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Abstract Background Taxane is one of the first line treatments of lung cancer. In order to identify novel single nucleotide polymorphisms (SNPs) that might contribute to taxane response, we performed a genome-wide association study (GWAS) for two taxanes, paclitaxel and docetaxel, using 276 lymphoblastoid cell lines (LCLs), followed by genotyping of top candidate SNPs in 874 lung cancer patient samples treated with paclitaxel. Methods GWAS was performed using 1.3 million SNPs and taxane cytotoxicity IC50 values for 276 LCLs. The association of selected SNPs with overall survival in 76 small or 798 non-small cell lung cancer (SCLC, NSCLC) patients were analyzed by Cox regression model, followed by integrated SNP-microRNA-expression association analysis in LCLs and siRNA screening of candidate genes in SCLC (H196) and NSCLC (A549) cell lines. Results 147 and 180 SNPs were associated with paclitaxel or docetaxel IC50s with p-values -4 in the LCLs, respectively. Genotyping of 153 candidate SNPs in 874 lung cancer patient samples identified 8 SNPs (p-value PIP4K2A, CCT5, CMBL, EXO1, KMO and OPN3, genes within 200 kb up-/downstream of the 3 SNPs that were associated with SCLC overall survival (rs1778335, rs2662411 and rs7519667), significantly desensitized H196 to paclitaxel. SNPs rs2662411 and rs1778335 were associated with mRNA expression of CMBL or PIP4K2A through microRNA (miRNA) hsa-miR-584 or hsa-miR-1468. Conclusions GWAS in an LCL model system, joined with clinical translational and functional studies, might help us identify genetic variations associated with overall survival of lung cancer patients treated paclitaxel.

Objectives: Interstitial lung disease (ILD) is the most common cause of death in patients with systemic sclerosis (SSc), although disease behavior is highly heterogeneous. While a usual interstitial pneumonia (UIP) pattern is associated with worse survival in other ILDs, its significance in SSc-ILD is unclear. We sought to assess the prognostic utility of a deep-learning HRCT algorithm of UIP probability in SSc-ILD., Methods: Patients with SSc-ILD were included if HRCT images, concomitant lung function tests, and follow-up data were available. We used the Systematic Objective Fibrotic Imaging analysis Algorithm (SOFIA), a convolution neural network algorithm which provides probabilities of a UIP pattern on HRCT images. These were converted into the Prospective Investigation of Pulmonary Embolism Diagnosis (PIOPED)-based UIP probability categories. Decline in lung function was assessed by mixed-effect model analysis and relationship with survival by Cox proportional hazards analysis., Results: 522 patients were included in the study. 19.5% were classified as UIP not in the differential, 53.5% as low probability of UIP, 25.7% as intermediate probability of UIP, and 1.3% as high probability of UIP. A higher likelihood of UIP probability expressed as PIOPED categories was associated with worse baseline FVC, as well as with decline in FVC (p= 0.008), and worse 15-year survival (p= 0.001), both independently of age, gender, ethnicity, smoking history, and baseline FVC or Goh et al. staging system., Conclusion: A higher probability of a SOFIA-determined UIP pattern is associated with more advanced ILD, disease progression, and worse survival, suggesting that it may be a useful prognostic marker in SSc-ILD., (© The Author(s) 2024. Published by Oxford University Press on behalf of the British Society for Rheumatology. All rights reserved. For permissions, please email: journals.permissions@oup.com.)

The use of steroids in fibrotic interstitial lung diseases is founded on limited evidence. This modified Delphi survey sheds light on current clinical practices. Given the risks of steroids, clinical trials are needed to evaluate efficacy and harm. https://bit.ly/3VkgvbS., Competing Interests: Conflicts of interest: M. Funke-Chambour declares grants or contracts from Boehringer Ingelheim, consultancy and speaker's fees from Boehringer Ingelheim, Roche, MSD, AstraZeneca, Pfizer, GSK and Sanofi, outside this project. P. Suter has nothing to disclose. G.R. Jenkins declares grants to his institution from AstraZeneca, Biogen, Galecto, GlaxoSmithKline, Nordic Biosciences, RedX and Pliant, consultancy fees from AbbVie, Apollo Therapeutics, AstraZeneca, Brainomix, Bristol Myers Squibb, Chiesi, Cohbar, Daewoong, GlaxoSmithKline, Resolution Therapeutics and Pliant, honoraria from Boehringer Ingelheim, Chiesi, Roche, PatientMPower and AstraZeneca, payment for expert testimony from Pinsent Masons LLP, and participation on data safety or advisory board for Boehringer Ingelheim, Galapagos and Vicore, outside this project. L. Kawano-Dourado declares grants paid to their institution from Boehringer Ingelheim and Bristol Myers Squibb, and consultancy fees from Boehringer Ingelheim, outside this project. C.J. Ryerson declares grants from Boehringer Ingelheim paid to his institution, consultancy fees from Boehringer Ingelheim, Pliant Therapeutics, AstraZeneca, Trevi Therapeutics and Veracyte, payment or honoraria for lectures or presentations from Hoffmann-La Roche and Boehringer Ingelheim, and payment for expert testimony and travel support from Boehringer Ingelheim, outside this project. A.U. Wells reports consultancy fees from Roche, and payment or honoraria for lectures or presentations from Boehringer Ingelheim and Veracyte, and is President of the World Association of Sarcoidosis and Other Granulomatous Diseases, outside of the submitted work. M. Kreuter declares grants or contracts and royalties or licenses from Boehringer Ingelheim and Roche, consulting fees from Nichtraucherhelden/Sanero, Boehringer Ingelheim, Roche, BMS, GSK and AstraZeneca, and a leadership or fiduciary role for the European Respiratory Society, outside this project. K.A. Johannson declares grants paid to her institution from the University Hospital Foundation and the Three Lakes Foundation, consulting fees from Boehringer Ingelheim, Roche, Pliant Therapeutics and Brainomix, payment or honoraria for lectures or presentations and travel support from Boehringer Ingelheim, participation on a Data Safety Monitoring Board for the PFOX trial, and a stock option from Thyron SAB, outside this project., (Copyright ©The authors 2024.)

Rationale: Shared symptoms and genetic architecture between coronavirus disease (COVID-19) and lung fibrosis suggest severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection may lead to progressive lung damage. Objectives: The UK Interstitial Lung Disease Consortium (UKILD) post-COVID-19 study interim analysis was planned to estimate the prevalence of residual lung abnormalities in people hospitalized with COVID-19 on the basis of risk strata. Methods: The PHOSP-COVID-19 (Post-Hospitalization COVID-19) study was used to capture routine and research follow-up within 240 days from discharge. Thoracic computed tomography linked by PHOSP-COVID-19 identifiers was scored for the percentage of residual lung abnormalities (ground-glass opacities and reticulations). Risk factors in linked computed tomography were estimated with Bayesian binomial regression, and risk strata were generated. Numbers within strata were used to estimate posthospitalization prevalence using Bayesian binomial distributions. Sensitivity analysis was restricted to participants with protocol-driven research follow-up. Measurements and Main Results: The interim cohort comprised 3,700 people. Of 209 subjects with linked computed tomography (median, 119 d; interquartile range, 83-155), 166 people (79.4%) had more than 10% involvement of residual lung abnormalities. Risk factors included abnormal chest X-ray (risk ratio [RR], 1.21; 95% credible interval [CrI], 1.05-1.40), percent predicted Dl CO less than 80% (RR, 1.25; 95% CrI, 1.00-1.56), and severe admission requiring ventilation support (RR, 1.27; 95% CrI, 1.07-1.55). In the remaining 3,491 people, moderate to very high risk of residual lung abnormalities was classified at 7.8%, and posthospitalization prevalence was estimated at 8.5% (95% CrI, 7.6-9.5), rising to 11.7% (95% CrI, 10.3-13.1) in the sensitivity analysis. Conclusions: Residual lung abnormalities were estimated in up to 11% of people discharged after COVID-19-related hospitalization. Health services should monitor at-risk individuals to elucidate long-term functional implications.

