We have developed a method to longitudinally classify subjects into two or more prognostic groups using longitudinally observed values of markers related to the prognosis. We assume the availability of a training data set where the subjects' allocation into the prognostic group is known. The proposed method proceeds in two steps as described earlier in the literature. First, multivariate linear mixed models are fitted in each prognostic group from the training data set to model the dependence of markers on time and on possibly other covariates. Second, fitted mixed models are used to develop a discrimination rule for future subjects. Our method improves upon existing approaches by relaxing the normality assumption of random effects in the underlying mixed models. Namely, we assume a heteroscedastic multivariate normal mixture for random effects. Inference is performed in the Bayesian framework using the Markov chain Monte Carlo methodology. Software has been written for the proposed method and it is freely available. The methodology is applied to data from the Dutch Primary Biliary Cirrhosis Study.

• MeSH
• Liver Cirrhosis, Biliary drug therapy   MeSH
• Biomarkers analysis   MeSH
• Cholagogues and Choleretics therapeutic use   MeSH
• Discriminant Analysis MeSH
• Data Interpretation, Statistical MeSH
• Ursodeoxycholic Acid therapeutic use   MeSH
• Humans MeSH
• Linear Models MeSH
• Longitudinal Studies MeSH
• Computer Simulation MeSH
• Disease Progression MeSH
• Check Tag
• Humans MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH

Patients with alcohol use disorder (AUD) who seek treatment show highly variable outcomes. A precision medicine approach with biomarkers responsive to new treatments is warranted to overcome this limitation. Promising biomarkers relate to prefrontal control mechanisms that are severely disturbed in AUD. This results in reduced inhibitory control of compulsive behavior and, eventually, relapse. We reasoned here that prefrontal dysfunction, which underlies vulnerability to relapse, is evidenced by altered neuroelectric signatures and should be restored by pharmacological interventions that specifically target prefrontal dysfunction. To test this, we applied our recently developed biocompatible neuroprosthesis to measure prefrontal neural function in a well-established rat model of alcohol addiction and relapse. We monitored neural oscillations and event-related potentials in awake alcohol-dependent rats during abstinence and following treatment with psilocybin or LY379268, agonists of the serotonin 2A receptor (5-HT2AR), and the metabotropic glutamate receptor 2 (mGluR2), that are known to reduce prefrontal dysfunction and relapse. Electrophysiological impairments in alcohol-dependent rats are reduced amplitudes of P1N1 and N1P2 components and attenuated event-related oscillatory activity. Psilocybin and LY379268 were able to restore these impairments. Furthermore, alcohol-dependent animals displayed a dominance in higher beta frequencies indicative of a state of hyperarousal that is prone to relapse, which particularly psilocybin was able to counteract. In summary, we provide prefrontal markers indicative of relapse and treatment response, especially for psychedelic drugs.

• MeSH
• Alcoholism * drug therapy   physiopathology   MeSH
• Amino Acids MeSH
• Bridged Bicyclo Compounds, Heterocyclic * pharmacology   MeSH
• Biomarkers MeSH
• Evoked Potentials drug effects   MeSH
• Rats MeSH
• Disease Models, Animal * MeSH
• Prefrontal Cortex * drug effects   physiopathology   metabolism   MeSH
• Psilocybin * pharmacology   MeSH
• Receptors, Metabotropic Glutamate MeSH
• Animals MeSH
• Check Tag
• Rats MeSH
• Male MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH

The mechanical stability of epithelial cells, which protect organisms from harmful external factors, is maintained by hemidesmosomes via the interaction between plectin 1a (P1a) and integrin α6β4. Binding of calcium-calmodulin (Ca(2+)-CaM) to P1a together with phosphorylation of integrin β4 disrupts this complex, resulting in disassembly of hemidesmosomes. We present structures of the P1a actin binding domain either in complex with the N-ter lobe of Ca(2+)-CaM or with the first pair of integrin β4 fibronectin domains. Ca(2+)-CaM binds to the N-ter isoform-specific tail of P1a in a unique manner, via its N-ter lobe in an extended conformation. Structural, cell biology, and biochemical studies suggest the following model: binding of Ca(2+)-CaM to an intrinsically disordered N-ter segment of plectin converts it to an α helix, which repositions calmodulin to displace integrin β4 by steric repulsion. This model could serve as a blueprint for studies aimed at understanding how Ca(2+)-CaM or EF-hand motifs regulate F-actin-based cytoskeleton.

