The effectiveness of physical activity interventions in improving higher education students' mental health: A systematic review.

Traditional interventions aiming to improve students' mental health and quality of life include meditation or canine therapy. The development of physical activity-related interventions has increased over the past decade. We aimed to review all studies using physical activity for improving the mental health and quality of life in higher education students whilst describing the interventions, measurements and effectiveness. A systematic search of six electronic databases including: ProQuest, MEDLINE, Embase, CINAHL, SPORTDiscus and CENTRAL, was conducted following PRISMA guidelines. Randomized or non-randomized controlled trial physical activity-related interventions involving higher education students aiming to improve their mental health and quality of life were included. Searches yielded 58 articles with interventions involving martial arts, sport, mind-body exercises and anaerobic exercises. Psychological measures varied across studies including the State Trait Anxiety Inventory, Beck Depression Inventory and the Perceived Stress Scale. Over half of the studies included in this review (n = 36) were effective in improving students' mental health or quality of life. Findings from our review suggest that interventions aiming to be effective in improving students' mental health quality of life should aim to deliver moderate-vigorous intensity exercises such as dance or Pilates. This systematic review was based on a published protocol in PROSPERO (registration number: CRD42022325975).

Exploring the feasibility of a cluster pilot randomised control trial to improve children's 24-hour movement behaviours and dietary intake: Happy homework.

We aimed to explore the feasibility, acceptability, and potential efficacy of Happy Homework (HH); an 8-week home-focussed intervention, with the purpose of encouraging children's positive dietary behaviours and engagement in positive physical activity (PA) and sleep behaviours. We randomised four Scottish schools (n = 71 participants; 5 classrooms) to either the HH intervention (n = 2) or usual curriculum control group (n = 2). HH consisted of movement and dietary-focused parent and child tasks. Primary outcome measures were intervention feasibility, acceptability, and potential efficacy. Secondary outcomes were objectively measured PA via ActiGraph GT3X+, sedentary behaviours (SBs) and sleep duration via activPAL4™ accelerometers and dietary behaviours, fruit and vegetable consumption and screen-time via questionnaires. After controlling for pre-test levels, post intervention stepping time and sleep duration were significantly greater for the HH group in comparison to the control group. The HH group reported eating more fruit and vegetables at post-test than the control group. Participants also reported the intervention to be enjoyable and motivating. These findings provide promising evidence that given a greater sample size, better retention and the prioritisation of health and wellbeing homework, HH could enhance children's health and wellbeing.

Relationship Between Parent and Child Physical Activity Using Novel Acceleration Metrics.

Background: Raw acceleration data lend themselves to innovative metrics in which comparisons can be made across populations. This study examined whether the intensity gradient (IG) or average acceleration (AvAcc) was associated with body mass index (BMI) in children and adults and explored parent-child associations between time spent in physical activity intensities and BMI. Methods: Parent-child dyads (n = 90) wore a wrist-mounted ActiGraph GT3X+ monitor for ≥ 3 valid days (i.e., ≥ 16 hrs each day). Participants' BMI was calculated. Results: Girls' BMI-z scores were positively associated with parent BMI and inactive time. Parent BMI was negatively associated with boys' inactive time. Parental inactive time predicted girls' BMI-z scores. The metrics were not associated with parent BMI, and no independent effects were observed between the metrics. Associations between boys' IG and BMI z-score were observed. No independent effects were observed between the metrics and no significant associations were observed between boys' BMI z-score and AvAcc. Girls' AvAcc and IG were not associated with BMI z-score and no independent effects were observed between the metrics. Conclusion: Findings support previous research which suggests that parent's activity does not influence their child's activity.

Feasibility and Acceptability of a Classroom-Based Active Breaks Intervention for 8-12-Year-Old Children.

