•Muscles of cod from Norwegian Sea are infected with Anisakis and Pseudoterranova nematodes.•The prevalence of infection with Anisakis was higher than that with Pseudoterranova.•Higher prevalence was ...recorded: Anisakis in FAO IIa1; Pseudoterranova in FAO IIa2.•Anisakis prefer belly flaps, Pseudoterranova also dorsal and caudal fillet.•A P. decipiens / P. krabbei hybrid was detected.
Atlantic cod (Gadus morhua) is an important commercial fish species on the world market. The aim of our studies was to explore the presence, intensity of infection and distribution of the zoonotic nematodes of the different genera of Anisakidae in the muscle tissue of G. morhua from the Norwegian Sea. Cod from fishing areas FAO IIa1 (n = 50) and FAO IIa2 (n = 56) were sampled in March 2017. The unskinned flesh of each fish was examined using a white-light transilluminator. Collected parasites were identified to the genus level, and a subsample was identified using molecular methods. We found a higher prevalence of infection with Anisakis than with Pseudoterranova in the musculature of cod from both fishing areas. In FAO IIa1, a lower prevalence of infection with Pseudoterranova was recorded (14%) than in FAO IIa2 (∼39%). However, the intensity of infection was higher (53) in FAO IIa2 than in FAO IIa1 (8 parasites per fish). The opposite was found with Anisakis (prevalence 88% in FAO IIa1 and ∼55% in FAO IIa2, intensity up to 30 and up to 25 parasites per fish respectively). Most Anisakis larvae were present in the belly flaps (predominantly the left side), while Pseudoterranova spp. were dispersed with descending frequency in belly flaps, dorsal fillet and caudal fillet. Molecular identification revealed the presence of A. simplex (s.s.), P. decipiens (s.s.) and P. krabbei in both areas, and a hybrid of P. decipiens and P. krabbei in FAO IIa2.
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Translational regulation is a fundamental step in gene expression with critical roles in biological processes within a cell. Here, we describe a protocol to assess translation activity in mammalian ...cells by incorporation of O-propargyl-puromycin (OP-Puro). OP-Puro is a puromycin analog that is incorporated into newly synthesized proteins and is detected by click chemistry reaction. We use OP-Puro labeling to assess translation activity between different cell types or cells under different growth conditions by confocal microscopy and flow cytometry.
For complete details on the use and execution of this protocol, please refer to Hsu et al. (2021) and Hsu et al. (2022).
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•Protocol for the measurement of global translation activity at the single-cell level•Incorporation of OP-Puro into newly synthesized proteins in living cells•Fluorescent labeling of incorporated OP-Puro using click chemistry reaction•Confocal microscopy and flow cytometry to define translation activity
Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.
Translational regulation is a fundamental step in gene expression with critical roles in biological processes within a cell. Here, we describe a protocol to assess translation activity in mammalian cells by incorporation of O-propargyl-puromycin (OP-Puro). OP-Puro is a puromycin analog that is incorporated into newly synthesized proteins and is detected by click chemistry reaction. We use OP-Puro labeling to assess translation activity between different cell types or cells under different growth conditions by confocal microscopy and flow cytometry.
Affective disorders include unipolar disorder (UP) (depression episodes) and bipolar disorder (BP) (depression and mania episodes). Currently, no biological markers are known that can help to ...differentiate these disorders. However, recent studies have suggested that psychiatric disorders can be connected with small, non-coding RNA, like microRNA.
The objective of this study was to analyse the expression level of three microRNAs (miR-499, miR-708, miR-1908) in bipolar and unipolar disorder during depression episodes and after entering the remission state.
The group consisted of adult women only, 17 UP (age: 50±17) and 15 BP (age: 33±13) patients. The expression level of miRNAs was investigated by RT-qPCR with the TaqMan assay.
Our study has shown a lower expression level of miR-499 (p=0.008), miR-708 (p=0.02) and miR-1908 (p=0.004) in depression episodes of the bipolar disorder patients in comparison to remission state.
We have not found similar differences in unipolar disorder and between those types in acute phase of depression and during remission.
Obtained results indicate that miRNAs: miR-499, miR-708 and miR-1908 are the most promising candidates for biomarkers of depression episodes of bipolar disorder.
Bipolar disorder (BD) is one of the most disabling psychiatric illnesses. Over half of BD patients experienced early onset of the disease, and in most cases, it begins with a depressed mood episode. ...Up to 50% of adolescents initially diagnosed with major depressive disorder (MDD) convert to bipolar spectrum disorder. Diagnostic tools or biomarkers to facilitate the prediction of diagnosis conversion from MDD to BD are still lacking. Our study aimed to find biomarkers of diagnosis conversion in young patients with mood disorders. We performed a 2-year follow-up study on 69 adolescent patients diagnosed with MDD or BD. The control group consisted of 31 healthy youths. We monitored diagnosis change from MDD to BD. Impulsiveness was assessed using Barratt Impulsiveness Scale (BIS-11) and defense mechanisms using Defense Style Questionnaire (DSQ-40). According to the immunological hypothesis of mood disorders, we investigated baseline cytokines levels either in depressive or hypomanic/manic episodes. We correlated interleukin 8 (IL-8) and Tumor Necrosis Factor-alpha (TNF-alpha) levels with clinical factors. We detected higher IL-8 and TNF-alpha in patients in hypomanic/manic compared to depressed episodes. We found correlations of cytokine levels with immature defense style. We did not discover predictors of diagnosis conversion from MDD to BD.
