Designing resilience: a contribution to the City Resilience Framework

In a time of rapid social and demographic change, growing exposure to risk factors due to climate change, resource shortages, migration strain and adverse economic conditions are sources of stress and continuous instability in urban contexts. Arup conceived the City Resilience Framework for the Rockefeller Foundation. ‘Housing’ is the only form of construction included within this framework. It is one of the action areas in its strategy for improving urban resilience. This article seeks to provide a methodological contribution, hypothesizing requisites for resilience, preventative procedures and ex-post validation that moves beyond an emergency approach to foster the resilience of residential construction and its measurability in all phases of the construction process

LPS-stimulated human macrophages displayed sex differences in estrogen receptors α and β

Macrophages play a key role in immunity, inflammation, and atherosclerosis. Moreover, several evidences demonstrate that 17-β-estradiol (E2) plays a key role in inflammation and atherosclerosis through estrogen receptors (ERs) ERα and ERβ, processes that display sex differences. It has been largely demonstrated that male tissues express active ERs, but there is still lack of knowledge on their role in inflammation in males. Macrophages, which have ERα and ERβ, are a good model to evaluate the role of ER levels and activation in inflammation. The aim of our work was to evaluate the ability of lipopolysaccharide (LPS) to modulate, in a sex-specific way, the expression and the activation status of ERα and ERβ in blood monocytes-derived macrophages (MDMs) from healthy men and women. MDMs were isolated from blood of 7 adult men and 7 adult and fertile women (aged 21 - 35 years), and cultured. After 10 days of culture, MDMs were incubated with 100 ng / ml LPS for 24 h and lysed for the analysis of ERα, ERβ, P- ERα, p38 and P-p38 expression by Western Blotting. We found that in basal conditions, the expression of ERα and ERβ was significantly higher in female MDMs than in male ones. Importantly, LPS stimulation up-regulated ERα and ERα phosphorylation in both sexes, but this regulation was more pronounced in male MDMs. Moreover, LPS down-regulated ERβ level only in female MDMs. The expression of p38 and P-p38 proteins, used as marker of ERβ activity, did not display any sex differences. Finally, the ratios between ERα / ERβ and P-ERα / ERα were significantly higher in male than in female MDMs. Our findings show, for the first time, that LPS can modulate the activation of ERα but not that of ERβ, identifying a critical role of the subtype ERα in inflammatory responses mediated by LPS, at least in human MDMs. These results represent a starting point in understanding the influence of sex in the relationship between LPS and ERα

Gender differences in estrogenic compounds effect on human EPCs

Endothelial dysfunction has been defined as an “integrated risk factor”, and several gender difference have been reported in endothelial function. Several evidences suggest that endothelial progenitor cells (EPCs) are an important endogenous system that maintains integrity and vascular homeostasis. Their function is regulated by estrogens and estrogen receptors (ERs), but the effect of estrogenic compounds such as bisphenol A (BPA) and (R,R)-5,11-diethyl-5,6,11,12-tetrahydro-2,8-chrysenediol (THC) on human EPCs is unknown. BPA is used in plastic industry and BPA exposure is associated to abnormalities such as obesity, diabetes, disorders of the reproductive and immune systems, to endothelial dysfunction, and oxidative stress. This endocrine disruptor binds to ERα and ERβ with higher affinity for ERβ. THC is a specific agonist of ERα with a stronger ERβ antagonist activity. Therefore, the present work aimed to analyze if BPA and THC influence in a sex-specific manner the migration of human EPCs, an essential process in endothelial regeneration after vascular injury. EPCs were isolated from healthy adult men and women aged between 18 and 30 years, using a magnetic positive selection with the CD34 MicroBeads, a well-established marker of human progenitor cells. EPCs were also characterized for acetylated LDL Dil- (acLDL) and isothiocyanate (FITC)-conjugated with Ulex europaeus agglutinin I (lectin) uptake. The expression of ERα and ERβ was analysed by Western Blotting, while the migration assay was performed with the transwell chemotaxis assay. Male and female EPCs expressed both classical ERs: ERα was higher, but not significantly, in female cells, while ERβ was similarly expressed in both sexes. Male and female EPCs did not differ in basal migration. 17-β-estradiol (10-9 M e 10-10 M) significantly inhibited migration in female EPCs but not in male ones. Moreover, both 10-5 M THC and BPA (10-8 M) were able to bock migration only in female cells. Considering that BPA has a ERα and a prevalent ERβ agonist activity while THC has ERα agonistic activity and a prevalent ERβ antagonist activity, our data show that the effect on migration observed in female EPCs is mediated by ERα. Our data demonstrate that estrogenic compound have a sexual divergent effect on human EPCs, improving our knowledge on the gender differences observed in the pathophysiology of endothelial function