Purpose: This study explored the feasibility of conducting a classroom-based active breaks intervention on sedentary behavior (SB), physical activity (PA) and attention in 8-12-year-old children. Methods: Eight schools were randomized on a 1:1 basis to the control or intervention. Teachers selected 10 cards detailing an activity break at random. Children then undertook each of the ten activity breaks for 30 s, three times per day for 6 weeks. School and participant recruitment, attrition rates, percentage of outcome measures collected, and acceptability were used to explore the feasibility of the study. Mixed effects models were undertaken to examine intervention effects upon measures of PA, SB and attention. Results: Two hundred and thirty-nine consent forms were issued and 153 were returned (64%). Of the 153 consents, 146 children (95%) were measured at baseline, and 117 participated in the follow-up measures (80%) 6 weeks later suggesting the intervention was acceptable for the participants. From teacher interviews, it was noted that the intervention was feasible to implement, and teachers observed positive classroom behavior changes in children. Inclusion rates for outcome measures ranged from 49% to 66%. Significant, intervention effects were observed for sitting time (B = -27.19; 95%CI: -36.84, -17.17), standing time (B = 23.51; 95%CI 14.1, 32.45) and the number of sit to stand transitions (B = 16.1; 95%CI 4.7, 26.79). Conclusion: Findings suggest that it was feasible and acceptable to implement an active breaks intervention within the classroom setting. Future work should consider the effectiveness of implementing this intervention across a full academic year.

The Effects of Socioeconomic Status on Parent and Child Moderate-to-Vigorous Physical Activity and Body Mass Index.

Purpose: Physical inactivity and overweight status has been linked to low socioeconomic status (SES) in youth. Parents are known to influence both their child's weight and physical activity (PA). The relationship between parent and child PA is of interest to many researchers; however, previous research typically relies on self-reported measures. The purpose of this study was to examine the relationship between parent and child moderate-to-vigorous PA (MVPA) and body mass index (BMI) in a sample of children (4-11 years old) using wrist-worn accelerometers and to explore mediating processes by which SES influences child MVPA and BMI through their parents MVPA and BMI. Methods: Parent and child dyads (n = 174) wore an ActiGraph GT3X+ accelerometer on their non-dominant wrist for 7 days. Mediation analyses were conducted to understand the indirect relationships between SES and child MVPA and BMI. Results: Weekend parent and child MVPA was significantly related (p < .01). Parent and child BMIs were also significantly related (p < .001). There was a significant negative direct effect of SES on child BMI (p < .05). Additionally, we observed a significant negative indirect effect of SES on child BMI via their parents BMI (B = -.04, SE .02, 95% CI = -.07 to -.01). Conclusions: Whilst parent and child MVPA were significantly related during the weekend, there were no associations between SES and MVPA. Future interventions aiming to improve health outcomes in children should consider the influence SES can have as well as parental activity on children's weekend MVPA.

Segmented sedentary time and physical activity patterns throughout the week from wrist-worn ActiGraph GT3X+ accelerometers among children 7-12 years old.

Background: This study examined the volume and patterns of physical activity (PA) and sedentary time (ST) across different segments of the week among boys and girls. Methods: A total of 188 children aged 7-12 years wore a wrist-mounted ActiGraph GT3X+ accelerometer for 7 days. Time spent in PA and ST was calculated using ActiLife software. The mean number of minutes of light PA, moderate PA, vigorous PA, moderate-to-vigorous PA (MVPA), and ST were calculated per weekday (before school, during school, and after school) and per weekend day (morning and afternoon-evening). Results: After school represented the greatest accumulation of ST compared with before school and during school segments. Boys engaged in 225.4 min/day of ST (95% confidence interval (CI): 216-235), and girls engaged in 222.2 min/day of ST (95%CI: 213-231). During school, boys engaged in significantly more MVPA than girls (46.1 min/day (95%CI: 44-48) vs. 40.7 min/day (95%CI: 39-43)). Across the whole weekday, boys participated in significantly more MVPA than girls (103.9 min/day (95%CI: 99-109) vs. 95.7 min/day (95%CI: 90-101)). The weekend afternoon-evening segment represented the larger accumulation of ST, where boys were significantly more sedentary than girls (367.5 min/day (95%CI: 353-382) vs. 339.8 min/day (95%CI: 325-355), respectively). Conclusion: Our findings suggest that children are highly sedentary and spend little of their time in school in MVPA, especially girls. Routine breaks in school elicit increases in light PA and MVPA. Future work should consider the use of more active breaks within school time to encourage PA and reduce ST.

An Insight Into the Involvement of Mothers of Low Socioeconomic Status in Scottish Primary School Health Education Activities.

