25 research outputs found
Macrophage phenotype and function in primary lung cancer
Background: Lung cancer is one of the most commonly reported cancers, and is known to be associated with a poor prognosis. The role of macrophages in lung cancer, including alveolar macrophages (AMs), tumour-associated macrophages (TAMs) and monocytes, is multifaceted and the literature shows conflicting roles. Aims: (I) To characterise the M1/M2 macrophage populations within the AM population in patients with non-small lung carcinoma (NSCLC) compared to controls; (II) To evaluate the M1/M2 monocyte populations and Th1/Th2 cytokine serum levels in patients with NSCLC compared to controls; (III) To use quantitative proteomics to investigate the up-regulation of novel proteins in bronchoalveolar lavage (BAL) fluid from patients with lung adenocarcinoma to identify new potential biomarkers expressed by AMs; and (IV) To characterise the M1/M2 macrophage populations within TAMs in different subtypes of NSCLC compared to non-tumour tissues. Methods and materials: AMs were obtained from patients with NSCLC and controls and analysed for surface marker differences using flow cytometry. The mRNA expression of different cytokines was measured using qPCR. Freshly prepared peripheral blood mononuclear cell (PBMC) samples were also obtained from patients with NSCLC and from controls. Flow cytometry was performed to investigate the expression of M1/M2 markers on classical monocytes. The Th1/Th2 cytokine levels were analysed in serum samples using the cytometric bead array (CBA) and the Bio-Plex systems. In addition, BAL fluid samples from subjects with and without lung adenocarcinoma were analysed using quantitative proteomics. Finally, TAM subsets from non-tumour and tumour tissues were analysed using immunohistochemistry (IHC). Results: The expression of M2 markers (CD163), CD71 and CD44 was greater on AMs from NSCLC patients compared to controls. However, there were no significant differences in the expression of M1 markers (HLA-DR) and CD11b. The mRNA expression levels of IL-10 and MMP-9 were increased in AMs from NSCLC patients compared to controls. There were no significant differences in the expression of HLA-DR, CD163, CD11b, CD11c, CD71 and CD44 markers on classical monocytes in NSCLC patients compared to controls. The Th1/Th2 cytokine levels in serum revealed no significant difference between NSCLC patients and controls. However, the level of IL-1β, IL-4, IL-6 and IL-8 was significantly increased in serum of large cell lung carcinoma patients compared to controls. In addition, 1,100 proteins were identified and 33 of these were found to be consistently over-expressed in the BAL fluid of adenocarcinoma samples compared to controls. Finally, the expression of CD68 and CD163 (M2 marker) was significantly increased in all NSCLC subtypes compared to non-tumour tissues. In contrast, the expression of iNOS (M1 marker) was significantly decreased in the tumour tissue of patients with adenocarcinoma and squamous cell lung carcinoma compared to non-tumour tissue. Conclusion: the outcome of this work suggests that the presence of NSCLC is associated with an alteration in macrophage phenotype and function. This study revealed a number of specific proteins (e.g. CD163) and cytokines (e.g. IL-1β) that were identified to be associated with NSCLC patients and we suggest may be used as novel prognostic and/or diagnostic biomarkers in patients with NSCLC
Hydration water and ionic aggregation in aqueous solutions of imidazolium-based protic ionic liquids
Water molecules, present as additive or as contaminant of Protic Ionic Liquids (PILs), can compete for the hydrogen bond sites leading to important modifications of the local order of these liquids and to the modulation of their physical–chemical properties. In this work, aqueous solutions of a set of N-methylimidazolium-based PILs [MIM][X] (X = NO3-- , TfO-- , HSO4-- , and Cl-- ) were investigated by deep UV Resonance Raman (UVRR) spectroscopy in the water-rich domain where ionic aggregates and bulk-like water coexist. A differential method was used to analyze the OH stretching profile to extract the so-called solute-correlated (SC) spectrum, which is particular informative of the hydration features of the PILs. Moreover, specific bands of the cation, sensitive to the hydrogen bonding, were comparatively investigated. The progressive evolution from solvent-separated ion pairs (SIP) and/or solvent-shared ion pairs (SSIP) to contact ion pairs (CIP) and/or larger ionic aggregates can be monitored as a function of the hydration level, in the water-rich domain. Our approach showed that, in the highly diluted regime, the hydration environment around the [MIM] cation does not depend on the type of anion. Moreover, [MIM][NO3] and [MIM][TfO] showed cation-water (ionic) H-bonds at the NH site stronger than the cation–anion (double-ionic) ones. The analysis of SC Raman spectra points out the formation of cation–anion Hbonds (through the CH ring groups), stronger than cation-water ones, upon PILs concentration increase, especially evident in the case of [MIM][Cl]. The H-bond strength between the anion and hydration water is found to decrease following the order: [Cl] ~ [HSO4] > [NO3] > [TfO]. Chloride ions tend to perturb a larger number of water molecules than the other anions. The number of perturbed water molecules decreases at increasing PIL concentration, showing a larger dependence for [MIM][Cl], consistently with its larger propensity to form ionic aggregates. The unique response of [MIM][Cl] to hydration found by analyzing SC-UVRR data is related to the synergy of different factors such as the anion reduced size (higher charge density), spherical symmetry, and high H-bond basicity