Abstract Introduction Studies have not given yet a clear answer what is the genetic background of suicidal predisposition. The associations between polymorphisms of the TPH1 and 5-HTTLPR genes and ...violent suicidal behavior was revealed with the least inconsistencies. Method We selected 10 “strong candidate genes” and 35 SNPs, SLC6A4 and ACP1 for replication study. We searched associations between precisely described suicidal phenotype in 825 affective patients and polymorphisms of selected neurobiological pathways genes as well as their interactions that constitute suicidal risk. Results The results confirm the role of TPH1, TPH2, 5HT2A, CRHR1 and ACP1 variants in the risk of suicidal behavior. Limitations In our study we analyzed limited number of candidate genes and only one of them is linked to lithium mechanism of action. We had no data on pharmacological treatment of investigated patients and its relation to the time of suicide attempt. Conclusion Our results indicate that polymorphisms of various signaling pathways are involved in the pathogenesis of suicidal behavior. Non-genetic factors are also involved in the risk of suicidal attempts.
Lithium has been the most important mood stabilizer used for the treatment of bipolar disorder and prophylaxis of manic and depressive episodes. Despite long use in clinical practice, the exact ...molecular mechanisms of lithium are still not well identified. Previous experimental studies produced inconsistent results due to different duration of lithium treatment and using animals without manic-like or depressive-like symptoms. Therefore, we aimed to analyze the gene expression profile in three brain regions (amygdala, frontal cortex and hippocampus) in the rat model of mania and depression during chronic lithium administration (2 and 4 weeks). Behavioral changes were verified by the forced swim test, open field test and elevated maze test. After the experiment, nucleic acid was extracted from the frontal cortex, hippocampus and amygdala. Gene expression profile was done using SurePrint G3 Rat Gene Expression whole transcriptome microarrays. Data were analyzed using Gene Spring 14.9 software. We found that chronic lithium treatment significantly influenced gene expression profile in both mania and depression models. In manic rats, chronic lithium treatment significantly influenced the expression of the genes enriched in olfactory and taste transduction pathway and long non-coding RNAs in all three brain regions. We report here for the first time that genes regulating olfactory and taste receptor pathways and long non-coding RNAs may be targeted by chronic lithium treatment in the animal model of mania.
Melatonin is a neurohormone that maintains the circadian rhythms of the body. By regulating the secretion of other hormones and neurotransmitters, it acts as a pleiotropic modulator that affects, for ...example, reproductive, immune, cardiovascular, sleep, and wake systems and mood. Thus, synthetic melatonin has become an essential component in the treatment of depressive disorders. Although we know the pathway of melatonin action in the brain, we lack comprehensive cross-sectional studies on the periphery of depressed patients. This study aimed to comprehensively analyze the differences between healthy control subjects (
= 84) and unipolar and bipolar depression patients (
= 94), including an analysis of the melatonin pathway at the level of the genes and serum biomarkers. An innovative approach is a pilot study based on gene expression profiling carried out on clinical and cell culture models using agomelatine and melatonin. We confirmed the melatonin biosynthesis pathway's molecular regulation dysfunctions, with a specific pattern for unipolar and bipolar depression, at the AANAT gene, its polymorphisms (rs8150 and rs3760138), and examined the serum biomarkers (serotonin, AANAT, ASMT, and melatonin). The biological pathway analysis uncovered pathways and genes that were uniquely altered after agomelatine treatment in a clinical model and melatonin treatment in a cell culture model. In both models, we confirmed the immunomodulatory effect of melatonin agents in depression.
The accurate assessment of suicide risk in psychiatric, especially affective disorder diagnosed patients, remains a crucial clinical need. In this study, we applied temperament and character ...inventory (TCI), Barratt impulsiveness scale 11 (BIS-11), PEBL simple reaction time (SRT) test, continuous performance task (CPT), and Iowa gambling task (IGT) to seek for variables linked with attempted suicide in bipolar affective disorder group (
n
= 60; attempters
n
= 17). The main findings were: strong correlations between self-report tool scores and objective parameters in CPT; the difference between attempters and non-attempters was found in the number of correctly responded trials in IGT; only one parameter differed between attempters and non-attempters in BPI diagnosis; and no significant differences between suicide attempters and non-attempters in TCI, BIS-11, and SRT were found. These justify the conclusion that impulsivity itself is not a strong predictor, and used as a single variable might not be sufficient to indicate the high suicide risk group among bipolar patients.
Anisakidae nematodes, especially
,
, and
have dispersed throughout the Baltic Sea over the last decade. Despite the fact that salmon,
, is a popular choice among consumers and therefore one of the ...most valuable Baltic fish species, information about the level of infection of salmon liver with these zoonotic nematodes is sparse. In 2020, a total of 120 salmon livers were inspected for the presence of parasites showing that 13% of salmon livers were infected with
. Furthermore, a single
sp. larva was detected in one salmon liver, representing a host-parasite system that has never previously been reported in the Baltic Sea.
The activation of CD8+ T‐cells requires the uptake of exogenous polypeptide antigens and proteolytic processing of these antigens to octamer or nonamer peptides, which are loaded on MHC‐I complexes ...and presented to the T‐cell. By using an azide as a bioorthogonal protecting group rather than as a ligation handle, masked antigens were generated—antigens that are not recognized by their cognate T‐cell unless they are deprotected on the cell using a Staudinger reduction.
Antigen cross‐presentation is the uptake of exogenous antigens followed by processing in the MHC‐I complex, which is normally reserved for endogenous antigens. This process may now be studied by using masked epitopes: Azides are exploited as bioorthogonal protecting groups to generate antigens that are not recognized by their cognate T‐cell unless they are deprotected using an on‐cell Staudinger reduction.
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