Are SMAD2/4/7 genetic variants associated with rheumatoid arthritis susceptibility and severity?

OBJECTIVES: SMADs play one of the key roles in the TGFß signalling pathway. Therefore, through their involvement in the immune response as well as in the fibrosis process, these proteins appear to take on one of the essential functions in the pathogenesis of autoimmune connective tissue diseases such as RA. This study aimed to investigate the association of selected SNPs in SMAD2/4/7 with RA risk in the Caucasian population and disease course in RA patients. METHODS: The study was conducted on 647 patients with established RA and 496 unrelated healthy controls (HCs). All patients fulfilled the American College of Rheumatology Diagnostic classification criteria for RA (ACR 1987). The analysis has been conducted using TaqMan genotyping assay. Transcript-inferred pathogenicity score (TraP-score) has been evaluated by TrapScore. PredictSNP.2 has been used to predict the effect of amino acid substitutions. RESULTS: The present study revealed in SMAD4 a significantly higher frequency of AG rs12456284 (under codominant model OR=0.62 p=0.027 and overdominant model OR=0.59 p=0.016) and GA rs10502913 (under codominant model OR=0.65 p=0.050 and overdominant OR=0.64 p=0.033) genotypes in healthy subjects in comparison to RA patients. Additionally, very strong LD has been noted between these two genetic variants (D'=0.95 r2=0.90). Moreover, bioinformatic analysis classified rs12456284 as deleterious change with 94% prediction accuracy. SMAD2 rs1792666 and SMAD7 rs3736242 showed to have the highest association with disease course. SMAD4 rs10502913, SMAD7 rs3736242, and SMAD7 rs4464148 were associated with the concentration of creatinine. CONCLUSIONS: Our results suggested that rs12456284 and rs10502913 in SMAD4 may have a potential protective effect against RA. Particularly, SMAD2 rs1792666 and SMAD7 rs3736242 seem to be significantly associated with diseases course in RA patients in the Caucasian population.

Artificial light at night (ALAN) affects behaviour, but does not change oxidative status in freshwater shredders.

Artificial light at night (ALAN) alters circadian rhythms in animals and therefore can be a source of environmental stress affecting their physiology and behaviour. The impact of ALAN can be related to the increased light level, but also to the spectral composition of night lighting. Previous research showed that many species can be particularly sensitive to the LED light, but it is unclear if they respond to its broad spectrum or specifically to the blue light wavelength. In this study, we tested whether dim ALAN (2 lx) differing in the spectral quality (warm white LED, blue LED, high-pressure sodium HPS light) modifies behaviour and changes oxidative status in two nocturnal freshwater shredder species: Dikerogammarus villosus and Gammarus jazdzewskii (Gammaroidea, Amphipoda). Our experiment revealed that ALAN, irrespective of its spectral quality, did not affect the oxidative stress markers in cells (the level of reactive oxygen species and lipid peroxidation). However, ALAN changed the gammarid behaviour in a species-specific manner, which can potentially reduce the fitness of the shredders. Dikerogammarus villosus avoided all types of light compared to darkness. Therefore, confined to the shelter, D. villosus may have fewer opportunities to forage and/or mate. Gammarus jazdzewskii was sensitive only to the narrow-spectrum blue light, but did not respond to the HPS and white LED light. Avoidance is a typical response of gammarids to natural light, thus the disruption of this behaviour in the presence of common ALAN sources can increase the predation risk in this species. To summarize, behavioural modifications induced by ALAN seem more pronounced than changes in physiology and can constitute the main driver of disturbances in the processing of organic matter in freshwater ecosystems by invertebrate shredders.

Disruptive effect of artificial light at night on leaf litter consumption, growth and activity of freshwater shredders.

