Fanconi anemia (FA) is a DNA repair syndrome generated by mutations in any of the 22 FA genes discovered to date
. Mutations in FANCA account for more than 60% of FA cases worldwide
. Clinically, FA ...is associated with congenital abnormalities and cancer predisposition. However, bone marrow failure is the primary pathological feature of FA that becomes evident in 70-80% of patients with FA during the first decade of life
. In this clinical study (ClinicalTrials.gov, NCT03157804 ; European Clinical Trials Database, 2011-006100-12), we demonstrate that lentiviral-mediated hematopoietic gene therapy reproducibly confers engraftment and proliferation advantages of gene-corrected hematopoietic stem cells (HSCs) in non-conditioned patients with FA subtype A. Insertion-site analyses revealed the multipotent nature of corrected HSCs and showed that the repopulation advantage of these cells was not due to genotoxic integrations of the therapeutic provirus. Phenotypic correction of blood and bone marrow cells was shown by the acquired resistance of hematopoietic progenitors and T lymphocytes to DNA cross-linking agents. Additionally, an arrest of bone marrow failure progression was observed in patients with the highest levels of gene marking. The progressive engraftment of corrected HSCs in non-conditioned patients with FA supports that gene therapy should constitute an innovative low-toxicity therapeutic option for this life-threatening disorder.
Aging is considered the single most significant risk factor for the majority of common malignances including lung cancer. Together immunosenescence, changes occurring with aging in the immune system, ...and inflammaging, characterizes by a chronic, subclinical accumulation of pro-inflammatory factors, are suggested to stand at the origin of most of the diseases of the elderly, such as cancer. The aim of this study was to determine associations among lymphocyte subpopulations, pro-inflammatory cytokines and epidermal growth factor (EGF) in patients diagnosed with non-small cell lung cancer (NSCLC). Forty-six advanced NSCLC patients were enrolled. Sixteen patients with newly diagnosed and before treatment and 30 patients after first-line platinum-based chemotherapy. Peripheral blood subpopulations were studied by flow cytometry and serum concentrations of soluble factors by ELISA. The frequency of naïve CD4
+
T cells, naïve B cells and central memory CD8
+
T cells were significantly lower in NSCLC patients after chemotherapy, while effector memory CD4
+
T cells and terminally differentiated CD8
+
T cells were significantly higher. IL-1β and TNFα significantly correlated among them before and after platinum-based chemotherapy. Terminally differentiated T cells expressing CD57
+
significantly correlated with TNFα and IL-1β. For the first time, associations between EGF serum levels and terminally differentiated CD4
+
T cells, and memory B cells were detected. This study confirms the association among terminally differentiated lymphocytes and pro-inflammatory cytokines in patients diagnosed with lung cancer, reinforcing the interconnection between terminally differentiated lymphocytes and pro-inflammatory cytokines.
Clinical trial registration number: RPCEC00000205,
http://registroclinico.sld.cu/
Background and AimsThe aquatic moss Fontinalis antipyretica requires a slow rate of dehydration to survive a desiccation event. The present work examined whether differences in the dehydration rate ...resulted in corresponding differences in the production of reactive oxygen species (ROS) and therefore in the amount of cell damage.MethodsIntracellular ROS production by the aquatic moss was assessed with confocal laser microscopy and the ROS-specific chemical probe 2,7-dichlorodihydrofluorescein diacetate. The production of hydrogen peroxide was also quantified and its cellular location was assessed.Key ResultsThe rehydration of slowly dried cells was associated with lower ROS production, thereby reducing the amount of cellular damage and increasing cell survival. A high oxygen consumption burst accompanied the initial stages of rehydration, perhaps due to the burst of ROS production.ConclusionsA slow dehydration rate may induce cell protection mechanisms that serve to limit ROS production and reduce the oxidative burst, decreasing the number of damaged and dead cells due upon rehydration.
Near-infrared spectroscopy (NIRS) is an accurate, fast and safe technique whose full potential remains to be exploited. Lichens are a paradigm of symbiotic association, with extraordinary properties, ...such as abiotic stress tolerance and adaptation to anhydrobiosis, but subjacent mechanisms await elucidation. Our aim is characterizing the metabolomic NIRS fingerprints of
and
thalli, and of the cultured phycobionts
and
. Thalli collected in an air-dry state and fresh cultivated phycobionts were directly used for spectra acquisition in reflectance mode. Thalli water peaks were associated to the solvation shell (1354 nm) and sugar-water interactions (1438 nm). While northern-southern orientation related with two hydrogen bonded (S
) water, the site was related to one hydrogen bonded (S
). Water, lipids (saturated and unsaturated), and polyols/glucides contributed to the profiles of lichen thalli and microalgae.