Background: Classroom lighting, usually bright fluorescent light, can significantly influence the learning environment and emotions of students., Objective: To assess the emotional impact of classroom lighting on students during an academic year., Methods: This study used an ABAB withdrawal research design in the following manner: in phase A, the baseline condition, classroom lighting was provided by conventional overhead white fluorescent classroom lights; in phase B, the intervention condition, the conventional overhead white fluorescent classroom lights were covered with fabric filters (thin, translucent, creamy-colored plastic sheets) that were attached to the lighting fixture frame with magnetic discs. The filters produced softer light in the classroom than the fluorescent lights. Each phase lasted for at least 2 weeks. During each phase, students rated 18 pairs of words from the Mehrabian and Russell pleasure, arousal, and dominance semantic differential scale at least four times to assess the emotional impact of the lighting conditions., Results: For all three emotional behaviors, the mean score of the filtered fluorescent light phase was significantly greater than the mean score of the baseline unfiltered fluorescent lighting phase, indicating more positive emotional responses. Students also noted they experienced fewer headaches and found it easier to see the whiteboard at the front of the classroom when the light filters were in place., Conclusion: The light filters exerted a positive impact on the students' emotions. Students preferred the filtered lighting to fluorescent lighting. This study supports the installation of filters over fluorescent lights in a college classroom., Competing Interests: None.

The purpose of this study is to explore physiological and psychological changes immediately after a short-term visit to urban parks in an uncontrolled condition. Sixty park visitors completed a short questionnaire evaluating their subjective well-being (SWB) and donated a saliva sample immediately before and after their park visit. In addition, participants wore a pedometer to track their physical activity level during the park visit. Results indicated the levels of all biomarkers (cortisol, dehydroepiandrosterone [DHEA], and cortisol/DHEA ratio) decreased while SWB scores increased after the park visit. The duration of park visit was identified as a key variable significantly associated with cortisol reduction, and changes in SWB scores after the park visit were significantly associated with changes in cortisol level. A park visit duration of 21.8 min has the predictive ability to discriminate park visitors who exhibit reductions in cortisol levels after the park visit from those who do not.

Importance: Low social competence is one of the most complex and resistant challenges faced by adolescents with autism spectrum disorder (ASD). Occupational therapy practitioners have recognized the potential benefits of using the arts, including training in magic tricks, as a therapeutic medium to improve and enhance clients' psychosocial well-being., Objective: To describe the efficacy of a virtual magic trick training program (MTTP) to enhance social skills and self-esteem in adolescents with ASD., Design: Pragmatic, nonrandomized, wait-list controlled trial with 1-mo follow-up., Setting: Participants' homes., Participants: Seventeen adolescents (ages 9-15 yr) with ASD participated in the MTTP, 9 in the first cohort and 8 in the second (wait-list control) cohort., Intervention: Participants received magic trick training from occupational therapy students in pairs via videoconferencing in 45-min sessions, 3 days/wk, for 3 wk., Outcomes and Measures: Participants completed the Rosenberg Self-Esteem Scale and the Social Skills Improvement System before and after participating in the program., Results: Between-cohort effect sizes (Cohen's d) were 0.58 for social skills and 0.66 for self-esteem, considered moderate effects. Combining the two cohorts revealed significant improvements in social skills and self-esteem, with gains maintained at 1-mo follow-up., Conclusions and Relevance: The results of this study support our hypothesis that adolescents with ASD who participate in the 3-wk virtual MTTP can experience enhanced social skills and self-esteem. What This Article Adds: Learning magic tricks through individual coaching from occupational therapy students in a virtual environment can enhance the social skills and self-esteem of adolescents with ASD., (Copyright © 2023 by the American Occupational Therapy Association, Inc.)

The Transition to Wellness Program is an occupational therapy (OT) based program that was developed in response to the COVID-19 pandemic. For OT students, fieldwork is a required element of their training. In March 2020, these were being halted due to lockdowns and the risk to students in healthcare settings. Facing significant delays to graduation for 60 students, including some unable to be placed into healthcare settings due to underlying conditions, the development of a telehealth program was conceived to address this, while still meeting the accreditation clinical/health/wellness standards for OT fieldwork. A group of students, with faculty oversight, developed the theoretical basis, tools, policies and procedures for the program. An incremental launch allowed testing and refinement of the program to meet the educational needs of the students and the wellness needs of the clients. At its launch, the program supported 10 of that cohorts' 60 students to continue to meet the required academic standards and progress through their fieldwork towards graduation. Each student got to experience the "telehealth" model in providing OT wellness interventions to individuals in the local community. Future work will focus on continuing to broaden the reach of the program and measure the outcomes., Competing Interests: Conflicts of Interest: All authors have completed the ICMJE uniform disclosure form (available at https://mhealth.amegroups.com/article/view/10.21037/mhealth-21-35/coif). The series “mHealth: Innovations on the Periphery” was commissioned by the editorial office without any funding or sponsorship. The authors have no other conflicts of interest to declare., (2022 mHealth. All rights reserved.)

This study seeks to examine the effect of the characteristics of park users, park characteristics, and the neighborhood environment on park visits and the resultant perceived health status of residents living in Alabama, US. One hundred and eighty-nine participants, representing residents from 49 counties in Alabama, were recruited via a random-digit dial method to complete a telephone survey related to park visits. Data collection spanned 11 months. Results from the path analysis indicated a few significant direct effects: park location and body mass index (BMI) were the only two factors significantly associated with park visits, with race being marginally significant; and work status was significantly associated with perceived health. Our findings suggest park proximity, and high BMI are related to park visits and work status is related to perceived health among Alabamians. It is recommended that more neighborhood parks are needed for promoting Alabamians to visit them more frequently.

Background and Objective: A proportion of patients with fibrotic hypersensitivity pneumonitis (fHP) follow a progressive disease course despite immunosuppressive treatment. Little is known about predictors of mortality in fHP. We aimed to investigate the impact of short-term lung function changes in fHP on mortality., Methods: Baseline demographics for 145 consecutive patients with a multi-disciplinary team diagnosis of fHP, as well as baseline and 1-year follow-up of lung function, baseline echocardiographic findings, bronchoalveolar lavage (BAL) cellularity and all-cause mortality were recorded. Changes in forced vital capacity (FVC) ≥ 5% and ≥10%, and diffusion capacity of the lung for carbon monoxide (DLCO) ≥ 10% and ≥15% at 1 year were calculated. Cox proportional hazards analysis was performed to test for associations with mortality., Results: Baseline lung function severity, age, presence of honeycombing on computed tomography (CT) and echocardiographic pulmonary arterial systolic pressure (PASP) ≥ 40 mm Hg were associated with early mortality, while BAL lymphocytosis was associated with improved survival. A decline in FVC ≥ 5% (hazard ratio [HR]: 3.10, 95% CI: 2.00-4.81, p < 0.001), FVC ≥ 10% (HR: 3.11, 95% CI: 1.94-4.99, p < 0.001), DLCO ≥ 10% (HR: 2.80, 95% CI: 1.78-4.42, p < 0.001) and DLCO ≥ 15% (HR: 2.92, 95% CI: 1.18-4.72, p < 0.001) at 1 year was associated with markedly reduced survival on univariable and multivariable analyses after correcting for demographic variables, disease severity, honeycombing on CT and treatment, as well as BAL lymphocytosis and PASP ≥ 40 mm Hg on echocardiography, in separate models., Conclusion: Worsening in FVC and DLCO at 1 year, including a marginal decline in FVC ≥ 5% and DLCO ≥ 10%, is predictive of markedly reduced survival in fHP., (© 2022 The Authors. Respirology published by John Wiley & Sons Australia, Ltd on behalf of Asian Pacific Society of Respirology.)

