Emotional functions in transsexuals after the first step in physical transformation

Osoby transseksualne muszą stawić czoła wielu problemom zdrowotnym, społecznym i biurokratycznym związanym z podjęciem decyzji o transformacji. Liczne wyzwania występują zarówno przed, w trakcie jak i po zabiegach medycznych. W dążeniu do poprawy jakości życia osób transseksualnych pożądane wydaje się zatem zbadanie funkcji psychologicznych w trakcie postępowania medycznego. Obecne badania prowadzono w celu określenia kondycji emocjonalnej tej grupy pacjentów i obejmowały one 28 transseksualistów w trakcie transformacji z płci biologicznej F/M oraz dwie grupy kontrolne; mężczyzn i kobiet w podobnym wieku. Jako metody badania wykorzystano następujące skale psychometryczne: CECS (Skala Kontroli Emocjonalnej M. Watson i S. Greer w polskiej adaptacji Z. Juczyńskiego), ISCL (polska adaptacja Inwentarza Lęku dla Dorosłych w adaptacji T. Sosnowskiego ) i GSES (polska adaptacja Z. Juczyńskiego i K. Wrześniewskiego Skali Uogólnionej Samoskuteczności R. Schwarzer, M. Jerusalem). Wyniki uzyskane w grupie badanych osób transseksualnych (F/M) porównano z wynikami analogicznych testów przeprowadzonych w grupach kontrolnych. Pacjenci transseksualni F/M prezentowali wyniki bardzo podobne do grupy mężczyzn w zakresie poczucia samoskuteczności, oraz stanu i cechy lęku, podczas gdy ich subiektywna ocena kontroli lęku była bardziej zbliżona do grupy kontrolnej kobiet i istotnie statystycznie niższa niż w grupie kontrolnej męskiej. Transsexuals have to face multiple medical, social and bureaucratic problems. These problems are not only encountered before the transformation, but also during and after medical procedures. In the search for improvement of transsexual individuals’ quality of life during therapy, it seems desirable to supplement hormonal treatments with psychological explorations. This study was conducted with the aim of defining emotional conditions and included 28 transsexual female-to-male (F/M) patients and two gender-divided control groups (males and females) of similar age. The following psychometric scales were used: CECS (Courtauld Emotional Control Scale constructed by M. Watson and S. Greer in the Polish Adaptation by Z. Juczyński), ISCL (the Polish Adaptation of the State-Trait Anxiety Inventory for Adults by T. Sosnowski), and GSES (the Polish Adaptation of the R. Schwarzer, M. Jerusalem Generalized Self-Efficacy Scaleby Z. Juczyński and K. Wrześniewski). Transsexual F/M patients appeared very similar to males in the male control group in terms of their subjective selfefficacy and state-trait anxiety, while their subjective belief of anxiety and fear control was more comparable to that of the female controls. It was also found to be statistically significantly lower than in the male controls

The circadian clock protein REVERBα inhibits pulmonary fibrosis development

Pulmonary inflammatory responses lie under circadian control; however, the importance of circadian mechanisms in the underlying fibrotic phenotype is not understood. Here, we identify a striking change to these mechanisms resulting in a gain of amplitude and lack of synchrony within pulmonary fibrotic tissue. These changes result from an infiltration of mesenchymal cells, an important cell type in the pathogenesis of pulmonary fibrosis. Mutation of the core clock protein REVERBα in these cells exacerbated the development of bleomycin-induced fibrosis, whereas mutation of REVERBα in club or myeloid cells had no effect on the bleomycin phenotype. Knockdown of REVERBα revealed regulation of the little-understood transcription factor TBPL1. Both REVERBα and TBPL1 altered integrinβ1 focal-adhesion formation, resulting in increased myofibroblast activation. The translational importance of our findings was established through analysis of 2 human cohorts. In the UK Biobank, circadian strain markers (sleep length, chronotype, and shift work) are associated with pulmonary fibrosis, making them risk factors. In a separate cohort, REVERBα expression was increased in human idiopathic pulmonary fibrosis (IPF) lung tissue. Pharmacological targeting of REVERBα inhibited myofibroblast activation in IPF fibroblasts and collagen secretion in organotypic cultures from IPF patients, thus suggesting that targeting of REVERBα could be a viable therapeutic approach

Characterization of the Adaptor Protein XB130, a Tyrosine Kinase Substrate and a Novel Component of the Lamellipodia

Adaptor proteins play a vital role in the propagation of cellular signals. Although they lack endogenous catalytic activity, they contain a variety of protein binding modules, which enable them to promote specific and efficient interactions with their binding partners. They form integrative platforms for a variety of molecules (e.g. lipids, tyrosine kinases, cytoskeletal and signaling proteins), and thereby link and coordinate key functions such as cell growth, motility and shape determination. Our laboratory has recently cloned a novel, 130 kDa adaptor protein, named XB130, as a structural homolog of actin-filament-associated-protein (AFAP-110), a stress fiber-binding Src substrate. However, the molecular interactions and functions of this novel adaptor remained to be elucidated. To characterize the function of XB130 we asked two general questions: (1) Is XB130 involved in the signal transduction pathways of tyrosine kinases? And (2) Is XB130 capable of regulating the cytoskeleton and/or is it regulated by the cytoskeleton? To address these questions first we investigated the tissue distribution of XB130 and discovered that it is abundantly expressed in thyroid. Therefore we asked whether it is a target of the thyroid-specific tyrosine kinase, RET/PTC, a genetically rearranged, constitutively active enzyme that plays a pathogenic role in papillary thyroid cancer. We found that XB130 is a RET/PTC substrate that couples RET/PTC signaling to phosphatidylinositol 3-kinase (PI3K) activation through its phosphorylation dependent interaction with the regulatory subunit p85 of PI3K. XB130 plays an important role in PI3K signaling, as downregulation of XB130 in TPC1 papillary thyroid cancer cells, harboring the RET/PTC1 kinase, strongly reduced Akt activity and concomitantly inhibited cell cycle progression and survival in suspension. In the second part we demonstrate that XB130 is a novel Rac- and cytoskeleton-regulated protein that exhibits high affinity to lamellipodial (branched) F-actin and impacts motility and invasiveness of tumor cells. In conclusion, my work characterized a novel adaptor protein and assigned two well-defined pathophysiological functions to it in the context of thyroid cancer cells.Ph