, with higher desiccation tolerance, shows higher S
water than
. In contrast, fresh phycobionts are dominated by free water. Whereas
shows higher solvation water content,
possesses more unsaturated lipids. Aquaphotomics demonstrates the involvement of strongly hydrogen bonded water conformations, polyols/glucides, and unsaturated/saturated fatty acids in the dehydration process, and supports a "rubbery" state allowing enzymatic activity during anhydrobiosis.
Salinity is a complex trait that affects growth and productivity in many crops, including rice. Mutation induction, a useful tool to generate salt tolerant plants, enables the analysis of plants with ...similar genetic background, facilitating the understanding of the salt tolerance mechanisms. In this work, we generated three salt tolerant mutant lines by irradiation of a salt-sensitive cultivar plants and screened M2 plants at seedling stage in the presence of high salinity. These three lines,
, and
, showed different responses to salinity, but exhibited similar phenotype to wild type plants, except
that displayed shorter height when grown in the absence of salt. Under salt conditions, all three mutants and the parental line showed similar reduction in yield, although relevant differences in other physiological parameters, such as Na
accumulation in healthy leaves of
, were registered. Microarray analyses of gene expression profiles in roots revealed the occurrence of common and specific responses in the mutants. The three mutants showed up-regulation of responsive genes, the activation of oxido-reduction process and the inhibition of ion transport. The participation of jasmonate in the plant response to salt was evident by down-regulation of a gene coding for a jasmonate O-methyltransferase. Genes dealing with lipid transport and metabolism were, in general, up-regulated except in
, that also exhibited down-regulation of genes involved in ion transport and Ca
signal transduction. The two most tolerant varieties,
and
, displayed lower levels of transcripts involved in K
uptake. The physiological study and the description of the expression analysis evidenced that the three lines showed different responses to salt:
showed a high Na
content in leaves,
presented an inhibition of lipid metabolism and ion transport and
differs in both features in the response to salinity. The analysis of these salt tolerant mutants illustrates the complexity of this trait evidencing the breadth of the plant responses to salinity including simultaneous cooperation of alternative or complementary mechanisms.
Near-infrared spectroscopy (NIRS) is an accurate, fast and safe technique whose full potential remains to be exploited. Lichens are a paradigm of symbiotic association, with extraordinary properties, ...such as abiotic stress tolerance and adaptation to anhydrobiosis, but subjacent mechanisms await elucidation. Our aim is characterizing the metabolomic NIRS fingerprints of Ramalina farinacea and Lobarina scrobiculata thalli, and of the cultured phycobionts Trebouxia lynnae and Trebouxia jamesii. Thalli collected in an air-dry state and fresh cultivated phycobionts were directly used for spectra acquisition in reflectance mode. Thalli water peaks were associated to the solvation shell (1354 nm) and sugar-water interactions (1438 nm). While northern-southern orientation related with two hydrogen bonded (Ssub.2) water, the site was related to one hydrogen bonded (Ssub.1). Water, lipids (saturated and unsaturated), and polyols/glucides contributed to the profiles of lichen thalli and microalgae. R. farinacea, with higher desiccation tolerance, shows higher Ssub.2 water than L. scrobiculata. In contrast, fresh phycobionts are dominated by free water. Whereas T. jamesii shows higher solvation water content, T. lynnae possesses more unsaturated lipids. Aquaphotomics demonstrates the involvement of strongly hydrogen bonded water conformations, polyols/glucides, and unsaturated/saturated fatty acids in the dehydration process, and supports a "rubbery" state allowing enzymatic activity during anhydrobiosis.
Detailed examination of the absorption spectra from dark ocean samples allowed us to identify and deconvolve two distinct chromophores centered at 302 nm (UV) and 415 nm (visible) from the ...exponential decay curve characteristic of humic substances. The UV chromophore was ubiquitous in intermediate and deep waters, and it has been proposed as the secondary absorption peak of nitrate. The visible chromophore was prominent at the central and intermediate water masses of the North Pacific, and it has been proposed as cytochrome c. Subtraction of the modeled absorption spectra of the two chromophores from the measured absorption spectrum of the samples leads to a spectral slope overestimation by 13.3 ± 6.0% for S275–295 and 14.8 ± 10.6% for S350–400. To only consider the chromophoric fraction of DOM, the absorption spectra of nitrate should be subtracted in samples with a NO3−:a302 ratio > 70 µM m.