School-based health activities that involve parents are more likely to be effective for child health and well-being than activities without a parent component. However, such school-based interventions tend to recruit the most motivated parents, and limited evidence exists surrounding the involvement of hard-to-reach parents with low socioeconomic status (SES). Mothers remain responsible for the majority of family care; therefore, this study investigated mothers with low SES to establish the reasons and barriers to their involvement in school-based health activities and to propose strategies to increase their involvement in those activities. Interviews were conducted with mothers with low SES, who were typically not involved in school-based health activities (n = 16). An inductive-deductive approach to hierarchical analysis revealed that there are several barriers resulting in mothers being less involved, particularly due to issues surrounding the schools' Parent Councils and the exclusivity of school-based events. Efforts made by the school to promote health activities and involve parents in such activities were revealed, alongside recommendations to improve on these practices. The findings offer multiple ways in which future school-based health interventions can recruit and involve mothers with low SES.

Measuring the impact of an interdisciplinary learning project on nursing, architecture and landscape design students' empathy.

DOMESTIC VIOLENCE AND REFUGE SERVICES IN AUSTRALIA: In Australia and internationally, domestic violence is a major cause of homelessness for women and children and yet provision for accommodation for this user-group is not well documented or understood. When designing emergency accommodation, the concerns, preferences, and perspectives of individuals who access refuge services must be sought in order to create spaces that are conducive to the needs of this diverse and vulnerable group. An empathic 'lens' can provide meaningful insights that can inform the design of refuge services specifically targeted at addressing these needs. This paper describes an authentic interdisciplinary learning experience for nursing, architecture and landscape students at a university in Sydney, Australia, and presents the results of a study designed to measure the impact of this initiative on participants' empathy towards women and children who access refuge services as a result of homelessness and/or domestic violence. Empathy levels were measured using the Comprehensive State Empathy Scale, a validated measurement tool. AN AUTHENTIC INTERDISCIPLINARY LEARNING EXPERIENCE: The learning experience consisted of collaborative meetings with stakeholders from the refuge sector, fieldwork, individual research, exchanging ideas and problem-solving in teams. Students then developed design guides for refuges that demonstrated their emerging understanding of the specific needs and perspectives of the issues faced by women and children who had experienced violence and found themselves homeless. Pre-post Comprehensive State Empathy Scale results indicated that the empathy levels of nursing and landscape students increased and those of architecture students decreased, however, these results were not statistically significant. BUILDING EMPATHY IN TEACHING AND LEARNING: The significance of the results from this study include an ability to compare the changes in empathy in students working collaboratively on a project and to ascertain possible reasons for this using a validated measurement tool. As empathy is one of the strongest negative correlates of prejudice, authentic teaching and learning activities, such as the one described in this paper, have the potential to positively impact the lived experience of women and children leaving situations of domestic violence.

The use of the intensity gradient and average acceleration metrics to explore associations with BMI z-score in children.

This study used the intensity gradient (IG) and average acceleration metrics to describe children's activity profiles and explore associations with body mass index (BMI) z-score. Two hundred and forty-six children (n = 138 girls) aged 9.6 ± 1.4 years wore a wrist-mounted ActiGraph wGT3X-BT accelerometer for 7 days on their non-dominant wrist. Physical activity (PA) metrics captured included: the IG which describes the intensity distribution of accelerations across the 24 h monitoring period; average acceleration which provides a measure of the volume of activity; total moderate-to-vigorous PA (MVPA) and inactive time. Acceleration was averaged over 5s epochs. Finally, BMI z-score was calculated for each participant. Average acceleration was negatively associated with BMI z-score (p < 0.05) independent of age and gender but not IG. The IG was negatively associated with BMI z-score independent of potential correlates and average acceleration. Total MVPA was not associated with BMI-z score. The IG and average acceleration metrics may be used to explore the independent or cumulative effects of the volume and intensity distribution of activity upon measures of health and well-being in children to inform specific activity recommendations.

A feasibility study with process evaluation of a teacher led resource to improve measures of child health.

