Reciprocal interference between the NRF2 and LPS signaling pathways on the immune-metabolic phenotype of peritoneal macrophages

The metabolic and immune adaptation to extracellular signals allows macrophages to carry out specialized functions involved in immune protection and tissue homeostasis. Nuclear factor erythroid 2-related factor 2 (NRF2) is a transcription factor that coordinates cell redox and metabolic responses to stressors. However, the individual and concomitant activation of NRF2 and inflammatory pathways have been poorly investigated in isolated macrophages. We here took advantage of reporter mice for the transcriptional activities of NRF2 and nuclear factor-kB (NF\u3baB), a key transcription factor in inflammation, and observe a persisting reciprocal interference in the response of peritoneal macrophages to the respective activators, tert-Butylhydroquinone (tBHQ) and lipopolysaccharide (LPS). When analyzed separately by gene expression studies, these pathways trigger macrophage-specific metabolic and proliferative target genes that are associated with tBHQ-induced pentose phosphate pathway (PPP) with no proliferative response, and with opposite effects observed with LPS. Importantly, the simultaneous administration of tBHQ + LPS alters the effects of each individual pathway in a target gene-specific manner. In fact, this co-treatment potentiates the effects of tBHQ on the antioxidant enzyme, HMOX1, and the antibacterial enzyme, IRG1, respectively; moreover, the combined treatment reduces tBHQ activity on the glycolytic enzymes, TALDO1 and TKT, and decreases LPS effects on the metabolic enzyme IDH1, the proliferation-related proteins KI67 and PPAT, and the inflammatory cytokines IL-1\u3b2, IL-6, and TNF\u3b1. Altogether, our results show that the activation of NRF2 redirects the metabolic, immune, and proliferative response of peritoneal macrophages to inflammatory signals, with relevant consequences for the pharmacological treatment of diseases that are associated with unopposed inflammatory responses

ER alpha-independent NRF2-mediated immunoregulatory activity of tamoxifen

Sex differences in immune-mediated diseases are linked to the activity of estrogens on innate immunity cells, including macrophages. Tamoxifen (TAM) is a selective estrogen receptor modulator (SERM) used in estrogen receptor-alpha (ERα)-dependent breast cancers and off-target indications such as infections, although the immune activity of TAM and its active metabolite, 4-OH tamoxifen (4HT), is poorly characterized. Here, we aimed at investigating the endocrine and immune activity of these SERMs in macrophages. Using primary cultures of female mouse macrophages, we analyzed the expression of immune mediators and activation of effector functions in competition experiments with SERMs and 17β-estradiol (E2) or the bacterial endotoxin LPS. We observed that 4HT and TAM induce estrogen antagonist effects when used at nanomolar concentrations, while pharmacological concentrations that are reached by TAM in clinical settings regulate the expression of VEGFα and other immune activation genes by ERα- and G protein-coupled receptor 1 (GPER1)-independent mechanisms that involve NRF2 through PI3K/Akt-dependent mechanisms. Importantly, we observed that SERMs potentiate cell phagocytosis and modify the effects of LPS on the expression of inflammatory cytokines, such as TNFα and IL1β, with an overall increase in cell inflammatory phenotype, further sustained by potentiation of IL1β secretion through caspase-1 activation

Studia Lichenologica in Italy. I. New records of red-listed species

We present new data concerning lichen species of the Italian flora and confirm 36 national redlisted species

Microclimatic Alteration after Logging Affects the Growth of the Endangered Lichen Lobaria pulmonaria

Microclimatic conditions are important in determining lichen distribution at small scale, and may determine whether the species persist when the surrounding environmental conditions have drastically changed. This is the case with forest management, since a sudden variation of microclimatic conditions (increase of solar radiation, temperature, wind and a reduction of humidity) may occur after logging. In this study, the combined effect of forest logging and microclimatic conditions on the growth probabilities and growth rates of the model species Lobaria pulmonaria was assessed in mixed oak stands. To this purpose, 800 fragments of L. pulmonaria (<1 cm) were transplanted in logged and unlogged stands for two years. Young and adult fragments were positioned on Turkey oak boles according to distance from the ground (100 and 50 cm) and aspect (north and south). The results, evaluated by generalized linear mixed models on a yearly basis, highlighted differences in growth—particularly on isolated trees in the logged stand. South-exposed samples in the logged stand showed a low probability of growth, while samples transplanted north in the unlogged stand showed higher growth probabilities. However, the highest annual growth coefficients corresponded to south-exposed samples 50 cm from the ground in the unlogged stand. In general, higher growth rates were observed in young thallus fragments when compared with adult ones. Beyond confirming the importance of microclimate for lichen ecology, these results could be implemented in conservation actions to preserve L. pulmonaria populations in logged forests

