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In a context of international concern about early adult mental health service provision, this study identifies characteristics and service outcomes of young people with attention-deficit hyperactivity disorder (ADHD) reaching the child and adolescent mental health service (CAMHS) transition boundary (TB) in Ireland. The iTRACK study invited all 60 CAMHS teams in Ireland to participate; 8 teams retrospectively identified clinical case files for 62 eligible young people reaching the CAMHS TB in all 4 Health Service Executive Regions. A secondary case note analysis identified characteristics, co-morbidities, referral and service outcomes for iTRACK cases with ADHD (n = 20). Two-thirds of young people with ADHD were on psychotropic medication and half had mental health co-morbidities, yet none was directly transferred to public adult mental health services (AMHS) at the TB. Nearly half were retained in CAMHS, for an average of over a year; most either disengaged from services (40%) and/or actively refused transfer to AMHS (35%) at or after the TB. There was a perception by CAMHS clinicians that adult services did not accept ADHD cases or lacked relevant service/expertise. Despite high rates of medication use and co-morbid mental health difficulties, there appears to be a complete absence of referral to publicly available AMHS for ADHD youth transitioning from CAMHS in Ireland. More understanding of obstacles and optimum service configuration is essential to ensure that care is both available and accessible to young people with ADHD., (© 2017 John Wiley & Sons Australia, Ltd.)

Objective: The primary objectives of this study were to evaluate the pharmacokinetics (PK) and tolerability of single and multiple doses of vortioxetine in children and adolescents with a depressive or anxiety disorder and to provide supportive information for appropriate dosing regimens for pediatric clinical trials., Methods: This prospective, open-label, multinational, multisite, multiple-dose trial enrolled 48 patients (children and adolescents; 1:1 ratio) divided into 8 cohorts (4 adolescent and 4 child), with each cohort including 6 patients. The cohorts in each age group were assigned to receive one of four dosing regimens: vortioxetine 5, 10, 15, or 20 mg q.d. for 14 days. The total treatment period lasted 14-20 days with patients in the higher dose cohorts uptitrated over 2-6 days. Plasma samples for PK analysis were obtained on the first and last days of dosing., Results: Among children and adolescents, respectively, 62% and 92% had depression and 58% and 33% had anxiety disorder. Comorbid attention-deficit/hyperactivity disorder (ADHD) was present in 50% of children and 38% of adolescents. After 14 days q.d. at the target dose, the PK of vortioxetine concentrations was generally proportional to the dose in both age groups. Exposure, as assessed by maximum plasma concentrations and area under the plasma concentration-time curve from time 0 to 24 hours, was 30%-40% lower in adolescents than in children. There was no significant relationship between sex, height, or ADHD diagnosis and PK parameters. Most adverse events were mild in severity and consistent with those seen in adults., Conclusion: The results suggest that the dosages of vortioxetine evaluated (5-20 mg q.d.; approved for treatment in adults) and the uptitration schedule used are appropriate for pediatric efficacy and safety trials.

Retention of a positively valued illness identity contributes to poor outcomes for individuals with eating disorders (EDs). Consequently, dis-identification from the illness identity and the adoption of a recovery identity are vital for successful recovery. While social identity processes have been shown to influence ED maintenance, their role in recovery is rarely considered. This study explores how a sense of shared identity helps individuals with EDs manage their condition and promotes recovery. Transcripts from 18 online support sessions involving 75 participants were thematically analysed. Our findings suggest that the illness identity initially operates as a social identity that forms the basis for connections with similar others. For those wishing to recover, identity-based support is then perceived to be more effective than that found outside the group. Online interactions also facilitate construction of a new shared recovery identity which promotes a shift from the illness identity as a primary source of definition and endorses group norms of illness disclosure and treatment engagement. While in the clinical literature, ED identity is seen as problematic and interventions are targeted at challenging an individual's self-concept, we suggest that interventions could instead harness identity resources to support a transition to a recovery identity., (© 2016 The British Psychological Society.)

Introduction: A significant barrier to recovery for individuals with co-morbid eating disorders and type 1 diabetes is the way in which group members self-categorise. Nonetheless, identity issues are neglected during the recovery process. The aim of this paper is to explore how group memberships (and the associated identities) both contribute to and hinder recovery in this cohort., Method: Transcripts from five online focus groups with 13 members of an online support group for individuals with 'Diabulimia' were thematically analysed., Results: Findings suggested that those with whom one shares a recovery identity can be well placed to provide psychological resources necessary for successful recovery although such connections can be damaging if group norms are not managed. Members recognised that other important relationships (including family and friends and health professionals) are also key to recovery; these other group memberships (and the associated identities) can be facilitated through the recovery identity group membership, which allows for external validation of the recovery identity, provides encouragement to disclose the illness to supportive others, and provides information to facilitate positive service interactions., Conclusions: While clinical interventions typically focus on eliminating disordered behaviours, we suggest that these should also include strengthening important group memberships that promote recovery.

There is increasing acceptance that children are not unaware of when they are targets of discrimination. However, discrimination as a consequence of socio-economic disadvantage remains understudied. The aim of this study was to examine the impact of perceived discrimination on well-being, perceptions of safety and school integration amongst children growing up within socio-economically disadvantaged communities in Limerick, Ireland. Mediation analysis was used to explore these relationships and to examine the potential role of parental support and community identity in boys and girls in the 6th to 9th year of compulsory education (N = 199). Results indicate perceived discrimination contributed to negative outcomes in terms of school integration, perceptions of safety and levels of well-being. Age and gender differences were observed which disadvantaged boys and younger children. All negative outcomes were buffered by parental support. Community identity also protected young people in terms of feelings of school integration and risk but not in terms of psychological well-being. Findings are discussed in terms of the different role of family and community supports for children negotiating negative social representations of their community., (© 2015 The British Psychological Society.)

Objectives: This study examines aspects of healthcare professionals' knowledge and attitudes about eating disorders (EDs), which might impede the effective detection or treatment of EDs in Ireland., Methods: A total of 1,916 healthcare professionals were invited to participate in a web-based survey. Participants were randomly allocated to view one of five vignettes depicting a young person with symptoms consistent with anorexia nervosa, bulimia nervosa, binge-eating disorder, depression or type 1 diabetes. Study-specific questions examined participants' responses to the vignettes and ED knowledge and experience., Results: In total, 171 clinicians responded (9% response rate). Participants had an average of 15.8 years of clinical experience (s.d.=9.2) and included psychiatrists, GPs, psychologists and counsellors. Although participants' knowledge of EDs was moderately good overall, responses showed poor recognition of the symptoms of EDs compared with depression [χ 2 (4, n=127)=20.17, p<0.001]. Participants viewed EDs as chronic disorders that primarily affected females. Participants believed that clinicians like working with patients with depression and diabetes more than with AN patients [F (4,101)=5.11, p=0.001]. Among the professionals surveyed, psychiatrists were the most knowledgeable about EDs [F (4,82)=9.18, p<0.001], and were more confident in their ability to diagnose and treat EDs than professionals of all other disciplines, except psychologists [F (4,85)=8.99, p<0.001]. Psychiatrists were also the most pessimistic about ED patients' long-term life prospects [χ 2 (4, n=65)=15.84, p=0.003]., Conclusions: This study recommends that specific attention should be given to EDs in professional educational programmes across healthcare disciplines. This training should not be restricted to improving healthcare professionals' knowledge of EDs, but should also strive to increase service-providers' awareness of how their own potentially stigmatising attitudes can undermine engagement with treatment.