Background: Children with attention-deficit/hyperactivity disorder (ADHD) have been reported to experience low self-esteem. Magic, a performing art, when used therapeutically, has been shown to enhance self-esteem in children with neurodevelopmental disorders. However, there is a paucity of evaluation studies using this intervention applied to children with ADHD., Purpose: This single-group study aims to describe the effectiveness of a virtual summer magic camp program to enhance self-esteem in children and adolescents with ADHD., Method: Six children with ADHD, age 8.6-14.3 years, who participated in a virtual magic camp program for children with disabilities, were included in this study. The camp met 3 days a week, about 45 min to an hour each session, over four consecutive weeks for a total of 9 to 12 hours. Participants completed the assessments before the beginning of the camp and post-camp. In addition, participants and their parents were individually interviewed to explore their camp experience at the post-camp data point., Results: A Wilcoxon signed-rank test indicated that the median ranks of the self-esteem scores at post-magic camp, median=21.5, were significantly higher than the median ranks of the self-esteem scores at baseline, median=19.5, with z =2.23, p =.026. The effect size of the virtual magic camp on self-esteem was .64, which is considered moderate. Findings were validated by the participants describing gains in self-esteem after participation in the magic camp and by the parents' statements regarding the positive impact on their child's psychological well-being., Discussion: This study supported the hypothesis that children with ADHD who participated in a four-week virtual magic camp experienced enhanced self-esteem. Future studies should investigate the holistic impact of magic camps on children and adolescents with ADHD and include measures that tap into other psychosocial attributes such as social functioning, social skills, and self-efficacy.

Previous studies have examined the quantitative results of using an organized approach of teaching and learning magic tricks-a therapeutic magic camp-but set aside the qualitative aspect. The purpose of this qualitative study is to explore the interpretation of the lived experience of children with hemiparesis after they have participated in a therapeutic magic camp. Six children, aged between 11 and 14 years old, with hemiparesis who completed the magic camp program 3 months prior were invited to participate in an in-person individual semistructured interview. The interviews were audio-recorded and transcribed verbatim for content analysis. Results from the initial qualitative analysis yielded categories of enjoyment, positive social and learning experience from participation in the camp, increased confidence, and self-motivation to use the affected upper limb. These categories were grouped into two themes: "having fun with others while learning" and "helping my arm perform." Having fun with others while learning was an amalgamation of fun, and positive social and learning experience. Helping my arm perform reflected a sense of increased competence and self-belief, and a willingness and motivation of the children to attempt challenging daily activities. The lived experiences of children with hemiplegia who participated in an organized delivery of learning to perform magic tricks yield-ed positive outcomes. These themes are consistent with the limited literature that is available suggesting that further research is needed to confirm the efficacy of the therapeutic magic camp intervention., Competing Interests: CONFLICT OF INTEREST No potential conflict of interest relevant to this article was reported., (Copyright © 2021 Korean Society of Exercise Rehabilitation.)

This study aimed to explore the amenable factors contributing to the improvement in subjective well-being (SWB) immediately after a short-term visit to an urban park in an uncontrolled condition. Ninety-four park visitors from three urban parks completed a short questionnaire evaluating SWB (with two components: affect and life satisfaction) immediately before and after their park visit. In addition, their level of physical activity was tracked by wearing an accelerometer during the park visit. Results indicated a significant improvement in SWB, affect, and life satisfaction scores of park visitor participants from before and after their visit. Duration of park visit was bivariately associated with SWB scores, and independently associated with the improvement in life satisfaction scores, controlling for parks and age, after the visit; a 20.5-min park visit predicted the highest overall accuracy (64%) improvement in life satisfaction. It is recommended that design of the park space should attract visitors to stay for at least 20 min in the park.

This single-group open trial was designed to evaluate the effectiveness of a two-week magic camp as a means of hand-arm motor skills training to improve upper limb motor function (unilateral and bilateral) in children with hemiparesis. Seven children with hemiparesis participated in a magic camp program which met 3 days a week, 4 hours each day, for two consecutive weeks for a total of 24 hours. Participants completed three assessments at the beginning of the camp, post-camp, and at a three-month follow-up: the Jebsen Hand Function Test (JHFT), Children's Hand Experience Questionnaire (CHEQ), and a box opening task that required coordination of both upper limbs. A Wilcoxon signed-rank test revealed significant improvement in JHFT composite scores of the affected limb at post-camp ( p = .04) and three-month follow-up ( p = .04). In addition, a significant improvement in the number of activities performed using two hands from baseline to three-month follow-up was observed ( p = .03). This pattern of improvement was also observed in the speed of completion for the box opening task. The improvement in motor function seems related to the participants' continuing performing daily activities with the affected hand and two hands after the magic camp.

Experts around the world support the integration of health promotion and wellness (HPW) services into traditional health care services. If successfully executed, the addition of HPW services would reduce rates of death and disability and significantly reduce health care costs. While all health care providers should be engaged in providing HPW services, many believe that physical therapists (PTs) and occupational therapists (OTs) are uniquely positioned to provide these services. However, research suggests that clinicians in both fields may fall short in doing so. Likewise, research indicates that entry-level educational programs inadequately prepare PT and OT students to be HPW practitioners. The overall purpose of this paper is to provide recommendations to educators for preparing PT and OT students and clinicians to better meet the HPW needs of the clients and patients they serve., Competing Interests: The authors declare no conflict of interest.

Background: To identify physical measures that predict maximal handgrip strength (MHGS) and provide evidence for identifying lack of sincerity of effort when assessing upper extremity weakness., Objective: This study investigated anthropometric measurements associated with MHGS of healthy young adults., Methods: A convenience sample of 150 healthy adults ages 19 to 34 years old completed the MHGS assessment, which was measured using a Jamar dynamometer according to the protocol of the American Society of Hand Therapists, for both dominant and non-dominant hands. Several anthropometric data were collected, which included height, body weight, forearm length, forearm circumference, hand length, and hand width., Results: Multivariable linear regression analysis indicated gender and hand width were uniquely and significantly associated with participants' MHGS for dominant and non-dominant hand and accounted for more than 60% of the variance, with R2 = 0.60, P < 0.001 for the dominant hand model and R2 = 0.64, P < 0.001 for the non-dominant hand model., Conclusions: Among the forearm and hand anthropometric measures, hand width is the best predictor of MHGS in both the non-dominant and dominant hands for healthy young adults.