Previous school-based interventions have produced positive effects upon measures of children's health and wellbeing but such interventions are often delivered by external experts which result in short-term effects. Thus, upskilling and expanding the resources available to classroom teachers could provide longer-term solutions. This paper presents a feasibility study of an online health resource (Healthy Schools Resource: HSR) developed to assist primary school teachers in the delivery of health-related education. Four schools (n = 2 intervention) participated in this study. Study feasibility was assessed by recruitment, retention and completion rates of several outcomes including height, weight, waist circumference, blood pressure and several metabolic markers including HDL-cholesterol, triglycerides, glucose and dietary knowledge following a 10-12-week intervention period. The process evaluation involved fidelity checks of teachers' use of the HSR and post-intervention teacher interviews. A total of 614 consent forms were issued and 267 were returned (43%), of which, 201 confirmed consent for blood sampling (75%). Retention of children participating in the study was also high (96%). Of the 13 teachers who delivered the intervention to the children, four teachers were excluded from further analyses as they did not participate in the fidelity checks. Overall, teachers found the online resource facilitative of teaching health and wellbeing and several recommendations regarding the resource were provided to inform further evaluations. Recruitment and retention rates suggest that the teacher led intervention is feasible and acceptable to both teachers, parents and children. Initial findings provide promising evidence that given a greater sample size, a longer intervention exposure period and changes made to the resource, teachers' use of HSR could enhance measures of health and wellbeing in children.

Diagnostic performance of Body Mass Index, Waist Circumference and the Waist-to-Height Ratio for identifying cardiometabolic risk in Scottish pre-adolescents.

BACKGROUND: Limited studies have examined the diagnostic performance of body mass index (BMI), waist circumference (WC) or waist-to-height ratio (WHtR) for identifying cardiometabolic risk (increased clustered glucose, triglycerides, mean arterial pressure and inv-HDL-cholesterol) in pre-adolescent youth. AIM: To compare the utility of BMI, WC and WHtR as predictors of cardiometabolic risk (CMR) in Scottish pre-adolescent children. SUBJECTS AND METHODS: A cross-sectional analysis of 223 Scottish children (55.2% boys, mean age =8.4 years) was undertaken. BMI, WC and WHtR were used as exposure variables within multivariate logistic regression analysis and ROC analysis to examine the utility of these anthropometrical indices in identifying those at cardiometabolic risk. RESULTS: Individuals with an elevated WHtR, WC and BMI were 3.51 (95% CI = 1.71-7.23; p < .001); 2.34 (95% CI = 1.35-4.06; p = .002) and 2.59 (95% CI = 1.42-4.73; p = .002) times more likely to be at cardiometabolic risk, respectively. The areas under the curves [AUC] to identify children with cardiometabolic risk were significant and similar among anthropometric indices (AUC's = 0.60-0.65). When stratified by BMI, both WC and WHtR demonstrated a fair-to-good ability for identifying those at cardiometabolic risk (AUC = 0.75-0.81). CONCLUSIONS: Findings suggest that the combination of BMI with either WC or WHtR may provide an added benefit in the assessment of cardiometabolic risk amongst pre-adolescents.

The activity and expression of chitinase in the equine lung and its activity in normal horses and animals with recurrent airway obstruction.

Recurrent airway obstruction (RAO) is a chronic inflammatory condition in equine lung, which may share a common immunological basis with human asthma, in which dysregulated Th2 responses occur. Mammals express chitinases and chitinase-like proteins, two of which are active enzymes, chitotriosidase and acidic mammalian chitinase (AMCase). Both enzymes are upregulated in a range of inflammatory conditions, including asthma. We investigated the activity of chitinase in bronchoalveolar lavage fluid from horses with and without RAO in response to organic dust challenges. No significant differences were found in activity, although in one study RAO animals had elevated chitinase activity that fell short of statistical significance. The pH optimum and pH lability of the activity was consistent with the presence of chitotriosidase. RT-PCR amplification of the mRNA encoding chitotriosidase and AMCase in normal equine lung showed that chitotriosidase, but not AMCase, is expressed in trachea, bronchi, and peripheral lung tissue. The gene for chitotriosidase was identified from the Equus caballus (horse) genome 1.1 database and its similarity to the same genes from other species was determined. The results of this study indicate that the involvement of chitotriosidase in RAO is uncertain.

Chitotriosidase is the primary active chitinase in the human lung and is modulated by genotype and smoking habit.