Service user involvement is essential for quality care in the Child and Adolescent Mental Health Services (CAMHS). This study was conducted to explore adolescents' and parents' experiences of CAMHS in relation to accessibility, approachability and appropriateness. This study used a descriptive qualitative design, and focus groups and single interviews were conducted with adolescents (n = 15) and parents (n = 32) from three mental health clinics. Data were analysed using thematic analysis. Accessing mental health services was a challenging experience for many parents and adolescents due to knowledge deficit, lack of information and limited availability of specialist services. Some parents and adolescents reported positive experiences while others reported negative experiences. They expressed a need for more information, involvement in decision making, flexible scheduling of appointments, school support and parent support groups. The nature and quality of the relationship with staff was critical to positive experience with the service; therefore, frequent changes of medical staff was disruptive. Mental health nurses can play a vital role in ensuring continuity, assessing adolescents' participation preferences and advocating for their information needs with other members of the interdisciplinary team., (© 2015 John Wiley & Sons Ltd.)

Aggressive behaviors can be classified into proactive and reactive functions, though there is disagreement about whether these are distinct constructs. Data suggest that proactive and reactive aggression have different etiologies, correlates, and response to treatment. Several rating scales are available to characterize aggressive behavior as proactive or reactive; one commonly used scale was originally developed for teacher ratings, referred to here as the Antisocial Behavior Scale (ABS). However, no data are available on the psychometric properties of the ABS for parent ratings. This study examined the factor structure and convergent/divergent validity of the parent-rated ABS among 168 children aged 6-12 years with attention-deficit hyperactivity disorder, a disruptive behavior disorder, and severe aggression enrolled in a randomized clinical trial. Multidimensional item response theory was used to confirm the original factor structure. The proactive and reactive factors were distinct but moderately correlated; the algorithm items exhibited acceptable fit on the original factors. The non-algorithm items caused theoretical problems and model misfit. Convergent and divergent validity of the scale was explored between the ABS and other parent-report measures. Proactive and reactive aggression showed differential correlates consistent with expectations for externalizing symptoms. The subscales were correlated weakly or not at all with most non-externalizing symptoms, with some exceptions. Thus, the original factor structure was supported and we found preliminary evidence for the validity of the scale, though the results suggest that the constructs measured by the ABS may not be totally distinct from general behavior problems in this clinical sample.

Educating undergraduate nurses in the 21st century provides some very realistic challenges. Decreased government health budgets, increased student numbers and higher patient acuities have resulted in a reduction in the availability and quality of clinical placements. Simulated nursing practice is an innovative strategy designed to address these concerns. A simulation programme was designed for first year undergraduate nursing students to help prepare them for clinical placement. The aim of this research is to assess student perspectives and learning from the newly introduced simulation programme. This study is a descriptive design with Kolb's experiential theory providing a theoretical framework. 158 first year students taking part in a four day simulation programme chose to complete a questionnaire on programme completion. Students responded to five statements using a likert scale and categories developed and refined for the remaining four questions. Students reported significant learning in the areas of basic clinical skills and clinical documentation and collaborative care. 100% of students recommended the programme continue., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

Background/aims: To determine the ophthalmic manifestations of HIV in a cohort of long-term survivors of perinatally acquired HIV., Methods: Twenty-two patients with perinatally acquired HIV who were aged ≥12 years were prospectively studied at a university clinic. They underwent complete ophthalmic examinations and fundus photography. Their medical histories, medications and CD4 counts were abstracted from the medical records. To evaluate for keratoconjunctivitis sicca, both HIV patients and 44 healthy controls (matched by age, gender and contact lens wear) underwent Schirmer testing and ocular surface staining., Results: Nine male and 13 female HIV patients with mean age of 16.6 years (SD, 3.4) were examined. Of the 22 HIV patients, 21 had been treated with highly active antiretroviral therapy (HAART). Only one patient had a CD4 count nadir of <200 cells/µL. The mean visual acuity of the eyes of the HIV subjects was 20/22 (SD, 1.6 lines). No patient had cytomegalovirus retinitis. Four of the 22 (18%) HIV patients had strabismus. HIV subjects and controls had similar rates of abnormal Schirmer (9% and 14%, p=0.62) and ocular staining scores (p=0.29)., Conclusions: In the post-HAART era, long-term survivors of perinatally acquired HIV exhibited little vision-threatening disease, but had a high prevalence of strabismus., (Published by the BMJ Publishing Group Limited. For permission to use (where not already granted under a licence) please go to http://group.bmj.com/group/rights-licensing/permissions.)

Objective: In this study, we evaluated parent and child characteristics as predictors and moderators of response in the four-site Treatment of Severe Childhood Aggression (TOSCA) study., Methods: A total of 168 children with severe aggression, disruptive behavior disorder, and attention-deficit/hyperactivity disorder (ADHD) were enrolled in a 9-week trial of basic treatment (n=84, stimulant+parent training+placebo) versus augmented treatment (n=84, stimulant+parent training+risperidone). In the initial report, augmented treatment surpassed basic treatment in reducing the primary outcome of disruptive behavior (D-Total) scores. In the current study, we evaluated parent (income, education, family functioning, employment) and child variables (intelligence quotient [IQ], aggression type, comorbid symptomatology) as predictors or moderators, using linear mixed models and the MacArthur guidelines., Results: Higher scores on ADHD symptom severity and callous/unemotional traits predicted better outcome on D-Total regardless of treatment assignment. Two moderators of D-Total were found: Higher anger/irritability symptoms and lower mania scores were associated with faster response, although not better overall effect at endpoint, in the augmented but not the basic group. Several variables moderated response on secondary outcomes (ADHD severity and prosocial behavior), and were characterized by faster response, although not better outcome, in the augmented but not in the basic group. Maternal education moderated outcome on the measure of positive social behavior; children of mothers with less education benefited more from augmented treatment relative to basic than those with more education., Conclusion: Although these findings require validation, they tentatively suggest that augmented treatment works equally well across the entire sample. Nevertheless, certain child characteristics may be useful indicators for the speed of response to augmented treatment.