Sex is a risk factor for development of cardiotoxicity, induced by the anti-cancer drug, doxorubicin (DOX), in humans. To explore potential mechanisms underlying differential susceptibility to DOX between sexes, 8-week old male and female B6C3F 1 mice were dosed with 3mg/kg body weight DOX or an equivalent volume of saline via tail vein once a week for 6, 7, 8, and 9 consecutive weeks, resulting in 18, 21, 24, and 27mg/kg cumulative DOX doses, respectively. At necropsy, one week after each consecutive final dose, the extent of myocardial injury was greater in male mice compared to females as indicated by higher plasma concentrations of cardiac troponin T at all cumulative DOX doses with statistically significant differences between sexes at the 21 and 24mg/kg cumulative doses. A greater susceptibility to DOX in male mice was further confirmed by the presence of cytoplasmic vacuolization in cardiomyocytes, with left atrium being more vulnerable to DOX cardiotoxicity. The number of TUNEL-positive cardiomyocytes was mostly higher in DOX-treated male mice compared to female counterparts, showing a statistically significant sex-related difference only in left atrium at 21mg/kg cumulative dose. DOX-treated male mice also had an increased number of γ-H2A.X-positive (measure of DNA double-strand breaks) cardiomyocytes compared to female counterparts with a significant sex effect in the ventricle at 27mg/kg cumulative dose and right atrium at 21 and 27mg/kg cumulative doses. This newly established mouse model provides a means to identify biomarkers and access potential mechanisms underlying sex-related differences in DOX-induced cardiotoxicity., (Published by Elsevier Inc.)

Identification of early biomarkers of cardiotoxicity could help initiate means to ameliorate the cardiotoxic actions of clinically useful drugs such as doxorubicin (DOX). Since DOX has been shown to target mitochondria, transcriptional levels of mitochondria-related genes were evaluated to identify early candidate biomarkers in hearts of male B6C3F1 mice given a weekly intravenous dose of 3mg/kg DOX or saline (SAL) for 2, 3, 4, 6, or 8 weeks (6, 9, 12, 18, or 24 mg/kg cumulative DOX doses, respectively). Also, a group of mice was pretreated (intraperitoneally) with the cardio-protectant, dexrazoxane (DXZ; 60 mg/kg) 30 min before each weekly dose of DOX or SAL. At necropsy a week after the last dose, increased plasma concentrations of cardiac troponin T (cTnT) were detected at 18 and 24 mg/kg cumulative DOX doses, whereas myocardial alterations were observed only at the 24 mg/kg dose. Of 1019 genes interrogated, 185, 109, 140, 184, and 451 genes were differentially expressed at 6, 9, 12, 18, and 24 mg/kg cumulative DOX doses, respectively, compared to concurrent SAL-treated controls. Of these, expression of 61 genes associated with energy metabolism and apoptosis was significantly altered before and after occurrence of myocardial injury, suggesting these as early genomics markers of cardiotoxicity. Much of these DOX-induced transcriptional changes were attenuated by pretreatment of mice with DXZ. Also, DXZ treatment significantly reduced plasma cTnT concentration and completely ameliorated cardiac alterations induced by 24 mg/kg cumulative DOX. This information on early transcriptional changes during DOX treatment may be useful in designing cardioprotective strategies targeting mitochondria., (Published by Elsevier Inc.)

This study investigated self-reported prevalence of and factors affecting clinicians' use of standardized assessments when evaluating clients for power mobility devices (PMDs), and explored assessments clinicians typically use when carrying out PMD evaluation. An e-mail survey was sent to assistive technology professionals listed in the Rehabilitation Engineering and Assistive Technology Society of North America directory. Three hundred fifty-four respondents, qualified to conduct formal power mobility skills assessments, completed the online survey. Of those, 122 (34.5%) respondents reported that they were aware of the presence of standardized performance-based power mobility skills assessments, but only 28 (7.9%) used these assessments in their practice. Multivariate analysis revealed that the odds of the respondents who use the standardized assessments were 18 times higher for those who were aware of the presence of these assessments than those who were not (adjusted odds ratio [OR] OR = 17.85, P < 0.0001). The odds of using the standardized assessment for respondents who did not identify themselves as occupational or physical therapists were five times higher than those who were therapists (adjusted OR = 0.20, P < 0.0001). This survey revealed that the assistive technology practitioners who recommend PMDs mainly use non-standardized mobility skills assessments.

This qualitative study explored the role of sensory characteristics embedded in the built environment and whether they support or hinder people with visual impairment in their use of public spaces. An online survey link was e-mailed to the presidents and committee members of each state's chapters and associations of the National Federation of the Blind in the United States, resulting in 451 direct invitations to participate. Written responses of the survey questions from 48 respondents with visual impairment were analyzed. Three main themes: Barriers, Supporters, and Context-Dependence emerged from the respondents' experience of multisensory characteristics within the built environment. The four subthemes subsumed in Barriers were: (1) Population specific design, (2) Extreme sensory backgrounds, (3) Uneven ground surfaces and objects, and (4) Inconsistent lighting. For Supporters, respondents provided specific examples of various sensory characteristics in built environments, including audible cues and echoes, smells, tactile quality of the ground surface, and temperature. Context-Dependence referred to the effects of sensory characteristics embedded in public spaces depending on one's vision condition, the proximity to the sensory cues and the purpose of the activities one was performing at that moment. Findings provide occupational therapy practitioners an in-depth understanding of the transactional relationship between embedded sensory characteristics in the built environment, occupations, and people with visual impairment in order to make appropriate modifications or removal of barriers that affect occupational performance and engagement. Suggestions for occupational therapists as well as architects, designers, planners, policy makers/legislators related to functional sensory cues in the design of built environments were provided to increase accessibility in the use of public spaces by people with visual impairment.

This study investigated the differences in the quality of park play spaces between an affluent and a non-affluent community in a large US Southeastern metropolitan area. Two cities were purposefully selected to reflect differences in household income and race/ethnicity characteristics. Using the Playable Space Quality Assessment Tool (PSQAT), all parks (n = 11, with six in the affluent city, and five in the non-affluent city) in these two cities were evaluated. The data were analyzed across three aspects of environmental features of the PSQAT: Location, Play Value and Care and Maintenance between parks in the two cities. The Mann-Whitney U test was used to test the study hypotheses. Results indicated significant differences between parks in the two cities in all three aspects of the PSQAT with p-values ≤ 0.03 and effect sizes of > 0.65, suggesting that the affluent city had parks of a higher quality than the non-affluent city. Significant disparity in Play Value (p = 0.009) in parks between these two communities suggests that children and young people are likely to have different experiences of the play spaces in their locality and therefore may experience different physical and psychological health benefits.

Two laboratories independently examined how the amount of starting material influences DNA extraction efficiency and, ultimately, the detection of low-level presence of genetically engineered (GE) traits in commercialized grains. GE traits from one maize, two canola, and two soybean samples were used as prototypical models in the study design as well as two commonly used DNA extraction methods, a small scale (0.1 and 0.2 g samples) and a large scale (1.0 and 2.0 g samples). The DNA samples were fortified (spiked) at 0.1 and 0.01% (w/w) levels. The amount of DNA recovery varied between the two laboratories, although a sufficient amount of DNA was obtained to perform replicate PCR analysis by both laboratories. Reliable detection of all five events was achieved by both laboratories at 0.1% level using either small-scale or large-scale DNA extractions. Reliable detection of the GE events was achieved at 0.01% level for soybean and canola but not for maize. Variability was observed among the two laboratories in terms of the Ct values generated. There was no difference between small-scale and large-scale DNA extraction methods for qualitative PCR detections of all five GE events.