Sex differences in redox state, autophagy and lysosomal function

Modifications of the normal redox state are connected with numerous diseases and conditions such as cardiovascular diseases, diabetes mellitus and its complications, liver diseases, and aging which are characterized by numerous sex differences. Evidences suggest that redox state might be different in males and females. Autophagy is crucial for the maintaining cellular homeostasis process whereby cytoplasmic components are delivered to lysosomes for degradation. Alteration in constitutive autophagy is implicated in many pathological conditions, including heart diseases, diabetes mellitus and its complications, and liver diseases. A cross-talk between ROS and autophagy has been described. The current study was conducted to investigate the influence of sex on lipid and protein oxidation and autophagic response in the heart, the liver and the kidney obtained from young adult healthy male and female rats. 7 week old Sprague-Dawley rats were used to obtain heart, liver and kidneys. Malondialdehyde (MDA), and carbonylated proteins were measured by spectrophotometric methods for redox state assessment. The autophagy biomarkers Beclin-1, and microtubule-associated protein 1 light chain 3 (LC3), the mammalian target of rapamycin (mTOR; checkpoint in autophagic process), and the lysosomal associated membrane protein (LAMP-1; biomarker of lysosomes) were evaluated by Western blotting. Immunofluorescence analysis was also performed for LC3 and LAMP-1 colocalization. In the heart, Beclin-1, LC3-II/LC3-I were higher in males than in females suggesting that male heart have a major constitutive autophagy and this was linked with higher levels of carbonyl groups, indicating that protein oxidation could play a role. In the liver, it was found that LAMP-1 was higher in males and greatly colocalized with LC3 indicating a larger number of autophagolysosomes. None of the above parameters was significantly different in the kidneys of both sexes with the exception of MDA, which was significantly higher in females. These results suggest that the sex determinant affects the autophagy process and oxidation of proteins and lipids in an organ specific manner. It seems that the protein oxidation is more linked with constitutive autophagy, at least in cardiac ventricles, in comparison with lipid peroxidation

Autophagic process is sexually different in VSMCs from human male and female neonates

Sex has largely been neglected in cellular studies. Autophagy is a sophisticatedly reg- ulated homeostatic mechanism, which ensures cell’s constituent turnover. Under physiological conditions, autophagy levels are usually low (constitutive autophagy), but can be induced by numerous cellular stresses such as starvation. The influence of sex on autophagy has been studied either in vitro or in vivo, and previous our findings demonstrated the existence of sex differences in rat heart and HUVECs. Vascular smooth muscle cells (VSMCs) are a good experimental model for studying the physiopatholo- gy of the cardiovascular system, in which autophagy plays an important physiological role. Therefore, we investigated the occurrence of sexual dimorphism in constitutive and starvation-induced autophagy between the VSMCs obtained from human umbilical cord arteries (HUASMCs) of male (MHUASMCs) and female (FHUASMCs) neonates. HUASMCs were isolated from the umbilical cord of healthy and normal weight male and female neonates. The expression of oestrogen receptor (ER-α and ER-β) and the primary molecules involved in autophagic process [the mammalian target of rapamycin (mTOR), beclin-1 and microtubule-associated protein 1 light chain 3 (LC3)] were analysed by western blotting. Both cell types expressed both isoforms of the oestrogen receptor (ER): ER-β was higher expressed in the MHUASMCs than the FHUASMCs while ER-α was similarly expressed in both sexes. The level of constitutive autophagy, measured as LC3II/I ratio was higher in FHUASMCs than in MHUASMCs, while male cells had an higher expression of Beclin-1, indicating that constitutive autophagy was at least partly beclin-1-independent. mTOR activity, a regulator of autophagy, did not vary between the sexes, indicating that the observed differences could not be attributed to this central pathway. Starvation promoted autophagy in both MHUASMCs and FHUASMCs, but the increase was more pro- nounced in FHUASMCs. Our results show that sex-differences start in utero and are parameter-specific, suggesting that HUASMCs of both sexes are necessary in in vitro studies to elevate the quality and translational value of research results. The observed differences in the autophagic process could help to emeliorate our knowledge on sex-differences observed in cardiovascular diseases