Inducing mutations in the mouse genome with the chemical mutagen ethylnitrosourea

When compared to other model organisms whose genome is sequenced, the number of mutations identified in the mouse appears extremely reduced and this situation seriously hampers our understanding of mammalian gene function(s). Another important consequence of this shortage is that a majority of human genetic diseases still await an animal model. To improve the situation, two strategies are currently used: the first makes use of embryonic stem cells, in which one can induce knockout mutations almost at will; the second consists of a genome-wide random chemical mutagenesis, followed by screening for mutant phenotypes and subsequent identification of the genetic alteration(s). Several projects are now in progress making use of one or the other of these strategies. Here, we report an original effort where we mutagenized BALB/c males, with the mutagen ethylnitrosourea. Offspring of these males were screened for dominant mutations and a three-generation breeding protocol was set to recover recessive mutations. Eleven mutations were identified (one dominant and ten recessives). Three of these mutations are new alleles (Otop1mlh, Foxn1sepe and probably rodador) at loci where mutations have already been reported, while 4 are new and original alleles (carc, eqlb, frqz, and Sacc). This result indicates that the mouse genome, as expected, is far from being saturated with mutations. More mutations would certainly be discovered using more sophisticated phenotyping protocols. Seven of the 11 new mutant alleles induced in our experiment have been localized on the genetic map as a first step towards positional cloning.Universidade de São Paulo Instituto de Ciências Biomédicas Departamento de ImunologiaUniversidade Federal de Minas Gerais Instituto de Ciências Biológicas Departamento de Biologia GeralUniversidade Federal de São Paulo (UNIFESP) Departamento de Bioquímica Disciplina de Biologia MolecularInstitut Pasteur Département de Biologie du DéveloppementUNIFESP, Depto. de Bioquímica Disciplina de Biologia MolecularSciEL

Testing an expanded set of sustainable forest management indicators in Mediterranean coppice area

Although coppice forests represent a significant part of the European forest area, especially across southern Countries, they received little attention within the Sustainable Forest Management (SFM) processes and scenarios, whose guidelines have been mainly designed to high forests and national scale. In order to obtain “tailored” information on the degree of sustainability of coppices on the scale of the stand, we evaluated (i) whether the main coppice management options result in different responses of the SFM indicators, and (ii) the degree to which the considered SFM indicators were appropriate in their application at stand level. The study considered three different management options (Traditional Coppice TC, coppice under Natural Evolution NE, and coppice under Conversion to high forest by means of periodical thinning CO). In each of the 43 plots considered in the study, which covered three different European Forest Types, we applied a set of eighteen “consolidated” SFM indicators, covering all the six SFM Criteria (FOREST EUROPE, 2020) and, additionally, tested other sixteen novel indicators shaped for agamic forests and/or applicable at stand level. Results confirmed that several consolidated indicators related to resources status (Growing stock and Carbon stock), health (Defoliation and Forest damage), and socio-economic functions (Net revenue, Energy and Accessibility) were highly appropriate for evaluating the sustainability of coppice at stand level. In addition, some novel indicators related to resources status (Total above ground tree biomass), health (Stand growth) and protective functions (Overstorey cover and Understorey cover) proved to be highly appropriate and able to support the information obtained by the consolidated ones. As a consequence, a subset of consolidated SFM indicators, complemented with the most appropriate novel ones, may represent a valid option to support the evaluation of coppice sustainability at stand level. An integrated analysis of the SFM indicators showed that NE and CO display significant higher environmental performances as compared with TC. In addition, CO has positive effects also on socio-economic issues, while TC -which is an important cultural heritage and a silvicultural option that may help to keep local communities engaged in forestry – combines high wood harvesting rates with dense understory cover. Overall, each of the three management options showed specific sustainability values; as a consequence, their coexistence at a local scale and in accordance with the specific environmental conditions and the social-economic context, is greatly recommended since it may fulfill a wider array of sustainability issues

