Current view of the immunopathogenesis in inflammatory bowel disease and its implications for therapy

Although the aetiology of inflammatory bowel disease (IBD) remains unknown, the pathogenesis is gradually being unravelled, seeming to be the result of a combination of environmental, genetic, and immunological factors in which an uncontrolled immune response within the intestinal lumen leads to inflammation in genetically predisposed individuals. Multifactorial evidence suggests that a defect of innate immune response to microbial agents is involved in IBD. This editorial outlines the immunopathogenesis of IBD and their current and future therapy. We present IBD as a result of dysregulated mucosal response in the intestinal wall facilitated by defects in epithelial barrier function and the mucosal immune system with excessive production of cytokines growth factors, adhesion molecules, and reactive oxygen metabolites, resulting in tissue injury. Established and evolving therapies are discussed in the second part of this editorial and at the end of this section we review new therapies to modulate the immune system in patients with IBD.

Effects of natural mineral-rich water consumption on the expression of sirtuin 1 and angiogenic factors in the erectile tissue of rats with fructose-induced metabolic syndrome

Consuming a high-fructose diet induces metabolic syndrome （MS）-Iike features, including endothelial dysfunction. Erectile dysfunction is an early manifestation of endothelial dysfunction and systemic vascular disease. Because mineral deficiency intensifies the deleterious effects of fructose consumption and mineral ingestion is protective against MS, we aimed to characterize the effects of 8weeks of natural mineral-rich water consumption on the structural organization and expression of vascular growth factors and receptors on the corpus cavernosum （CC） in 10% fructose-fed Sprague-Dawley rats （FRUCT）. Differences were not observed in the organization of the CC either on the expression of vascular endothelial growth factor （VEGF） or the components of the angiopoietins/Tie2 system. However, opposing expression patterns were observed for VEGF receptors （an increase and a decrease for VEGFR1 and VEGFR2, respectively） in FRUCT animals, with these patterns being strengthened by mineral-rich water ingestion. Mineral-rich water ingestion （FRUCTMIN） increased the proportion of smooth muscle cells compared with FRUCT rats and induced an upregulatory tendency of sirtuin I expression compared with the control and FRUCT groups. Western blot results were consistent with the dual immunofluorescence evaluation. Plasma oxidized low-density lipoprotein and plasma testosterone levels were similar among the experimental groups, although a tendency for an increase in the former was observed in the FRUCTMIN group. The mineral-rich water-treated rats presented changes similar to those observed in rats treated with MS-protective polyphenol-rich beverages or subjected to energy restriction, which led us to hypothesize that the effects of mineral-rich water consumption may be more vast than those directly observed in this study.

Steatosis recovery after treatment with a balanced sunflower or olive oil-based diet: Involvement of perisinusoidal stellate cells

AIM： To analyze the relationship between perisinusoidal stellate cell （PSC） activation and the dietary fat quantity and composition in the treatment of hepatic steatosis. METHODS： Using an experimental rat model of steatosis based on the intake of a hyperlipidic diet （14% fat as olive oil or sunflower oil, HL-O and HL-S, respectively）, we analyzed the liver＇s capability of recovery after the treatment with a normal-lipidic diet （5% fat as olive oil or sunflower oil, NL-O and NL-S, respectively） by immunoo/tochemical and Western blot analysis of glial fibrillary acidic protein （GFAP） expression in PSCs, collagen quantification and serum aminotransferase determination. RESULTS： The fatty infiltration in the steatotic livers decreased after the treatment with both NL diets, indicating liver recovery. This decrease was accompanied with a lower collagen deposition and aminotransferase level as well as changes in the PSC population that increased the GFAP expression. The above-mentioned effects were more pronounced in animals fed on NL-O based diet. CONCLUSION： Treatment with a balanced diet enriched in olive oil contributes to the liver recovery from a steatotic process. The PSC phenotype is a marker of this hepatic-recovery model.