Human umbilical endothelial cells (HUVECs) and sex differences

HUVECs are worldwide used to study the endothelial physiology and pathology that might be involved in sex and gender differences detected at the cardiovascular level. The present work characterised the phenotype of HUVECs in terms of morphology, proliferative and migratory capacity and in the gene expression of oestrogen and androgen receptors and nitric oxide synthase 3 (NOS3) to evaluated if they are sexually dimorphic. Moreover, autophagic process was analysed in male and female HUVECs (MHUVECs and FHUVECs), as autophagy is influenced by sex. Umbilical cords were obtained from healthy, normal weight, male and female neonates born to healthy non-obese and non-smoking women. HUVECs morphology was analysed by electron microscopy, and their function was investigated by proliferation, viability, wound healing and chemotaxis assays. Real-time PCR was used to evaluate gene expression for oestrogen and androgen receptors and for NOS3, while the expression of the primary molecules involved in autophagic process [(Akt, the mammalian target of rapamycin (mTOR), beclin-1 and microtubule-associated protein 1 light chain 3 (LC3)] and NOS3 were analysed by western blotting. FHUVECs showed significantly higher proliferation and migration rate, and NOS3 mRNA and protein expression than MHUVECs. Conversely, beclin-1 and the LC3-II/LC3-I ratio were higher in MHUVECs than in FHUVECs, indicating a higher autophagy in male cells as also indicated by ultrastructural analysis showing a buildup of autophagic vacuoles at different stages in MHUVECs. The expression of oestrogen and androgen receptor genes, the protein expression of Akt, mTOR, and cellular size and shape were not influenced by sex. Male and female neonates did not differ in body weight, but the weight of male babies was positively associated with the weight of the mother, suggesting that the weight of the mother may exert a different influence on male and female babies. Our findings indicate that sex differences exist from prenatal life and are parameter- specific, suggesting that a better quality of the research on the endothelium in vitro can be obtained by analyzing HUVECs of both sexes as well as its translational value. Moreover, the sex differences observed in HUVECs could help the diseases of adulthood because endothelial dysfunction has a key role in cardiovascular diseases, diabetes mellitus, neurodegeneration and immune diseases

Human umbilical endothelial cells (HUVECs) have a sex: characterisation of the phenotype of male and female cells

