Autoantibodies against 21-hydroxylase and side-chain cleavage enzyme in autoimmune Addison's disease are mainly immunoglobulin G1

OBJECTIVE: Immunoglobulin G (IgG) antibodies to the steroidogenic enzymes 21-hydroxylase (21OH) and side-chain cleavage enzyme (SCC) are important diagnostic markers for autoimmune Addison's disease and autoimmune polyendocrine syndromes (APS) types I and II. The characterization of autoantibody (IgG) subclasses may reveal information on how tIssue destruction takes place; therefore, IgG subtypes of anti-21OH and anti-SCC antibodies from sera of patients with Addison's disease, APS I and APS II were determined using recombinant 21OH and SCC. METHODS: SCC(51-521) and his-SCC(51-521) were expressed by pET-scc in the Escherichia coli strain BL21 Star (DE3) and inclusion bodies were purified. Full-length, human 21OH fused to an N-terminal 6x histidine affinity tag was expressed in insect cells by using the baculovirus expression system bac-to-bac. Western blots were used to investigate the IgG subtype(s) of the autoantibodies against 21OH and SCC in patients and healthy blood donors. RESULTS: All anti-SCC positive sera (n=10) contained autoantibodies of the IgG1 subclass, while four out of ten also contained IgG3. All anti-21OH positive sera (n=16) had autoantibodies exclusively against IgG1. Sera from 20 healthy subjects did not show any reactivity against 21OH or SCC. CONCLUSIONS: The finding of a predominating IgG1 response against 21OH and SCC may suggest that T helper (Th) cells of the Th1 subclass are involved in destruction of the adrenal cortex in patients with autoimmune Addison's disease.     

Workplace Bullying and Suicidal Ideation: A 3-Wave Longitudinal Norwegian Study

Objectives. We examined whether victimization from bullying is related to an increased risk of suicidal ideation over time and whether suicidal ideation is related to subsequent bullying.Methods. In a longitudinal study (2005–2010), we used well-established single-item measures to assess victimization from bullying and suicidal ideation. We used latent Markov models to determine forward and reverse relationships between variables at 3 time points with 2 or 3 years between the measurement points among a randomized nationwide sample of 1846 employees in Norway.Results. Victimization from bullying was associated with subsequent suicidal ideation (odds ratio = 2.05; 95% confidence interval = 1.08, 3.89). Suicidal ideation at baseline was not related to subsequent victimization from workplace bullying.Conclusions. Workplace bullying may be a precursor to suicidal ideation, whereas suicidal ideation seems to have no impact on subsequent risk of being bullied. Regulations against bullying should be integrated into work-related legislation and public health policies.Suicide is a leading cause of death around the globe. Estimates show that more than 800 000 people take their own lives every year.1 In Norway (with a population of 5 165 802) there are about 530 reported suicides every year.2 In the United States, 12 suicide deaths per 100 000 people were reported in 2010, making suicide the 10th leading cause of death among Americans.3 Altogether, 1719 employees committed suicide in US workplaces between 2003 and 2010.4Although psychiatric disorders are involved in the majority of suicide attempts,5 most psychiatric patients do not commit suicide. A psychiatric disorder alone is, therefore, an insufficient condition for suicide.6 To identify other risk factors, we must look beyond the presence of a psychiatric syndrome and understand the underlying factors of suicide and suicidal ideation. Among many potential causes, exposure to workplace bullying has been proposed to be an important predictor of both suicidal ideation and actual suicide.7,8To date, bullying as an antecedent to suicide has been examined only with anecdotal evidence9,10 and cross-sectional research designs.11,12 Einarsen et al.13 established that severely bullied workers were 6 times more likely than nonbullied workers to report suicidal ideations. Sterud et al.8 found that workplace bullying was positively associated with suicidal ideation in a nationwide sample of 1022 Norwegian ambulance personnel. Bullying was more strongly associated with suicidal ideation than were gender, neuroticism, anxiety, somatic complaints, depersonalization, and job dissatisfaction.Cross-sectional research cannot provide adequate evidence for anything more than that suicidal ideation is a correlate of bullying. To understand the directional nature of the association, longitudinal research is needed. In this representative, longitudinal study, we contribute to the literature by examining whether victimization from bullying is related to increased risk of suicidal ideation over time and whether suicidal ideation is related to subsequent bullying.Workplace bullying refers to a situation in which 1 or several individuals persistently perceive themselves to be on the receiving end of negative actions from superiors or co-workers and in which the targets find it difficult to defend themselves against these actions.14,15 Following this definition, there are 3 main characteristics of workplace bullying: (1) an employee becomes the target of systematic negative and unwanted social behavior in the workplace; (2) the exposure occurs over a prolonged period, often with ever more escalating intensity and frequency in the attacks; and (3) targets feel they cannot easily escape the situation or stop the unwanted treatment. This third characteristic, the feeling of being victimized by the harassment, distinguishes bullying from other forms of mistreatment in the workplace.15 Globally, about 11% of workers perceive themselves as victims of bullying,16 and 5% of the Norwegian working population is victimized by bullying at any time.17The interpersonal theory of suicide (ITS)5 provides a theoretical foundation for how exposure to such bullying may be related to suicide. The theory posits that fundamental to suicidal ideation and behavior is that an individual has both the desire and the ability to die by suicide.18 With regard to the desire to die, displayed through suicidal ideation, the ITS asserts that when people over a prolonged period perceive themselves to be socially alienated from others and simultaneously feel that they are a burden on others, they develop a desire for death.19 As for the ability to commit suicide, displayed through suicidal behavior, the ITS proposes that people who are repeatedly exposed to painful and provocative events will lose any fear of pain, injury, and death and thereby be able to overcome the instinct of self-preservation.Because of its focus on persistent exposure to painful events and social alienation, the ITS strongly suggests that repeated and long-term exposure to negative treatment and social exclusion from one’s peers or supervisors at work constitutes a risk factor for suicidal ideation and behavior.Although previous research has assumed bullying to be an antecedent to suicidal ideation, it is possible that the established cross-sectional association reflects a relationship in which suicidal ideation is a precursor to bullying. Two different mechanisms can explain such a reverse association. First, employees with suicidal ideation may report less favorable work characteristics because their distress makes them evaluate their work environment increasingly more negatively.20 Second, employees with suicidal ideations may elicit aggressive behavior in others because their psychological state creates aversive feelings among co-workers and supervisors.21,22To provide better indications of how workplace bullying is related to suicidal ideation, we investigated direct forward and reverse associations with longitudinal data. We tested the following hypotheses:

• Hypothesis 1: Victimization from bullying is associated with an increased risk of later suicidal ideation.
• Hypothesis 2: Suicidal ideation is associated with an increased risk of later victimization from bullying.

     

Comparing the adrenaline-Pirquet test with international PPD tuberculin tests

Several tuberculins, strengths and setting methods are in everyday use. We wanted to compare the Norwegian reference adrenaline Pirquet test with the internationally recommended Mantoux PPD test and Rhoditest. In responders of a random sample of young adults, with randomization of test subjects, we intra-individually compared the adrenaline-Pirquet (aP) test with Norwegian-produced synthetic medium tuberculin (SMT) with either the Mantoux test with PPD 2 tuberculin units (TU) (M2), the Mantoux-PPD 5 TU (M5) or the PPD 2 TU-Rhoditest (Rh). The criteria for a positive reaction were > or = 4mm for the aP test, > or = 10 mm for the M2 test, > or = 6 mm for the M5 test and > or = 2 mm for the Rh test. Strongly positive reactions were defined as aP test > or = 10 mm and M2/M5 test > or = 15 mm. One of the tuberculin tests was positive while the comparison test was negative in 14% of the M2 test group (n = 236), 15% in the M5 test group (n = 162) and 20% in the Rh test group (n = 187). The three PPD tests had positive reactions 3-10 times as often, with a simultaneous negative aP test, than vice versa. Strongly positive reactions occurred in 7% of the aP tests (> or = 10 mm), 32% of the M2 tests (> or = 15 mm) and in 41% of the M5 tests (> or = 15 mm). Receiver operating characteristic (ROC) curves gave the best agreement, with aP test >3mm compared with the M2 and the Rh tests. Regression equations are presented for transformations of the Norwegian reference method of adrenaline-Pirquet results to internationally recommended PPD tests. The international recommendations, globalization in general and the skill of Norwegian public health nurses in performing intra-dermal BCG suggest a future shift to the PPD 2 TU Mantoux test in Norway. Due to the lack of sensitivity and specificity of all tuberculin tests they might be used in targeted tuberculin testing and not as a general screening procedure in a low-incidence tuberculosis area with BCG-vaccinated inhabitants.     