• MeSH
• Hemidesmosomes chemistry   MeSH
• Hydrophobic and Hydrophilic Interactions MeSH
• Integrin beta4 chemistry   MeSH
• Protein Interaction Domains and Motifs MeSH
• Calmodulin chemistry   MeSH
• Rats MeSH
• Crystallography, X-Ray MeSH
• Humans MeSH
• Models, Molecular MeSH
• Molecular Sequence Data MeSH
• Mice MeSH
• Cell Line, Tumor MeSH
• Plectin chemistry   MeSH
• Amino Acid Sequence MeSH
• Protein Structure, Tertiary MeSH
• Protein Binding MeSH
• Animals MeSH
• Check Tag
• Rats MeSH
• Humans MeSH
• Mice MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH

The COVID-19 outbreak has raised questions about how vulnerable groups experience the pandemic. Research that focuses on the view of individuals with pre-existing mental health conditions is still limited, and so are cross-country comparative surveys. We gathered our sample of qualitative data during the first lockdown after governmental measures against the spread of the SARS-CoV-2 virus came into force in Austria, Czechia, Germany, and Slovakia. A total of n = 1690 psychotherapists from four middle European countries answered the question of how the COVID-19 pandemic was addressed in sessions by their patients during the early stage of unprecedented public health conditions. We employed a descriptive qualitative methodology to determine themes following levels of the social-ecological model (SEM) regarding how the COVID-19 pandemic affected patients. At the public policy level, stressful environmental conditions concerned the governmental mitigation efforts. At the level of community/society, reported key themes were employment, restricted access to educational and health facilities, socioeconomic consequences, and the pandemic itself. Key themes at the interpersonal level regarded forced proximity, the possibility of infection of loved ones, childcare, and homeschooling. Key themes at the individual level were the possibility of contracting COVID-19, having to stay at home/isolation, and a changing environment. Within the SEM framework, adaptive and maladaptive responses to these stressors were reported, with more similarities than differences between the countries. A quantification of word stems showed that the maladaptive reactions predominated.

• MeSH
• COVID-19 * epidemiology   MeSH
• Communicable Disease Control MeSH
• Humans MeSH
• Pandemics MeSH
• Psychotherapists MeSH
• SARS-CoV-2 MeSH
• Check Tag
• Humans MeSH
• Publication type
• Journal Article MeSH

Serotonin (5-HT) receptors have been suggested to play key roles in psychosis, cognition, and mood via influence on neurotransmitters, synaptic integrity, and neural plasticity. Specifically, genetic evidence indicates that 5-HT(1A), 5-HT(2A), and 5-HT(2C) receptor single-nucleotide polymorphisms (SNPs) are related to psychotic symptoms, cognitive disturbances, and treatment response in schizophrenia. Data from animal research suggest the role of 5-HT in cognition via its influence on dopaminergic, cholinergic, glutamatergic, and GABAergic function. This article provides up-to-date findings on the role of 5-HT receptors in endophenotypic variations in schizophrenia and the development of newer cognition-enhancing medications, based on basic science and clinical evidence. Imaging genetics studies on associations of polymorphisms of several 5-HT receptor subtypes with brain structure, function, and metabolism suggest a role for the prefrontal cortex and the parahippocampal gyrus in cognitive impairments of schizophrenia. Data from animal experiments to determine the effect of agonists/antagonists at 5-HT(1A), 5-HT(2A), and 5-HT(2C) receptors on behavioral performance in animal models of schizophrenia based on the glutamatergic hypothesis provide useful information. For this purpose, standard as well as novel cognitive tasks provide a measure of memory/information processing and social interaction. In order to scrutinize mixed evidence for the ability of 5-HT(1A) agonists/antagonists to improve cognition, behavioral data in various paradigms from transgenic mice overexpressing 5-HT(1A) receptors provide valuable insights. Clinical trials reporting the advantage of 5-HT(1A) partial agonists add to efforts to shape pharmacologic perspectives concerning cognitive enhancement in schizophrenia by developing novel compounds acting on 5-HT receptors. Overall, these lines of evidence from translational research will facilitate the development of newer pharmacologic strategies for the treatment of cognitive disturbances of schizophrenia.