Objective: The ITRACK study explored the process and predictors of transition between Child and Adolescent Mental Health Services (CAMHS) and Adult Mental Health Services (AMHS) in the Republic of Ireland., Method: Following ethical approval, clinicians in each of Ireland's four Health Service Executive (HSE) areas were contacted, informed about the study and were invited to participate. Clinicians identified all cases who had reached the transition boundary (i.e. upper age limit for that CAMHS team) between January and December 2010. Data were collected on clinical and socio-demographic details and factors that informed the decision to refer or not refer to the AMHS, and case notes were scrutinised to ascertain the extent of information exchanged between services during transition., Results: A total of 62 service users were identified as having crossed the transition boundary from nine CAMHS [HSE Dublin Mid-Leinster (n=40, 66%), HSE South (n=18, 30%), HSE West (n=2, 3%), HSE Dublin North (n=1, 2%)]. The most common diagnoses were attention deficit hyperactivity disorder (ADHD; n=19, 32%), mood disorders (n=16, 27%), psychosis (n=6, 10%) and eating disorders (n=5, 8%). Forty-seven (76%) of those identified were perceived by the CAMHS clinician to have an 'on-going mental health service need', and of these 15 (32%) were referred, 11 (23%) young people refused and 21 (45%) were not referred, with the majority (12, 57%) continuing with the CAMHS for more than a year beyond the transition boundary. Young people with psychosis were more likely to be referred [χ 2 (2, 46)=8.96, p=0.02], and those with ADHD were less likely to be referred [χ 2 (2, 45)=8.89, p=0.01]. Being prescribed medication was not associated with referral [χ 2 (2, 45)=4.515, p=0.11]. In referred cases (n=15), there was documented evidence of consent in two cases (13.3%), inferred in another four (26.7%) and documented preparation for transition in eight (53.3%). Excellent written communication (100%) was not supported by face-to-face planning meetings (n=2, 13.3%), joint appointments (n=1, 6.7%) or telephone conversations (n=1, 6.7%) between corresponding clinicians., Conclusions: Despite perceived on-going mental health (MH) service need, many young people are not being referred or are refusing referral to the AMHS, with those with ADHD being the most affected. CAMHS continue to offer on-going care past the transition boundary, which has resource implications. Further qualitative research is warranted to understand, in spite of perceived MH service need, the reason for non-referral by the CAMHS clinicians and refusal by the young person.

Aim: Ensuring a seamless transition from child to adult mental health services poses challenges for services worldwide. This is an important process in the ongoing care of young people with mental illness; therefore, it is incumbent on all countries to probe their individual structures to assess the quality of mental health service delivery to this vulnerable cohort. To date, there have been no published studies on the transition from Child to Adult Mental Health Services in the Republic of Ireland. To this end, a nationwide survey of transition policies of community mental health teams in both services was conducted in order to compare best practice guidelines for transition with current process and experience in clinical practice., Method: Structured interviews were conducted with 57 consultant psychiatrists (representing 32 CAMHS teams and 25 AMHS teams) to obtain information on annual transition numbers, existing transition policies and operational practice from the professional perspective., Results: Numbers of young people considered suitable for transfer to adult services (M = 7.73, SD = 9.86, n = 25) were slightly higher than numbers who actually transferred (M = 4.50, SD = 3.33, n = 20). There is a lack of standardized practice nationwide regarding the service transition boundary, an absence of written transition policies and protocols, and minimal formal interaction between child and adult services., Conclusions: The findings suggest that there are critical gaps between current operational practice and best practice guidelines. Future studies will investigate the impact this has on the transition experiences of young people, their carers and health-care professionals., (© 2013 Wiley Publishing Asia Pty Ltd.)

: Clinician-scientists bridge the gap between basic research and patient care. At the 2012 Annual Meeting, a symposium highlighting the application of cutting-edge optometric research within the anterior segment was held to present and discuss some of the recent basic scientific advances that will both shape and guide the development of future clinical care practices. This article summarizes this work, bringing together four experts, all clinician-scientists in the field of cornea and ocular surface. Collectively, this work provides new insights to clinicians and researchers alike, as well as brings forth a greater appreciation of the impact of ongoing optometric bench research in advancing clinical care.

Purpose: Dry eye is commonly associated with autoimmune diseases such as Sjögren's syndrome (SS), in which exocrinopathy of the lacrimal gland leads to aqueous tear deficiency and keratoconjunctivitis sicca (KCS). KCS is among the most common and debilitating clinical manifestations of SS that is often recalcitrant to therapy. We established mice deficient in the autoimmune regulator (Aire) gene as a model for autoimmune-mediated aqueous-deficient dry eye. In Aire-deficient mice, CD4+ T cells represent the main effector cells and local signaling via the interleukin-1 (IL-1/IL-1R1) pathway provides an essential link between autoreactive CD4+ T cells and ocular surface disease. In the current study, we evaluated the efficacy of topical administration of IL-1R1 antagonist (IL-1RA) anakinra in alleviating ocular surface damage resulting from aqueous-deficient dry eye in the setting of autoimmune disease., Methods: We compared the effect of commercially available IL-1R1 antagonist, anakinra (50 μg/mL concentration) to that of carboxymethylcellulose (CMC) vehicle control as a treatment for dry eye. Age-matched, Aire-deficient mice were treated three times daily with anakinra or CMC vehicle for 14 days using side-by-side (n = 4 mice/group) and paired-eye (n = 5) comparisons. We assessed (1) ocular surface damage with lissamine green staining; (2) tear secretion with wetting of phenol-red threads; (3) goblet cell (GC) mucin glycosylation with lectin histochemistry; (4) immune cell infiltration using anti-F4/80, CD11c, and CD4 T cell antibodies; and (5) gene expression of cornified envelope protein, Small Proline-Rich Protein-1B (SPRR1B) with real-time quantitative polymerase chain reaction., Results: Aire-deficient mice treated with anakinra experienced significant improvements in ocular surface integrity and tear secretion. After 7 days of treatment, lissamine green staining decreased in eyes treated with anakinra compared to an equivalent increase in staining following treatment with CMC vehicle alone. By day 14, lissamine green staining in anakinra-treated eyes remained stable while eyes treated with CMC vehicle continued to worsen. Accordingly, there was a progressive decline in tear secretion in eyes treated with the CMC vehicle compared to a progressive increase in the anakinra-treated eyes over the 2-week treatment period. Aberrant acidification of GC mucins and pathological keratinization of the ocular surface were significantly reduced in anakinra-treated eyes. Significantly fewer Maackia amurensis leukoagglutinin positive goblet cells were noted in the conjunctiva of anakinra-treated eyes with a corresponding decrease in the expression of the pathological keratinization marker, SPRR1B. Finally, there was a downward trend in the infiltration of each immune cell type following anakinra treatment, but the cell counts compared to eyes treated with the vehicle alone were not significantly different., Conclusions: IL-1R antagonist, anakinra, demonstrates therapeutic benefits as a topical treatment for aqueous-deficient dry eye in a spontaneous mouse model of autoimmune KCS that mimics the clinical characteristics of SS. Targeting the IL-1/IL-1R1 signaling pathway through topical administration of IL-1RA may provide a novel option to improve ocular surface integrity, increase tear secretion, and restore the normal glycosylation pattern of GC mucins in patients with SS.

