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OBJECTIVE: The aim of our study was to evaluate obstetric outcome of women affected by idiopathic infertility showing persistently positive antiphospholipid antibodies (aPL). METHODS: : From 2000 consecutive patients undergoing ART, we selected 151 (7.55%) clinical records of patients affected by idiopathic infertility undergoing ICSI and showing positive aPL. RESULTS: Persistently positive aPL were found in 64/151 (42.38%) of the patients: in 34/64 (53.12%) at medium/high titers (group A) and in 30/64 (46.87%) at low titers (group B). Primary or secondary antiphospholipid syndrome (APS) was diagnosed in 25% of the patients, whereas 37.5% women showed clinical and/or laboratory features suggestive of APS, but not fulfilling clinical or laboratory classification criteria. Idiopathic infertility was the sole symptom in 31.25%. In 55% of these infertile patients, a history of recurrent failures of assisted reproductive techniques (ART) was also observed. Eighty-eight percent (88.88%) of women became pregnant and 77.77% gave birth. During pregnancy, an increase of aPL values was observed in 29.41% women of group B. CONCLUSIONS: A careful selection of patients allowed us to confirm that women affected by idiopathic infertility show a high prevalence of aPL, suggesting that these autoantibodies can also affect conception. Considering pregnancy complications and thrombotic risk related to ovarian stimulation, measuring aPL can represent a valid tool to identify among infertile women undergoing ART those at higher risk of pregnancy complications potentially life-threatening for mother and the fetus. In such patients, an accurate diagnosis and an adequate therapy are related to a better ART outcome.
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Síndrome Antifosfolípido , Infertilidad Femenina , Lupus Eritematoso Sistémico , Complicaciones del Embarazo , Anticuerpos Antifosfolípidos/inmunología , Síndrome Antifosfolípido/complicaciones , Síndrome Antifosfolípido/diagnóstico , Síndrome Antifosfolípido/inmunología , Femenino , Humanos , Infertilidad Femenina/inmunología , Lupus Eritematoso Sistémico/complicaciones , Masculino , Embarazo , Complicaciones del Embarazo/diagnósticoRESUMEN
Systemic auto-inflammatory diseases (SAID) are a group of rare inherited conditions characterized by a dysregulation of the immune system and associated with recurrent episodes of fever and systemic inflammation. Patients with NLRP12 variants develop a rare autosomal dominant condition known as familial cold-induced autoinflammatory syndrome (FCAS2, OMIM #611762) that has been related to several different clinical manifestations including autoimmunity and immune deficiencies. In past years, several new variants have been described; however, their clinical relevance is sometimes uncertain, especially when they have been detected in healthy subjects. To our knowledge 61 patients with NLRP12 variants have been reported so far in the literature. Here we report the case of a 33-year-old woman with a history of recurrent fever and symmetric and additive poly-arthritis, fulfilling diagnostic criteria for RA, who was found to harbour two variants in the NLRP12 gene (OMIM *609648) and provide a review of the literature on similar cases.
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Síndromes Periódicos Asociados a Criopirina/genética , Péptidos y Proteínas de Señalización Intracelular/genética , Mutación , Adulto , Femenino , Humanos , LinajeRESUMEN
Monocytes are a heterogeneous cell population distinguished into three subsets with distinctive phenotypic and functional properties: "classical" (CD14++CD16-), "intermediate" (CD14++CD16+), and "nonclassical" (CD14+CD16++). Monocyte subsets play a pivotal role in many inflammatory systemic diseases including atherosclerosis (ATS). Only a low number of studies evaluated monocyte behavior in patients affected by cardiovascular diseases, and data about their role in acute aortic dissection (AAD) are lacking. Thus, the aim of this study was to investigate CD14++CD16-, CD14++CD16+, and CD14+CD16++ cells in patients with Stanford-A AAD and in patients with carotid artery stenosis (CAS). Methods. 20 patients with carotid artery stenosis (CAS group), 17 patients with Stanford-A AAD (AAD group), and 17 subjects with traditional cardiovascular risk factors (RF group) were enrolled. Monocyte subset frequency was determined by flow cytometry. Results. Classical monocytes were significantly increased in the AAD group versus CAS and RF groups, whereas intermediate monocytes were significantly decreased in the AAD group versus CAS and RF groups. Conclusions. Results of this study identify in AAD patients a peculiar monocyte array that can partly explain depletion of T CD4+ lymphocyte subpopulations observed in patients affected by AAD.
