Hypomethylation-induced regulatory programs in T cells unveiled by transcriptomic analyses.

Regulatory T cells (Tregs) are essential mediators of tolerance mitigating aberrant immune responses. While naturally occurring Treg (nTreg) development and function are directed by epigenetic events, induced Treg (iTreg) identity and mechanisms of action remain elusive. Mirroring the epigenetic circuits of nTregs, we and others have used hypomethylation agents (HAs) to ex vivo convert T cells into iTregs (HA-iTregs) and further showed that the suppressive properties of the HA-iTregs are predominantly confined in an emergent population, which de novo expresses the immunomodulatory molecule HLA-G, consequently providing a surface marker for isolation of the suppressive HA-iTreg compartment (G+ cells). We isolated the HA-induced G+ cells and their G- counterparts and employed high-throughput RNA-sequencing (RNA-seq) analyses to uncover the G+-specific transcriptomic changes guiding T cells toward a regulatory trajectory upon their exposure to HA. We found a distinct transcriptional upregulation of G+ cells accompanied by enrichment of immune-response-related pathways. Although single-cell RNA-seq profiling revealed regulatory G+ cells to have molecular features akin to nTregs, when assessed in conjunction with the comparative transcriptomic analysis and profiling of secreted cytokines against the non-suppressive G- cells, FOXP3 and other T-helper signatures appear to play a minor role in their suppressive phenotype. We found an ectopic expression of IDO-1 and CCL17/22 in G+ cells, denoting that in vitro exposure of T cells to HA may well unlock myeloid suppressor genes. This report provides transcriptional data shaping the molecular identity of a highly purified and potent HA-iTreg population and hints toward ectopic myeloid-specific molecular mechanisms mediating HA-iTreg function.

Emerging trends in domestic homicide/femicide in Greece over the period 2010-2021.

Temporal trends in epidemiological parameters of domestic homicide and femicide in Greece over the last decade have not yet been studied. We conducted this study to fulfill this purpose. Specifically, we conducted a retrospective epidemiological study using 11-year data from the official nationwide Hellenic Police Archives and statistically analyzed data regarding domestic homicide and femicide. Overall, 1370 records of homicides among which 236 domestic homicides were identified. The pattern emerging from the statistical results of the present study highlighted the phenomenon of femicide as the gravest current issue to be interpreted and addressed. Nationally, the average number of homicides was 114.2/year, among which 19.7 domestic homicides. However, in 2021, while a decrease was recorded in homicides in general to 89 incidents per year, domestic homicides skyrocketed to 34 cases, reaching the highest annual number ever nationally recorded. On average, domestic homicides account for 18.2% of all homicides in Greece. In 2021, however, this percentage rose to 38.2%. The number of male victims of domestic homicide has declined over the years, with a further decline in 2021, in stark contrast to the number of women escalating over time and even more sharply in 2021. The proportion of female victims of domestic homicides in Greece was fourfold higher on average. The fact that cases of domestic homicide and femicide have received a lot of media attention, the recent Greek financial crisis, as well as increased alcohol and drug consumption due to the COVID-19 pandemic constitute possible aggravating factors.

Pharmacological activation of the C5a receptor leads to stimulation of the ß-adrenergic receptor and alleviates cognitive impairment in a murine model of familial Alzheimer's disease.

Alzheimer's disease (AD) is a progressive neurodegenerative disease of the brain causing either familial or sporadic dementia. We have previously administered the modified C5a receptor agonist (EP67) for a short period to a transgenic mouse model of AD (5XFAD) and have observed not only reduction in ß-amyloid deposition and gliosis but also improvement in cognitive impairment. Inquiring, however, on the effects of EP67 in an already heavily burdened animal, thus representing a more realistic scenario, we treated 6-month-old 5XFAD mice for a period of 14 weeks. We recorded a significant decrease in both fibrillar and pre-fibrillar ß-amyloid as well as remarkable amelioration of cognitive impairment. Following proteomic analysis and pathway association, we postulate that these events are triggered through the upregulation of ß-adrenergic and GABAergic signaling. In summary, our results reveal how inflammatory responses can be employed in inducing tangible phenotype improvements even in advanced stages of AD.

A de novo SFMBT1 pathogenic variant identified in a boy with Poland syndrome.

