG/GGA/AGA/GAA haplotypes of the TNF gene. Thirty-two patients were given TNF antagonists. Nineteen of these patients were carriers of the GGG haplotype of the TNF gene, whereas 13 were carriers of other haplotypes; favourable responses as evidenced by the Sartorius score were registered in 15 (79%) and five (38%, P = 0·025), respectively. Carriage of the TLR4 gene alleles was not associated with any disease parameter. CONCLUSIONS: A significant role of SNPs at the promoter region of the TNF gene is indicated for susceptibility to HS and for response to TNF antagonists.
Asunto(s)
Hidradenitis Supurativa/genética , Polimorfismo de Nucleótido Simple/genética , Receptor Toll-Like 4/genética , Factor de Necrosis Tumoral alfa/genética , Adulto , Anticuerpos Monoclonales/uso terapéutico , Estudios de Casos y Controles , Fármacos Dermatológicos/uso terapéutico , Etanercept , Femenino , Predisposición Genética a la Enfermedad/genética , Genotipo , Haplotipos , Hidradenitis Supurativa/tratamiento farmacológico , Hidradenitis Supurativa/inmunología , Humanos , Inmunoglobulina G/uso terapéutico , Infliximab , Masculino , Reacción en Cadena de la Polimerasa , Regiones Promotoras Genéticas/genética , Estudios Prospectivos , Receptores del Factor de Necrosis Tumoral/uso terapéutico , Factor de Necrosis Tumoral alfa/antagonistas & inhibidoresRESUMEN
Regulatory T cells (T(regs) ) have an anti-inflammatory role. A former study in a limited number of patients found that absolute counts of T(regs) increase when infection by the new influenza H1N1 virus is complicated with pneumonia. These results generate the question if H1N1-related pneumonia is associated with a state of hypo-inflammation. A total of 135 patients were enrolled with blood sampling within less than 24 h from diagnosis; 23 with flu-like syndrome; 69 with uncomplicated H1N1-infection; seven with bacterial pneumonia; and 36 with H1N1-related pneumonia. T(regs) and CD14/HLA-DR co-expression were estimated by flow cytometry; concentrations of tumour necrosis factor-alpha (TNF-α), of interleukin (IL)-6 and of soluble triggering receptor expressed on myeloid cells-1 (sTREM-1) by an enzyme immunoassay; those of procalcitonin (PCT) by immuno-time-resolved amplified cryptate technology assay. Expression of human leucocyte antigen D-related (HLA-DR) on monocytes was similar between groups; absolute T(reg) counts were greater among patients with H1N1-related pneumonia than flu-like syndrome or H1N1-uncomplicated infection. Serum TNF-α of patients with bacterial pneumonia was greater than those of other groups, but IL-10 was similar between groups. Serum PCT was greater among patients with H1N1-related pneumonia and sTREM-1 among those with H1N1-related pneumonia. Regression analysis revealed that the most important factors related with the advent of pneumonia were the existence of underlying illnesses (P = 0·006) and of T(regs) equal to or above 16 mm(3) (P = 0·013). It is concluded that the advent of H1N1-related pneumonia is related to an early increase of the absolute T(reg) counts. This increase is probably not part of a hypo-inflammatory state of the host.
Asunto(s)
Subtipo H1N1 del Virus de la Influenza A/inmunología , Gripe Humana/inmunología , Neumonía Bacteriana/inmunología , Linfocitos T Reguladores/inmunología , Adulto , Anciano , Femenino , Citometría de Flujo , Antígenos HLA-DR/metabolismo , Humanos , Inmunofenotipificación , Gripe Humana/sangre , Gripe Humana/complicaciones , Interleucina-6/sangre , Receptores de Lipopolisacáridos/metabolismo , Masculino , Glicoproteínas de Membrana/sangre , Persona de Mediana Edad , Neumonía Bacteriana/sangre , Neumonía Bacteriana/complicaciones , Receptores Inmunológicos/sangre , Linfocitos T Reguladores/metabolismo , Receptor Activador Expresado en Células Mieloides 1 , Factor de Necrosis Tumoral alfa/sangre , Adulto JovenRESUMEN
S-Nitrosoalbumin (SNOALB) is the most abundant physiological circulating nitric oxide (NO) carrier regulating NO-dependent biological actions in humans. The mechanisms of its formation and biological actions are still incompletely understood. Nitrosation by authentic NO and S-transnitrosylation of the single sulfhydryl group located at Cys-34 of human albumin by the physiological S-nitroso compounds S-nitrosocysteine (SNOC) and S-nitrosoglutathione (GSNO) are two possible mechanisms. On a quantitative basis, we investigated by gas chromatography-mass spectrometry the contribution of these two mechanisms to SNOALB formation in human plasma and blood in vitro. GSNO and SNOC (0-100 microM) rapidly and efficiently (recovery=35%) S-transnitrosylated albumin to form SNOALB. NO (100 microM) S-nitrosated albumin to SNOALB at a considerably lower extent (recovery=5%). The putative NO-donating drugs glyceryl trinitrate and sodium nitroprusside (each 100 microM) failed completely in S-nitrosating albumin. Bubbling NO into human plasma and blood resulted in formation of SNOALB that inhibited ADP-induced platelet aggregation. Infusion of GS(15)NO in the rat resulted in formation of S(15)NOALB, [(15)N]nitrate and [(15)N]nitrite. Our results suggest that S-transnitrosylation of albumin by SNOC and GSNO could be a more favored mechanism for the formation of SNOALB in the circulation in vivo than S-nitrosation of albumin by NO itself.