Artificial light at night (ALAN) is a globally widespread phenomenon potentially affecting ecosystem processes, such as leaf litter breakdown, which is a source of organic matter in fresh waters. Here, we conducted a long-term experiment to test the effects of ALAN (2 lx) differing in spectral composition: white LEDs and high pressure sodium lamps (HPS) on leaf consumption, growth and activity of two macroinvertebrate species of shredders: Gammarus jazdzewskii and Dikerogammarus villosus (Crustacea, Amphipoda), compared to the undisturbed light-dark cycle. We also tested if the nocturnal illumination would influence the algal community colonising leaves, which is an important component of the leaf-shredder diet. We found that LED light increased the consumption of leaves by both species, which was nearly twice as high as in other treatments, and supressed the growth rate of G. jazdzewskii, whereas the growth of D. villosus was not affected by either light type. Moreover, D. villosus reduced its activity when exposed to ALAN of both types. As ALAN-induced changes in shredder growth and consumption were not associated with their increased activity or decreased food quality, we suggest that LED light may be a source of physiological stress for shredders, raising their energy expenditure, which was compensated by increased food intake. We have shown that LED illumination induces greater effects on wildlife than alternative, narrow wavelength spectrum light sources, such as HPS lamps, and may potentially alter the litter breakdown in aquatic ecosystems. It may accelerate the turnover of leaves by shredders, but on the other hand, it may negatively affect the fitness of macroinvertebrates and thus disturb the leaf processing over a longer term.

The role of autofluorescence, photodynamic diagnosis and Photodynamic therapy in malignant tumors of the duodenum.

This article presents the current state of knowledge and a review of the literature in terms of the prevalence, etiopathogenesis, differential diagnosis, management, prognosis, and treatment of malignant tumors of the duodenum. The role of autofluorescence and photodynamic diagnosis as an emerging treatment method for rarely o ccurring duodenal malignant neoplasms .. We selected publications which can be found in databases such as The National Center for Biotechnology Information, U.S. National Library of Medicine (PubMed), The American Chemical Society, The American Association of Pharmaceutical Sciences and The American Society for Photobiology and The Canada Institute for Scientific and Technical Information.

Comparison of treatment standards in Atopic Dermatitis management across selected geographies prior to emerging targeted therapies onset.

Atopic Dermatitis (AD) is a chronic inflammatory disease persisting predominantly in the pediatric population. Treatment is generally supervised by various medical specialists, including primary care practitioners, allergists, and dermatologists. This divergence in disease management allows various therapeutic approaches to be administered to patients by supervised physicians. This article covers etiology of the disease and summarizes dermatologic treatment standards of selected countries binding prior to the registration of dupilumab by both the European Medicines Agency (EMA) and Federal Drug Administration (FDA) in 2017. Before recent development in targeted therapies (small molecules and biologic agents), standards in AD treatment remained unchanged for years with extensive similarities across a sample group of countries in particular geographic and economic regions. The spectrum of available and popular therapeutic options can be categorized into three dominating groups: non-pharmacologic, pharmacologic, and systemic interventions. Their prescription, in principle, was historically driven by disease severity and previous treatment history. However, advances in targeted therapies may change AD management guidelines and medical care standards.

Epidemiology of atopic dermatitis in Europe.

Atopic dermatitis (AD) is a chronic inflammatory disease persisting predominantly in the pediatric population. Its development is most presumably multifactorial and a derivative of interplay between genetic, immunologic, and environmental causes. To the authors' knowledge, no multinational and systematic database of AD prevalence is established and maintained for Europe. Thus, epidemiologic data originating from the multinational studies was compiled to draw a picture of AD in both pediatric and adult populations in Europe. The outcomes of this exercise support the general observation that AD prevalence follows the latitudinal pattern with higher prevalence values in northern Europe and decreases progressively towards southern Europe. Noteworthy, the data shows significant differences on the country-level, with higher prevalence in municipal areas than rural. Finally, and unsurprisingly, the collected data reinforces the observation of AD prevalence being highest in pediatric populations in contrast to adults. Herein, data presented was additionally supplemented with the information on current standing on AD etiology.

Combined effects of nocturnal exposure to artificial light and habitat complexity on fish foraging.