BACKGROUND: Chitinolytic enzymes play important roles in the pathophysiology of allergic airway responses in mouse models of asthma. Acidic mammalian chitinase (AMCase) and chitotriosidase (CHIT1) have chitinolytic activity, but relatively little is known about their expression in human asthma. OBJECTIVE: We sought to determine the expression and activity of AMCase and CHIT1 in healthy subjects, subjects with asthma, and habitual smokers, taking account of the null 24-bp duplication in the CHIT1 gene. METHODS: We measured chitinase activity in bronchoalveolar lavage (BAL) fluid at multiple pHs by using a synthetic chitin substrate. We also determined AMCase and CHIT1 gene expression in epithelial brushings and BAL fluid macrophages by means of real time RT-PCR. Paired DNA samples were genotyped for the CHIT1 duplication. RESULTS: In all subgroups the pH profile of chitinase activity in BAL fluid matched that of CHIT1, but not AMCase, and chitinase activity was absent in subjects genetically deficient in active CHIT1. Although AMCase protein was detectable in lavage fluid, AMCase transcripts in macrophages were consistent with an isoform lacking enzymatic activity. Median chitinase activity in BAL fluid tended to be lower than normal in asthmatic subjects but was increased 7-fold in habitual smokers, where CHIT1 gene expression in macrophages was increased. CONCLUSIONS: Chitinase activity in the lung is the result of CHIT1 activity. Although AMCase protein is detectable in the lung, our data indicate that it is inactive. Chitinase activity is not increased in subjects with asthma and in fact tends to be decreased. The high levels of chitinase activity in habitual smokers result from upregulation of CHIT1 gene expression, especially in macrophages.

Genome-wide profiling identifies epithelial cell genes associated with asthma and with treatment response to corticosteroids.

Airway inflammation and epithelial remodeling are two key features of asthma. IL-13 and other cytokines produced during T helper type 2 cell-driven allergic inflammation contribute to airway epithelial goblet cell metaplasia and may alter epithelial-mesenchymal signaling, leading to increased subepithelial fibrosis or hyperplasia of smooth muscle. The beneficial effects of corticosteroids in asthma could relate to their ability to directly or indirectly decrease epithelial cell activation by inflammatory cells and cytokines. To identify markers of epithelial cell dysfunction and the effects of corticosteroids on epithelial cells in asthma, we studied airway epithelial cells collected from asthmatic subjects enrolled in a randomized controlled trial of inhaled corticosteroids, from healthy subjects and from smokers (disease control). By using gene expression microarrays, we found that chloride channel, calcium-activated, family member 1 (CLCA1), periostin, and serine peptidase inhibitor, clade B (ovalbumin), member 2 (serpinB2) were up-regulated in asthma but not in smokers. Corticosteroid treatment down-regulated expression of these three genes and markedly up-regulated expression of FK506-binding protein 51 (FKBP51). Whereas high baseline expression of CLCA1, periostin, and serpinB2 was associated with a good clinical response to corticosteroids, high expression of FKBP51 was associated with a poor response. By using airway epithelial cells in culture, we found that IL-13 increased expression of CLCA1, periostin, and serpinB2, an effect that was suppressed by corticosteroids. Corticosteroids also induced expression of FKBP51. Taken together, our findings show that airway epithelial cells in asthma have a distinct activation profile and identify direct and cell-autonomous effects of corticosteroid treatment on airway epithelial cells that relate to treatment responses and can now be the focus of specific mechanistic studies.

Epithelial mucin stores are increased in the large airways of smokers with airflow obstruction.

BACKGROUND: Habitual cigarette smoking is associated with chronic mucus hypersecretion, but the relationship between mucus abnormalities and airflow obstruction in smokers is uncertain. METHODS: We collected bronchial biopsy samples and epithelial brushings from 24 smokers with and without airflow obstruction and 19 nonsmoking healthy control subjects. Epithelial mucin stores, mucin immunostains, and goblet cell morphology were quantified in bronchial biopsy samples using stereology, and mucin gene expression was quantified in epithelial brushings using real-time reverse transcriptase-polymerase chain reaction. RESULTS: Goblet cell size and number were higher than normal in smokers (both p < 0.05), leading to a 2.2-fold increase in the volume of stored mucin in the epithelium per surface area of basal lamina (1.94 +/- 0.31 microm(3)/microm(2) vs 4.32 +/- 0.55 microm(3)/microm(2) in control subjects vs smokers, p = 0.001). The increase in stored mucin occurred because of an increase in MUC5AC (p = 0.018) and despite a decrease in MUC5B (p < 0.0001). Stored mucin was significantly higher in the subgroup of smokers with airflow obstruction (p = 0.029) and correlated with FEV(1)/FVC even when controlling for diffusing capacity as a measure of emphysema (p = 0.034). CONCLUSIONS: Epithelial mucin stores are increased in habitual smokers because of goblet cell hypertrophy and hyperplasia, and the pattern of mucin gene expression is abnormal. The highest epithelial mucin stores are found in smokers with airflow obstruction, suggesting a mechanistic link between epithelial mucin dysregulation and airflow obstruction.