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Disección Aórtica/inmunología , Estenosis Carotídea/inmunología , Monocitos/inmunología , Enfermedad Aguda , Anciano , Disección Aórtica/sangre , Estenosis Carotídea/sangre , Femenino , Citometría de Flujo , Proteínas Ligadas a GPI/análisis , Humanos , Receptores de Lipopolisacáridos/análisis , Masculino , Persona de Mediana Edad , Monocitos/clasificación , Receptores de IgG/análisis , Estudios RetrospectivosRESUMEN
BACKGROUND: Atherosclerosis is a multifactorial disease characterized by an immune-inflammatory remodeling of the arterial wall. Treg and Th17 subpopulations are detectable inside atherosclerotic plaque; however, their behavior in symptomatic carotid artery stenosis (CAS) is not fully elucidated. The aim of this study was to evaluate Th17 and Treg subsets and their ratio in patients affected by symptomatic and asymptomatic CAS. METHODS: 14 patients with symptomatic CAS (CAS-S group), 41 patients with asymptomatic CAS (CAS-A group), 32 subjects with traditional cardiovascular risk factors (RF group), and 10 healthy subjects (HS group) were enrolled. Th17 and Treg frequency was determined by flow cytometry and by histology and immunohistochemistry. Interleukin (IL)-10, IL-17, and metalloproteinase (MMP)-12 levels were measured by ELISA. RESULTS: Th17 were significantly increased in CAS-A versus RF and versus HS. Tregs were significantly increased in CAS-S versus CAS-A. Tregs/Th17 ratio was significantly reduced in CAS-A versus RF and versus HS, whereas it was significantly increased in CAS-S versus CAS-A. CONCLUSIONS: The results of this study suggest that Th17 are related to the late stages of CAS but not to plaque instability. Moreover, Treg expansion seems to represent a specific cellular pattern displayed by patients with symptomatic CAS and associated with brain injury. KEY MESSAGES Tregs expansion seems to represent a specific cellular pattern displayed by patients with symptomatic CAS and associated with CD4+ effector depletion and brain ischemic injury. Th17 lymphocytes are related to the late stages of CAS but not to plaque instability.
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Linfocitos T CD4-Positivos/metabolismo , Estenosis Carotídea/inmunología , Subunidad alfa del Receptor de Interleucina-2/metabolismo , Linfocitos T Reguladores/metabolismo , Células Th17/metabolismo , Anciano , Anciano de 80 o más Años , Estenosis Carotídea/metabolismo , Femenino , Humanos , Interleucina-10/metabolismo , Interleucina-17/metabolismo , Masculino , Metaloproteinasa 12 de la Matriz/metabolismo , Persona de Mediana EdadRESUMEN
OBJECTIVE: The aim of this study was to evaluate the levels of matrix metalloproteinase-12 (MMP-12) and tissue inhibitors of metalloproteinases (TIMP)-1, TIMP-2, TIMP-3, and TIMP-4 in patients with symptomatic and asymptomatic critical carotid artery stenosis (CAS). METHODS: We enrolled 10 patients affected by symptomatic CAS within 12 hours from onset of stroke (S group) and 30 patients with asymptomatic CAS (CAS group); 31 patients matched for age, sex, and traditional cardiovascular risk factors were used as controls (RF group). Serum levels of MMP-12, TIMP-1, TIMP-2, TIMP-3, and TIMP-4 were assessed by Luminex. RESULTS: MMP-12 levels were significantly higher both in the S and CAS groups than in the RF group (P < .001). We found a significant decrease of all TIMPs in the CAS group compared with the RF group, whereas a significant increase was observed in the S group compared with the CAS group. A significant increase of TIMP-3 and TIMP-4 levels was observed in the S group compared with all other groups. CONCLUSION: MMP-12 is related to critical CAS both symptomatic and asymptomatic, being mainly released in the late stage of plaque development. Moreover, we suggest that a specific pattern of matrix degrading enzyme inhibitors arises during the early phases of stroke.
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Estenosis Carotídea/sangre , Metaloproteinasa 12 de la Matriz/sangre , Inhibidor Tisular de Metaloproteinasa-1/sangre , Inhibidor Tisular de Metaloproteinasa-2/sangre , Inhibidor Tisular de Metaloproteinasa-3/sangre , Inhibidores Tisulares de Metaloproteinasas/sangre , Anciano , Biomarcadores/sangre , Femenino , Humanos , Masculino , Persona de Mediana Edad , Accidente Cerebrovascular/sangre , Inhibidor Tisular de Metaloproteinasa-4RESUMEN
Acute aortic dissection (AAD) is a life-threatening disease with an incidence of about 2.6-3.6 cases per 100,000/year. Depending on the site of rupture, AAD is classified as Stanford-A when the ascending aortic thoracic tract and/or the arch are involved, and Stanford-B when the descending thoracic aorta and/or aortic abdominal tract are targeted. It was recently shown that inflammatory pathways underlie aortic rupture in both type A and type B Stanford AAD. An immune infiltrate has been found within the middle and outer tunics of dissected aortic specimens. It has also been observed that the recall and activation of macrophages inside the middle tunic are key events in the early phases of AAD. Macrophages are able to release metalloproteinases (MMPs) and pro-inflammatory cytokines which, in turn, give rise to matrix degradation and neoangiogenesis. An imbalance between the production of MMPs and MMP tissue inhibitors is pivotal in the extracellular matrix degradation underlying aortic wall remodelling in dissections occurring both in inherited conditions and in atherosclerosis. Among MMPs, MMP-12 is considered a specific marker of aortic wall disease, whatever the genetic predisposition may be. The aim of this review is, therefore, to take a close look at the immune-inflammatory mechanisms underlying Stanford-A AAD.
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Aneurisma de la Aorta Torácica/metabolismo , Disección Aórtica/enzimología , Disección Aórtica/patología , Inflamación/enzimología , Inflamación/patología , Metaloproteasas/metabolismo , Animales , Aneurisma de la Aorta Torácica/complicaciones , Aneurisma de la Aorta Torácica/epidemiología , HumanosRESUMEN
The aim of this study was to evaluate fibroblast growth factor (FGF)-23 serum levels and its tissue expression in patients with critical carotid artery stenosis (CAS). We selected 35 patients with critical CAS undergoing carotid thromboendoarterectomy. In each patient, FGF-23 serum levels were evaluated just prior to the surgery (t0) and 30 min (t1) thereafter. Moreover, macrophage cytokines were measured at baselines. Carotid artery specimens were used for immune histochemistry. On the basis of the histology, the patients were divided into 2 groups: A with complicated plaque and B with uncomplicated plaque. Twenty complicated plaques (57.14%, group A,) and 15 uncomplicated (42.86%, group B) were evaluated: calcifications were present in 16/20 (80%) complicated plaques and in 6/15 (40%) uncomplicated plaques. An inflammatory infiltrate was observed in 26/35 carotid samples: 18/26 (69.23%) complicated and 8/26 (30.76%) uncomplicated. FGF-23(+) cells were present in 17/20 complicated (85%) and in 8 uncomplicated (53%) plaques. The double-staining immunofluorescence confirmed that macrophage cells (CD68(+)) were also positive for FGF-23 staining. Serum levels of FGF-23 were significantly higher in group A versus group B at t0 (p < 0.05) and t1 (p 0.0047). Moreover, in group A patients a significant increase of FGF-23 serum levels was observed at t1 in comparison with t0 (p 0.0011). Our results suggest that FGF-23 acts in the late phases of atherosclerotic disease and may potentially represent a marker of complications in critical CAS.
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Estenosis Carotídea/metabolismo , Factores de Crecimiento de Fibroblastos/metabolismo , Placa Aterosclerótica/metabolismo , Anciano , Biomarcadores/metabolismo , Estenosis Carotídea/patología , Estenosis Carotídea/cirugía , Estudios de Casos y Controles , Endarterectomía Carotidea , Femenino , Factor-23 de Crecimiento de Fibroblastos , Humanos , Masculino , Persona de Mediana Edad , Placa Aterosclerótica/patología , Placa Aterosclerótica/cirugía , Factores de RiesgoRESUMEN
Among polymeric polycations, chitosan has emerged as a powerful carrier for gene delivery. Only a few studies have focused on the stability of the chitosan/DNA complex under storage, although this is imperative for nanomedicinal applications. Here, we synthesized polyelectrolyte complexes at a charge ratio of 10 using 50 kDa chitosan and plasmid (p)DNA that encodes a GFP reporter. These preparations were stable up to 3 months at 4 °C and showed reproducible transfection efficiencies in vitro in HEK293 cells. In addition, we developed a methodology that increases the in vitro transfection efficiency of chitosan/pDNA complexes by 150% with respect to standard procedures. Notably, intracellular pDNA release and transfected cells peaked 5 days following transection of mitotically active cells. These new developments in formulation technology enhance the potential for polymeric nanoparticle-mediated gene therapy.
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Quitosano/metabolismo , ADN/metabolismo , Técnicas de Transferencia de Gen , Plásmidos , Transfección/métodos , Línea Celular , Estabilidad de Medicamentos , Células Epiteliales/metabolismo , Humanos , Reproducibilidad de los Resultados , Temperatura , Factores de Tiempo , Transformación GenéticaRESUMEN
BACKGROUND: The study evaluated macrophage cytokines and macrophage metalloprotease (MMP)-12 levels in patients with Stanford-A acute aortic dissection (AAD) and in patients with critical carotid artery stenosis (CAS) compared with patients matched for age, sex, and traditional cardiovascular risk factors (RF). The aim was to identify possible early serum markers of risk for atherosclerotic complications. MATERIALS AND METHODS: We selected 65 patients: 23 AAD patients, 21 CAS patients, 21 RF, and 10 healthy subjects (HS). In each patient and control serum, levels of interleukin (IL)-6, IL-8, tumour necrosis factor (TNF)-α, monocyte chemoattractant protein (MCP)-1, vascular endothelial growth factor (VEGF), and MMP-12 were assessed by ELISA. RESULTS: A significant increase of MMP-12, IL-6, and IL-8 levels in AAD versus CAS was found. Moreover, MMP-12 was shown to be significantly higher in AAD versus RF, but not in CAS versus RF. A significant increase of IL-6, IL-8, MCP-1, TNF-α, and VEGF levels was observed both in AAD and CAS versus RF. CONCLUSIONS: The results suggest that MMP-12 may be considered to be a specific marker of Stanford-A AAD. Furthermore, the study confirmed that in AAD and CAS macrophage cytokines play a key role in the progression of the atherosclerotic disease towards complications.
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Enfermedades de la Aorta/sangre , Metaloproteinasa 12 de la Matriz/sangre , Enfermedad Aguda , Anciano , Anciano de 80 o más Años , Enfermedades de la Aorta/enzimología , Enfermedades de la Aorta/patología , Aterosclerosis/sangre , Aterosclerosis/enzimología , Aterosclerosis/patología , Biomarcadores/sangre , Estenosis Carotídea/sangre , Estenosis Carotídea/enzimología , Estenosis Carotídea/patología , Estudios de Casos y Controles , Femenino , Humanos , Activación de Macrófagos , Macrófagos/enzimología , Macrófagos/patología , Masculino , Persona de Mediana EdadRESUMEN
Pseudomonas aeruginosa is the most common pathogen for chronic lung infection in cystic fibrosis (CF) patients. About 80% of adult CF patients have chronic P. aeruginosa infection, which accounts for much of the morbidity and most of the mortality. Both bacterial genetic adaptations and defective innate immune responses contribute to the bacteria persistence. It is well accepted that CF transmembrane conductance regulator (CFTR) dysfunction impairs the airways-epithelium-mediated lung defence; however, other innate immune cells also appear to be affected, such as neutrophils and macrophages, which thus contribute to this infectious pathology in the CF lung. In macrophages, the absence of CFTR has been linked to defective P. aeruginosa killing, increased pro-inflammatory cytokine secretion, and reduced reactive oxygen species (ROS) production. To learn more about macrophage dysfunction in CF patients, we investigated the generation of the oxidative burst and its impact on bacterial killing in CF macrophages isolated from peripheral blood or lung parenchyma of CF patients, after P. aeruginosa infection. Our data demonstrate that CF macrophages show an oxidative response of similar intensity to that of non-CF macrophages. Intracellular ROS are recognized as one of the earliest microbicidal mechanisms against engulfed pathogens that are activated by macrophages. Accordingly, NADPH inhibition resulted in a significant increase in the intracellular bacteria survival in CF and non-CF macrophages, both as monocyte-derived macrophages and as lung macrophages. These data strongly suggest that the contribution of ROS to P. aeruginosa killing is not affected by CFTR mutations.
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Fibrosis Quística/microbiología , Fibrosis Quística/patología , Macrófagos/microbiología , Macrófagos/patología , Viabilidad Microbiana , Pseudomonas aeruginosa/fisiología , Especies Reactivas de Oxígeno/metabolismo , Adolescente , Adulto , Separación Celular , Inhibidores Enzimáticos/farmacología , Femenino , Humanos , Espacio Intracelular/microbiología , Pulmón/patología , Macrófagos/efectos de los fármacos , Macrófagos/enzimología , Masculino , Viabilidad Microbiana/efectos de los fármacos , Persona de Mediana Edad , NADPH Oxidasas/antagonistas & inhibidores , NADPH Oxidasas/metabolismo , Óxido Nítrico Sintasa de Tipo II/antagonistas & inhibidores , Óxido Nítrico Sintasa de Tipo II/metabolismo , Fenotipo , Infecciones por Pseudomonas/patología , Pseudomonas aeruginosa/efectos de los fármacos , Estallido Respiratorio/efectos de los fármacos , Adulto JovenRESUMEN
Vertebrate-like T2AG3 telomeres in tlc1-h yeast consist of short double-stranded regions and long single-stranded overhang (G-tails) and, although based on Tbf1-capping activity, they are capping deficient. Consistent with this idea, we observe Y' amplification because of homologous recombination, even in the presence of an active telomerase. In these cells, Y' amplification occurs by different pathways: in Tel1(+) tlc1h cells, it is Rad51-dependent, whereas in the absence of Tel1, it depends on Rad50. Generation of telomeric G-tail, which is cell cycle regulated, depends on the MRX (Mre11-Rad50-Xrs2) complex in tlc1h cells or is MRX-independent in tlc1h tel1Δ mutants. Unexpectedly, we observe telomere elongation in tlc1h lacking Rad51 that seems to act as a telomerase competitor for binding to telomeric G-tails. Overall, our results show that Tel1 and Rad51 have multiple roles in the maintenance of vertebrate-like telomeres in yeast, supporting the idea that they may participate to evolutionary conserved telomere protection mechanism/s acting at uncapped telomeres.
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Péptidos y Proteínas de Señalización Intracelular/fisiología , Proteínas Serina-Treonina Quinasas/fisiología , Recombinasa Rad51/fisiología , Proteínas de Saccharomyces cerevisiae/fisiología , Homeostasis del Telómero , Telómero/metabolismo , Proteínas de Unión al ADN/genética , Eliminación de Gen , Péptidos y Proteínas de Señalización Intracelular/genética , Proteínas Serina-Treonina Quinasas/genética , Recombinasa Rad51/metabolismo , Recombinación Genética , Saccharomyces cerevisiae/genética , Saccharomyces cerevisiae/metabolismo , Proteínas de Saccharomyces cerevisiae/genética , Proteínas de Saccharomyces cerevisiae/metabolismo , Telomerasa/antagonistas & inhibidoresRESUMEN
Bovine tuberculosis (bTB) is an emerging disease among wild animals in many parts of the world. Wildlife reservoir hosts may thus represent a potential source of infection for livestock and humans. We investigated the role played by the Sicilian black pig, an autochthonous free- or semi-free-ranging domestic pig breed, as a potential source of bTB infection in an area where bTB prevalence in cattle is high. We initially performed a preliminary field study to assess the occurrence of bTB in such animals. We sampled 119 pigs at abattoir and found 6.7% and 3.4% of them to be affected by gross tuberculous-like lesions (TBL) and Mycobacterium bovis culture positive, respectively. We then proceeded to investigate the dissemination and characteristics of lesions in a second field study performed on 100 animals sampled from infected herds. Here, tissues collected at the abattoir were examined macroscopically, microscopically, and by culture tests. Most pigs with TBL showed generalized lesions in both gross and histological examinations (53% and 65.5%, respectively). Head lymph nodes were the most frequently affected in both localized and generalized TB cases observed macroscopically and microscopically. M. bovis was the most frequently isolated etiologic agent. The molecular characterization of isolates from both field studies by spoligotyping and analysis of 12 mycobacterial interspersed repetitive-unit-variable number tandem repeat (MIRU-VNTR) loci, followed by their comparison to isolates of cattle origin, suggested a potential transmission of mycobacteria from domestic animals to black pigs and vice versa. Our findings, along with ethological, ecological, and management considerations, suggest that the black pig might act as a bTB reservoir in the ecosystem under study. However, additional studies will be necessary to establish the true epidemiological significance of the Sicilian black pig.
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Reservorios de Enfermedades , Mycobacterium bovis/aislamiento & purificación , Sus scrofa/microbiología , Enfermedades de los Porcinos/epidemiología , Tuberculosis Bovina/epidemiología , Animales , Técnicas de Tipificación Bacteriana , Bovinos , Granuloma/microbiología , Granuloma/patología , Cabeza/microbiología , Cabeza/patología , Ganglios Linfáticos/microbiología , Ganglios Linfáticos/patología , Tipificación de Secuencias Multilocus , Mycobacterium bovis/genética , Sicilia/epidemiología , Porcinos , Enfermedades de los Porcinos/diagnóstico , Enfermedades de los Porcinos/microbiología , Tuberculosis Bovina/diagnóstico , Tuberculosis Bovina/microbiologíaRESUMEN
Burkholderia cenocepacia is an important human pathogen in patients with cystic fibrosis (CF). Non-clinical reservoirs may play a role in the acquisition of infection, so it is important to evaluate the pathogenic potential of environmental B. cenocepacia isolates. In this study, we investigated the interactions of two environmental B. cenocepacia strains (Mex1 and MCII-168) with two bronchial epithelial cell lines, 16HBE14o(-) and CFBE41o(-), which have a non-CF and a CF phenotype, respectively. The environmental strains showed a significantly lower level of invasion into both CF and non-CF cells in comparison with the clinical B. cenocepacia LMG16656(T) strain. Exposure of polarized CFBE41o(-) or 16HBE14o(-) cells to the environmental strains resulted in a significant reduction in transepithelial resistance (TER), comparable with that observed following exposure to the clinical strain. A different mechanism of tight junction disruption in CF versus non-CF epithelia was found. In the 16HBE41o(-) cells, the environmental strains resulted in a drop in TER without any apparent effect on tight junction proteins such as zonula occludens-1 (ZO-1). In contrast, in CF cells, the amount of ZO-1 and its localization were clearly altered by the presence of both the environmental strains, comparable with the effect of LMG16656. This study demonstrates that even if the environmental strains are significantly less invasive than the clinical strain, they have an effect on epithelial integrity comparable with that of the clinical strain. Finally, the tight junction regulatory protein ZO-1 appears to be more susceptible to the presence of environmental strains in CF cells than in cells which express a functional cystic fibrosis transmembrane regulator (CFTR).
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Infecciones por Burkholderia/patología , Burkholderia cenocepacia/patogenicidad , Fibrosis Quística/microbiología , Células Epiteliales/microbiología , Bronquios/citología , Línea Celular , Humanos , Proteínas de la Membrana/metabolismo , Fosfoproteínas/metabolismo , Rizosfera , Uniones Estrechas/microbiología , Zea mays/microbiología , Proteína de la Zonula Occludens-1RESUMEN
Chronic inflammation of the lung, as a consequence of persistent bacterial infections by several opportunistic pathogens represents the main cause of mortality and morbidity in cystic fibrosis (CF) patients. Mechanisms leading to increased susceptibility to bacterial infections in CF are not completely known, although the involvement of cystic fibrosis transmembrane conductance regulator (CFTR) in microbicidal functions of macrophages is emerging. Tissue macrophages differentiate in situ from infiltrating monocytes, additionally, mature macrophages from different tissues, although having a number of common activities, exhibit variation in some molecular and cellular functions. In order to highlight possible intrinsic macrophage defects due to CFTR dysfunction, we have focused our attention on in vitro differentiated macrophages from human peripheral blood monocytes. Here we report on the contribution of CFTR in the bactericidal activity against Pseudomonas aeruginosa of monocyte derived human macrophages. At first, by real time PCR, immunofluorescence and patch clamp recordings we demonstrated that CFTR is expressed and is mainly localized to surface plasma membranes of human monocyte derived macrophages (MDM) where it acts as a cAMP-dependent chloride channel. Next, we evaluated the bactericidal activity of P. aeruginosa infected macrophages from healthy donors and CF patients by antibiotic protection assays. Our results demonstrate that control and CF macrophages do not differ in the phagocytic activity when infected with P. aeruginosa. Rather, although a reduction of intracellular live bacteria was detected in both non-CF and CF cells, the percentage of surviving bacteria was significantly higher in CF cells. These findings further support the role of CFTR in the fundamental functions of innate immune cells including eradication of bacterial infections by macrophages.
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Regulador de Conductancia de Transmembrana de Fibrosis Quística/fisiología , Macrófagos/inmunología , Pseudomonas aeruginosa/inmunología , Adolescente , Adulto , Femenino , Humanos , Masculino , Microscopía Confocal , Microscopía Fluorescente , Persona de Mediana EdadRESUMEN
The importance of probiotics in human nutrition has been gaining recognition in recent years. These organisms have been shown to promote human health by enhancing immunological and digestive functions and fighting respiratory tract infections. We propose an improved in vitro model for the study of probiotic antimicrobial activity against enteropathogens, by attempting to re-create, in a common culture medium, environmental growth conditions comparable to those present in the small intestine. A preliminary experiment was carried out in order to find a culture medium able to support both probiotics and pathogens. This was done with the aim of obtaining correct assessment of the interaction under shared growth conditions. BHI medium was selected as the common culture medium and was therefore used in antimicrobial activity assays. The interactions between Salmonella 1344 and Lactobacillus rhamnosus and Lactobacillus reuteri were then assessed at different pH and oxygen availability conditions mimicking the small intestinal environment. L. rhamnosus GG ATCC 53103 (LGG) had the strongest antimicrobial effect, in particular under anaerobic conditions and at lower pH levels. Its antagonistic activity involved both lactic acid and secreted non-lactic acid molecules. Our findings suggest that each probiotic strain has an optimum range of action and should therefore be thoroughly investigated to optimize its use.
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Antibiosis , Bacillus/fisiología , Lacticaseibacillus rhamnosus/fisiología , Limosilactobacillus reuteri/fisiología , Probióticos , Salmonella typhimurium/crecimiento & desarrollo , Anaerobiosis , Bacteriocinas/metabolismo , Medios de Cultivo , Concentración de Iones de Hidrógeno , Mucosa Intestinal/microbiología , Intestino Delgado/microbiología , Estrés OxidativoRESUMEN
The present report describes a rare case of generalized bovine-type tuberculosis in a slaughtered 4-year-old ewe discovered during routine surveillance at an abattoir. A postmortem examination revealed lesions in the ewe's thoracic and abdominal cavities, ranging from encapsulated, mineralized foci to extensive, soft, caseous tissue. Lesions in the lungs, liver, and lymph nodes were consistent with mycobacterial infection. A histopathological examination detected granulomatous lesions in all tissue samples. The presence of Mycobacterium tuberculosis complex genome was confirmed through a polymerase chain reaction (PCR) analysis of tissues, using IS6110 primers, followed by a nucleotide sequence analysis of PCR products. Acid-fast bacteria, characterized as Mycobacterium bovis, were isolated from lesions following 38 days of incubation.