Due to the widespread use of artificial light, freshwater ecosystems in urban areas at night are often subjected to light of intensities exceeding that of the moonlight. Nocturnal dim light could modify fish behaviour and benefit visual predators because of enhanced foraging success compared to dark nights. However, effects of nocturnal light could be mitigated by the presence of structured habitats providing refuges for prey. We tested in laboratory experiments whether nocturnal light of low intensity (2 lx) increases foraging efficiency of the Eurasian perch (Perca fluviatilis) on invertebrate prey (Gammarus fossarum). The tests were conducted at dusk and night under two light regimes: natural cycle with dark nights and disturbed cycle with artificially illuminated nights, in habitats differing in structural complexity: sand and woody debris. We found that nocturnal illumination significantly enhanced the consumption of gammarids by fish compared to dark nights. In addition, the perch was as effective predator in illuminated nights (2 lx) as at dusk (10 lx). Woody debris provided an effective refuge only in combination with undisturbed darkness, but not in illuminated nights. Our results suggest that nocturnal illumination in aquatic ecosystems may contribute to significant reductions in invertebrate population sizes through fish predation. The loss of darkness reduces the possibility of using shelters by invertebrates and hence the effects of elevated light levels at night could not be mitigated by an increased habitat complexity.

Neuropeptide Y receptor interactions regulate its mitogenic activity.

Neuropeptide Y (NPY) is a multifunctional neurotransmitter acting via G protein-coupled receptors - Y1R, Y2R and Y5R. NPY activities, such as its proliferative effects, are mediated by multiple receptors, which have the ability to dimerize. However, the role of this receptor interplay in NPY functions remains unclear. The goal of the current study was to identify NPY receptor interactions, focusing on the ligand-binding fraction, and determine their impact on the mitogenic activity of the peptide. Y1R, Y2R and Y5R expressed in CHO-K1 cells formed homodimers detectable on the cell surface by cross-linking. Moreover, Y1R and Y5R heterodimerized, while no Y2R/Y5R heterodimers were detected. Nevertheless, Y5R failed to block internalization of its cognate receptor in both Y1R/Y5R and Y2R/Y5R transfectants, indicating Y5R transactivation upon stimulation of the co-expressed receptor. These receptor interactions correlated with an augmented mitogenic response to NPY. In Y1R/Y5R and Y2R/Y5R transfectants, the proliferative response started at picomolar NPY concentrations, while nanomolar concentrations were needed to trigger proliferation in cells transfected with single receptors. Thus, our data identify direct and indirect heterotypic NPY receptor interactions as the mechanism amplifying its activity. Understanding these processes is crucial for the design of treatments targeting the NPY system.

Cell Surface Protein Detection to Assess Receptor Internalization.

The migration of membrane receptors upon exposure to different stimulants/inhibitors is of great importance. Among others, the internalization of membrane receptors affects their accessibility to ligands and cell responsiveness to environmental cues. Experimentally, receptor internalization can be used as a measure of their activation. In our studies, we employed this approach to explore cross-talk between a seven transmembrane domain receptor for neuropeptide Y (NPY), Y5R, and a tyrosine kinase receptor for brain-derived neurotrophic factor (BDNF), TrkB. To this end, we measured the internalization of Y5R upon stimulation with the TrkB ligand, BDNF. Upon treatment with BDNF, the cells were exposed to a membrane impermeable, biotinylation reagent that selectively labels surface proteins. Subsequently, the biotinylated membrane proteins were affinity-purified on columns with avidin resins and analyzed by Western blot. Differences in the fraction of receptors present on the cell surface of control and ligand-treated cells served as a measure of their internalization and response to particular stimuli.

Hypoxia shifts activity of neuropeptide Y in Ewing sarcoma from growth-inhibitory to growth-promoting effects.

Ewing sarcoma (ES) is an aggressive malignancy driven by an oncogenic fusion protein, EWS-FLI1. Neuropeptide Y (NPY), and two of its receptors, Y1R and Y5R are up-regulated by EWS-FLI1 and abundantly expressed in ES cells. Paradoxically, NPY acting via Y1R and Y5R stimulates ES cell death. Here, we demonstrate that these growth-inhibitory actions of NPY are counteracted by hypoxia, which converts the peptide to a growth-promoting factor. In ES cells, hypoxia induces another NPY receptor, Y2R, and increases expression of dipeptidyl peptidase IV (DPPIV), an enzyme that cleaves NPY to a shorter form, NPY3-36. This truncated peptide no longer binds to Y1R and, therefore, does not stimulate ES cell death. Instead, NPY3-36 acts as a selective Y2R/Y5R agonist. The hypoxia-induced increase in DPPIV activity is most evident in a population of ES cells with high aldehyde dehydrogenase (ALDH) activity, rich in cancer stem cells (CSCs). Consequently, NPY, acting via Y2R/Y5Rs, preferentially stimulates proliferation and migration of hypoxic ALDHhigh cells. Hypoxia also enhances the angiogenic potential of ES by inducing Y2Rs in endothelial cells and increasing the release of its ligand, NPY3-36, from ES cells. In summary, hypoxia acts as a molecular switch shifting NPY activity away from Y1R/Y5R-mediated cell death and activating the Y2R/Y5R/DPPIV/NPY3-36 axis, which stimulates ES CSCs and promotes angiogenesis. Hypoxia-driven actions of the peptide such as these may contribute to ES progression. Due to the receptor-specific and multifaceted nature of NPY actions, these findings may inform novel therapeutic approaches to ES.