Quantitative real-time polymerase chain reaction (qPCR) is a technology commonly used for the detection and quantification of genetically engineered (GE) traits in grains and oilseeds. The method involves measuring copy numbers of taxon-specific, endogenous control genes exposed to the same manipulations as trait-specific target genes. Accurate DNA quantification is essential for successful and predictable results with qPCR. A systematic study of seven different DNA quantification methods, incorporating different chemistries and different instrumentation, were evaluated on corn and soy DNA that was extracted using two distinct extraction methods. A time course study showed that corn and soy DNA was stable under typical laboratory storage conditions. CornCTAB and cornQiagen DNA extracts produced statistically similar quantification values when measured by picogreen PG(TD700), PG(Lum20/20), Hoescht(TD700), and Hoescht(Lum20/20) methods, suggesting that these methods can be used interchangeably to quantify DNA in corn samples prior to initiation of qPCR. Soy(Qiagen) provided greater stochastic measurement variability when quantification methods were compared, whereas soyCTAB had statistically significant differences when a PG method was compared to a Hoescht method of DNA quantification. Finally, agarose gel electrophoresis data revealed more pronounced degradation for Qiagen-extracted DNA compared with CTAB extracts in both corn and soy. Consequently, Ct values generated by qPCR suggested that absolute copy numbers of PCR amplifiable targets were not concordant between Qiagen and CTAB DNA extracts. Understanding measurement uncertainty from component steps used in qPCR can contribute toward harmonizing methods for the detection of GE traits in grains and oilseeds.

Eggs of the helminth Schistosoma mansoni accumulate in the colon following infection and generate Th2-biassed inflammatory granulomas which become down- modulated in size as the infection proceeds to chronicity. However, although CD4+CD25+FoxP3+ regulatory T cells (T(regs)) are known to suppress Th1-mediated colitis, it is not clear whether they control Th2-associated pathologies of the large intestine which characterise several helminth infections. Here we used a novel 3D-multiphoton confocal microscopy approach to visualise and quantify changes in the size and composition of colonic granulomas at the acute and chronic phases of S. mansoni infection. We observed decreased granuloma size, as well as reductions in the abundance of DsRed+ T cells and collagen deposition at 14 weeks (chronic) compared to 8 weeks (acute) post-infection. Th2 cytokine production (i.e. IL-4, IL-5) in the colonic tissue and draining mesenteric lymph node (mLN) decreased during the chronic phase of infection, whilst levels of TGF-β1 increased, co-incident with reduced mLN proliferative responses, granuloma size and fibrosis. The proportion of CD4+CD25+FoxP3+T(regs): CD4+ cells in the mLN increased during chronic disease, while within colonic granulomas there was an approximate 4-fold increase. The proportion of CD4+CD25+FoxP3+T(regs) in the mLN that were CD103+ and CCR5+ also increased indicating an enhanced potential to home to intestinal sites. CD4+CD25+ cells suppressed antigen-specific Th2 mLN cell proliferation in vitro, while their removal during chronic disease resulted in significantly larger granulomas, partial reversal of Th2 hypo-responsiveness and an increase in the number of eosinophils in colonic granulomas. Finally, transfer of schistosome infection-expanded CD4+CD25+T(regs) down-modulated the development of colonic granulomas, including collagen deposition. Therefore, CD4+CD25+FoxP3+T(regs) appear to control Th2 colonic granulomas during chronic infection, and are likely to play a role in containing pathology during intestinal schistosomiasis.

Infection of the mammalian host by schistosome larvae occurs via the skin, although nothing is known about the development of immune responses to multiple exposures of schistosome larvae, and/or their excretory/secretory (E/S) products. Here, we show that multiple (4x) exposures, prior to the onset of egg laying by adult worms, modulate the skin immune response and induce CD4(+) cell hypo-responsiveness in the draining lymph node, and even modulate the formation of hepatic egg-induced granulomas. Compared to mice exposed to a single infection (1x), dermal cells from multiply infected mice (4x), were less able to support lymph node cell proliferation. Analysis of dermal cells showed that the most abundant in 4x mice were eosinophils (F4/80(+)MHC-II(-)), but they did not impact the ability of antigen presenting cells (APC) to support lymphocyte proliferation to parasite antigen in vitro. However, two other cell populations from the dermal site of infection appear to have a critical role. The first comprises arginase-1(+), Ym-1(+) alternatively activated macrophage-like cells, and the second are functionally compromised MHC-II(hi) cells. Through the administration of exogenous IL-12 to multiply infected mice, we show that these suppressive myeloid cell phenotypes form as a consequence of events in the skin, most notably an enrichment of IL-4 and IL-13, likely resulting from an influx of RELMα-expressing eosinophils. We further illustrate that the development of these suppressive dermal cells is dependent upon IL-4Rα signalling. The development of immune hypo-responsiveness to schistosome larvae and their effect on the subsequent response to the immunopathogenic egg is important in appreciating how immune responses to helminth infections are modulated by repeated exposure to the infective early stages of development.

Biotechnology-derived varieties of canola, cotton, corn and soybean are being grown in the USA, Canada and other predominantly grain exporting countries. Although the amount of farmland devoted to production of biotechnology-derived crops continues to increase, lingering concerns that unintended consequences may occur provide the EU and most grain-importing countries with justification to regulate these crops. Legislation in the EU requires traceability of grains/oilseeds, food and feed products, and labelling, when a threshold level of 0.9% w/w of genetically engineered trait is demonstrated to be present in an analytical sample. The GE content is routinely determined by quantitative PCR (qPCR) and plant genomic DNA provides the template for the initial steps in this process. A plethora of DNA extraction methods exist for qPCR applications. Implementing standardized methods for detection of genetically engineered traits is necessary to facilitate grain marketing. The International Organization for Standardization draft standard 21571 identifies detergent-based methods and commercially available kits that are widely used for DNA extraction, but also indicates that adaptations may be necessary depending upon the sample matrix. This review assesses advantages and disadvantages of various commercially available DNA extraction kits, as well as modifications to published cetyltrimethylammonium bromide methods. Inhibitors are a major obstacle for efficient amplification in qPCR. The types of PCR inhibitors and techniques to minimize inhibition are discussed. Finally, accurate quantification of DNA for applications in qPCR is not trivial. Many confounders contribute to differences in analytical measurements when a particular DNA quantification method is applied and different methods do not always provide concordant results on the same DNA sample. How these differences impact measurement uncertainty in qPCR is considered.

An online survey was conducted by the International Life Sciences Institute, Food Biotechnology Committee, on the use of qualitative and quantitative polymerase chain reaction (PCR) assays for cauliflower mosaic virus 35S promoter and Agrobacterium tumefaciens Tnos DNA sequence elements for the detection of genetically engineered (GE) crop plant material. Forty-four testing laboratories around the world completed the survey. The results showed the widespread use of such methods, the multiplicity of published and in-house methods, and the variety of reference materials and calibrants in use. There was an interest on the part of respondents in validated quantitative assays relevant to all GE events that contain these two genetic elements. Data are presented by testing two variations each of five published real-time quantitative PCR methods for 35S detection on eight maize reference materials. The results showed that two of the five methods were not suitable for all the eight reference materials, with poor linear regression parameters and multiple PCR amplification products for some of the reference materials. This study demonstrates that not all 35S methods produce satisfactory results, emphasizing the need for method validation.

