Biomarkers of NRF2 signalling: Current status and future challenges.

The cytoprotective transcription factor NRF2 regulates the expression of several hundred genes in mammalian cells and is a promising therapeutic target in a number of diseases associated with oxidative stress and inflammation. Hence, an ability to monitor basal and inducible NRF2 signalling is vital for mechanistic understanding in translational studies. Due to some caveats related to the direct measurement of NRF2 levels, the modulation of NRF2 activity is typically determined by measuring changes in the expression of one or more of its target genes and/or the associated protein products. However, there is a lack of consensus regarding the most relevant set of these genes/proteins that best represents NRF2 activity across cell types and species. We present the findings of a comprehensive literature search that according to stringent criteria identifies GCLC, GCLM, HMOX1, NQO1, SRXN1 and TXNRD1 as a robust panel of markers that are directly regulated by NRF2 in multiple cell and tissue types. We assess the relevance of these markers in clinically accessible biofluids and highlight future challenges in the development and use of NRF2 biomarkers in humans.

Proximity extension assay-based proteomics studies in neurodegenerative disorders and multiple sclerosis.

Neurodegenerative diseases impact the structure and operation of the nervous system, causing progressive and irreparable harm. Efforts for distinguishing neurodegenerative diseases in their early stages are continuing. Despite several biomarkers being identified, there is always search for more accurate and abundant ones. Additionally, it can be difficult to pinpoint the precise neurodegenerative disorder affecting a patient as the symptoms of these conditions frequently overlap. Numerous studies have shown that pathological changes occur years before clinical signs appear. Therefore, it is crucial to discover blood-based biomarkers for neurodegenerative diseases for easier and earlier diagnosis. Proximity extension assay is a unique proteomics method that uses antibodies linked to oligonucleotides for quantifying proteins with real-time PCR. Proximity extension assay can identify even low-quantity proteins using a small volume of specimens with increased sensitivity compared to conventional methods. In this article, we reviewed the employment of proximity extension assay technology to detect biomarkers or protein profiles for several neurodegenerative diseases.

Melatonin Alters the miRNA Transcriptome of Inflammasome Activation in Murine Microglial Cells.

Systemic inflammation can have devastating effects on the central nervous system via its resident immune cells, the microglia. One of the primary mediators of this inflammation is inflammasomes, multiprotein complexes that trigger a release of inflammatory proteins when activated. Melatonin, a hormone with anti-inflammatory effects, is an attractive candidate for suppressing such inflammation. In this study, we have investigated how melatonin alters the microRNA (miRNA) transcriptome of microglial cells. For that purpose, we have performed RNA sequencing on a lipopolysaccharide and adenosine triphosphate (LPS + ATP) induced NLR family pyrin domain containing 3 (NLRP3) inflammasome activation model in the N9 mouse microglial cell line, with and without melatonin pre-treatment. We have identified 136 differentially expressed miRNAs in cells exposed to LPS + ATP compared to controls and 10 differentially expressed miRNAs in melatonin pre-treated cells compared to the inflammasome group. We have identified miR-155-3p as a miRNA that is upregulated with inflammasome activation and downregulated with melatonin treatment. We further confirmed this pattern of miR-155-3p expression in the brains of mice injected intraperitoneally with LPS. Moreover, an overexpression study with miRNA-155-3p mimic supported the idea that the protective effects of melatonin in NLRP3 inflammasome activation are partly associated with miRNA-155-3p inhibition.

Targeting NLRP3 Inflammasome With Nrf2 Inducers in Central Nervous System Disorders.