Dipeptidyl peptidases as survival factors in Ewing sarcoma family of tumors: implications for tumor biology and therapy.

Ewing sarcoma family of tumors (ESFT) is a group of aggressive pediatric malignancies driven by the EWS-FLI1 fusion protein, an aberrant transcription factor up-regulating specific target genes, such as neuropeptide Y (NPY) and its Y1 and Y5 receptors (Y5Rs). Previously, we have shown that both exogenous NPY and endogenous NPY stimulate ESFT cell death via its Y1 and Y5Rs. Here, we demonstrate that this effect is prevented by dipeptidyl peptidases (DPPs), which cleave NPY to its shorter form, NPY(3-36), not active at Y1Rs. We have shown that NPY-induced cell death can be abolished by overexpression of DPPs and enhanced by their down-regulation. Both NPY treatment and DPP blockade activated the same cell death pathway mediated by poly(ADP-ribose) polymerase (PARP-1) and apoptosis-inducing factor (AIF). Moreover, the decrease in cell survival induced by DPP inhibition was blocked by Y1 and Y5R antagonists, confirming its dependence on endogenous NPY. Interestingly, similar levels of NPY-driven cell death were achieved by blocking membrane DPPIV and cytosolic DPP8 and DPP9. Thus, this is the first evidence of these intracellular DPPs cleaving releasable peptides, such as NPY, in live cells. In contrast, another membrane DPP, fibroblast activation protein (FAP), did not affect NPY actions. In conclusion, DPPs act as survival factors for ESFT cells and protect them from cell death induced by endogenous NPY. This is the first demonstration that intracellular DPPs are involved in regulation of ESFT growth and may become potential therapeutic targets for these tumors.

Chronic stress, combined with a high-fat/high-sugar diet, shifts sympathetic signaling toward neuropeptide Y and leads to obesity and the metabolic syndrome.

In response to stress, some people lose while others gain weight. This is believed to be due to either increased beta-adrenergic activation, the body's main fat-burning mechanism, or increased intake of sugar- and fat-rich "comfort foods." A high-fat, high-sugar (HFS) diet alone, however, cannot account for the epidemic of obesity, and chronic stress alone tends to lower adiposity in mice. Here we discuss how chronic stress, when combined with an HFS diet, leads to abdominal obesity by releasing a sympathetic neurotransmitter, neuropeptide Y (NPY), directly into the adipose tissue. In vitro, when "stressed" with dexamethasone, sympathetic neurons shift toward expressing more NPY, which stimulates endothelial cell (angiogenesis) and preadipocyte proliferation, differentiation, and lipid-filling (adipogenesis) by activating the same NPY-Y2 receptors (Y2Rs). In vivo, chronic stress, consisting of cold water or aggression in HFS-fed mice, stimulates the release of NPY and the expression of Y2Rs in visceral fat, increasing its growth by 50% in 2 weeks. After 3 months, this results in metabolic syndrome-like symptoms with abdominal obesity, inflammation, hyperlipidemia, hyperinsulinemia, glucose intolerance, hepatic steatosis, and hypertension. Remarkably, local intra-fat Y2R inhibition pharmacologically or via adenoviral Y2R knock-down reverses or prevents fat accumulation and metabolic complications. These studies demonstrated for the first time that chronic stress, via the NPY-Y2R pathway, amplifies and accelerates diet-induced obesity and the metabolic syndrome. Our findings also suggest the use of local administration of Y2R antagonists for treatment of obesity and NPY-Y2 agonists for fat augmentation in other clinical applications.