Poland syndrome is a rare developmental disorder characterized by unilateral, complete or partial, absence of the pectoralis major (and often minor) muscle, accompanied with ipsilateral hand malformations. To date, no clear genetic cause has been associated with Poland syndrome, although familial cases have been reported. We report the employment of trio exome investigation and the identification of a heterozygous de novo pathogenic variant in the SFMBT1 gene, a transcription factor associated with transcriptional repression during development, in a 14-yr-old boy with Poland syndrome. We further demonstrate by means of cDNA sequencing and western blot analysis that this variant results in SFMBT1 exon 10 skipping and a lower concentration of the SFMBT1 wild-type protein. To our knowledge, the heterozygous pathogenic SFMBT1 variant identified in association with this condition is novel as it has not been elsewhere described in the literature and it can be incorporated to the limited reported cases published.

Enhanced biodegradation and valorization of drilling wastewater via simultaneous production of biosurfactants and polyhydroxyalkanoates by Pseudomonas citronellolis SJTE-3.

Pseudomonas citronellolis SJTE-3 was isolated as a highly efficient microorganism for biodegradation and valorization of drilling fluids (DF) wastewater. The strain metabolised DF and oily mud exhibiting up to 93%, 86%, 85% and 88% of chemical oxygen demand (COD), n-dodecane, n-tetradecane and naphthalene removal efficiency respectively. Enhanced bioconversion was enabled through production of biosurfactants that reduced the surface tension of water by 53% and resulted in 43.3% emulsification index (E24), while synthesizing 24% of dry cell weight (DCW) as medium-chain-length polyhydroxyalkanoates (PHA). Expression from the main pathways for alkanes and naphthalene biodegradation as well as biosurfactants and PHA biosynthesis revealed that although the alkanes and naphthalene biodegradation routes were actively expressed even at stationary phase, PHA production was stimulated at late stationary phase and putisolvin could comprise the biosurfactant synthesized. The bioconversion of toxic petrochemical residues to added-value thermoelastomers and biosurfactants indicate the high industrial significance of P. citronellolis SJTE-3.

FAK displacement from focal adhesions: a promising strategy to target processes implicated in cancer progression and metastasis.

BACKGROUND: Focal adhesion kinase (FAK) is a non-receptor tyrosine kinase that is overexpressed or activated in several advanced-stage solid cancers. It is known to play both kinase-dependent and -independent roles in promoting tumor progression and metastasis. Numerous inhibitors, targeting either the enzymatic or scaffolding activities of FAK have been generated, with varying degree of success. Here, we describe a novel approach to site-specifically target both kinase-dependent and -independent FAK functions at focal adhesions (FAs), the primary sites at which the kinase exerts its activity. METHODS: We took advantage of the well-characterized interactions between the paxillin LD motifs and the FAK FAT domain and generated a polypeptide (LD2-LD3-LD4) expected to compete with interactions with paxillin. Co-immunoprecipitation experiments were performed to examine the interaction between the LD2-LD3-LD4 polypeptide and FAK. The effects of LD2-LD3-LD4 in the localization and functions of FAK, as well as FA composition, were evaluated using quantitative immunofluorescence, cell fractionation, FA isolation and Western Blot analysis. Live cell imaging, as well as 2-D migration and cell invasion assays were used to examine the effects on FA turnover and tumor cell migration and invasion. RESULTS: Expression of the LD2-LD3-LD4 polypeptide prevents FAK localization at FAs, in a controlled and dose-dependent manner, by competing with endogenous paxillin for FAK binding. Importantly, the LD2-LD3-LD4 peptide did not otherwise affect FA composition or integrin activation. LD2-LD3-LD4 inhibited FAK-dependent downstream integrin signaling and, unlike existing inhibitors, also blocked FAK's scaffolding functions. We further show that LD2-LD3-LD4 expression markedly reduces FA turnover and inhibits tumor cell migration and invasion. Finally, we show that dimers of a single motif, linked through a flexible linker of the proper size, are sufficient for the displacement of FAK from FAs and for inhibition of tumor cell migration. This work raises the possibility of using a synthetic peptide as an antimetastatic agent, given that effective displacement of FAK from FAs only requires dimers of a single LD motif linked by a short flexible linker. CONCLUSION: In conclusion, these results suggest that FAK displacement from FAs is a promising new strategy to target critical processes implicated in cancer progression and metastasis. Video abstract.