The NLRP3 inflammasome is an intracellular multiprotein complex that plays an essential role in the innate immune system by identifying and eliminating a plethora of endogenous and exogenous threats to the host. Upon activation of the NLRP3 complex, pro-inflammatory cytokines are processed and released. Furthermore, activation of the NLRP3 inflammasome complex can induce pyroptotic cell death, thereby propagating the inflammatory response. The aberrant activity and detrimental effects of NLRP3 inflammasome activation have been associated with cardiovascular, neurodegenerative, metabolic, and inflammatory diseases. Therefore, clinical strategies targeting the inhibition of the self-propelled NLRP3 inflammasome activation are required. The transcription factor Nrf2 regulates cellular stress response, controlling the redox equilibrium, metabolic programming, and inflammation. The Nrf2 pathway participates in anti-oxidative, cytoprotective, and anti-inflammatory activities. This prominent regulator, through pharmacologic activation, could provide a therapeutic strategy for the diseases to the etiology and pathogenesis of which NLRP3 inflammasome contributes. In this review, current knowledge on NLRP3 inflammasome activation and Nrf2 pathways is presented; the relationship between NLRP3 inflammasome signaling and Nrf2 pathway, as well as the pre/clinical use of Nrf2 activators against NLRP3 inflammasome activation in disorders of the central nervous system, are thoroughly described. Cumulative evidence points out therapeutic use of Nrf2 activators against NLRP3 inflammasome activation or diseases that NLRP3 inflammasome contributes to would be advantageous to prevent inflammatory conditions; however, the side effects of these molecules should be kept in mind before applying them to clinical practice.

Dimethyl Fumarate Alleviates NLRP3 Inflammasome Activation in Microglia and Sickness Behavior in LPS-Challenged Mice.

NLRP3 inflammasome activation contributes to several pathogenic conditions, including lipopolysaccharide (LPS)-induced sickness behavior characterized by reduced mobility and depressive behaviors. Dimethyl fumarate (DMF) is an immunomodulatory and anti-oxidative molecule commonly used for the symptomatic treatment of multiple sclerosis and psoriasis. In this study, we investigated the potential use of DMF against microglial NLRP3 inflammasome activation both in vitro and in vivo. For in vitro studies, LPS- and ATP-stimulated N9 microglial cells were used to induce NLRP3 inflammasome activation. DMF's effects on inflammasome markers, pyroptotic cell death, ROS formation, and Nrf2/NF-κB pathways were assessed. For in vivo studies, 12-14 weeks-old male BALB/c mice were treated with LPS, DMF + LPS and ML385 + DMF + LPS. Behavioral tests including open field, forced swim test, and tail suspension test were carried out to see changes in lipopolysaccharide-induced sickness behavior. Furthermore, NLRP3 and Caspase-1 expression in isolated microglia were determined by immunostaining. Here we demonstrated that DMF ameliorated LPS and ATP-induced NLRP3 inflammasome activation by reducing IL-1ß, IL-18, caspase-1, and NLRP3 levels, reactive oxygen species formation and damage, and inhibiting pyroptotic cell death in N9 murine microglia via Nrf2/NF-κB pathways. DMF also improved LPS-induced sickness behavior in male mice and decreased caspase-1/NLRP3 levels via Nrf2 activation. Additionally, we showed that DMF pretreatment decreased miR-146a and miR-155 both in vivo and in vitro. Our results proved the effectiveness of DMF on the amelioration of microglial NLRP3 inflammasome activation. We anticipate that this study will provide the foundation consideration for further studies aiming to suppress NLRP3 inflammasome activation associated with in many diseases and a better understanding of its underlying mechanisms.

Melatonin Attenuates LPS-Induced Acute Depressive-Like Behaviors and Microglial NLRP3 Inflammasome Activation Through the SIRT1/Nrf2 Pathway.

