A Brief Mindfulness-Based Family Psychoeducation Intervention for Chinese Young Adults With First Episode Psychosis: A Study Protocol

Family psychoeducation (FPE) has been recommended as a major component in the treatment of psychosis. Many previous studies have implemented an intensive program design that often only emphasized improvements in patients’ illness outcomes but the benefits for caregivers were limited. There have been calls for a time-limited but cost-effective FPE program to mitigate the looming reality of the suffering of people with psychosis and their families. A Brief Mindfulness-Based Family Psychoeducation for psychosis program is developed to reduce caregivers’ burden and promote young adult’s recovery. A randomized controlled trial will be conducted to compare this intervention with an ordinary FPE intervention. Both arms will involve six sessions, with a total contact time of 12 h. 300 caregivers of young adults who have experienced first episode psychosis within last 3 years will be recruited. Program effectiveness will be assessed by comparing outcomes measuring the caregivers’ burden, mental health symptoms, positive well-being, and the young adult’s mental health symptoms during the study and at 9-month post-randomization. The role of expressed emotions, interpersonal mindfulness, and non-attachment in mediating these outcomes will be explored. An additional qualitative approach Photovoice is selected to explore the complex family experiences and the benefits of mindfulness from the caregivers’ personal perspectives.Trial Registration: The trial is registered with the United States Clinical Trials Registry (ClinicalTrials.gov): NCT03688009

Mitochondrial Uncoupling Protein-2 (UCP2) Mediates Leptin Protection Against MPP+ Toxicity in Neuronal Cells

Mitochondrial dysfunction is involved in the pathogenesis of neurodegenerative diseases, including Parkinson’s disease (PD). Uncoupling proteins (UCPs) delink ATP production from biofuel oxidation in mitochondria to reduce oxidative stress. UCP2 is expressed in brain, and has neuroprotective effects under various toxic insults. We observed induction of UCP2 expression by leptin in neuronal cultures, and hypothesize that leptin may preserve neuronal survival via UCP2. We showed that leptin preserved cell survival in neuronal SH-SY5Y cells against MPP+ toxicity (widely used in experimental Parkinsonian models) by maintaining ATP levels and mitochondrial membrane potential (MMP); these effects were accompanied by increased UCP2 expression. Leptin had no effect in modulating reactive oxygen species levels. Stable knockdown of UCP2 expression reduced ATP levels, and abolished leptin protection against MPP+-induced mitochondrial depolarization, ATP deficiency, and cell death, indicating that UCP2 is critical in mediating these neuroprotective effects of leptin against MPP+ toxicity. Interestingly, UCP2 knockdown increased UCP4 expression, but not of UCP5. Our findings show that leptin preserves cell survival by maintaining MMP and ATP levels mediated through UCP2 in MPP+-induced toxicity

Human germline heterozygous gain-of-function STAT6 variants cause severe allergic disease

STAT6 (signal transducer and activator of transcription 6) is a transcription factor that plays a central role in the pathophysiology of allergic inflammation. We have identified 16 patients from 10 families spanning three continents with a profound phenotype of early-life onset allergic immune dysregulation, widespread treatment-resistant atopic dermatitis, hypereosinophilia with esosinophilic gastrointestinal disease, asthma, elevated serum IgE, IgE-mediated food allergies, and anaphylaxis. The cases were either sporadic (seven kindreds) or followed an autosomal dominant inheritance pattern (three kindreds). All patients carried monoallelic rare variants in STAT6 and functional studies established their gain-of-function (GOF) phenotype with sustained STAT6 phosphorylation, increased STAT6 target gene expression, and TH2 skewing. Precision treatment with the anti-IL-4Rα antibody, dupilumab, was highly effective improving both clinical manifestations and immunological biomarkers. This study identifies heterozygous GOF variants in STAT6 as a novel autosomal dominant allergic disorder. We anticipate that our discovery of multiple kindreds with germline STAT6 GOF variants will facilitate the recognition of more affected individuals and the full definition of this new primary atopic disorder

Robust estimation of bacterial cell count from optical density

Optical density (OD) is widely used to estimate the density of cells in liquid culture, but cannot be compared between instruments without a standardized calibration protocol and is challenging to relate to actual cell count. We address this with an interlaboratory study comparing three simple, low-cost, and highly accessible OD calibration protocols across 244 laboratories, applied to eight strains of constitutive GFP-expressing E. coli. Based on our results, we recommend calibrating OD to estimated cell count using serial dilution of silica microspheres, which produces highly precise calibration (95.5% of residuals <1.2-fold), is easily assessed for quality control, also assesses instrument effective linear range, and can be combined with fluorescence calibration to obtain units of Molecules of Equivalent Fluorescein (MEFL) per cell, allowing direct comparison and data fusion with flow cytometry measurements: in our study, fluorescence per cell measurements showed only a 1.07-fold mean difference between plate reader and flow cytometry data

Characterization of the 5'flanking region of mitochondrial uncoupling protein 4 (UCP 4) and its relationship with nuclear factor-kappa B(NF-KB) in MPP+ -induced toxicity

published_or_final_versionMedicineDoctoralDoctor of Philosoph

Assessment of cellular estrogenic activity based on estrogen receptor-mediated reduction of soluble-form catechol-O-methyltransferase (COMT) expression in an ELISA-based system.

Xenoestrogens are either natural or synthetic compounds that mimic the effects of endogenous estrogen. These compounds, such as bisphenol-A (BPA), and phthalates, are commonly found in plastic wares. Exposure to these compounds poses major risk to human health because of the potential to cause endocrine disruption. There is huge demand for a wide range of chemicals to be assessed for such potential for the sake of public health. Classical in vivo assays for endocrine disruption are comprehensive but time-consuming and require sacrifice of experimental animals. Simple preliminary in vitro screening assays can reduce the time and expense involved. We previously demonstrated that catechol-O-methyltransferase (COMT) is transcriptionally regulated by estrogen via estrogen receptor (ER). Therefore, detecting corresponding changes of COMT expression in estrogen-responsive cells may be a useful method to estimate estrogenic effects of various compounds. We developed a novel cell-based ELISA to evaluate cellular response to estrogenicity by reduction of soluble-COMT expression in ER-positive MCF-7 cells exposed to estrogenic compounds. In contrast to various existing methods that only detect bioactivity, this method elucidates direct physiological effect in a living cell in response to a compound. We validated our assay using three well-characterized estrogenic plasticizers - BPA, benzyl butyl phthalate (BBP), and di-n-butyl phthalate (DBP). Cells were exposed to either these plasticizers or 17β-estradiol (E2) in estrogen-depleted medium with or without an ER-antagonist, ICI 182,780, and COMT expression assayed. Exposure to each of these plasticizers (10(-9)-10(-7)M) dose-dependently reduced COMT expression (p<0.05), which was blocked by ICI 182,780. Reduction of COMT expression was readily detectable in cells exposed to picomolar level of E2, comparable to other in vitro assays of similar sensitivity. To satisfy the demand for in vitro assays targeting different cellular components, a cell-based COMT assay provides useful initial screening to supplement the current assessments of xenoestrogens for potential estrogenic activity

Three common plasticizers (bisphenol-A (BPA), benzyl butyl phthalate (BBP), and di-n-butyl phthalate (DBP)) dose-dependently decreased COMT expression via estrogen receptor (ER) in MCF-7 cells.

<p>Cells were treated with graded doses of each of the three compounds, in the absence or presence of ICI 182,780 (ER antagonist). The resultant changes of soluble-COMT (S-COMT; 23kDa) protein expression were determined by Western blotting. Equal loading of samples were normalized by actin (42kDa). Results are mean ± SEM of four separate experiments (n=4). Statistical significance at the level of *p<0.05 or **p<0.01, as compared to untreated controls. ^## p<0.01, as compared between the two designated treatment groups.</p

Bioenergetic characterization of SH-SY5Y cells stably overexpressing UCP4.

<p>(a) Human UCP4 protein expression level was significantly higher in UCP4-overexpressing cells than vector controls. (b) Total intracellular ATP level in UCP4-overexpressing cells was higher than vector control cells, as determined by luciferase bioluminescent assay in total cell lysates. (c) Rate of respiration in vector control and UCP4-overexpressing cells. Under normal culture condition, oxygen consumption rate of UCP4-overexpressing cells was higher than vector controls. Results are expressed as mean ± SEM based on at least three independent trials. ** represents statistical significance at p<0.01, * represents p<0.05, compared to the vector control cells.</p

Level of proton leak and integrity of isolated mitochondria from UCP4-overexpressing and vector control cells.

<p>(a) Oxygraphs showing changes in dissolved oxygen in isolated mitochondria suspension from these cells under substrates-induced respiration. Maximum rate of oxygen consumption was recorded for 2 min after addition of substrates containing 5 mM glutamate & malate, 5 mM succinate, and 0.5 mM ADP. ATP synthase inhibitor, oligomycin (2.5 µg/ml) was then added to block ATP synthesis by Complex V. The rate of oxygen consumption with oligomycin was recorded for an additional 2 min, indicating Complex V-insensitive proton leak. Numbers in brackets represent the rate of oxygen consumption in µmol O₂/min/mg mitochondrial protein. (b) The level of proton leak was defined as the ratio of the rate of oxygen consumption in the presence of oligomycin to the rate of substrates-stimulated oxygen consumption (<i>state 3</i> respiration). Results are expressed as mean ratios ± SEM based on at least three independent trials. ** represents statistical significance at p<0.01, compared to the vector control. (c) Representative oxygraphs reflecting integrity of the outer mitochondrial membrane of isolated mitochondria isolated from both vector control and UCP4-overexpressing cells. Exogenous cytochrome c (Cyt <i>c</i>) did not further increase mitochondrial oxygen consumption. Intact and functional mitochondria were demonstrated by <i>state 3</i> respiration as stimulated by addition of substrates (glutamate/malate/succinate) and ADP.</p

Classical <i>state 4</i> and <i>state 3</i> respiration rate in UCP4-overexpressing and control mitochondria.

<p>Classical <i>state 4</i> and <i>state 3</i> respiration rate in UCP4-overexpressing and control mitochondria.</p