Genotyping-Guided Discovery of Persiamycin A From Sponge-Associated Halophilic Streptomonospora sp. PA3
Microbial natural products have been a cornerstone of the pharmaceutical industry, but the supply of novel bioactive secondary metabolites has diminished due to extensive exploration of the most easily accessible sources, namely terrestrialStreptomycesspecies. The Persian Gulf is a unique habitat for marine sponges, which contain diverse communities of microorganisms including marine Actinobacteria. These exotic ecosystems may cradle rare actinomycetes with high potential to produce novel secondary metabolites. In this study, we harvested 12 different species of sponges from two locations in the Persian Gulf and isolated 45 symbiotic actinomycetes to assess their biodiversity and sponge-microbe relationships. The isolates were classified intoNocardiopsis(24 isolates),Streptomyces(17 isolates) and rare genera (4 isolates) by 16S rRNA sequencing. Antibiotic activity tests revealed that culture extracts from half of the isolates displayed growth inhibitory effects against seven pathogenic bacteria. Next, we identified five strains with the genetic potential to produce aromatic polyketides by genotyping ketosynthase genes responsible for synthesis of carbon scaffolds. The combined data led us to focus onStreptomonosporasp. PA3, since the genus has rarely been examined for its capacity to produce secondary metabolites. Analysis of culture extracts led to the discovery of a new bioactive aromatic polyketide denoted persiamycin A and 1-hydroxy-4-methoxy-2-naphthoic acid. The genome harbored seven gene clusters involved in secondary metabolism, including a tetracenomycin-type polyketide synthase pathway likely involved in persiamycin formation. The work demonstrates the use of multivariate data and underexplored ecological niches to guide the drug discovery process for antibiotics and anticancer agents
Reseña sobre espectroscopia de rompimiento inducida por láser
Se presenta una reseña sobre la espectroscopia de rompimiento inducida por láser (LIBS). Asimismo, se presenta un recuento histĂłrico sobre dicha tĂ©cnica, su funcionamiento e instrumentaciĂłn y una descripciĂłn de su configuraciĂłn experimental en general. LIBS ha crecido significativamente convirtiĂ©ndose en una tĂ©cnica rápida, fiable y adecuada para la detecciĂłn y análisis espectral de los elementos constituyentes de la materia. El análisis de los elementos se puede hacer de manera simultánea e incluso en tiempo real de forma independiente de la naturaleza, origen y estado de agregaciĂłn de las muestras. Se discuten los alcances y limitaciones de la tĂ©cnica, asĂ como sus potenciales aplicaciones en diversos campos tales como fĂsica, quĂmica, análisis de materiales, biomedicina, biotecnologĂa, genĂłmica, entre otros
Isolation of ASR7 Actinomycete Isolated from S12 Demospongia Marine Sponge and Study of Its Antibacterial Activity
Background and Aims: Marine Actinomycetes are gram-positive bacteria that sometimes are free, saprophytic or plant and animal-associated, including marine sponges. More than 75% of antibiotics and antimicrobial compounds are produced by actinomycetes. In recent years, due to the need for new drugs, marine microorganisms have been considered as new sources of potential production of significant metabolites. The purpose of this study is isolation and identification of marine sponge-associated Actinomycete and investigation of its antibacterial activity.
Materials and Methods: The Actinomycete was isolated from the marine Sponge collected from the depths of coastal waters in Bushehr and screened for antibacterial activity on pathogenic microorganisms of Escherichia coliŘŚ Bacillus cereusŘŚ Klebsiella spp.ŘŚ Salmonella spp. and Proteus spp. using a Disk Diffusion Method. For molecular identification, genomic DNA was first extracted from isolate and then, the16S rDNA gene was amplified by PCR and Sequenced. The results were analyzed using bioinformatic programs, Bioedit and MEGA6.
Results: In this study, based on phylogeny studies, it was determined that the isolate belonged to thegenus Streptomyces, and biochemical studies showed that all tests except catalase and gram were negative; antibacterial activity study showed significant activity against three pathogenic bacteria, E. coli, Bacillus cereus and Salmonella spp. It was more active against Salmonella spp. (around 16mm inhibition zone diameter).
Conclusions: The results showed that depths of the Bushehr coastal waters have marine sponge associated actinomycetes, which are a source of secondary metabolites with biological activity
Vitellogenin Gene Expression and Sex Steroid Levels as Biomarkers in Yellowfin Seabream (Acanthopagrus latus) Exposed to Bisphenol-A
Background: The egg yolk precursor protein vitellogenin (VTG) has proven to be a useful biomarker, used to identify organisms exposed to estrogenic compounds.
Methods: We investigated variations in the VTG gene expression pattern and plasma sex steroid hormones concentrations in the yellowfin Seabream, Acanthopagrus latus, (A. latus) by various doses of bisphenol-A (BPA) exposure for 7 and 14 days. We developed a quantitative real time polymerase chain reaction (RT-PCR) assay for the expression of VTG gene in A. latus. The dose-response pattern of VTG gene expression in A. latus exposed to various doses of BPA was characterized. In order to design RT-PCR primers specific to A. latus VTG, a partial sequence of the VTG gene was obtained.