Inflammation is a crucial component of various stress-induced responses that contributes to the pathogenesis of major depressive disorder (MDD). Depressive-like behavior (DLB) is characterized by decreased mobility and depressive behavior that occurs in systemic infection induced by Lipopolysaccharide (LPS) in experimental animals and is considered as a model of exacerbation of MDD. We assessed the effects of melatonin on behavioral changes and inflammatory cytokine expression in hippocampus of mice in LPS-induced DLB, as well as its effects on NLR Family Pyrin Domain Containing 3 (NLRP3) inflammasome activation, oxidative stress and pyroptotic cell death in murine microglia in vitro. Intraperitoneal 5 mg/kg dose of LPS was used to mimic depressive-like behaviors and melatonin was given at a dose of 500 mg/kg for 4 times with 6 h intervals, starting at 2 h before LPS administration. Behavioral assessment was carried out at 24 h post-LPS injection by tail suspension and forced swimming tests. Additionally, hippocampal cytokine and NLRP3 protein levels were estimated. Melatonin increased mobility time of LPS-induced DLB mice and suppressed NLRP3 expression and interleukin-1ß (IL-1ß) cleavage in the hippocampus. Immunofluorescence staining of hippocampal tissue showed that NLRP3 is mainly expressed in ionized calcium-binding adapter molecule 1 (Iba1) -positive microglia. Our results show that melatonin prevents LPS and Adenosine triphosphate (ATP) induced NLRP3 inflammasome activation in murine microglia in vitro, evidenced by inhibition of NLRP3 expression, Apoptosis-associated speck-like protein containing a CARD (ASC) speck formation, caspase-1 cleavage and interleukin-1ß (IL-1ß) maturation and secretion. Additionally, melatonin inhibits pyroptosis, production of mitochondrial and cytosolic reactive oxygen species (ROS) and nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) signaling. The beneficial effects of melatonin on NLRP3 inflammasome activation were associated with nuclear factor erythroid 2-related factor 2 (Nrf2) and Silent information regulator 2 homolog 1 (SIRT1) activation, which were reversed by Nrf2 siRNA and SIRT1 inhibitor treatment.

Transmission ratio distortion is frequent in Arabidopsis thaliana controlled crosses.

The equal probability of transmission of alleles from either parent during sexual reproduction is a central tenet of genetics and evolutionary biology. Yet, there are many cases where this rule is violated. The preferential transmission of alleles or genotypes is termed transmission ratio distortion (TRD). Examples of TRD have been identified in many species, implying that they are universal, but the resolution of species-wide studies of TRD are limited. We have performed a species-wide screen for TRD in over 500 segregating F2 populations of Arabidopsis thaliana using pooled reduced-representation genome sequencing. TRD was evident in up to a quarter of surveyed populations. Most populations exhibited distortion at only one genomic region, with some regions being repeatedly affected in multiple populations. Our results begin to elucidate the species-level architecture of biased transmission of genetic material in A. thaliana, and serve as a springboard for future studies into the biological basis of TRD in this species.

Hepatitis B Virus and Hepatitis D Virus Recurrence in Patients Undergoing Liver Transplantation for Hepatitis B Virus and Hepatitis B Virus Plus Hepatitis D Virus.

BACKGROUND: In this study, we retrospectively analyzed the recurrence of hepatitis B virus (HBV) and hepatitis D virus (HDV) infection after liver transplantation for HBV and HBV+HDV co-infection. METHODS: Data from patients infected with HBV and HBV+HDV who underwent liver transplantation between March 2003 and June 2013 at the Liver Transplantation Institute of Inonu University were analyzed retrospectively. A total of 255 patients were included in the study. Group 1 (n = 127) comprised patients who underwent liver transplantation because of HBV, and group 2 (n = 128) comprised patients who underwent liver transplantation because of HBV+HDV. A positive HDV antibody serologic test result was taken to indicate liver disease caused by HBV+HDV. RESULTS: Thirteen of 255 were positive for the HBs Ag (5.1%). Nine (7.1%) and 4 (3.1%) patients were positive for the HBs Ag in groups 1 and 2, respectively (7.1%); the difference was not significant (P = .150). No HDV recurrence was detected in either group. The average time to HBs Ag seroconversion in 13 patients was 7.8 months after surgery (6.34 months in group 1 and 11.1 months in group 2). CONCLUSIONS: In our study, recurrence rate of HBV after liver transplantation is not statistically different than the recurrence rate of HBV+HDV co-infection. A low recurrence rate was achieved by the prophylaxis protocol in use at our center. There is no standard protocol for prevention of HBV and HDV recurrence; therefore, we need new studies.

Torsion of wandering spleen.