Transcript signatures of lymphocytic bronchitis in lung allograft biopsy specimens.

BACKGROUND: Rejection and obliterative bronchiolitis are barriers to sustained graft function in recipients of transplanted lungs. Early detection is hindered by inadequate tests and an incomplete understanding of the molecular events preceding or accompanying graft deterioration. METHODS: Hypothesizing that genes involved in immune responses and tissue remodeling produce biomarkers of rejection, we measured the expression of 192 selected genes in 72 sets of biopsy specimens from human lung allografts. Gene transcripts were quantified using a 2-step, multiplex, real-time polymerase chain reaction approach in endobronchial and transbronchial biopsy specimens from transplant recipients without acute infections undergoing routine surveillance bronchoscopy. RESULTS: Comparisons of histopathology in parallel biopsy specimens identified 6 genes correlating with rejection as manifested by lymphocytic bronchitis, a suspected harbinger of obliterative bronchiolitis. For example, beta2-defensin and collagenase transcripts in inflamed bronchi increased 37-fold and 163-fold, respectively. By contrast, these transcripts did not correlate with acute rejection in transbronchial specimens. Further, no correspondence was noted between histopathologic bronchitis and parenchymal rejection when endobronchial and transbronchial samples were obtained from the same patient. CONCLUSIONS: Our highly sensitive method permits quantitation of many gene transcripts simultaneously in small, bronchoscopically acquired biopsy specimens of allografts. Transcript signatures obtained by this approach suggest that airway and alveolar responses to rejection differ and that endobronchial biopsy specimens assess lymphocytic bronchitis and chronic rejection but are not proxies for transbronchial biopsy specimens. Further, they reveal changes in airway expression of the specific genes involved in host defense and remodeling and suggest that the measurement of transcripts correlating with lymphocytic bronchitis may be diagnostic adjuncts to histopathology.

c-Kit immunophenotyping and metalloproteinase expression profiles of mast cells in interstitial lung diseases.

Diverse interstitial lung diseases (ILD) demonstrate mesenchymal infiltration by an abundance of activated mast cells whose role in parenchymal fibrogenesis remains unclear. Since mast cells differentiate in a dynamic, tissue-specific manner via signals transduced by c-Kit receptor, we examined the effect of ILD microenvironments on c-Kit expression and metalloproteinase phenotypes of mesenchymal mast cell populations. Immunohistochemical and flow cytometric analyses characterized surface expression of c-Kit on mast cells in tissues obtained from patients with idiopathic pulmonary fibrosis, systemic sclerosis, sarcoidosis, and lymphangioleiomyomatosis, thus identifying a unique immunophenotype not shared by normal lung mast cells. Isolation of c-Kit+/FcepsilonRI+/CD34- mast cells via immunocytometric sorting of heterogeneous cell populations from mechanically disaggregated lung tissues permitted analysis of gene expression patterns by two-step real-time polymerase chain reaction. Transcriptional profiling identified expression of c-Kit and the neutral serine proteases, tryptase and chymase, establishing the identity of sorted populations as mature mast cells. Mast cells harvested from ILD tissues demonstrated characteristic metalloproteinase phenotypes which included expression of matrix metalloproteinase (MMP)-1 and a disintegrin and metalloproteinase (ADAM)-9, -10, and -17. Immunohistochemical co-localization guided by gene profiling data confirmed expression of chymase, MMP-1, and ADAM-17 protein in subpopulations of mast cells in remodelling interstitium. Gene profiling of harvested mast cells also showed increased transcript copy numbers for TNFalpha and CC chemokine receptor 2, which play critical roles in lung injury. We conclude that ILD microenvironments induce unique c-Kit receptor and metalloproteinase mast cell phenotypes.

c-Maf and JunB mediation of Th2 differentiation induced by the type 2 G protein-coupled receptor (VPAC2) for vasoactive intestinal peptide.

Vasoactive intestinal peptide and its G protein-coupled receptors, VPAC(1) and VPAC(2), regulate critical aspects of innate and adaptive immunity. T cell VPAC(2)Rs mediate changes in cytokine generation, which potently increase the Th2/Th1 ratio and consequently shift the effector responses toward allergy and inflammation. To examine mechanisms of VPAC(2) promotion of the Th2 phenotype, we analyzed controls of IL-4 transcription in CD4 T cells from T cell-targeted VPAC(2) transgenic (Tg), VPAC(2) knockout, and wild-type (WT) mice. c-maf and junB mRNA, protein, and activity were significantly up-regulated to a higher level in TCR-stimulated CD4 T cells from Tg mice compared with those from knockout and WT C57BL/6 mice. In contrast, GATA3, T-bet, and NFATc levels were identical in WT and Tg CD4 T cells. Vasoactive intestinal peptide binding to VPAC(2) on CD4 T cells specifically induces an up-regulation of the Th2-type transcription factors c-Maf and JunB, which consequently enhances IL-4 and IL-5 production, leading to a Th2-type phenotype.

Hyperplasia of smooth muscle in mild to moderate asthma without changes in cell size or gene expression.

Bronchial hyperresponsiveness in mild to moderate asthma may result from airway smooth muscle cell proliferation or acquisition of a hypercontractile phenotype. Because these cells have not been well characterized in mild to moderate asthma, we examined the morphometric and gene expression characteristics of smooth muscle cells in this subgroup of patients with asthma. Using bronchial biopsies from 14 subjects with mild to moderate asthma and 15 control subjects, we quantified smooth muscle cell morphology by stereology and the expression of a panel of genes related to a hypercontractile phenotype of airway smooth muscle, using laser microdissection and two-step real-time polymerase chain reaction. We found that airway smooth muscle cell size was similar in both groups, but cell number was nearly twofold higher in subjects with asthma (p = 0.03), and the amount of smooth muscle in the submucosa was increased 50-83% (p < 0.005). Gene expression profiling in smooth muscle cells showed no difference in the expression of genes encoding phenotypic markers in cells from healthy subjects and subjects with asthma (all p > 0.1). We conclude that airway smooth muscle proliferation is a pathologic characteristic of subjects with mild to moderate asthma. However, smooth muscle cells in mild to moderate asthma do not show hypertrophy or gene expression changes of a hypercontractile phenotype observed in vitro.

Phylogenetic analysis and rapid identification of Candida dubliniensis based on analysis of ACT1 intron and exon sequences.

The phylogenetic position of Candida dubliniensis has previously been established on the basis of the sequence of rRNA genes. In order to confirm the relationship between C. dubliniensis and other yeast species, particularly Candida albicans, using non-rRNA gene sequences the ACT1 gene was chosen for analysis. Three overlapping fragments that together span the entire C. dubliniensis ACT1 gene (CdACT1) were amplified from a recombinant phage isolated from a genomic DNA lambda library using PCR. These were cloned and used to determine the contiguous sequence of the gene. Analysis of the sequence data revealed the presence of a 1131 bp ORF interrupted by a single 632 bp intron at the 5' extremity of the gene. Comparison of the CdACT1 sequence with the C. albicans homologue (CaACT1) revealed that although the exons are 97.9% identical the introns are only 83.4% identical. Phylogenetic trees generated using ACT1 exon and intron sequences from a range of yeast species unequivocally confirmed the phylogenetic position of C. dubliniensis as a unique taxon within the genus Candida. Analysis of the ACT1-associated intron sequences from 10 epidemiologically unrelated C. dubliniensis isolates from disparate geographical locations showed a very low level of intraspecies sequence variation. In order to develop an accurate and rapid method to identify C. dubliniensis from primary isolation plates the significant divergence between the C. dubliniensis and C. albicans ACT1 intron sequences was exploited by designing C. dubliniensis-specific PCR primers. Using a rapid boiling method to produce template DNA directly from colonies from primary isolation plates in 10 min, these primers were used in a blind test with 122 isolates of C. dubliniensis, 53 isolates of C. albicans, 10 isolates of C. stellatoidea and representative isolates of other clinically relevant Candida and other yeast species. Only the C. dubliniensis isolates yielded the C. dubliniensis-specific 288 bp amplimer. Use of this technique on colonies suspected to be C. dubliniensis allows their correct identification as C. dubliniensis in as little as 4 h.