Tyrosine Phosphorylation of Caspase-8 Abrogates Its Apoptotic Activity and Promotes Activation of c-Src.

Src family tyrosine kinases (SFKs) phosphorylate caspase-8A at tyrosine (Y) 397 resulting in suppression of apoptosis. In addition, the phosphorylation of caspase-8A at other sites including Y465 has been implicated in the regulation of caspase-8 activity. However, the functional consequences of these modifications on caspase-8 processing/activity have not been elucidated. Moreover, various Src substrates are known to act as potent Src regulators, but no such role has been explored for caspase-8. We asked whether the newly identified caspase-8 phosphorylation sites might regulate caspase-8 activation and conversely, whether caspase-8 phosphorylation might affect Src activity. Here we show that Src phosphorylates caspase-8A at multiple tyrosine sites; of these, we have focused on Y397 within the linker region and Y465 within the p12 subunit of caspase-8A. We show that phosphomimetic mutation of caspase-8A at Y465 prevents its cleavage and the subsequent activation of caspase-3 and suppresses apoptosis. Furthermore, simultaneous phosphomimetic mutation of caspase-8A at Y397 and Y465 promotes the phosphorylation of c-Src at Y416 and increases c-Src activity. Finally, we demonstrate that caspase-8 activity prevents its own tyrosine phosphorylation by Src. Together these data reveal that dual phosphorylation converts caspase-8 from a pro-apoptotic to a pro-survival mediator. Specifically, tyrosine phosphorylation by Src renders caspase-8 uncleavable and thereby inactive, and at the same time converts it to a Src activator. This novel dynamic interplay between Src and caspase-8 likely acts as a potent signal-integrating switch directing the cell towards apoptosis or survival

Hyperosmotic stress regulates the distribution and stability of myocardin-related transcription factor, a key modulator of the cytoskeleton

Hyperosmotic stress initiates several adaptive responses, including the remodeling of the cytoskeleton. Besides maintaining structural integrity, the cytoskeleton has emerged as an important regulator of gene transcription. Myocardin-related transcription factor (MRTF), an actin-regulated coactivator of serum response factor, is a major link between the actin skeleton and transcriptional control. We therefore investigated whether MRTF is regulated by hyperosmotic stress. Here we show that hypertonicity induces robust, rapid, and transient translocation of MRTF from the cytosol to the nucleus in kidney tubular cells. We found that the hyperosmolarity-triggered MRTF translocation is mediated by the RhoA/Rho kinase (ROK) pathway. Moreover, the Rho guanine nucleotide exchange factor GEF-H1 is activated by hyperosmotic stress, and it is a key contributor to the ensuing RhoA activation and MRTF translocation, since siRNA-mediated GEF-H1 downregulation suppresses these responses. While the osmotically induced RhoA activation promotes nuclear MRTF accumulation, the concomitant activation of p38 MAP kinase mitigates this effect. Moderate hyperosmotic stress (600 mosM) drives MRTF-dependent transcription through the cis-element CArG box. Silencing or pharmacological inhibition of MRTF prevents the osmotic stimulation of CArG-dependent transcription and renders the cells susceptible to osmotic shock-induced structural damage. Interestingly, strong hyperosmolarity promotes proteasomal degradation of MRTF, concomitant with apoptosis. Thus, MRTF is an osmosensitive and osmoprotective transcription factor, whose intracellular distribution is regulated by the GEF-H1/RhoA/ROK and p38 pathways. However, strong osmotic stress destabilizes MRTF, concomitant with apoptosis, implying that hyperosmotically induced cell death takes precedence over epithelial-myofibroblast transition, a potential consequence of MRTF-mediated phenotypic reprogramming

Tyrosine phosphorylation of caspase-8A in multiple sites follows Src expression.

<p>A) Caspase-8A structure and SFK tyrosine phosphorylation sites predicted by GPS2.1. B) HEK293 cells were co-transfected with inactive C377S mutant of GFP-caspase-8A and Y527F Src for 24 hours then lysates immunoprecipitated using an anti-GFP antibody. GFP-caspase-8A IP was subjected to SDS-PAGE, and the appropriate band (80 kDa) was excised and sent for LC-MS/MS analysis. C) Mutagenesis was performed at Y397/Y380 and Y465/Y448 sites to generate phosphomimetic (tyrosine to glutamic acid) and non-phosphorylatable (tyrosine to phenylalanine) caspase-8A/B mutants.</p