Wandering spleen is characterized by ectopic localization of spleen owing to the lack or weakening of the major splenic ligaments. In present study, two cases with torsion of wandering spleen were reported. The first case was a 30-year-old female who was admitted to emergency department with acute abdominal pain and vomiting. Abdominal Ultrasonography and computed tomography showed a round solid hypodense mass that was located in the left hypochondriac region of abdomen. At laparotomy, the patient was found to have torsion of a wandering spleen with complete infarction and pancreatic tail infarction. Splenectomy and distal pancreatectomy were performed. The second patient was a 19-year-old female. She was admitted to emergency department with abdominal pain. Axial computed tomography (CT) showed pelvic mass that indicated a possibility of a wandering spleen. The wandering spleen was removed with its long pedicle because of infarction. Torsion of wandering spleen must be considered in differential diagnosis of acute abdomen when a palpable painful abdominal mass is present on physical examination, and the spleen is absent in its normal anatomical location on radiological examination (Fig. 4, Ref. 8). Full Text (Free, PDF) www.bmj.sk.

Novel antifoam for fermentation processes: fluorocarbon-hydrocarbon hybrid unsymmetrical bolaform surfactant.

As foaming appears as a problem in chemical and fermentation processes that inhibits reactor performance, the eminence of a novel fluorocarbon-hydrocarbon unsymmetrical bolaform (FHUB: OH(CH2)11N+(C2H4)2(CH2)2(CF2)5CF3 I-) surfactant as an antifoaming agent as well as a foam-reducing agent was investigated and compared with other surfactants and a commercial antifoaming agent. The surface elasticity of FHUB was determined as 4 mN/m, indicating its high potential on thinning of the foam film. The interactions between FHUB and the microoganism were investigated in a model fermentation process related with an enzyme production by recombinant Escherichia coli, in V = 3.0 dm3 bioreactor systems with V(R) = 1.65 dm3 working volume at air inlet rate of Q(o)/V(R) = 0.5 dm3 dm(-3) min(-1) and agitation rate of N = 500 min(-1) oxygen transfer conditions, at T = 37 degrees C, pH(o) = 7.2, and C(FHUB) = 0 and 0.1 mM, in a glucose-based defined medium. As FHUB did not influence the metabolism, specific enzyme activity values obtained with and without FHUB were close to each other; however, because of the slight decrease in oxygen transfer coefficient, slightly lower volumetric enzyme activity and cell concentrations were obtained. However, when FHUB is compared with widely used silicon oil based Antifoam A, with the use of the FHUB, higher physical oxygen transfer coefficient (K(L)a) values are obtained. Moreover, as the amount required for the foam control is very low, minute changes in the working volume of the bioreactor were obtained indicating the high potential of the use of FHUB as an antifoaming agent as well as a foam-reducing agent.

Product spin-orbit state resolved dynamics of the H+H2O and H+D2O abstraction reactions.

The product state-resolved dynamics of the reactions H+H(2)O/D(2)O-->OH/OD((2)Pi(Omega);v',N',f )+H(2)/HD have been explored at center-of-mass collision energies around 1.2, 1.4, and 2.5 eV. The experiments employ pulsed laser photolysis coupled with polarized Doppler-resolved laser induced fluorescence detection of the OH/OD radical products. The populations in the OH spin-orbit states at a collision energy of 1.2 eV have been determined for the H+H(2)O reaction, and for low rotational levels they are shown to deviate from the statistical limit. For the H+D(2)O reaction at the highest collision energy studied the OD((2)Pi(3/2),v'=0,N'=1,A') angular distributions show scattering over a wide range of angles with a preference towards the forward direction. The kinetic energy release distributions obtained at 2.5 eV also indicate that the HD coproducts are born with significantly more internal excitation than at 1.4 eV. The OD((2)Pi(3/2),v'=0,N'=1,A') angular and kinetic energy release distributions are almost identical to those of their spin-orbit excited OD((2)Pi(1/2),v'=0,N'=1,A') counterpart. The data are compared with previous experimental measurements at similar collision energies, and with the results of previously published quasiclassical trajectory and quantum mechanical calculations employing the most recently developed potential energy surface. Product OH/OD spin-orbit effects in the reaction are discussed with reference to simple models.