• MeSH
• Serotonin Receptor Agonists pharmacology   therapeutic use   MeSH
• Serotonin Antagonists pharmacology   therapeutic use   MeSH
• Buspirone pharmacology   therapeutic use   MeSH
• Behavior drug effects   MeSH
• Isoindoles pharmacology   therapeutic use   MeSH
• Cognition Disorders * drug therapy   metabolism   physiopathology   MeSH
• Humans MeSH
• Serotonin Plasma Membrane Transport Proteins genetics   metabolism   MeSH
• Brain physiopathology   drug effects   MeSH
• Piperazines pharmacology   therapeutic use   MeSH
• Polymorphism, Genetic * MeSH
• Pyrimidines pharmacology   therapeutic use   MeSH
• Receptor, Serotonin, 5-HT1A * genetics   metabolism   MeSH
• Schizophrenia * drug therapy   metabolism   physiopathology   MeSH
• Animals MeSH
• Check Tag
• Humans MeSH
• Animals MeSH
• Publication type
• Research Support, Non-U.S. Gov't MeSH
• Review MeSH

Protein import and genome replication are essential processes for mitochondrial biogenesis and propagation. The J-domain proteins Pam16 and Pam18 regulate the presequence translocase of the mitochondrial inner membrane. In the protozoan Trypanosoma brucei, their counterparts are TbPam16 and TbPam18, which are essential for the procyclic form (PCF) of the parasite, though not involved in mitochondrial protein import. Here, we show that during evolution, the 2 proteins have been repurposed to regulate the replication of maxicircles within the intricate kDNA network, the most complex mitochondrial genome known. TbPam18 and TbPam16 have inactive J-domains suggesting a function independent of heat shock proteins. However, their single transmembrane domain is essential for function. Pulldown of TbPam16 identifies a putative client protein, termed MaRF11, the depletion of which causes the selective loss of maxicircles, akin to the effects observed for TbPam18 and TbPam16. Moreover, depletion of the mitochondrial proteasome results in increased levels of MaRF11. Thus, we have discovered a protein complex comprising TbPam18, TbPam16, and MaRF11, that controls maxicircle replication. We propose a working model in which the matrix protein MaRF11 functions downstream of the 2 integral inner membrane proteins TbPam18 and TbPam16. Moreover, we suggest that the levels of MaRF11 are controlled by the mitochondrial proteasome.

• MeSH
• DNA, Mitochondrial * genetics   metabolism   MeSH
• Mitochondrial Proteins metabolism   genetics   MeSH
• Mitochondria metabolism   genetics   MeSH
• Evolution, Molecular MeSH
• Protozoan Proteins * metabolism   genetics   MeSH
• DNA Replication * MeSH
• Trypanosoma brucei brucei * metabolism   genetics   MeSH
• Publication type
• Journal Article MeSH

In Type 1 Diabetes Mellitus (T1DM), leukocyte infiltration of the pancreatic islets and the resulting immune-mediated destruction of beta cells precede hyperglycemia and clinical disease symptoms. In this context, the role of the pancreatic endothelium as a barrier for autoimmunity- and inflammation-related destruction of the islets is not well studied. Here, we identified Robo4, expressed on endothelial cells, as a regulator of pancreatic vascular endothelial permeability during autoimmune diabetes. Circulating levels of Robo4 were upregulated in mice subjected to the Multiple Low-Dose Streptozotocin (MLDS) model of diabetes. Upon MLDS induction, Robo4-deficiency resulted in increased pancreatic vascular permeability, leukocyte infiltration to the islets and islet apoptosis, associated with reduced insulin levels and faster diabetes development. On the contrary, in vivo administration of Slit2 in mice modestly delayed the emergence of hyperglycaemia and ameliorated islet inflammation in MLDS-induced diabetes. Thus, Robo4-mediated endothelial barrier integrity reduces insulitis and islet destruction in autoimmune diabetes. Our findings highlight the importance of the endothelium as gatekeeper of pancreatic inflammation during T1DM development and may pave the way for novel Robo4-related therapeutic approaches for autoimmune diabetes.