Energy restriction and exercise modulate angiopoietins and vascular endothelial growth factor expression in the cavernous tissue of high-fat diet-fed rats

The purpose of the current study was to evaluate the effect of a high-fat （HF） diet, energy restriction and exercise on the expression of vascular endothelial growth factor （VEGF）, angiopoietin （Ang） I and 2, and their receptors in rat corpus cavernosum （CC）. Male Wistar rats were fed adlibitum with an H F diet for 8 or 16 weeks. After 8 weeks of the H F diet, a group of rats was subjected to energy restriction with or without exercise for 8 weeks. Control animals had free access to standard diet for the same period. After euthanasia, blood was collected and the penises removed for immunofluorescence assays （VEGF, VEGF receptor （VEGFR） I and 2, Angl, Ang2 and Tie2） and semiquantification of VEGF, VEGFR 1, VEGFR2, Angl, Ang2, Tie2, endothelial nitric oxide synthase （eNOS） and Aktlphospho-Akt by Western blotting. HF diet-fed rats exhibited lower high-density lipoprotein cholesterol （HDL-c） levels, higher systolic blood pressure and an increased atherogenic index. A significant increase in Ang2 expression in the CC was verified and coupled to a decrease in VEGF and VEGFRs. The Akt pathway was activated by the HF diet. Energy restriction and exercise increased eNOS expression and restored most HF diet-induced modifications except for VEGFR2 expression. These results emphasize the role of diet on vascular function regulation, demonstrating that cavernous imbalance of VEGF/VEGFRs and Angs/Tie2 systems occurs before serum lipid changes and obesity onset, antedating structural atherosclerotic features.

A comparative study of Epipactis atrorubens in two different forest communities of the Middle Urals, Russia

The objective of this study was to compare ecophysiological and morphological parameters of a regionally endangered orchid species,Epipactis atrorubens(Hoffm.ex Bernh.)Bess.,growing in two forest communities(on serpentine and granite outcrops)of the Middle Urals,Russia.Biodiversity,dominance,and phytocoenosis studies showed the colonization of a wide range of plant species on both sites.The physicochemical properties of the soil,chemical composition and morphological features of E.atrorubens,growing under technogenic conditions(asbestos deposits),on serpentine outcrops and in the natural environment of the granite massif were studied for the first time.The serpentine substrate differed from the granite one by its greater stoniness,circumneutral pH and lower contents of available nitrogen and phosphorus.Extremely high concentrations of magnesium were found in the serpentine soil,some 79 times higher than in the granite substrate.High concentrations of nickel(94 times),chromium(59 times),cobalt(17 times),and iron(4 times)were found in the serpentine substrate,higher than in the granite substrate.The differences between the sites for available metal contents and for root and shoot metal contents were significantly less.Concentrations of most of the metals in the roots were higher than in the shoots.Despite higher metal concentrations and lower nitrogen and phosphorus levels in serpentine soils,E.atrorubens had a larger population and greater viability compared to those growing on granite.Plants on serpentine outcrops were characterized by the formation of a larger number of fruits,greater root lengths and thicker leaf blades,compared to plants on granites.The well-developed orchid mycorrhizae contributed to the survival of this species under unfavorable serpentine conditions.Hence,serpentine outcrops formed due to the mining of asbestos could be a suitable substrate for the light-demanding E.atrorubens due to its capacity to adapt to dry,rocky,nutrient-depleted soils and limited competition from other plants.