Cross section for the H+H2O abstraction reaction: experiment and theory.

The absolute value of the cross section for the abstraction reaction between fast H atoms and H2O has been determined experimentally at a mean collision energy of 2.46 eV. The OH population distribution at the same mean energy has also been determined. The new measurements are compared with state-of-the-art quantum mechanical and quasiclassical scattering calculations on the most recently developed potential energy surface.

[The effect of toluene and ammonia on the invasive process and immunological indices in experimental trichinelliasis in mice]. / Vliianie toluola i ammiaka na invazionnyi protsess i immunologicheskie pokazateli pri éksperimental'nom trikhinelleze u myshei.

Rap mice were exposed to toluene (T) inhalation for 10 days before invasion with 20 Trichinella spiralis larvae per g body weight (moderate infection). This resulted in diminished number of intestinal parasites in the presence of greater number of mast cells in the peritoneal exudate, higher IgE production, enhanced cell adhesion to trichinella larvae and of migration of splenic lymphocytes. Simultaneous inhalation of T and ammonia diminished the immune stimulating effect of the former. The number of intestinal trichinella was 1.5 times more but still twice less than in controls. Inhalation of T during the first 10 days of infection stimulated the immune response only in mice given 5 larvae per g. In those given 20 or 60 larvae per g, the immune response was suppressed and 40 and 100% of mice perished respectively. The exposure to T during 30-39 days of infection of mice given 35 larvae per g (the intensive infection) resulted in 50% death of the animals without significant changes in immune response. Simultaneous therapy with mebendazole (75 mg/kg) provided 100% survival in the presence of suppressed immune response. 100% of mice of the same group not exposed to T but treated with mebendazole died. The toxic and immunomodulating effects of T differ in intact and infected mice due to the dense, the stage of infection and to chemotherapy.

[The role of genetic and ecological factors in development of resistant phenotype in mice during experimental trichinelliasis]. / Znachenie geneticheskikh i ékologicheskikhh faktorov v formirovanii fenotipa rezistentnosti u myshei pri éksperimental'nom trikhinelleze.

The role of genetic (genes for wool, eyes colour, X and Y chromosomes, H-2 haplotype) and ecological (contamination intensity leading to strengthening of illness) factors in forming phenotype of resistance in mice under experimental trichinelliasis is studied. The experiments have been performed on male and female mice CBA/Ca, BALB/c, C57/BL/6J, DBA/2J, C3HA/Mv, CC57BR/Mv, CC577W/Mv and hybrids /C57BL/6J x CBA/Ca/F having 19-25 g mass each, which were infected with trichinella larvae of Byelorussian laboratory "strains" in dosage of 5, 20, 35 and 70 units per 1 g body mass. The intensity of intestine invasion, the level of free histamine in the liver, the index of inhibition of spleen leucocyte migration on the 7th day and intensity of muscle invasion on the 30th day after contamination were defined. The analysis of the results of the study allowed to formulate the following points of regularity of trichinella resistance phenotype formation in mice: 1. The phenotype is formed as the result of interaction of genetic and ecological factors. 2. Mice of different genotypes form different phenotypes of resistance to trichinella. 3. Mice of the same genotypes form different phenotypes of resistance to intestine and muscle trichinella. The problems of inheritance and mechanisms of resistance are discussed.

[Antibody-dependent cell-mediated immune reactions in experimental trichinelliasis in mice]. / Antitelozavisimye kletochno-oposredovannye immunnye reaktsii pri éksperimental'nom trikhinelleze myshei.

The experiments on mice experimentally infected with Trichinella larvae have established that in mild infection adhesive and killer cell reactions developed on day 14, reached their maximum on days 21-30 and began to attenuate on day 45. The cells from animals with moderate infection showed a higher immune activity which remained unchanged till day 90 of infection. The cells from mice with severe infection showed inhibition of immune activity which was most marked on days 21 and 30 of the infection. Eosinophiles and neutrophils are characterized by IgE-dependent cell adherence and killing activity, the involvement of macrophages in the responses seems to be insignificant.