• MeSH
• Apoptosis MeSH
• Insulin-Secreting Cells metabolism   pathology   MeSH
• Cell Line MeSH
• Diabetes Mellitus, Type 1 metabolism   pathology   MeSH
• Endothelial Cells metabolism   pathology   MeSH
• Diabetes Mellitus, Experimental metabolism   pathology   MeSH
• Capillary Permeability * MeSH
• Cells, Cultured MeSH
• Humans MeSH
• Mice, Inbred C57BL MeSH
• Mice MeSH
• Receptors, Cell Surface blood   genetics   metabolism   MeSH
• Animals MeSH
• Check Tag
• Humans MeSH
• Mice MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH

FOXO transcription factors are critical regulators of cell homeostasis and steer cell death, differentiation and longevity in mammalian cells. By combined pharmacophore-modeling-based in silico and fluorescence polarization-based screening we identified small molecules that physically interact with the DNA-binding domain (DBD) of FOXO3 and modulate the FOXO3 transcriptional program in human cells. The mode of interaction between compounds and the FOXO3-DBD was assessed via NMR spectroscopy and docking studies. We demonstrate that compounds S9 and its oxalate salt S9OX interfere with FOXO3 target promoter binding, gene transcription and modulate the physiologic program activated by FOXO3 in cancer cells. These small molecules prove the druggability of the FOXO-DBD and provide a structural basis for modulating these important homeostasis regulators in normal and malignant cells.

• MeSH
• DNA chemistry   genetics   metabolism   MeSH
• Transcription, Genetic drug effects   MeSH
• Gene Knockdown Techniques MeSH
• HEK293 Cells MeSH
• Small Molecule Libraries chemistry   metabolism   pharmacology   MeSH
• Nucleic Acid Conformation MeSH
• Humans MeSH
• Magnetic Resonance Spectroscopy MeSH
• Models, Molecular MeSH
• Cell Line, Tumor MeSH
• Promoter Regions, Genetic genetics   MeSH
• Forkhead Box Protein O3 chemistry   genetics   metabolism   MeSH
• Protein Domains MeSH
• Molecular Docking Simulation MeSH
• Gene Expression Profiling methods   MeSH
• Protein Binding MeSH
• Binding Sites genetics   MeSH
• Check Tag
• Humans MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH

OBJECTIVE: This study examined associations between physical activity (PA) and neuropsychiatric symptoms (NPS) in older adults free of dementia. METHODS: This cross-sectional study included 3,222 individuals ≥70 years of age (1,655 men; mean±SD age=79.2±5.6; cognitively unimpaired, N=2,723; mild cognitive impairment, N=499) from the population-based Mayo Clinic Study of Aging. PA (taken as a presumed predictor) in midlife (i.e., when participants were 50-65 years of age) and late life (i.e., the year prior to assessment) was assessed with a self-reported, validated questionnaire; PA intensity and frequency were used to calculate composite scores. NPS (taken as presumed outcomes) were assessed with the Neuropsychiatric Inventory Questionnaire, Beck Depression Inventory (BDI-II), and Beck Anxiety Inventory (BAI). Regression analyses included midlife and late-life PA in each model, which were adjusted for age, sex, education, apolipoprotein E ɛ4 status, and medical comorbidity. RESULTS: Higher late-life PA was associated with lower odds of having apathy (OR=0.89, 95% CI=0.84-0.93), appetite changes (OR=0.92, 95% CI=0.87-0.98), nighttime disturbances (OR=0.95, 95% CI=0.91-0.99), depression (OR=0.94, 95% CI=0.90-0.97), irritability (OR=0.93, 95% CI=0.89-0.97), clinical depression (OR=0.92, 95% CI=0.88-0.97), and clinical anxiety (OR=0.90, 95% CI=0.86-0.94), as well as lower BDI-II (β estimate=-0.042, 95% CI=-0.051 to -0.033) and BAI (β estimate=-0.030, 95% CI=-0.040 to -0.021) scores. Higher midlife PA was associated only with higher BDI-II scores (β estimate=0.011, 95% CI=0.004 to 0.019). Sex modified the associations between PA and NPS. CONCLUSIONS: Late-life PA was associated with a lower likelihood of clinical depression or anxiety and subclinical NPS. These findings need to be confirmed in a cohort study.