Testing an expanded set of sustainable forest management indicators in Mediterranean coppice area

Although coppice forests represent a significant part of the European forest area, especially across southern Countries, they received little attention within the Sustainable Forest Management (SFM) processes and scenarios, whose guidelines have been mainly designed to high forests and national scale. In order to obtain “tailored” information on the degree of sustainability of coppices on the scale of the stand, we evaluated (i) whether the main coppice management options result in different responses of the SFM indicators, and (ii) the degree to which the considered SFM indicators were appropriate in their application at stand level. The study considered three different management options (Traditional Coppice TC, coppice under Natural Evolution NE, and coppice under Conversion to high forest by means of periodical thinning CO). In each of the 43 plots considered in the study, which covered three different European Forest Types, we applied a set of eighteen “consolidated” SFM indicators, covering all the six SFM Criteria (FOREST EUROPE, 2020) and, additionally, tested other sixteen novel indicators shaped for agamic forests and/or applicable at stand level. Results confirmed that several consolidated indicators related to resources status (Growing stock and Carbon stock), health (Defoliation and Forest damage), and socio-economic functions (Net revenue, Energy and Accessibility) were highly appropriate for evaluating the sustainability of coppice at stand level. In addition, some novel indicators related to resources status (Total above ground tree biomass), health (Stand growth) and protective functions (Overstorey cover and Understorey cover) proved to be highly appropriate and able to support the information obtained by the consolidated ones. As a consequence, a subset of consolidated SFM indicators, complemented with the most appropriate novel ones, may represent a valid option to support the evaluation of coppice sustainability at stand level. An integrated analysis of the SFM indicators showed that NE and CO display significant higher environmental performances as compared with TC. In addition, CO has positive effects also on socio-economic issues, while TC -which is an important cultural heritage and a silvicultural option that may help to keep local communities engaged in forestry – combines high wood harvesting rates with dense understory cover. Overall, each of the three management options showed specific sustainability values; as a consequence, their coexistence at a local scale and in accordance with the specific environmental conditions and the social-economic context, is greatly recommended since it may fulfill a wider array of sustainability issue

The frequency of CD127low expressing CD4+CD25high T regulatory cells is inversely correlated with human T lymphotrophic virus type-1 (HTLV-1) proviral load in HTLV-1-infection and HTLV-1-associated myelopathy/tropical spastic paraparesis

<p>Abstract</p> <p>Background</p> <p>CD4⁺CD25^highregulatory T (T_Reg) cells modulate antigen-specific T cell responses, and can suppress anti-viral immunity. In HTLV-1 infection, a selective decrease in the function of T_Regcell mediated HTLV-1-tax inhibition of FOXP3 expression has been described. The purpose of this study was to assess the frequency and phenotype of T_Regcells in HTLV-1 asymptomatic carriers and in HTLV-1-associated neurological disease (HAM/TSP) patients, and to correlate with measures of T cell activation.</p> <p>Results</p> <p>We were able to confirm that HTLV-I drives activation, spontaneous IFNγ production, and proliferation of CD4+ T cells. We also observed a significantly lower proportion of CTLA-4⁺T_Regcells (CD4⁺CD25^highT cells) in subjects with HAM/TSP patients compared to healthy controls. Ki-67 expression was negatively correlated to the frequency of CTLA-4⁺T_Regcells in HAM/TSP only, although Ki-67 expression was inversely correlated with the percentage of CD127^lowT_Regcells in healthy control subjects. Finally, the proportion of CD127^lowT_Regcells correlated inversely with HTLV-1 proviral load.</p> <p>Conclusion</p> <p>Taken together, the results suggest that T_Regcells may be subverted in HAM/TSP patients, which could explain the marked cellular activation, spontaneous cytokine production, and proliferation of CD4⁺T cells, in particular those expressing the CD25^highCD127^lowphenotype. T_Regcells represent a potential target for therapeutic intervention for patients with HTLV-1-related neurological diseases.</p