Cell distributions and functions of Toll-like receptor 4 studied by fluorescent gene constructs

Bacterial lipopolysaccharide (LPS) is recognized in mammals by a receptor complex composed of CD14, Toll-like receptor 4 (TLR4), and MD-2. The detailed mechanisms of how TLR4 transmits the signal from the outside to the inside of the cell remain to be elucidated. One way of studying TLR4 signaling mechanisms is to construct chimeras of TLR molecules C-terminally fused to fluorescent proteins and stably express these constructs in cells. Such constructs are functional when transfected into HEK293 epithelial cells. Confocal microscopy of TLR4 expression in live cells demonstrated pronounced expression on the plasma membrane as well in the Golgi apparatus. Studies were performed to clarify whether expression of TLR4 in the Golgi was necessary for LPS stimulation. Rapid recycling of TLR4/CD14/MD-2 complexes between the Golgi and the plasma membrane was a prominent phenomenon. In agreement with other types of plasma membrane receptors, aggregation of TLR4 by immobilized TLR4 antibodies was sufficient to induce signaling. Also, pharmacological disruption of the Golgi did not inhibit LPS induced NF-kappaB activation. Furthermore, LPS stimulation recruited the adapter molecule, MyD88, to the inside of the plasma membrane. Thus, LPS signaling commences on the plasma membrane and is independent of trafficking to the Golgi.     

Pancreas Transplantation With Enteroanastomosis to Native Duodenum Poses Technical Challenges—But Offers Improved Endoscopic Access for Scheduled Biopsies and Therapeutic Interventions

To facilitate endoscopic access for rejection surveillance and stenting of the pancreas, we have abandoned the duodenojejunostomy (DJ) in favor of duodenoduodenostomy (DD) in pancreas transplantation (PTx). From September 2012 to September 2013 we performed 40 PTx with DD; 20 solitary‐PTx (S‐PTx) and 20 simultaneous pancreas and kidney transplantation (SPK). We compared the outcomes with results from 40 PTx‐DJ (10 S‐PTx and 30 SPK) from the preceding era. The DD‐enteroanastomoses were performed successfully. Endoscopic pancreas biopsies (endoscopic ultrasound examination [EUS]) yielded representative material in half of the cases. One exocrine fistula was treated by endoscopic stenting. PTxs‐DD were associated with a higher rate of thrombosis compared to PTx‐DJ (23% vs. 5%) and reoperations (48% vs. 30%), as well as inferior graft survival (80% vs. 88%). Time on waiting list, HLA A + B mismatches and reoperations were associated with graft loss. Only recipient age remained an independent predictor of patient death in multivariate analysis. PTx‐DD showed a higher rate of thrombosis and inferior results, but facilitated a protocol biopsy program by EUS that was feasible and safe. Given that technical difficulties can be solved, the improved endoscopic access might confer long‐term benefits, yet this remains to be proven.     

The effect of educational level on the incidence of asthma and respiratory symptoms

Few studies have examined the impact of socioeconomic status on the incidence of asthma and respiratory symptoms. Between 1985 and 1996/97, we conducted an 11-years community cohort study with 2819 subjects, aged 15-70 years at baseline, in Western Norway. We examined the cumulative incidence of asthma and respiratory symptoms by educational level (primary, secondary, and university), as well as estimating the odds ratios (ORs) of educational level on the incidences, after adjustment for sex, age, hay fever, smoking habits, pack years, and occupational exposure. For all respiratory symptoms, the incidences decreased with increasing educational level. The cumulative incidence of asthma was 5.3%, 4.1%, and 1.8%, respectively, for those with a primary educational level, secondary educational level, and university level. Subjects with a primary educational level had adjusted ORs (95% CI) from 1.4 (0.9, 2.3) for the incidence of chronic cough to 2.5 (1.6, 4.0) for the incidence of dyspnea grade 2, compared to those with a university level education. The adjusted OR (95% CI) for the incidence of asthma was 2.1 (1.01, 4.4) in subjects with a primary educational level, and 2.0 (1.04, 3.6) in subjects with a secondary educational level, compared to subjects with a university educational level. In conclusion, subjects with a lower educational level had a higher risk of developing asthma and respiratory symptoms, after adjustment for sex, age, hay fever, smoking, and occupational exposure.