Background: Acne is a common chronic inflammatory dermatosis of the pilosebaceous unit. It is characterized by seborrhoea, comedone formation and an inflammatory response consistent with defective cellular immunity to Propionibacterium acnes., Objectives: The objective of this study was to investigate the immune reactivity of patients with acne compared with healthy controls by examining the response of peripheral blood mononuclear cells (PBMCs) to stimulation with P. acnes. Particular focus was placed upon measuring the production of interleukin (IL)-10, which has an established immunoregulatory role., Patients and Methods: Venous blood was collected from 47 patients with acne and 40 age- and sex-matched healthy controls with no prior history of acne. PBMCs were cultured and their cytokine response to P. acnes investigated., Results: Proinflammatory IL-8 and tumour necrosis factor (TNF)-alpha secretion from PBMCs was higher in patients with acne when stimulated with P. acnes. In contrast, a statistically significant reduction in PBMC secretion of anti-inflammatory IL-10 in patients with acne was identified. The impaired production of IL-10 by PBMCs from patients with acne was confined to CD14+ cells presumed to be monocytes. The ability of CD14 cells from patients with acne to phagocytose P. acnes bacteria was also observed to be defective but the addition of exogenous IL-10 to PBMC cultures restored phagocytic activity., Conclusions: These data suggest that patients with acne have a proinflammatory cytokine milieu and crucially are unable to contain early inflammatory changes due to a specific defect in immunosurveillance, namely low monocyte IL-10 production. Our observations raise the possibility that acne therapeutics might profitably target IL-10 both as a regulator of proinflammatory cytokines and in augmenting the CD14+ cell phagocytic response.

The agricultural biotechnology industry routinely utilizes real-time quantitative PCR (RT-qPCR) for the detection of biotechnology-derived traits in plant material, particularly for meeting the requirements of legislative mandates that rely upon the trace detection of DNA. Quantification via real-time RT-qPCR in plant species involves the measurement of the copy number of a taxon-specific, endogenous control gene exposed to the same manipulations as the target gene prior to amplification. The International Organization for Standardization (ISO 21570:2005) specifies that the copy number of an endogenous reference gene be used for normalizing the concentration (expressed as a % w/w) of a trait-specific target gene when using RT-qPCR. For this purpose, the copy number of a constitutively expressed endogenous reference gene in the same sample is routinely monitored. Real-time qPCR was employed to evaluate the predictability and performance of commonly used endogenous control genes (starch synthase, SSIIb-2, SSIIb-3; alcohol dehydrogenase, ADH; high-mobility group, HMG; zein; and invertase, IVR) used to detect biotechnology-derived traits in maize. The data revealed relatively accurate and precise amplification efficiencies when isogenic maize was compared to certified reference standards, but highly variable results when 23 nonisogenic maize cultivars were compared to an IRMM Bt-11 reference standard. Identifying the most suitable endogenous control gene, one that amplifies consistently and predictably across different maize cultivars, and implementing this as an internationally recognized standard would contribute toward harmonized testing of biotechnology-derived traits in maize.

Background: Most patients with cystic fibrosis (CF) die of respiratory failure due to chronic infection and destructive neutrophilic inflammation., Objective: To identify potential therapeutic targets by characterising the neutrophil stimulating mediators in the CF airway., Methods: Spontaneously expectorated CF sputum was extracted in phosphate buffered saline for assays of neutrophil chemotaxis, intracellular calcium mobilisation and cell shape change. Mediators were purified by ion exchange, C(18) reversed phase and size exclusion chromatography., Results: A pool of CF sputum contained considerable neutrophil stimulating activity but neutralisation of interleukin (IL)8/CXCL8 had little inhibitory effect on neutrophil chemotactic (10149 (2023) migrating cells vs 8661 (2597) at 62 mg sputum/ml; NS) or shape change (% forward scatter increase 46 (8) vs 38 (5) at 19 mg sputum/ml; p<0.05) responses. Furthermore, the CF sputum pool induced an elevation in intracellular calcium ions even after desensitisation of the neutrophils to IL8. Chromatography identified contributions to the neutrophil shape change inducing activity from IL8, other CXC chemokines, leukotriene (LT) B(4) and two formyl peptides. There was also suggestive evidence for contributions from platelet activating factor (PAF) and C5a. Using non-chromatographed individual sputum samples, anti-IL8 alone did have an inhibitory effect on neutrophil chemotaxis (median inhibition 41%; p = 0.0002). However, even in this experiment, there were clearly significantly important, non-IL8 mediated, effects of CF sputum on neutrophils, and an inhibitor cocktail of anti-IL8 plus CXCR2, LTB(4), formyl peptide, PAF and C5a receptor antagonists inhibited chemotaxis by a median of 97% (p = 0.0002)., Conclusion: Many chemoattractants contribute to the neutrophil stimulating activity in CF sputum although the relative contribution of these mediators differs in different patients. Selective blockade of single mediators may not be sufficient to control neutrophil recruitment and activation in the CF airway.

Rationale: Dithiothreitol (DTT) is commonly used to liquefy induced sputum samples before assessment of cytology, but causes reduction of disulfide bonds and denaturation of proteins., Objectives: To process sputum supernatants containing DTT to enable quantification of cytokines and chemokines., Methods: A standard solution of 22 pooled chemokines and cytokines was incubated with DTT at the concentrations used during sputum liquefaction and then dialyzed under 20 different denaturant and redox conditions., Measurements and Main Results: After incubation of the standard solution with DTT there was loss of detectable protein mediators on immunoassay, but optimized dialysis permitted recovery of chemokines to 96 +/- 4% and cytokines to 91 +/- 6%. Optimized dialysis of DTT supernatants from subjects with asthma covering a range of severities (n = 35) was performed in the presence of a cocktail of protease inhibitors and demonstrated significantly elevated levels of the chemokine CXCL10 (IFN-gamma-inducible protein-10), CXCL8 (IL-8), and CCL3 (macrophage inflammatory protein-1alpha); with lower but significantly elevated levels of CCL2 (monocyte chemotactic protein-1), CCL11 (eotaxin), and CCL5 (regulated on activation, normal T-cell expressed and secreted) in severe asthma. In sputum from subjects with severe asthma there were also significantly elevated levels of IL-4, IL-5, IL-13, tumor necrosis factor-alpha, IL-6, granulocyte-macrophage colony-stimulating factor, and IL-12(p40)., Conclusions: The technique of optimized dialysis and protease inhibition of sputum DTT supernatants aids the detection of chemokines and cytokines. The detection of elevated levels of particular sputum chemokines and cytokines in individual patients may provide a rationale for specific therapies.