Objectives: Attention deficit hyperactivity disorder (ADHD) persists into adulthood in ∼2/3 of patients, yet services for adults are lacking in Ireland. This may involve negative attitudes and beliefs as to the validity of ADHD or lack of knowledge and training in its treatment. The objectives of this study are to explore the views of Adult Psychiatrists regarding ADHD knowledge and the treatment options available and pursued in Ireland., Methods: A questionnaire was constructed based on the stated aims of the study, and was either posted, emailed or handed to 400 Consultants and Senior Registrars throughout the Republic of Ireland between February and December 2011. A total of 92 questionnaires were returned (23%); one was excluded from analysis due to insufficient information entered by the respondent., Results: Seventy-five per cent of respondents correctly estimated the prevalence rates of adult ADHD to be under 3%, but stated it is currently under-diagnosed (77%). Seventy-four per cent indicated that Adult ADHD should be a diagnostic category in the Diagnostic and Statistical Manual, 5th Edition (DSM V). Sixty-six per cent of respondents were willing to accept referrals of childhood ADHD for ongoing care and a similar number for new ADHD assessments (61%). Less than half (42%) surveyed had actually diagnosed ADHD and of these, only 33% felt confident in managing ADHD in their patients., Conclusions: Although there is a general willingness to offer services for new and existing ADHD cases and a recognition that Adult ADHD is valid and under-diagnosed, the low confidence levels when treating ADHD and the perception of under-diagnosis suggests a role for further training and links between child and adult services.

Schizophrenia is a complex mental disorder with unknown aetiology. Both candidate gene and genome-wide association (GWA) studies suggest that the human leukocyte antigen (HLA) system may play a part in development of the illness, but the causal HLA variant(s) remain(s) unclear. Previous studies showed that the DRB1*0101 and DRB1*13 alleles might be associated with a high risk of schizophrenia. Therefore, the present study was undertaken to test their association with the disease by genotyping seven DRB1-tagging single nucleotide polymorphisms (SNPs) in a British population. The results showed that, of the previously reported variants that were associated with schizophrenia, the DRB1*1303 allele was the only one marginally associated with a protective effect on the illness in our sample set (χ² = 4.138, P = 0.042, odds ratio (OR) = 0.42, 95 % confidence interval (CI) 0.27-0.66). Interestingly, a significant association was found for rs424232 (χ² = 9.404, P = 0.002, OR = 0.69, 95 % CI 0.54-0.88), which is a tag SNP for the DRB1*1303 allele and located near to the NOTCH4 gene that is a schizophrenia susceptibility locus confirmed by GWA studies. Analysis with the Haploview program demonstrated that rs424232 was in complete linkage disequilibrium with rs3130297 and rs3131296 present in the NOTCH4 locus. While we have failed to confirm association of the candidate alleles in the DRB1 gene with a high risk of schizophrenia, the present work suggests that the association signal detected in the HLA class II locus may extend a relatively long distance, and more work is needed in order to identify the true causal variants within this region or nearby.

This paper considers the impact of pubertal status and pubertal timing on disordered eating in Irish adolescents. 1190 boys and 1841 girls completed the Eating Attitudes Test-26, the Eating Disorder Inventory-III and self-report measures of pubertal status and pubertal timing. Regarding pubertal status, greater maturity in girls was associated with increased overall eating concerns, higher drive for thinness and higher levels of body dissatisfaction. In boys, greater maturity was associated with lower drive for thinness and lower body dissatisfaction. Regarding pubertal timing, early-maturing girls showed the most eating concerns, the highest drive for thinness, scored highest on bulimic symptoms and were the most dissatisfied with their bodies. In contrast, late-maturing boys had more bulimic symptoms and more dissatisfaction with their bodies than on-time peers. The findings suggest that puberty itself is a risk factor for disordered eating for girls rather than boys; however, pubertal timing is a risk factor for both., (Copyright © 2012 John Wiley & Sons, Ltd and Eating Disorders Association.)

Objective: We propose new classification criteria for Sjögren's syndrome (SS), which are needed considering the emergence of biologic agents as potential treatments and their associated comorbidity. These criteria target individuals with signs/symptoms suggestive of SS., Methods: Criteria are based on expert opinion elicited using the nominal group technique and analyses of data from the Sjögren's International Collaborative Clinical Alliance. Preliminary criteria validation included comparisons with classifications based on the American–European Consensus Group (AECG) criteria, a model-based “gold standard”obtained from latent class analysis (LCA) of data from a range of diagnostic tests, and a comparison with cases and controls collected from sources external to the population used for criteria development., Results: Validation results indicate high levels of sensitivity and specificity for the criteria. Case definition requires at least 2 of the following 3: 1) positive serum anti-SSA and/or anti-SSB or (positive rheumatoid factor and antinuclear antibody titer >1:320), 2) ocular staining score >3, or 3) presence of focal lymphocytic sialadenitis with a focus score >1 focus/4 mm2 in labial salivary gland biopsy samples. Observed agreement with the AECG criteria is high when these are applied using all objective tests. However, AECG classification based on allowable substitutions of symptoms for objective tests results in poor agreement with the proposed and LCA-derived classifications., Conclusion: These classification criteria developed from registry data collected using standardized measures are based on objective tests. Validation indicates improved classification performance relative to existing alternatives, making them more suitable for application in situations where misclassification may present health risks., (Copyright © 2012 by the American College of Rheumatology.)

The prevalence of congenital myopathies in the United States has not been examined. To address this, we determined the point prevalence of congenital myopathies in a well-defined pediatric population from Southeastern Michigan. The overall point prevalence was 1:26,000. Mutations in RYR1 were the most common cause of congenital myopathies at 1:90,000. Our data broadly agrees with estimates from previous European studies and provides the first estimate of the prevalence of congenital myopathies in the United States. Ann Neurol 2011;70:662-665., (Copyright © 2011 American Neurological Association.)

Schizophrenia is a severe psychotic illness with a heterogeneous presentation and a devastating impact on social and occupational function. Worldwide variations in schizophrenia incidence rates suggest that local conditions may modify disease risk. The human leukocyte antigen (HLA) region has been confirmed to be associated with schizophrenia by genome-wide association studies in populations across the world. While the presence of autoimmune processes in a subgroup of schizophrenia cases is contentious, the immune system could allow environmental exposures to lead to schizophrenia by generating improper immune response. To investigate this topic, we reviewed the current evidence of the relationship between schizophrenia and coeliac disease. Based on this review, we performed genetic analysis of the MYO9B gene and the IL-2/IL-21 locus by genotyping SNPs that have been previously associated with coeliac disease or schizophrenia in 223 families, 108 unrelated individuals with schizophrenia and 120 controls. Finding no evidence for association with these two loci in our study samples, we applied meta-analytic techniques to combine our findings with previous reports. This synthesis, in light of our review of previous reports, suggests a differing developmental trajectory for schizophrenia and coeliac disease. It is possible that these two conditions do not share any functional overlap., (Copyright © 2011 Wiley-Liss, Inc.)

There is a lack of clarity in the literature about what nursing research is or ought to be. There are ample guidelines and texts on how to do nursing research and on syllabus development and countless arguments on why nursing research is synonymous with qualitative approach. This suggests concerns within the profession that nursing research has been developing in a direction of its own that could have had very little influence on health care policy development. Some claim the opposite that nursing practice, in particular in North America, has been evidence-based for the past three decades. In this context, evidence-based is often interpreted as taking part in randomised clinical trials. In this paper, we use historical observations to identify areas of interest which include all issues related to education and training, expertise and working environment for future research.

Cigarette smoke (CS) accounts for nearly 90% of lung cancer deaths worldwide; however, an incomplete understanding of how CS initiates preneoplastic changes in the normal airway hinders early diagnosis. Short-term exposure to CS causes aberrant activation of epidermal growth factor receptor (EGFR) and canonical Wnt/beta-catenin signaling pathways in human bronchial epithelial (HBE) cells. We hypothesize that this response is elicited through the disruption of spatially segregated cell membrane proteins in the polarized airway epithelium. Using an in vitro model of highly differentiated HBE cells, we observed membrane characteristics consistent with the native airway, including the presence of a membrane mucin, MUC1, at the apical cell pole, beta-catenin at the apical-lateral membrane, and EGFR at the basolateral membrane. Following exposure to smoke, intercellular spaces enlarge and cilia disappear. This histopathology is accompanied by molecular events that include perinuclear trafficking of basolateral EGFR, EGFR phosphorylation, pEGFR-mediated phosphorylation of MUC1's cytoplasmic tail (CT), loss of E-cadherin/beta-catenin complexes at the adherens junctions (AJs), intracellular formation and nuclear shuffling of beta-catenin/MUC1-CT complexes, and, ultimately, up-regulation and nuclear localization of Wnt nuclear effector, Lef-1. In the presence of EGFR inhibitor, AG1478, CS-induced histopathology and molecular events were inhibited. These data point to EGFR as a portal through which CS mediates its damaging effects on AJ-mediated cell polarity and activation of canonical Wnt/beta-catenin signaling.

Plant-derived organic matter inputs are thought to be a key driver of soil bacterial community composition and associated soil processes. We sought to investigate the role of acid grassland vegetation on soil bacterial community structure by assessing bacterial diversity in combination with other soil variables in temporally and spatially distinct samples taken from a field-based plant removal experiment. Removal of aboveground vegetation resulted in reproducible differences in soil properties, soil respiration and bacterial diversity. Vegetated soils had significantly increased carbon and nitrogen concentrations and exhibited higher rates of respiration. Molecular analyses revealed that the soils were broadly dominated by Alphaproteobacterial and Acidobacterial lineages, with increased abundances of Alphaproteobacteria in vegetated soils and more Acidobacteria in bare soils. This field-based study contributes to a growing body of evidence documenting the effect of soil nutrient status on the relative abundances of dominant soil bacterial taxa, with Proteobacterial taxa dominating over Acidobacteria in soils exhibiting higher rates of C turnover. Furthermore, we highlight the role of aboveground vegetation in mediating this effect by demonstrating that plant removal can alter the relative abundances of dominant soil taxa with concomitant changes in soil CO(2)-C efflux.

Purpose: To use protein arrays to delineate the spectrum of angiogenic bioactive protein modulators that might be secreted and up-regulated by the corneal epithelium in response to killed bacterial products., Methods: Immortalized human corneal epithelial cells were grown in culture, serum starved, and exposed to heat-killed Pseudomonas aeruginosa in a dose-dependent manner. The resultant culture medium was screened by antibody arrays for 43 proteins that can modulate angiogenesis and immune and inflammatory processes. Parallel analysis was carried out on tears recovered in the open and closed eye phases (OTF and CTF) of the diurnal cycle., Results: Array analysis reveals that the immortalized cells constitutively secrete several proteins and up-regulate the secretion of IL-6, IL-8, and GRO in response to killed bacteria. Also evident was the emergence of a strong signal for GM-CSF and moderate/weak signals for MCP-1, MMP-9, Leptin, and INF gamma in a dose-dependent manner. Several of these proteins, including IL-6, IL-8, GRO, MMP-9, TIMP-1, and MCP-1, accumulate in the CTF. Other proteins are unique to tear fluid., Conclusions: Nine proteins were identified that are secreted by epithelium in response to killed bacteria that contribute to the innate and adaptive defense system through potentiating PMN and macrophage recruitment, activation, and opsonization in a cooperative manner. The vast majority of these proteins are angiogenic modulators, perhaps contributing to the imbalance between angiogenic and angiostatic processes and risk of corneal vascularization.