Results: The RT-PCR assay was effective in detecting increased VTG gene expression in A. latus exposed to BPA. It also demonstrated that the VTG expression was affected by BPA in a dose and time-dependent manner. Plasma testosterone (T) levels were decreased in the treated fish in comparison with those found in the control group, when they were exposed to 100 µg/g of BPA and 2 µg/g of E2. In contrast, the plasma levels of 17β-estradiol (E2) were significantly increased in a dose-dependent manner.
Conclusion: The results suggest that VTG mRNA quantification can provide a sensitive and early signal in the detection of estrogens in marine wildlife. It also indicated that BPA could lead to an imbalance of sex steroid hormones with potentially harmful consequences on sexually immature male A. latus
Screening, isolation and study of antifungal activity of marine actinomycetes from Deylam nearshore sediments
Background: Marine actinomycetes, gram positive bacteria, have been prolific sources of novel secondary metabolites with a range of biological antibacterial activities. Marine sediments are potential sources for isolation of novel actinomycetes yielding new products and are recognized as source of novel antibiotic. In this study, we reported the isolation, characterization and antifungal activities of 8 actinomycetes isolated from Deylam nearshore sediments.
Materials and Methods: The marine soil sediment samples were collected from Deylam nearshore at the depth of 10 cm. The treated samples were serially diluted and used starch casein agar as a culture medium. Morphological and biochemical characterization of isolated strain was carried out by using standard methods. Antifungal assay of the bacterial extracts was performed using standard well diffusion assay.
Results: In this study, 8 marine actinomycetes were isolated from Deylam near shore sediments according to their morphology. All of isolate was belonged to Streptomyces genus. Differential analyses results for catalase and Gram test were positive for all isolates, the positive isolates for TSI, simmon citrate and ornitin decarboxylase were 1, 2 and 5 respectively, all isolates were negative for lysine decarboxylase, VP, MR and indol test, SIM test results showed that all isolates were non-motile, one isolate was produced H2S and some isolates formed pigmented colony. Most isolates showed antifungal activity against tested pathogenic fungi.
Conclusion: Results of this investigation revealed that the marine actinomycetes of Deylam nearshore sediments were potent source of bioactive compounds with antifungal activity
Blood classical monocytes phenotype is not altered in primary non-small cell lung cancer
AIM: To evaluate the M1 and M2 monocyte phenotype in patients with non-small cell lung cancer (NSCLC) compared to controls. Also, to examine the expression of Th1 and Th2 cytokines in plasma of NSCLC vs controls. METHODS: Freshly prepared peripheral blood mononuclear cells samples were obtained from patients with NSCLC (lung adenocarcinoma and squamous cell lung carcinoma) and from non-cancer controls. Flow cytometry was performed to investigate M1 and M2 phenotypes in peripheral monocytes (classical monocytes CD14+, CD45+ and CD16-) using conventional surface markers. Th1 and Th2 cytokine production was also analysed in the plasma using cytometric bead array technique. RESULTS: There were no significant difference in expression of M1 (HLA-DR) and/or M2 markers (CD163 and CD36) markers on classical monocytes in patients with NSCLC compared to non-cancer controls. Expression of CD11b, CD11c, CD71 and CD44 was also shown to be similar in patients with NSCLC compared to noncancer controls. Th1 and Th2 cytokines [interleukin (IL)-1β, IL-2, IL-4, IL-5, IL-8, IL-10, IL-12 (p70), tumor necrosis factor (TNF)-α, TNF-β, and interferon-γ] analysis revealed no significant difference between patients with NSCLC and non-cancer controls. CONCLUSION: This study shows no alteration in peripheral monocyte phenotype in circulating classical monocytes in patients with NSCLC compared to noncancer controls. No difference in Th1 and Th2 cytokine levels were noted in the plasma of these patients
Quantitative proteomics of bronchoalveolar lavage fluid in lung adenocarcinoma
Background: The most commonly reported primary lung cancer subtype is adenocarcinoma, which is associated with a poor prognosis and short survival. Proteomic studies on human body fluids such as bronchoalveolar lavage fluid (BALF) have become essential methods for biomarker discovery, examination of tumor pathways and investigation of potential treatments. Aim: This study used quantitative proteomics to investigate the up-regulation of novel proteins in BALF from patients with primary lung adenocarcinoma in order to identify potential biomarkers. Materials and Methods: BALF samples from individuals with and without primary lung adenocarcinoma were analyzed using liquid chromatography-mass spectrometry. Results: One thousand and one hundred proteins were identified, 33 of which were found to be consistently overexpressed in all lung adenocarcinoma samples compared to non-cancer controls. A number of overexpressed proteins have been previously shown to be related to lung cancer progression including S100-A8, annexin A1, annexin A2, thymidine phosphorylase and transglutaminase 2. Conclusion: The overexpression of a number of specific proteins in BALF from patients with primary lung adenocarcinoma may be used as a potential biomarker for lung adenocarcinoma