Background: Rhinovirus infection is a major cause of asthma exacerbations. While rhinovirus infection is known to generate kinins in the upper respiratory track, little is known about the effect of rhinovirus on kinin generation in the lower airway. We previously identified human tissue kallikrein (hTK) as the principal lung kininogenase during allergic airway inflammation. In this report we investigate the effect of experimental rhinovirus infection on hTK activity in the airways of atopic subjects with and without asthma., Methods: Eight atopic subjects, 4 with asthma, underwent bronchoscopy with lavage. At least 1 month later, subjects were inoculated with rhinovirus, then underwent repeat bronchoscopy with lavage 4 and 18 days later. hTK mRNA was measured in nasal scrape samples by quantitative real-time PCR. hTK activity (chromogenic substrate assay) and IL-8 levels (ELISA) were assessed in the bronchoalveolar lavage fluid., Results: At day 4 after rhinovirus inoculation, nasal hTK mRNA was modestly increased in both the rhinitis (1.7-fold) and asthmatic (2.1-fold) groups. A doubling or greater increase in hTK activity after rhinovirus infection was observed in all 4 asthmatic subjects (mean 19-fold increase) but only in 1 of 4 atopic subjects without asthma (mean 2-fold increase). Rhinovirus infection also increased the IL-8 protein level in bronchoalveolar lavage fluid, which correlated with hTK activity (R = 0.82)., Conclusion: Experimental rhinovirus infection in allergic asthmatic subjects is accompanied by increased lower airway hTK activation, which parallels the appearance of IL-8. Rhinovirus-induced hTK activation may contribute to airway inflammation and asthmatic exacerbations., (2008 S. Karger AG, Basel.)

Homogeneity analysis was performed on four distinctive commercial lots, derived from the 2006 rice harvest in the United States. Lots that had previously been tested and suspected to have some level of LL601 were selected to determine lot homogeneity. LL601 infiltration in the lots was low and estimated to contain <0.01% (sigma = 0.026), 0.014% (sigma = 0.020), 0.054% (sigma = 0.043), and 0.074% (sigma = 0.031) LL601. Lots were analyzed statistically as a one-way classification, or one-factor experiment, to assess the presence of strata within the lot. A p value of 0.05 or lower is needed to declare statistical significance and would suggest significant differences among the samples. The data revealed p values ranging between 0.105 and 0.607. The calculated p values for all lots were greater than the critical value of 0.05. Samples taken from different locations throughout these four commercial lots did not show statistically significant stratifications within the lot.

Infectious cercariae of Schistosoma mansoni gain entry to the mammalian host through the skin where they induce a transient inflammatory influx of mononuclear cells. Some of these cells have antigen-presenting cell function (MHCII+) and have been reported to migrate to the skin-draining lymph nodes (sdLN) where they have the potential to prime CD4+ cells of the acquired immune response. Here, in mice exposed to vaccinating radiation-attenuated schistosome larvae, which induce high levels of protective immunity to challenge infection, we describe the parasite-induced migration of Langerhans cells (LCs) from the epidermal site of immunisation to the sdLN using a specific monoclonal antibody that recognises langerin (CD207). CD207+ cells with dendritic morphology were abundant in the epidermis at all times and their migration into the dermis was detected soon after vaccination. All CD207+ LCs were MHCII+ but not all MHCII+ cells in the skin were CD207+. LCs migrated from the dermis in enhanced numbers after vaccination, as detected in dermal exudate populations recovered after in vitro culture of skin biopsies. Elevated numbers of CD207+ LCs were also detected in the sdLN from 24h to 4 days after vaccination. However, compared with other dermal-derived antigen-presenting cells that were CD207-MHCII+ or CD207-CD11c+, the relative numbers of CD207+ cells in the dermal exudate population and in the sdLN were very small. Furthermore, the migration of CD207+ cells after exposure to 'protective' radiation-attenuated, compared with 'non-protective' normal cercariae, was similar in terms of numbers and kinetics. Together, these studies suggest that CD207+ LCs are only a minor component of the antigen-presenting cell population that migrates from the epidermis and they are unlikely to be important in the priming of protective CD4+ cells in the sdLN.

The mechanisms through which Schistosoma mansoni larvae induce Th1 rather than Th2 immune responses are not well understood. In this study, using CD154-/- mice exposed to radiation-attenuated S. mansoni larvae, we demonstrate roles for CD154/CD40 in the activation of skin-derived APCs and the development of Th1 cells in the skin-draining lymph nodes (sdLN). The presence of CD154 was important for optimal IL-12p40 and essential for Ag-specific IFN-gamma, but CD154 expression by wild-type CD4- cells was insufficient to rescue recall responses of CD4+ cells from CD154-/- mice. This defect is probably due to impaired CD40-dependent IL-12 production in vivo, because administration of anti-CD40 Ab, or rIL-12, restored IFN-gamma production by sdLN cells from CD154-/- mice. CD154 ligation of CD40 was not required for the migration of skin-derived APCs, but did have a limited role in their maturation (increased MHC II and CD86). Unexpectedly, although CD4 cells from CD154-/- mice were deficient in their ability to produce IFN-gamma, they produced significant amounts of IL-4 and IL-5 in the presence of skin-derived APCs from wild-type and CD154-/- mice. Thus, in contrast to IFN-gamma, the production of Th2-associated cytokines is (in this model) independent of CD154. We conclude that whereas the priming of Th1 responses soon after exposure to schistosome larvae is completely CD40/CD154 dependent, IL-4, IL-5, and IL-13 are independent of CD154, suggesting a dichotomy in the specific mechanisms that induce these cytokines by CD4+ cells in the sdLN.

Immunoassays for biotechnology engineered proteins are used by AgBiotech companies at numerous points in product development and by feed and food suppliers for compliance and contractual purposes. Although AgBiotech companies use the technology during product development and seed production, other stakeholders from the food and feed supply chains, such as commodity, food, and feed companies, as well as third-party diagnostic testing companies, also rely on immunoassays for a number of purposes. The primary use of immunoassays is to verify the presence or absence of genetically modified (GM) material in a product or to quantify the amount of GM material present in a product. This article describes the fundamental elements of GM analysis using immunoassays and especially its application to the testing of grains. The 2 most commonly used formats are lateral flow devices (LFD) and plate-based enzyme-linked immunosorbent assays (ELISA). The main applications of both formats are discussed in general, and the benefits and drawbacks are discussed in detail. The document highlights the many areas to which attention must be paid in order to produce reliable test results. These include sample preparation, method validation, choice of appropriate reference materials, and biological and instrumental sources of error. The article also discusses issues related to the analysis of different matrixes and the effects they may have on the accuracy of the immunoassays.

Schistosomes appear to have evolved several strategies to down-regulate the host's immune response in order to promote their own survival. For the host, down-regulation is also beneficial as it can limit the extent of pathology. It is widely accepted that schistosomes modulate the immune response during the chronic phase of infection after egg deposition has started. However, there is increasing evidence that modulation of the immune response can occur much earlier at the time infective cercariae penetrate the host skin. In this review, we explore the various lines of evidence that excretory/secretory (ES) molecules from cercariae down-regulate the host's immune response. We highlight the immunological factors that are produced and may be involved in regulating the immune system (e.g. IL-10, and eicosanoids), as well as speculating on possible mechanisms of immune modulation (e.g. mast-cell activation, T-cell apoptosis, and/or the skewed activation of antigen-presenting cells [APCs]). Finally, we draw attention to several molecules of schistosome origin that have the potential to stimulate the regulatory response (e.g. glycans) and link these to potential host receptors (e.g. TLRs and C-type lectins).