• MeSH
• Exercise MeSH
• Depression * psychology   MeSH
• Cognitive Dysfunction * diagnosis   MeSH
• Cohort Studies MeSH
• Humans MeSH
• Neuropsychological Tests MeSH
• Cross-Sectional Studies MeSH
• Aged, 80 and over MeSH
• Aged MeSH
• Aging MeSH
• Check Tag
• Humans MeSH
• Male MeSH
• Aged, 80 and over MeSH
• Aged MeSH
• Publication type
• Journal Article MeSH

The nucleotide-binding-domain (NBD)-and leucine-rich repeat (LRR)-containing (NLR) family, pyrin-domain-containing 3 (NLRP3) inflammasome drives pathological inflammation in a suite of autoimmune, metabolic, malignant, and neurodegenerative diseases. Additionally, NLRP3 gain-of-function point mutations cause systemic periodic fever syndromes that are collectively known as cryopyrin-associated periodic syndrome (CAPS). There is significant interest in the discovery and development of diarylsulfonylurea Cytokine Release Inhibitory Drugs (CRIDs) such as MCC950/CRID3, a potent and selective inhibitor of the NLRP3 inflammasome pathway, for the treatment of CAPS and other diseases. However, drug discovery efforts have been constrained by the lack of insight into the molecular target and mechanism by which these CRIDs inhibit the NLRP3 inflammasome pathway. Here, we show that the NAIP, CIITA, HET-E, and TP1 (NACHT) domain of NLRP3 is the molecular target of diarylsulfonylurea inhibitors. Interestingly, we find photoaffinity labeling (PAL) of the NACHT domain requires an intact (d)ATP-binding pocket and is substantially reduced for most CAPS-associated NLRP3 mutants. In concordance with this finding, MCC950/CRID3 failed to inhibit NLRP3-driven inflammatory pathology in two mouse models of CAPS. Moreover, it abolished circulating levels of interleukin (IL)-1β and IL-18 in lipopolysaccharide (LPS)-challenged wild-type mice but not in Nlrp3L351P knock-in mice and ex vivo-stimulated mutant macrophages. These results identify wild-type NLRP3 as the molecular target of MCC950/CRID3 and show that CAPS-related NLRP3 mutants escape efficient MCC950/CRID3 inhibition. Collectively, this work suggests that MCC950/CRID3-based therapies may effectively treat inflammation driven by wild-type NLRP3 but not CAPS-associated mutants.

• MeSH
• Cytokines antagonists & inhibitors   MeSH
• Furans pharmacology   MeSH
• HEK293 Cells MeSH
• Inflammasomes antagonists & inhibitors   MeSH
• Humans MeSH
• Lipopolysaccharides MeSH
• Macrophages drug effects   MeSH
• Disease Models, Animal MeSH
• Mice, Inbred C57BL MeSH
• Mice MeSH
• Cryopyrin-Associated Periodic Syndromes genetics   MeSH
• Drug Evaluation, Preclinical MeSH
• NLR Family, Pyrin Domain-Containing 3 Protein antagonists & inhibitors   genetics   MeSH
• Protein Domains MeSH
• Sulfonamides pharmacology   MeSH
• Animals MeSH
• Check Tag
• Humans MeSH
• Mice MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH