Can social inclusion be evaluated? Investigating the psychometric properties of the social inclusion intervention scale

The present study aims to validate a newly developed Social Inclusion Intervention Scale (SIIS) using Exploratory Factor Analysis and Confirmatory Factor Analysis. The participants were 128 children aged 45-84 month-old from local integrated preschools in Hong Kong. The factor structure of the SIIS fit the data well (RMSEA = .08, NFI = .92, and TLI = .95, CFI = .96, SRMR = .04), with good convergent validity (all CR values > .92, all AVE values > .61). The internal consistency was good across items (all α values > .91) and factors (all CR values > .92). Hence, the sample obtained from the clinical trials of this study showed a good model fit, which suggested that the SIIS is adequate in measuring social inclusion among preschool children in social inclusion intervention programmes. The implications of the two emerged themes of social inclusion from the findings, Relationships and Acceptance, are further discussed to ascertain how they shed light on the design of social inclusion intervention

Physiological and pathological roles of tissue plasminogen activator and its inhibitor neuroserpin in the nervous system

Although its roles in the vascular space are most well known, tissue plasminogen activator (tPA) is widely expressed in the developing and adult nervous system, where its activity is believed to be regulated by neuroserpin, a predominantly brain-specific member of the serpin family of protease inhibitors. In the normal physiological state, tPA has been shown to play roles in the development and plasticity of the nervous system. Ischemic damage, however, may lead to excess tPA activity in the brain and this is believed to contribute to neurodegeneration. In this article, we briefly review the physiological and pathological roles of tPA in the nervous system, which includes neuronal migration, axonal growth, synaptic plasticity, neuroprotection and neurodegeneration, as well as a contribution to neurological disease. We summarize the tPA’s multiple mechanisms of action and also highlight the contributions of the inhibitor neuroserpin to these processes

AAV-Mediated Overexpression of Neuroserpin in the Hippocampus Decreases PSD-95 Expression but Does Not Affect Hippocampal-Dependent Learning and Memory

<div><p>Neuroserpin is a serine protease inhibitor, or serpin, that is expressed in the nervous system and inhibits the protease tissue plasminogen activator (tPA). Neuroserpin has been suggested to play a role in learning and memory but direct evidence for such a role is lacking. Here we have used an adeno-associated virus (AAV) vector expression system to investigate the effect of neuroserpin on hippocampal-dependent learning and memory in the young adult rat. A FLAG-tagged neuroserpin construct was initially characterized by <i>in vitro</i> transcription/translation and transfection into HEK293 cells and shown to interact with tPA and be targeted to the secretory pathway. Targeted injection of a chimeric AAV1/2 vector expressing FLAG-neuroserpin resulted in localized overexpression in the dorsal hippocampus. Neuroserpin overexpression led to the appearance of an unstable neuroserpin:tPA complex in zymographic assays consistent with interaction with endogenous tPA <i>in vivo</i>. Rats overexpressing neuroserpin also showed a significant decrease in the levels of postsynaptic density protein 95, a major postsynaptic scaffolding protein. Three weeks after injection, a range of behavioural tests was performed to measure spatial and associative learning and memory, as well as innate and acquired fear. These tests provided no evidence of a role for neuroserpin in hippocampal-dependent learning and memory. In summary this study does not support a role for neuroserpin in hippocampal-dependent learning and memory in young adult rats but does suggest an involvement of neuroserpin in hippocampal synaptic plasticity.</p></div

Increased neuroserpin expression and unchanged PAI-1 and tPA levels in AAV-FLAG-NS-transduced hippocampi.

<p>(A,B) qRT-PCR of neuroserpin (A) and PAI-1 (B) transcripts from hippocampi, 61 days after injection with AAV-empty (n = 3), AAV-hrGFP (n = 3) or AAV-FLAG-NS (n = 3). (C) Western blot analysis of endogenous and FLAG-tagged neuroserpin and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) in hippocampal lysates (40 µg) from rats injected with AAV-empty (n = 3) or AAV-FLAG-NS (n = 3). (D) Western blot analysis of immunoreactive neuroserpin following N-linked deglycosylation of AAV-FLAG-NS hippocampal lysates (40 µg) with peptide-<i>N-</i>Glycosidase F (PNGaseF). (E) Quantitation of neuroserpin expression in panel C by measurement of the intensity of neuroserpin immunoreactive bands and normalization to GAPDH expression. (F) Quantitation of tPA transcripts from hippocampi injected with AAV-empty (n = 3), AAV-hrGFP (n = 3) or AAV-FLAG-NS (n = 3). (G) tPA enzymatic activity in the hippocampi lysates (50 µg) from rats injected with AAV-empty (n = 2) or AAV-FLAG-NS (n = 2). Lanes separated by dotted lines are from the same blot but with the lane order changed for presentation. All values are presented as the mean ± SEM. ***indicates <i>p</i>&lt;0.001; *indicates <i>p</i>&lt;0.05.</p

FLAG-tagged neuroserpin forms complexes with tPA and is secreted from transduced HEK293 cells.

<p>(A) Radiolabelled neuroserpins were incubated with or without tPA separated by a SDS-PAGE and imaged using a Fuji Phosphoimager (NS, neuroserpin; tPA, tissue plasminogen activator; RCL, reactive centre loop; sctPA, single-chain tPA; tctPA: two-chain tPA). (B) Lysate and medium samples collected from HEK293 cells that were either not transduced or transduced with AAV-FLAG-NS were separated by SDS-PAGE, followed by Western blotting with anti-NS and anti-FLAG antibodies.</p

Expression of proteins associated with synaptic plasticity in the hippocampi of AAV-injected rats.

<p>Hippocampal lysates (35 µg) from rats injected with AAV-empty (n = 3) or AAV-FLAG-NS (n = 3) were analyzed by SDS-PAGE and Western blotting. Each lane represents protein expression from the same rat. Blots were probed with antibodies for synapsin I (A), PSD-95 (B), NR1 (C), Tau-1 (D) and GAP-43 (E) and immunoreactive bands normalized to GAPDH expression. Values are presented as the mean ± SEM. *indicates <i>p</i>&lt;0.05.</p

Expression of endogenous neuroserpin and FLAG-tagged-neuroserpin in the rat hippocampus.

<p>Immunohistochemical localization of neuroserpin and FLAG-tagged neuroserpin in rat brain sections near the AAV injection sites 21days after unilaterally injection with AAV-empty (A–B) or AAV-FLAG-NS (C–J) and detected using antibodies against neuroserpin (A, C, E, G, I) or FLAG (B, D, F, H, J). Representative brain sections from rats injected with AAV-empty (n = 2) or AAV-FLAG-NS (n = 2) are shown. CA1, pyramidal cell layers in CA1 subfield; CA3, pyramidal cell layers in CA3 subfield; gcl, dentate gyrus granule cell layer; h, hilus. Scale bar, 100 µm.</p

Learning and memory and anxiety and fear levels were not affected by neuroserpin overexpression.

<p>(A) All groups showed a decrease in swimming duration in the Morris water maze across the 5-day acquisition phase of spatial navigation but there was no significant difference between treatment groups (AAV-empty, n = 8; AAV-hrGFP, n = 7; AAV-FLAG-NS, n = 8) (F_8,80 = 1.038, <i>p</i> = 0.4148). (B) In the Morris water maze there was no statistical significance in the time spent in Q1, the quadrant where the hidden platform was located, between the treatment groups (F_2,20 = 1.222, <i>p</i> = 0.3156). (C) The contextual fear conditioning test showed no difference between treatment groups in the percentage of time the rats displaying freezing behaviour before, immediately after, 2 h and 24 h after the stimulus (AAV-empty, n = 11; AAV-hrGFP, n = 11; AAV-FLAG-NS, n = 13) (F_6,96 = 1.218, <i>p</i> = 0.3039). (D) The elevated T-maze revealed no difference in innate and acquired fear between treatment groups (AAV-empty, n = 11; AAV-hrGFP, n = 11; AAV-FLAG-NS, n = 13). The average avoidance latency measured on day 1 and day 4 showed a significant treatment-by-inhibitory avoidance interaction (F_2,32 = 0.3816, <i>p</i> = 0.0326). A significant difference was seen between the control and hrGFP group at day 4 (<i>t</i> = 3.491, <i>p</i> = 0.01) but there was no difference between control and neuroserpin group. (E) There was no significant difference in the average escape latency from the open arm to the enclosed arm at day 1 and day 4 between the treatment groups (F_2,32 = 1.357, <i>p</i> = 0.2719). All measurements are presented as the mean ± SEM. **indicates <i>p</i>&lt;0.01.</p

Influences of Lifestyle Profiles and Problematic Internet Use on Mental Distress in University Students

Hong Kong university students suffer a high prevalence of mental distress, but their lifestyle behaviors are not well-understood. We aimed to examine the relationship between mental distress, lifestyle behaviors, and problematic Internet use of this student population during the summer holiday and term-time. A two-cohort contrast group survey study was conducted. Students were surveyed in July during the summer holiday and September during term-time. The General Health Questionnaire-12, Health Promoting Lifestyle Profile II, and Generalized Problematic Internet Use Scale 2 were administered to measure mental distress, lifestyle behaviors, and problematic Internet use. 949 students (summer=467; term-time=482; Mage±SD, 20.11±1.54) participated in the study. Students reported statistically significant lower mental distress and higher spiritual growth during the summer holiday compared to term-time. The results of Structural Equation Modelling (SEM) found that spiritual growth was directly associated with reduced mental distress and problematic internet use, and indirectly associated with reduced mental distress through a negative correlation with problematic internet use. Problematic internet use was positively correlated with mental distress. Physical activity was more associated with better mental health during the summer holiday and for female students. In contrast, Health management was associated with better mental health during term-time and for male students. In conclusion, spiritual growth supports mental health improvement and counters problematic internet use in university students in general, factors such as physical activity and health management show differential influences based on gender and time of year. Results may help inform the development of student support workshops in higher education

Comprehensive Evaluation of the MBT STAR-BL Module for Simultaneous Bacterial Identification and β-Lactamase-Mediated Resistance Detection in Gram-Negative Rods from Cultured Isolates and Positive Blood Cultures

Objective: This study evaluated the capability of a MALDI Biotyper system equipped with the newly introduced MBT STAR-BL module to simultaneously perform species identification and β-lactamase-mediated resistance detection in bacteremia -causing bacteria isolated from cultured isolates and patient-derived blood cultures (BCs).Methods: Two hundred retrospective cultured isolates and 153 prospective BCs containing Gram-negative rods (GNR) were collected and subjected to direct bacterial identification, followed by the measurement of β-lactamase activities against ampicillin, piperacillin, cefotaxime, ceftazidime, and meropenem using the MBT STAR-BL module. The results and turnaround times were compared with those of routine microbiological processing. All strains were also characterized by beta-lactamase PCR and sequencing.Results: Using the saponin-based extraction method, MALDI-TOF MS correctly identified bacteria in 116/134 (86.6%) monomicrobial BCs. The detection sensitivities for β-lactamase activities against ampicillin, piperacillin, third-generation cephalosporin and meropenem were 91.3, 100, 97.9, and 100% for cultured isolates, and 80.4, 100, 68.8, and 40% for monomicrobial BCs (n = 134) respectively. The overall specificities ranged from 91.5 to 100%. Furthermore, the MBT STAR-BL and conventional drug susceptibility test results were concordant in 14/19 (73.7%) polymicrobial cultures. Reducing the logRQ cut-off value from 0.4 to 0.2 increased the direct detection sensitivities for β-lactamase activities against ampicillin, cefotaxime and meropenem in BCs to 85.7, 87.5, and 100% respectively. The MBT STAR-BL test enabled the reporting of β-lactamase-producing GNR at 14.16 and 47.64 h before the interim and final reports of routine BCs processing, respectively, were available.Conclusion: The MALDI Biotyper system equipped with the MBT STAR-BL module enables the simultaneous rapid identification of bacterial species and β-lactamase-mediated resistance from BCs and cultured isolates. Adjustment of the logRQ cut-off value to 0.2 significantly increased the detection sensitivities for clinically important drug-resistant pathogens