Sensitive and accurate testing for trace amounts of biotechnology-derived DNA from plant material requires pure, high-quality genomic DNA as template for subsequent amplification using the polymerase chain reaction (PCR). Six methodologies were evaluated for extracting DNA from ground corn kernels spiked with 0.1% (m/m) CBH351 (StarLink) corn. DNA preparations were evaluated for purity and fragment size. Extraction efficiency was determined. The alcohol dehydrogenase gene (adh1) and the CBH351 (cry9C, 35S promoter) genes in the genomic DNA were detected using PCR. DNA isolated by two of the methods proved unsuitable for performing PCR amplification. All other methods produced some DNA preparations that gave false negative PCR results. We observed that cornstarch, a primary component of corn kernels, was not an inhibitor of PCR, while acidic polysaccharides were. Our data suggest that amplification of an endogenous positive control gene, as an indicator for the absence of PCR inhibitors, is not always valid. This study points out aspects of DNA isolation that need to be considered when choosing a method for a particular plant/tissue type.

Background: T cells of intraepithelial phenotype have previously been detected in bronchoalveolar lavage (BAL) fluid in a range of lung diseases; these cells express the adhesion molecule alpha(E)beta(7) integrin, CD103, the ligand for epithelial cell E-cadherin. In subjects with asthma CD4+ lymphocytes are the predominant T cell subtype found in bronchial biopsy specimens and in BAL fluid, whereas CD8+ lymphocytes have been shown to predominate in subjects with chronic obstructive pulmonary disease (COPD). The aim of this study was to analyse the expression of CD103, activation markers (CD25 and CD69), and chemokine receptors (CXCR3, CCR5 and CCR3) on CD4+ and CD8+ lymphocytes from sputum and peripheral blood of subjects with asthma, COPD, and healthy controls., Methods: T cell surface markers were assessed by immunofluorescence labelling and flow cytometry of gated lymphocytes among CD45+ leucocytes in sputum cell suspensions., Results: Sputum lymphocytes expressed higher levels of CD103 and CD69 than blood lymphocytes in all subject groups, with CD103 expressed at higher levels on CD8+ than on CD4+ cells. There were no detectable differences in numbers of CD4+ and CD8+ T cells between subjects with asthma, COPD and controls. The percentage of sputum lymphocytes expressing CXCR3 was lower in subjects with asthma or COPD than in healthy controls; CCR3 was not detectable on sputum or blood lymphocytes., Conclusions: Sputum T lymphocytes are predominantly of activated intraepithelial phenotype (CD103+ CD69+), and normal numbers of CD4+ and CD8+ T cell populations are found in the sputum of patients with asthma and COPD.

Background: Increased levels of bradykinin and IL-8 have been detected within the airways of individuals with active symptoms of allergic rhinitis and asthma., Objective: We sought to investigate the in vivo effect of bradykinin on the expression of the IL-8 receptors CXCR1 and CXCR2 in nasal cells., Methods: Nasal samples were obtained from patients with active allergic rhinitis; patients with mild, quiescent allergic rhinitis; and healthy control subjects. CXCR1 and CXCR2 mRNA expression in the nasal cells was measured by means of quantitative real-time RT-PCR in baseline samples from all subjects, as well as in samples obtained after in vivo bradykinin challenge in healthy control subjects and patients with mild allergic rhinitis. CXCR1 and CXCR2 cell-surface expression was also assessed by means of flow cytometry in nasal epithelial cells at baseline and after ex vivo bradykinin challenge., Results: No difference was seen in CXCR1 or CXCR2 mRNA expression between healthy control subjects and patients with quiescent allergic rhinitis at baseline; however, patients with active allergic rhinitis had increased baseline expression of both CXCR1 and CXCR2 mRNA. In vivo nasal bradykinin challenge significantly increased CXCR1 and CXCR2 mRNA expression in patients with quiescent allergic rhinitis but had no effect in healthy control subjects. Low levels of CXCR1 but not CXCR2 cell-surface expression was detected in nasal epithelial cells at baseline, and ex vivo bradykinin challenge induced CXCR2 cell-surface expression in nasal epithelial cells from patients with mild allergic rhinitis., Conclusion: This study demonstrates the in vivo regulation of chemokine receptors by means of bradykinin in human airway tissue in patients with allergic rhinitis.

Plasmin is the major enzyme that dissolves fibrin in the vasculature and the predominant source of its zymogen, plasminogen, is liver. However, plasmin has a broad substrate spectrum and, if present in other tissues, may perform additional functions. We tested the hypothesis that plasminogen is expressed broadly extrahepatically. A sensitive and specific isotopic quantitative RT-PCR assay was developed to detect plasminogen mRNA from total RNA isolated from C57BL/6J mice tissues. Plasminogen mRNA was detected in adrenal, kidney, brain, testis, heart, lung, uterus, spleen, thymus and gut. Of these tissues, adrenal had the highest plasminogen mRNA content. In situ hybridization was utilized to localize plasminogen mRNA expressing cell types. Besides hepatocytes, positive cells were identified in both adrenal and kidney medullae and cortexes. Plasminogen mRNA expression was detected in cerebral, hippocampal and cerebellar neurons. Plasminogen mRNA was detected in cells in early stages of spermatogenesis in testis, present in the cortex and medulla of the thymus and in splenic white and red pulps. Our results suggest that the plasminogen gene is expressed broadly in extrahepatic tissues. Thus, tissues separated by local anatomic barriers as well as tissues accessible to circulating plasminogen have the capacity to provide local sources of plasminogen.

Fumonisin B1, a fungal mycotoxin that grows on corn and other agricultural products, alters sphingolipid metabolism by inhibiting ceramide synthase. The precise mechanism of fumonisin B1 toxicity has not been completely elucidated; however, a central feature in the cytotoxicity is alteration of sphingolipid metabolism through interruption of de novo ceramide synthesis. An affinity column consisting of fumonisin B1 covalently bound to an HPLC column matrix was used to isolate a rat liver protein that consistently bound to the column. The protein was identified as argininosuccinate synthetase by protein sequencing. The enzyme-catalyzed formation of argininosuccinic acid from citrulline and aspartate by recombinant human and rat liver argininosuccinate synthetase was inhibited by fumonisin B1. Fumonisin B1 showed mixed inhibition against citrulline, aspartate, and ATP to the enzyme. Fumonisin B1 had a Ki' of approximately 6 mM with the recombinant human argininosuccinate synthase and a Ki' of 35 mM with a crude preparation of enzyme prepared from rat liver. Neither tricarballylic acid nor hydrolyzed fumonisin B1 inhibited recombinant human argininosuccinate synthetase. This is the first demonstration of fumonisin B1 inhibition of argininosuccinate synthethase, a urea cycle enzyme, which adds to the list of enzymes that are inhibited in vitro by fumonisin B1 (ceramide synthase, protein serine/threonine phosphatase). The extent of the inhibition of argininosuccinate synthetase in cells, and the possible role of this enzyme inhibition in the cellular toxicity of FB1, remains to be established.

Fumonisin B1 stimulates apoptosis in a variety of cell types and tissues. We examined the role of sphingolipid changes in fumonisin B1-stimulated apoptosis. Sphinganine accumulated rapidly, sphingosine levels remained unchanged, and ceramides decreased during fumonisin B1 exposure. Increased DNA fragmentation, decreased viability, and apoptotic morphology were observed in cells exposed to fumonisin B1, sphinganine, or N-acetylsphingosine. Co-exposure to N-acetylsphingosine or beta-chloroalanine, which blocks sphinganine accumulation, partially protected cells from fumonisin B1-induced apoptosis. These results illustrate three sphingolipid-dependent mechanisms for inducing apoptosis: accumulation of excess ceramide, accumulation of excess sphinganine, and depletion of ceramide or complex sphingolipids derived from ceramide.