Knockdown of apoptosis-inducing factor disrupts function of respiratory complex I

Recent findings suggest that apoptotic protein apoptosis-inducing factor (AIF) may also play an important non-apoptotic function inside mitochondria. AIF was proposed to be an important component of respiratory chain complex I that is the major producer of superoxide radical. The possible role of AIF is still controversial. Superoxide production could be used as a valuable measure of complex I function, because the majority of superoxide is produced there. Therefore, we employed superoxide-specific mitochondrial fluo- rescence dye for detection of superoxide production. We studied an impact of AIF knockdown on function of mitochondrial complex I by analyzing superoxide production in selected cell lines. Our results show that tumoral telomerase-positive (TP) AIF knockdown cell lines display significant increase in superoxide pro- duction in comparison to control cells, while a non-tumoral cell line and tumoral telomerase-negative cell lines with alternative lengthening of telomeres (ALT) show a decrease in superoxide production. According to these results, we can conclude that AIF knockdown disrupts function of complex I and therefore increases the superoxide production in mitochondria. The distinct effect of AIF depletion in various cell lines could result from recently discovered activity of telomerase in mitochondria of TP cancer cells, but this hypothesis needs further investigation.

Reevaluation of dialect boundaries in the Yellowhammer(Emberiza citrinella)based on citizen science data:Should we split or lump?

Dialects are a specific form of geographic variation of birdsong with relatively sharp boundaries between distinct song characteristics,which provide opportunities for focused studies of processes underlying the emergence of spatial patterns in vocalization.Several songbird species that exhibit dialects became models for such research,and for some of them large-scale datasets were assembled that included recordings provided by the public.Among them,the Yellowhammer(Emberiza citrinella,Emberizidae) is particularly prominent,as it has been recently a subject of dedicated citizen science projects focusing on its dialect distribution.The most successful,in terms of public participation as well as the number and density of obtained recordings,was the Dialects of Czech Yellowhammers(DCY) project,which assembled detailed data at the whole-country level.A mosaic of almost all common song variants known across Europe was observed in Czechia,but the results indicated that some of the traditionally recognized Yellowhammer dialects may not represent geographically clustered song variants,at least not in Central Europe.We quantitatively analysed variation(frequency and temporal characteristics and modulation) of the terminal song element in three dialects defined by arbitrary frequency thresholds in DCY.Multivariate analyses indicated that pooling these to two distinct groups reflects the variation in the songs,as well as their spatial distribution,better than retaining the current classification to three dialects or their finer splitting to even more categories.We provide simple measures that may be used for classification of these Yellowhammer song variants in Central Europe.However,we warn from indiscriminate transposing of results from one region to another,as that may lead to substantial biases.Future studies of birdsong variation will benefit from big data assembled by citizen scientists,but to maximise their usefulness for further dialect research,careful delineation of dialect boundaries is essential.

Contribution of mi RNAs to ion-channel remodelling in atrial fibrillation

Atrial fibrillation(AF) is the most commonly encountered clinical arrhythmia associated with pronounced mortality and morbidity, which are related to palpitations, fainting, congestive heart failure, and stroke. Prolonged episodes of AF promote AF persistence mainly due to electrical remodelling that alters ion-channel expression and/or function. Micro RNAs(mi RNAs), a new class of noncoding mR NAs of around 22 nucleotides in length, have recently emerged as one of the key players in the geneexpression regulatory networks. The potential roles of miR NAs in controlling AF have recently been investigated. Several recent studies have provided promising results for a better understanding of the molecular mechanisms of AF. In this review, we summarize the mechanism of miR NAs as regulators of ion-channel gene expression and their role in causing AF through electrical remodelling.

New aspects in celiac disease

Celiac disease (CD) is a common autoimmune disorder characterized by an immune response to ingested gluten and has a strong HLA association with HLA- DQ2 and HLA-DQ8 molecules, but human HLA-DQ risk factors do not explain the entire genetic susceptibility to gluten intolerance. CD is caused by the lack of immune tolerance (oral tolerance) to wheat gluten. In this sense, the expression of soluble HLA-G in CD is of special interest because the molecule plays an important role in the induction of immune tolerance. The enhanced expression of soluble HLA-G found in CD may be part of a mechanism to restore the gluten intolerance. In this editorial, we review recent progress in understanding CD in relation to its prevalence, diagnosis and possible mechanisms of pathogenesis.

Wheat streak mosaic virus： incidence in field crops, potential reservoir within grass species and uptake in winter wheat cultivars

Wheat streak mosaic virus（WSMV）has become a re-emerging pathogen in cereal crops in the Czech Republic.WSMV was first reported in the former Czechoslovakia in the early 1980s,and then no record of the virus was documented until 2009.The incidence of the virus was recorded in recent years in several winter wheat fields and many grass species.Here,we surveyed the incidence of WSMV in cereal crops.The results demonstrated the existence of the virus in winter wheat and volunteer wheat during each year of the monitoring period,which spanned from 2013–2016.Although the range of infected samples was low（6.4%of the total tested samples）,a high incidence of well-distributed virus was recorded.In at least six fields,the virus reached severe and potentially epidemic levels.In accordance with our previous report detailing WSMV infection of native grasses,we tested several grass species commonly grown in the Czech Republic.We found that some grass species acted as experimental hosts and possible reservoirs of the virus;these included Anthoxanthum odoratum（sweet vernal grass）,Arrhenatherum elatius（false oat-grass）,Lolium multiflorum（Italian rye-grass）,Bromus japonicus（Japanese chess）,Echinochloa crus-galli（barnyard grass）,Holcus lanatus（meadow soft grass）and Holcus mollis（creeping soft grass）.Some of these grass species are also important weeds of cereals,which may be the potential source of WSMV infection in cereal crops.Several widely used winter wheat cultivars were tested in the field after artificial inoculation with WSMV to evaluate virus titre by RT-qPCR.Overall,the tested cultivars had a low virus titre,which is associated with mild disease symptoms and may provide a good level of crop resistance to WSMV.

Associative learning in plants:light quality history may matter

The possibility of associative learning in plants is a topic of ongoing controversy.In one published study,growing pea plants were reported to associate two stimuli(airflow and light)and thereafter use one(airflow)as an indicator for the other(light),similar to dogs in Pavlov’s famous experiments.However,this observation could not be independently repeated.Here we examine a possible reason for the failure of a published reproduction attempt,which used substantially different light quality during plant cultivation prior to experimental treatments than in the original study.This could have resulted in dramatically different growth characteristics.While the relevance of the original report of plant associative learning remains questionable,greater attention should be paid to good documenting and standardizing the light conditions,in particular spectral quality,not only in studies of plant learning and memory,but also in other areas of experimental plant biology.

Gene regulatory networks in atrial fibrillation

Atrial fibrillation(AF)is the most frequent arrhythmogenic syndrome in humans.With an estimate incidence of1%-2%in the general population,AF raises up to almost10%-12%in 80+years.Thus,AF represents nowadays a highly prevalent medical problem generating a large economic burden.At the electrophysiological level,distinct mechanisms have been elucidated.Yet,despite its prevalence,the genetic and molecular culprits of this pandemic cardiac electrophysiological abnormality have remained largely obscure.Molecular genetics of AF familiar cases have demonstrated that single nucleotide mutations in distinct genes encoding for ion channels underlie the onset of AF,albeit such alterations only explain a minor subset of patients with AF.In recent years,analyses by means of genome-wide association studies have unraveled a more complex picture of the etiology of AF,pointing out to distinct cardiac-enriched transcription factors,as well as to other regulatory genes.Furthermore a new layer of regulatory mechanisms have emerged,i.e.,post-transcriptional regulation mediated by non-coding RNA,which have been demonstrated to exert pivotal roles in cardiac electrophysiology.In this manuscript,we aim to provide a comprehensive review of the genetic regulatory networks that if impaired exert electrophysiological abnormalities that contribute to the onset,and subsequently,on self-perpetuation of AF.

Prompt and Easy Activation by Specific Thioredoxins of Calvin Cycle Enzymes of Arabidopsis thaliana Associated in the GAPDH/CP12/PRK Supramolecular Complex

The Calvin cycle enzymes glyceraldehyde-3-phosphate dehydrogenase （GAPDH） and phosphoribulokinase （PRK） can form under oxidizing conditions a supramolecular complex with the regulatory protein CP12. Both GAPDH and PRK activities are inhibited within the complex, but they can be fully restored by reduced thioredoxins （TRXs）. We have investigated the interactions of eight different chloroplast thioredoxin isoforms （TRX f1, m1, m2, m3, m4, y1, y2, x） with GAPDH （A4, B4, and B8 isoforms）, PRK and CP12 （isoform 2）, all from Arabidopsis thaliana. In the complex, both A4-GAPDH and PRK were promptly activated by TRX f1, or more slowly by TRXs ml and m2, but all other TRXs were ineffective. Free PRK was regulated by TRX f1, m1, or m2, while B4- and Bs-GAPDH were absolutely specific for TRX fl. Interestingly, reductive activation of PRK caged in the complex was much faster than reductive activation of free oxidized PRK, and activation of A4-GAPDH in the complex was much faster （and less demanding in terms of reducing potential） than activation of free oxidized B4- or BB-GAPDH. It is proposed that CP12-assembled supramolecular complex may represent a reservoir of inhibited enzymes ready to be released in fully active conformation following reduction and dissociation of the complex by TRXs upon the shift from dark to low light. On the contrary, autonomous redox-modulation of GAPDH （B- containing isoforms） would be more suited to conditions of very active photosynthesis.

Potential role of the IL-33/ST2 axis in celiac disease

The IL-33/ST2 axis has been implicated in the pathogenesis of several tissue-specific autoimmune diseases. Celiac disease （CD） is the only autoimmune disease in which both the major genetic factors （HLA-DQ2/DQ8） and etiologic ones （dietary gluten） for susceptibility are known. We have measured serum levels and determined intestinal tissue expression of IL-33 and its receptor soluble ST2 in patients with CD to investigate their association with disease activity. Serum and tissue levels of both IL-33 and sST2 were significantly higher in patients with CD compared with those in control patients without CD. We show that toxic peptides extracted from barley and wheat gliadin significantly stimulate the production of IL-33 and ST2 in cultured peripheral blood mononuclear cell from celiac patients, strongly implicating the IL-33/ST2 axis in the pathogenesis of CD. The higher levels of IL-33 and its receptor ST2 in tissue and serum reflect an active inflammatory state and may represent a potential biomarker for disease activity. A better understanding of IL-33/ ST2 release, mode of action, and regulation will be crucial to develop therapeutics that target the IL-33/ST2 pathway to treat CD.

Dysregulation of the PD-1/PD-L1 pathway contributes to the pathogenesis of celiac disease

In this study,we focus on the alteration of the programmed cell death 1(PD-1)/PD-L1 pathway in celiac disease.We discuss the diverse roles of the PD-1 pathway in regulating immune responses and how this knowledge can improve celiac disease(CD)autoimmunity.The PD-1 and PD-L1 levels in the serum and in intestinal biopsies of CD patients may be relevant to the determination of a possible correlation between markers of the autoimmune response,inflammation,and disease activity.

The AFB1 auxin receptor controls the cytoplasmic auxin response pathway in Arabidopsis thaliana

The phytohormone auxin triggers root growth inhibition within seconds via a non-transcriptionalpathway.Among members of the TIR1/AFB auxin receptor family,AFB1 has a primary role in this rapidresponse. However, the unique features that confer this specific function have not been identified.Here we show that the N-terminal region of AFB1, including the F-box domain and residues thatcontribute to auxin binding,is essential and sufficient for its specific role in the rapid response. Substitutionof the N-terminal region of AFB1 with that of TIR1 disrupts its distinct cytoplasm-enriched localizationand activity in rapid root growth inhibition by auxin. Importantly, the N-terminal region of AFB1 isindispensable for auxin-triggered calcium influx, which is a prerequisite for rapid root growth inhibition.Furthermore, AFB1 negatively regulates lateral root formation and transcription of auxin-induced genes,suggesting that it plays an inhibitory role in canonical auxin signaling. These results suggest that AFB1may buffer the transcriptional auxin response, whereas it regulates rapid changes in cell growth thatcontributeto rootgravitropism.