Asunto(s)
Cisteína/análogos & derivados , Glutatión/análogos & derivados , S-Nitrosotioles , Albúmina Sérica Bovina/biosíntesis , Albúmina Sérica/metabolismo , Adenosina Difosfato/farmacología , Animales , Cisteína/farmacología , Relación Dosis-Respuesta a Droga , Cromatografía de Gases y Espectrometría de Masas/métodos , Glutatión/farmacología , Humanos , Técnicas In Vitro , Óxido Nítrico/metabolismo , Óxido Nítrico/farmacología , Donantes de Óxido Nítrico/farmacología , Nitroglicerina/farmacología , Nitroprusiato/farmacología , Compuestos Nitrosos/farmacología , Agregación Plaquetaria/efectos de los fármacos , Ratas , S-Nitrosoglutatión , Albúmina Sérica/química , Albúmina Sérica/efectos de los fármacos , Albúmina Sérica Bovina/farmacología , Compuestos de Sulfhidrilo/química , Compuestos de Sulfhidrilo/metabolismoRESUMEN
Epstein-Barr virus (EBV) is a known oncogenic virus associated with a wide variety of cancers, including nasopharyngeal carcinoma. Waldeyer's ring, a collection of lymphoid tissues, includes the nasopharynx, pharyngeal, and lingual tonsils. To determine if EBV plays a causative role in carcinomas arising from other tissues in Waldeyer's ring, we examined pharyngeal tonsillar carcinomas for evidence of EBV infection. As previously reported, DNA was extracted from 53 consecutive tonsil cancers, as well as from age- and gender-matched non-cancerous tonsillectomy specimens. Three different sets of primers for discrete exons of EBV were then used to determine if active or latent EBV infection was expressed in the extracted DNA using the polymerase chain reaction (PCR). All positive bands were then sequenced to confirm the presence of amplified EBV fragments. None of the samples showed evidence for active EBV infection. In primers demonstrating latent infection, 1 of 53 (1.9%) of tumors were positive, versus 6 of 53 (11.3%) of the controls. These results indicate that EBV expression is not increased in DNA from tonsil cancers and that EBV infection does not have a causal relationship with tonsil cancer.
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Carcinoma de Células Escamosas/química , ADN Viral/análisis , Herpesvirus Humano 4/genética , Neoplasias Tonsilares/química , Carcinoma de Células Escamosas/virología , Infecciones por Virus de Epstein-Barr/metabolismo , Femenino , Humanos , Masculino , Persona de Mediana Edad , Reacción en Cadena de la Polimerasa , Neoplasias Tonsilares/virologíaRESUMEN
BACKGROUND: Elevated serum levels of the cell adhesion molecule E-cadherin have been associated with the presence of tissue injury and inflammation. We compared soluble E-cadherin response during laparoscopic and open cholecystectomy. METHODS: The E-cadherin response to surgery was studied in 16 patients undergoing laparoscopic cholecystectomy and 12 patients undergoing open cholecystectomy. Serum E-cadherin levels were measured by an enzyme immunoassay (ELISA) preoperatively, 10 and 30 min after the commencement of surgery, and at 6 and 24 h following the operation. RESULTS: Serum E-cadherin levels decreased progressively during laparoscopic cholecystectomy; their concentrations at 24 h after surgery were significantly lower when compared with preoperative values. In the open cholecystectomy group, serum E-cadherin levels did not differ from preoperative values at any time point. Serum E-cadherin concentrations at 24 h after surgery and the cumulative E-cadherin response were significantly higher in the open cholecystectomy group than in the laparoscopic group. CONCLUSION: Compared with open cholecystectomy, the cumulative E-cadherin response is significantly reduced following laparoscopic cholecystectomy.
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Cadherinas/sangre , Colecistectomía Laparoscópica/efectos adversos , Colecistectomía Laparoscópica/métodos , Colecistectomía/efectos adversos , Colecistectomía/métodos , Femenino , Humanos , Inflamación/sangre , Inflamación/cirugía , Masculino , Persona de Mediana Edad , Estrés Fisiológico/sangre , Estrés Fisiológico/cirugía , Factores de TiempoRESUMEN
A gas chromatographic-mass spectrometric method for the determination of nitric oxide synthase activity is described. The method is based on the gas chromatographic-mass spectrometric measurement of L-[15N2]arginine-derived [15N]nitrite as its pentafluorobenzyl derivative in the negative-ion chemical ionization mode. Application of the method to the analysis of [15N]nitrite formation by purified neuronal nitric oxide synthase revealed K(M) values of 3.1 microM by Hanes and 4.6 microM by Lineweaver-Burk for L-[15N2]arginine. The corresponding Vmax values were 0.204 and 0.228 micromol [15N]nitrite min(-1) mg(-1) NOS, respectively. N(G)-Nitro-L-arginine and N(G),N(G)-dimethylarginine (asymmetric dimethylarginine) were identified by this method as the most potent enzyme inhibitors. Nitric oxide synthase activity was also assessed in vivo by i.v. injection of L-[15N2]arginine in a rat and determination of plasma [15N]nitrite and [15N]nitrate. The assay described in this work allows for accurate, specific and highly sensitive determination of nitric oxide synthase activity in vitro and in vivo.