Key Points
Two distinct chromophores have been detected in the dark global ocean
A chromophore centered at 302 nm is due to the secondary absorption peak of nitrate
A chromophore centered at 415 nm is presumably due to cytochrome c and can be a used as proxy for cumulative respiration
NO is a multifaceted molecule, key in functions such as abiotic stress tolerance and symbioses establishment and permanence. Lichens are complex symbiotic associations of microalgae, fungi and ...prokaryotes that release NO under stress conditions such as dessication-rehydration cycles and the presence of xenobiotics. NO synthase (NOS) oxidises l-arginine to produce NO in animals and some aquatic microalgae, while nitrate reductase (NR) reduces nitrate to NO in plants and fungi. Inhibition studies suggest that both activities might be present in thalli. Due to its multipartner compotition, our hypothesis is that Ramalina farinacea biosynthesises NO through both oxidative (NOS) and reductive (NR) enzymatic pathways. NR activity was quantified with a method optimised for lichens using NADH or/and NADPH, and NOS with a commercial kit in R. farinacea thalli and cultures of the isolated main symbionts: R. farinacea mycobiont, and Trebouxia jamesii and Trebouxia lynnae phycobionts. Inhibition studies in vitro were performed with L-NAME and tungstate. Immunodetection was carried out with specific polyclonal antibodies (anti-plant NADH-NR and anti-iNOS animal isoform). NADH-NR specific activity of R. farinacea is an order of magnitude higher than Arabidopsis thaliana's and in the range of the chlorophyte Ulva intestinalis. R. farinacea mycobiont possesses a canonical plant-like Moco-NR, while Trebouxia phycobionts' NR activity presents interesting peculiarities. NOS has not been immunodetected and NOS-like activity is inhibited by L-NAME only partially in T. jamesii. Despite NOS-like activity is very high in the isolated microalgae and fungus, it is strongly depressed in the holobiont. In summary, NR activity seems to be the main source of NO biosynthesis for the holobiont R. farinacea but it presents intriguing features that deserve further study.
•Symbiosis in the holobiont modifies NO biosynthetic activities of the partners.•NR activity appears to be the main NO biosynthesizing source in R. farinacea.•Both canonical and non-canonical NR activities are reported for algal symbionts.•Algal symbionts do not present a plant-like NR protein as does R. farinacea.•NOS-like activity in the isolated R. farinacea symbionts is relatively relevant.
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•The sexually reproducing lichen-forming fungus Ramalina fraxinea associates with at least seven different Trebouxia lineages.•These lineages belong to at least two species: T. ...decolorans and T. jamesii, which coexist within each analyzed thallus.•Some of these lineages are shared in different extent by several other lichen taxa.•The different algal lineages observed may be consequence of an incipient speciation within T. decolorans.•This study stresses the importance of coordinated ultrastructural and phylogenomic analyses in phycobiont diversity estimates.
The precise boundary delineations between taxa in symbiotic associations are very important for evolutionary and ecophysiological studies. Growing evidence indicates that in many cases, the use of either morphological characters or molecular markers results in diversity underestimation. In lichen symbioses, Trebouxia is the most common genus of lichen phycobionts, however, the diversity within this genus has been poorly studied and as such there is no clear species concept. This study constitutes a multifaceted approach incorporating aspects of ultrastructural characterization by TEM and phylogenomics to evaluate the morphological and genetic diversity of phycobionts within the sexually reproducing lichen Ramalina fraxinea in the context of Mediterranean and temperate populations. Results reveal an association with at least seven different Trebouxia lineages belonging to at least two species, T. decolorans and T. jamesii, and diverse combinations of such lineages coexisting within the same thallus depending on the analyzed sample. Some of these lineages are shared by several other non-related lichen taxa. Our findings indicate the existence of a highly diverse assemblage of Trebouxia algae associating with R. fraxinea and suggest a possible incipient speciation within T. decolorans rendering a number of lineages or even actual species. This study stresses the importance of coordinated ultrastructural and molecular analyses to improve estimates of diversity and reveal the coexistence of more than one Trebouxia species within the same thallus. It is also necessary to have clearer species delimitation criteria within the genus Trebouxia and microalgae in general.
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