Background: Human umbilical endothelial cells (HUVECs) are widely used to study the endothelial physiology and pathology that might be involved in sex and gender differences detected at the cardiovascular level. This study evaluated whether HUVECs are sexually dimorphic in their morphological, proliferative and migratory properties and in the gene and protein expression of oestrogen and androgen receptors and nitric oxide synthase 3 (NOS3). Moreover, because autophagy is influenced by sex, its degree was analysed in male and female HUVECs (MHUVECs and FHUVECs). Methods: Umbilical cords from healthy, normal weight male and female neonates born to healthy non-obese and non-smoking women were studied. HUVEC morphology was analysed by electron microscopy, and their function was investigated by proliferation, viability, wound healing and chemotaxis assays. Gene and protein expression for oestrogen and androgen receptors and for NOS3 were evaluated by real-time PCR and Western blotting, respectively, and the expression of the primary molecules involved in autophagy regulation [protein kinase B (Akt), mammalian target of rapamycin (mTOR), beclin-1 and microtubule-associated protein 1 light chain 3 (LC3)] were detected by Western blotting. Results: Cell proliferation, migration NOS3 mRNA and protein expression were significantly higher in FHUVECs than in MHUVECs. Conversely, beclin-1 and the LC3-II/LC3-I ratio were higher in MHUVECs than in FHUVECs, indicating that male cells are more autophagic than female cells. The expression of oestrogen and androgen receptor genes and proteins, the protein expression of Akt and mTOR and cellular size and shape were not influenced by sex. Body weights of male and female neonates were not significantly different, but the weight of male babies positively correlated with the weight of the mother, suggesting that the mother’s weight may exert a different influence on male and female babies. Conclusions: The results indicate that sex differences exist in prenatal life and are parameter-specific, suggesting that HUVECs of both sexes should be used as an in vitro model to increase the quality and the translational value of research. The sex differences observed in HUVECs could be relevant in explaining the diseases of adulthood because endothelial dysfunction has a crucial role in the pathogenesis of cardiovascular diseases, diabetes mellitus, neurodegeneration and immune disease.</br

Genetic divergence of HIV-1 B subtype in Italy over the years 2003\u20132016 and impact on CTL escape prevalence

HIV-1 is characterized by high genetic variability, with implications for spread, and immune-escape selection. Here, the genetic modification of HIV-1 B subtype over time was evaluated on 3,328 pol and 1,152 V3 sequences belonging to B subtype and collected from individuals diagnosed in Italy between 2003 and 2016. Sequences were analyzed for genetic-distance from consensus-B (Tajima-Nei), non-synonymous and synonymous rates (dN and dS), CTL escapes, and intra-host evolution over four time-spans (2003\u20132006, 2007\u20132009, 2010\u20132012, 2013\u20132016). Genetic-distance increased over time for both pol and V3 sequences (P < 0.0001 and 0.0003). Similar results were obtained for dN and dS. Entropy-value significantly increased at 16 pol and two V3 amino acid positions. Seven of them were CTL escape positions (protease: 71; reverse-transcriptase: 35, 162, 177, 202, 207, 211). Sequences with 653 CTL escapes increased from 36.1% in 2003\u20132006 to 54.0% in 2013\u20132016 (P < 0.0001), and showed better intra-host adaptation than those containing 642 CTL escapes (intra-host evolution: 3.0 7 10 123 [2.9 7 10 123\u20133.1 7 10 123] vs. 4.3 7 10 123 [4.0 7 10 123\u20135.0 7 10 123], P[LRT] < 0.0001[21.09]). These data provide evidence of still ongoing modifications, involving CTL escape mutations, in circulating HIV-1 B subtype in Italy. These modifications might affect the process of HIV-1 adaptation to the host, as suggested by the slow intra-host evolution characterizing viruses with a high number of CTL escapes

Towards a Muon Collider

A muon collider would enable the big jump ahead in energy reach that is needed for a fruitful exploration of fundamental interactions. The challenges of producing muon collisions at high luminosity and 10 TeV centre of mass energy are being investigated by the recently-formed International Muon Collider Collaboration. This Review summarises the status and the recent advances on muon colliders design, physics and detector studies. The aim is to provide a global perspective of the field and to outline directions for future work.Comment: 118 pages, 103 figure

Towards a muon collider

A muon collider would enable the big jump ahead in energy reach that is needed for a fruitful exploration of fundamental interactions. The challenges of producing muon collisions at high luminosity and 10 TeV centre of mass energy are being investigated by the recently-formed International Muon Collider Collaboration. This Review summarises the status and the recent advances on muon colliders design, physics and detector studies. The aim is to provide a global perspective of the field and to outline directions for future work