Purpose: Squamous metaplasia of the ocular surface epithelium in severe Sjögren syndrome (SS) dry eye has been implicated to be associated with chronic engagement of immune-mediated inflammation. While the detailed immunopathological mechanism underlying keratinization of the ocular muco-epithelium in this setting remains unclear, mice deficient in the autoimmune regulator gene (Aire) demonstrate SS-like pathological changes in the exocrine organs and ocular surface including squamous metaplasia. Using this murine model, we sought to determine the specific immune events that predict squamous metaplasia of the cornea in Aire deficiency., Methods: Lissamine green staining, goblet cell density, and corneal small proline-rich protein 1B (SPRR1B) were compared in Aire-sufficient and -deficient mice at 4, 8, and 16 weeks of age. Corneal, limbal and conjunctival infiltration of CD4(+) and CD8(+) T cells as well as CD11c(+) and MHC class II (I-A(d+)) dendritic cells (DCs) were examined at the same time points. Ordinary least squares regression was used to model SPRR1B's relationship with lissamine green staining, goblet cell density, and immune cell infiltration., Results: Lissamine green staining was present in Aire-deficient mice by four weeks of age and increased over time. Compared to Aire-sufficient controls, conjunctival goblet cell density (GCD) decreased and corneal SPRR1B increased in Aire-deficient mice with significant differences noted at both 8 and 16 weeks. Immune-mediated CD4(+) T cell infiltration of the conjunctiva and limbus peaked at eight weeks and then decreased. In contrast, corneal T cell infiltration continued to increase over time, reaching a maximum cell number at 16 weeks. CD11c(+) myeloid-derived DCs were found in the conjunctiva and limbus at all time points. As the mice aged, there was a notable increase in corneal CD11c(+) cell counts. Interestingly, the dynamic of activated MHC class II(+) DCs was nearly identical to that of CD4(+) T cells, peaking first in the limbus at eight weeks with maximum infiltration of the cornea by 16 weeks. Regression analysis showed that squamous metaplasia biomarker, SPRR1B, is strongly related to the lissamine green staining of the ocular surface. Corneal infiltration of activated DCs was most prognostic of corneal SPRR1B expression while the presence of precursor DCs, activated DCs, and CD4(+) T cells in the limbus were also significant predictors of SPRR1B., Conclusions: Aire-deficient mice represent a useful model to study Sjögren-like autoimmune-mediated ocular surface disease. Results of the current study suggest that squamous cell precursor protein, SPRR1B, provides an important readout to evaluate ocular surface damage and specific events related to immune-mediated inflammation. Results also define an appropriate time frame for interventional studies to develop more effective therapies for keratinizing ocular surface disease.

To evaluate the use of stationary phase protein array technology for tear analysis and to characterize the distribution of inflammatory mediators in normal and allergic tears in the open and closed eye states. Microcapillary tube collected Open (OTF) and closed eye tear fluid (CTF) samples from normals (N), from individuals with various active chronic ocular and other allergies (CA), as well as from an individual subsequent to unilateral induction of an acute allergic conjunctivitis were assayed using membrane arrays that were optimized to allow the detection of GM-CSF, ILs-1 alpha, 1 beta, 2-10, 12-13, INF gamma, MCP-1 and TNFalpha in clinical size samples. The protocol of a micro-well plate array specific for ILs-2, 4, 5, 8, 10, 12, 13, TNFalpha and INF gamma was modified to minimize the impact of tear matrix effects. This was used to carry out parallel analysis on selected samples. By optimizing the protocol as well as the composition of a membrane array it proved possible to significantly increase the signal-to-noise ratio and sensitivity of assay allowing for the detection of some inflammatory mediators into the sub-picogram range. This provided sufficient sensitivity to allow the assay of clinically obtainable size samples. Analysis revealed that OTF from most Ns contained a high level of IL-8 and faint signals if any for the other probed proteins. In contrast, OTF samples from most CA individuals with and without ocular symptoms exhibited to varying degrees detectable levels of most of the other probed entities. The difference between normal and pathological tears and the levels of signals became far more pronounced in the CTF compared to the OTF samples. Use of the micro-well plate assay kit without modification revealed two tear matrix effects that profoundly impact the ability to obtain meaningful ELISA data. Modifying the assay protocol reduces but does not eliminate these artifacts making it possible to approximate the concentration of many of the probed entities. The obtained data is consistent using both methodologies revealing elevated levels of IL-8 and other cytokines in approximately 60% of the OTF samples from the CA population. Other than a modest increase in IL-8, no change could be observed in the profile of OTF after induction of an acute allergic reaction.

Bacterial flagellin can interact with both Toll-like receptor 5 (TLR5) and the cell surface glycolipid, asialoGM1, to activate an innate immune response. The induction of mucin by flagellin in human lung epithelial cells (NCIH292) is dependent on asialoGM1 ligation, ATP receptor signaling, Ca2+ mobilization, and Erk1/2 activation. Conversely, the activation of NF-kappaB by flagellin is dependent on signaling through TLR5. These results prompted us to ask whether the flagellin-induced TLR5 signaling pathway was intersecting with or mutually independent of the nucleotide receptor pathway activated downstream of asialoGM1. Herein, we demonstrate that the release of ATP induced by flagellin is dependent on a Toll signaling cascade. Although Toll was able to activate NF-kappaB in the absence of extracellular ATP, Toll required ATP to activate Erk1/2. These results suggest interdependence between the asialoGM1 and TLR5 pathways and reveal a previously unsuspected role for autocrine extracellular ATP signaling in TLR signaling.

Mucus hypersecretion is a hallmark of asthma that contributes to airway obstruction. While the etiology is not well understood, hypersecretion has been linked to the presence of cytokines such as IL-4, IL-5, IL-9, and IL-13 in the inflamed airway. The presence of adenosine has also been noted in asthmatic airways, and adenosine-mediated signaling in mast cells has been implicated in the severe bronchoconstriction and inflammation prevalent in these patients (1, 2). Here we examine the possibility that adenosine also contributes to mucus hypersecretion by airway epithelial cells. Results in cultured airway epithelial cells showed that MUC2 mucin expression increased in response to adenosine. This appeared to be mediated by a pathway initiated at the adenosine A1 receptor that transduced signals through a Ca2+-activated Cl- channel and EGFR. That this signaling cascade is relevant to asthmatic hypersecretion was indicated by results showing that mucin induction by asthmatic tracheal aspirates was reduced by A1, CLCA1, and EGFR inhibitors. These results suggest that adenosine cooperates with inflammatory cytokines to stimulate mucin production in the asthmatic airway and supports the use of A1, CLCA1, and EGFR inhibitors in the treatment of asthma.

Recently developed 13CO2 pulse labelling and stable isotope probing (SIP) methods offer the potential to track 13C-labelled plant photosynthate into phylogenetic groups of microbial taxa in the rhizosphere, permitting an examination of the link between soil microbial diversity and carbon flow in situ. We tested the feasibility of this approach to detect functional differences in microbial communities utilising recently fixed plant photosynthate in moisture perturbed grassland turfs. Specifically, we addressed two questions: (1) How does moisture perturbation (three treatments; continual wetting, drying, and drying followed by rewetting) affect the assimilation of 13C-labelled exudates carbon into the soil microbial community?; (2) Can 13C deposited in soil from pulse-labelled plants be used to identify microbes utilising plant exudates using SIP methodologies? Net CO2 fluxes showed that prior to 13CO2 pulse labelling, all treatments were photosynthetically active, but differences were observed in night time respiration, indicating moisture treatments had impacted on net CO2 efflux. Measurements of pulse-derived 13C incorporated into soil RNA over 2 months showed that there was only evidence of 13C enrichment in the continuously wetted treatments. However, isotopic values represented only a 0.1-0.2 13C at.% increase over natural abundance levels and were found to be insufficient for the application of RNA-SIP. These findings reveal that in this experimental system, the microbial uptake of labelled carbon from plant exudates is low, and further optimisation of methodologies may be required for application of SIP to natural plant-soil systems where 13C tracer dilution is a consideration., (Copyright 2004 Elsevier B.V.)

Double-stranded (ds) RNA is a biologically active component of many viruses including rhinoviruses infecting the upper respiratory tract. Mucus production is a common symptom of such infections. Here, we show that mucin, the glycoprotein subunit of mucus gels, is transcriptionally upregulated in an NF-kappaB- and p38-dependent manner when homogeneous cultures of epithelial cells are exposed to dsRNA. Furthermore, upstream of p38 in this system, dsRNA stimulates the extracellular release of ATP and activation of cell surface ATP receptors, which are G protein-coupled. This results in the stimulation of phospholipase C and protein kinase C. These findings suggest that ATP receptor antagonists could be used to modulate mucus production induced by virus.

We describe the development of a novel mobile field laboratory, purposely designed for the automated capture and subsequent stable isotopic analyses of multiple gas samples. The multiple capture system is integrated into a mobile laboratory that is fully capable of measuring the concentration of carbon dioxide, methane and nitrous oxide trace gases in a flow-through system connected to a gas chromatograph fitted with both electron capture and flame ionisation detectors. The capture of gases is achieved by routing samples through a series of 135 mL gas flasks that are sealed by micro-solenoid valves triggered by a timing system. Trace gas light stable isotope ratio mass spectrometry can then be carried out on gas samples collected by the system (NERC (15)N Stable Isotope Facility). The excitingly unique potential of the system to the ecological research field is that it will allow the collection of cyclical data for three different trace gases both in real-time and in situ. We present data arising from the validation of this mobile system as well as a preliminary experimental assessment of this technique. This technique was used to measure delta(13)C in CO(2) and CH(4) in soil gases released from waterlogged cores and delta(13)C-CH(4) values were significantly depleted in wet cores compared with dry ones (p < 0.001)., (Copyright 2002 John Wiley & Sons, Ltd.)

Human lung cells exposed to pathogenic bacteria upregulate the production of mucin, the major macromolecular component of mucus. Generally this upregulation is beneficial for the host, however, in the lungs of cystic fibrosis patients, overproduction of mucin can lead to the plugging of pulmonary airways. Mucus plugging impedes airflow and creates an environment that is highly compartmentalized: those bacteria within the mucus layer are shielded from high doses of antibiotics whereas those outside the mucus are exposed. These conditions augment mutation rate and the development of drug resistance in bacteria that colonize the lungs of cystic fibrosis patients. While therapeutic inhibition of mucin induction would improve airflow and reduce antibiotic resistance in these patients, the challenge is to develop drugs that block excessive mucin production while leaving beneficial aspects of the response intact. To do this, we must understand the molecular mechanisms underlying mucin production. Here we review the signal transduction pathways that control mucin production in response to Gram-positive and Gram-negative bacteria.

The flagella of the Gram-negative bacterium Pseudomonas aeruginosa serve not only for motility but also to bind bacteria to the host cell glycolipid asialoGM1 (ASGM1) through the protein flagellin. This interaction triggers defensive responses in host cells. How this response occurs is unclear because ASGM1 lacks transmembrane and cytoplasmic domains and there is little information about the downstream effectors that connect ASGM1 ligation to the initiation of host defense responses. Here, we show that ASGM1 ligation promotes ATP release from the host cell, followed by autocrine activation of a nucleotide receptor. This response links ASGM1 to cytoplasmic signaling molecules and results in activation of phospholipase C, Ca(2+) mobilization, phosphorylation of a mitogen-activated protein kinase (Erk 1/2), and activation of mucin transcription. These results indicate that bacterial interaction with host cells can trigger autocrine nucleotide signaling and suggest that agents affecting nucleotide receptors may modulate host responses to bacteria.

Bordetella pertussis, the causative agent of whooping cough, has many well-studied virulence factors and a characteristic clinical presentation. Despite this information, it is not clear how B. pertussis interaction with host cells leads to disease. In this study, we examined the interaction of B. pertussis with a human bronchial epithelial cell line (BEAS-2B) and measured host transcriptional profiles by using high-density DNA microarrays. The early transcriptional response to this pathogen is dominated by altered expression of cytokines, DNA-binding proteins, and NFkappaB-regulated genes. This previously unrecognized response to B. pertussis was modified in similar but nonidentical fashions by the antiinflammatory agents dexamethasone and sodium salicylate. Cytokine protein expression was confirmed, as was neutrophil chemoattraction. We show that B. pertussis induces mucin gene transcription by BEAS-2B cells then counters this defense by using mucin as a binding substrate. A set of genes is described for which the catalytic activity of pertussis toxin is both necessary and sufficient to regulate transcription. Host genomic transcriptional profiling, in combination with functional assays to evaluate subsequent biological events, provides insight into the complex interaction of host and pathogen.

Mucin production is an evolutionarily ancient defense mechanism that is retained in mammals and operates at all mucosal surfaces to protect the host against pathogens and irritants. As in lower organisms, the mammalian mucosa (epithelium) produces mucin in response to diverse insults. Our studies aim to understand the intracellular signaling and gene regulation mechanisms mediating mucin production in response to clinically important insults. To date, we find that the signaling pathway triggered by each type of insult is distinct. Relatively common, however, is the involvement of the protein tyrosine kinase c-Src, the MAP kinase kinase MEK 1/2, and the transcription factor NF-kappaB. Basbaum C, Lemjabbar H, Longphre M, Li D, Gensch E, McNamara N. Control of mucin transcription by diverse injury-induced signaling pathways.

Purpose: The current study prospectively evaluates the response to daily and extended wear of disposable lenses with each patient acting as his/her own control. This paradigm allowed for elimination of individual variation as a confounding factor, as well as determining whether an intrinsic factor, other than lens wear, predisposed patients to complications., Methods: One hundred thirty-four patients were randomly assigned to contralateral daily wear (DW) or extended wear (EW) with Acuvue lenses for 1 year. All lenses were disposed of on a weekly basis. Bilateral bacteriology of lids, conjunctivae, lenses, and fingers was carried out on those patients presenting with complications., Results: The results after 1 year indicate a higher incidence of complications in the EW eye (peripheral infiltrative keratopathy, 12%; corneal striae, 3%). No significant clinical complications occurred in the DW eye. Microbiological cultures did not reveal any significant difference between those eyes with keratopathy and those of controls. The most common pathogen was Staphyloccocus epidermidis. No gram-negative organisms grew from any of the culture sites., Conclusions: The results of this study indicate that extended wearing schedules, and not the lens entity, predispose the patient to a higher risk of keratopathy with disposable contact lenses. No microbiological association with the adverse events could be determined, and no intrinsic patient factor could be demonstrated that would predict the likelihood of complications.

Purpose: To study the effect of long-term polymethyl methacrylate (PMMA) contact lens wear (CLW) on corneal endothelial morphology and corneal hydration control (CHC)., Methods: Sixteen long-term PMMA CL wearers (mean duration of wear 20.8 +/- 8.39 SD years; mean age 40.37 years +/- 10.47 SD) served as subjects and 14 age-matched, non-lens wearing subjects (mean age 41.29 years +/- 10.15 SD) served as controls. Corneal function was assessed by measuring the corneal thickness recovery rate (PRPH) after induced edema from a hypoxic stress load. Endothelial morphology was assessed by taking specular microscopy photographs, digitizing the images, and calculating the cell density, size, and coefficient of variation of cell size (CV)., Results: Morphologic analysis showed a significant decrease in mean endothelial cell density (2147 vs. 2865, p < 0.01), an increase in mean cell size (582 vs. 381, p = 0.001), and an increase in polymegethism (CV, 0.32 vs. 0.26, p < 0.01) for PMMA lens wearers compared to agematched controls. Corneal function as measured by PRPH was not significantly different for PMMA lens wearers compared to controls when taken as a whole. However, a subgroup of PMMA lens wearers (seven eyes, four subjects) did show a substantial reduction in PRPH. These subjects also showed a strong relation between reduced PRPH, morphology, and years of lens wear (r2 = 0.99)., Conclusions: Longterm PMMA CLW leads to increased polymegethism in most, if not all, cases and results in a substantial decrease in corneal function in some cases.

We evaluated the effect of diclofenac sodium (Voltaren) drops on patients with hypoxia-induced corneal edema. Thirty age- and sex-matched subjects were randomly assigned to one of three groups. Members of each group received masked solutions of either Voltaren, Voltaren vehicle, or a non-preserved lubricant (Cellufresh) every 6 hours for 24 hours and then hourly for 2 hours immediately prior to inducing corneal edema in the experimental eye. Bilateral ultrasonic pachymetry was performed prior to applying a thick contact lens and light patch on the experimental eye of all subjects for 3 hours. The fellow eye served as the control. Following lens removal, bilateral corneal thickness was measured every 30 minutes. The percentage change in corneal swelling for each subject and group was calculated. The findings were also normalized to the control eye to minimize diurnal and individual variability. The results were plotted both as percentage change from hour 0 and percentage change normalized to the control eye. Corneal swelling ranged from 9-11% in all 3 groups, with recovery at 2-3 hours. No significant difference was found among the three groups (P > 0.05, ANOVA). There was a slight trend toward reduced thickness in the Cellufresh group, but this was not statistically significant. Voltaren does not appear to have an effect on the hypoxia-induced corneal edema associated with the production of arachidonic acid pathway metabolites.

Extracts of tissue showed that prostatic carcinomas contain less leucine aminopeptidase activity than benign prostatic hyperplasia. This is true when activity is expressed as specific activity (P = 0.0033), specific activity/% epithelium (P = 0.0007), activity/wet weight of tissue (P = 0.0028), or activity/wet weight of tissue/% epithelium (P = 0.0005). Almost all histochemically demonstrable activity is located in the epithelium. Enzymatic activities in extracts and in histochemical preparations showed similar differences between carcinoma and benign prostatic hyperplasia and were related (R = -0.38, P = 0.0400) to Gleason's grades. The transurethrally resected prostate cancers studied contained no well-differentiated tumors and a high proportion of poorly differentiated tumors. Histochemical activity is absent in most prostatic carcinomas and decreased in others. This observation is particularly interesting in view of the growing knowledge of tumor suppressor genes.

Virus-like particles (80 mmu to 100 mmu in diameter) occur in the endoplasmic reticulum of germ cells in the ovaries of fetal guinea pigs, and are confined to this population of cells. Using these particles as a marker, the phagocytosis of degenerating germ cells by somatic cells in the cortex of the ovary was traced.

Comparison of the fine structural features of guinea pig adrenocortical cells as seen in thin sections with those revealed by freeze-fracture confirms the structural appearance of steroid-secreting cells as interpreted from thin sections and reveals significant new features of the membranous organelles. Smooth-surfaced endoplasmic reticulum appears as a network of tubules, interwoven or in parallel, and as cisternae, fenestrated and non-fenestrated. These elements are tightly packed in the deeper cortical cells, excluding other organelles from their domain. Tubules and fenestrated cisternae possess randomly distributed intramembranous particles on their PF faces, while closely packed non-fenestrated cisternae possess aggregates of particles interspersed with aparticulate regions on their PF faces. These differences in particle distribution suggest functional specialization among the various forms of reticulum. Mitochondria appear as elongated structures of varying shape. Freeze-fracture reveals that all their cristae have circular origins from the inner membrane. Sinuous tubules, which appear as tubules in section, and straight tubules, which appear as lamellae in section, arise from single sites. Flattened sac-like cristae may have multiple circular origins. Definite contact points seen between inner and outer membranes may facilitate passage of molecules, including steroids, into the mitochondrial compartments. Lysosomes and peroxisomes, which are easily identified in thin sections with the aid of cytochemistry, are difficult to identify with certainty by freeze-fracture. Single membrane-bound granules of slightly smaller diameter than mitochondria may represent lysosomes. Smaller granules interconnected with the tubular reticulum, as well as dilated regions of this organelle, may represent peroxisomes. Plasma membranes show no indication of tight junctions but do have abundant gap junctions which show a zonal differentiation: small gap junctions throughout the cortex, medium-sized regularly shaped gap junctions in zona fasciculata externa, and large irregular gap junctions in zona fasciculata interna and zona reticularis. The large junctions cover planar areas as well as surfaces of projections of one cell into another. Such junctions may allow passage of ions as well as of low-molecular-weight substances between the cells, facilitating or even amplifying the response to trophic hormone stimulation.

Isolated guinea pig adrenocortical cells were maintained in long-term culture in order to perform sequential experiments on the same cell populations. The cells produced fluorogenic steroids, shown by thin-layer chromatography to be at least aldosterone, cortisol, and corticosterone. In addition, they increased production of these steroids when treated with either ACTH or dibutyryl cyclic AMP. Of particular interest was the fact that cultures treated for the initial 24-hour culture period with ACTH maintained enhanced levels of secretion for several days in absence of hormone and had an enhanced response to ACTH later in the culture period. Such enhancement of secretion was not seen following early treatment with dibutyryl cyclic AMP. The fine structure of the ACTH-treated cells was consistent with increased steroidogenesis. They possessed more smooth-surfaced endoplasmic reticulum, larger mitochondrial crystal surfaces, and larger Golgi complexes than the cells in untreated cultures.