Targeting hormone-resistant breast cancer cells with docetaxel:a look inside the resistance

Aim:The study aims to analyze the effect of long-term incubation of ERα-positive MCF7 breast cancer cells with 4-hydroxytamoxifen(HT)on their sensitivity to tubulin polymerization inhibitor docetaxel.Methods:The analysis of cell viability was performed by the MTT method.The expression of signaling proteins was analyzed by immunoblotting and flow cytometry.ERαactivity was evaluated by gene reporter assay.To establish hormone-resistant subline MCF7,breast cancer cells were treated with 4-hydroxytamoxifen for 12 months.Results:The developed MCF7/HT subline has lost sensitivity to 4-hydroxytamoxifen,and the resistance index was 2.Increased Akt activity(2.2-fold)and decreased ERαexpression(1.5-fold)were revealed in MCF7/HT cells.The activity of the estrogen receptorαwas reduced(1.5-fold)in MCF7/HT.Evaluation of class Ⅲβ-tubulin expression(TUBB3),a marker associated with metastasis,revealed the following trends:higher expression of TUBB3 was detected in triple-negative breast cancer MDA-MB-231 cells compared to hormone-responsive MCF7 cells(P<0.05).The lowest expression of TUBB3 was found in hormone-resistant MCF7/HT cells(MCF7/HT<MCF7<MDA-MB-231,approximately 1:2:4).High TUBB3 expression strongly correlated with docetaxel resistance:IC_(50)value of docetaxel for MDA-MB-231 cells was greater than that for MCF7 cells,whereas resistant MCF7/HT cells were the most sensitive to the drug.The accumulation of cleaved PARP(a 1.6-fold increase)and Bcl-2 downregulation(1.8-fold)were more pronounced in docetaxel-treated resistant cells(P<0.05).The expression of cyclin D1 decreased(2.8-fold)only in resistant cells after 4 nM docetaxel treatment,while this marker was unchanged in parental MCF7 breast cancer cells.Conclusion:Further development of taxane-based chemotherapy for hormone-resistant cancer looks highly promising,especially for cancers with low TUBB3 expression.

Making the Genotypic Variation Visible:Hyperspectral Phenotyping in Scots Pine Seedlings

Hyperspectral reflectance contains valuable information about leaf functional traits,which can indicate a plant's physiological status.Therefore,using hyperspectral reflectance for high-throughput phenotyping of foliar traits could be a powerful tool for tree breeders and nursery practitioners to distinguish and select seedlings with desired adaptation potential to local environments.We evaluated the use of 2 nondestructive methods(i.e.,leaf and proximal/canopy)measuring hyperspectral reflectance in the 350-to 2,500-nm range for phenotyping on 1,788 individual Scots pine seedlings belonging to lowland and upland ecotypes of 3 different local populations from the Czech Republic.Leaf-level measurements were collected using a spectroradiometer and a contact probe with an internal light source to measure the biconical reflectance factor of a sample of needles placed on a black background in the contact probe field of view.The proximal canopy measurements were collected under natural solar light,using the same spectroradiometer with fiber optical cable to collect data on individual seedlings'hemispherical conical reflectance factor.The latter method was highly susceptible to changes in incoming radiation.Both spectral datasets showed statistically significant differences among Scots pine populations in the whole spectral range.Moreover,using random forest and support vector machine learning algorithms,the proximal data obtained from the top of the seedlings offered up to 83%accuracy in predicting 3 different Scots pine populations.We conclude that both approaches are viable for hyperspectral phenotyping to disentangle the phenotypic and the underlying genetic variation within Scots pine seedlings.