Cancer immunotherapy based on recombinant Salmonella enterica serovar Typhimurium aroA strains secreting prostate-specific antigen and cholera toxin subunit B.

Prostate cancer is the most common malignant tumor in men and is normally associated with increased serum levels of prostate-specific antigen (PSA). Therefore, PSA is one potential target for a prostate cancer vaccine. In this study we analyzed the functionality of new bacterial PSA vaccines, expressed and secreted via the hemolysin (HlyA) secretion system of Escherichia coli, the prototype of Type I secretion systems (T1SS) using an attenuated Salmonella enterica serovar Typhimurium aroA strain as carrier. The data demonstrate that a bacterial live vaccine encompassing T1SS in combination with cholera toxin subunit B can be successfully used for delivery of PSA to induce cytotoxic CD8+ T-cell responses resulting in an efficient prevention of tumor growth in mice.

Benevolent voices are not so kind: the functional significance of auditory hallucinations.

BACKGROUND: This study measures the impact of beliefs about auditory hallucinations on social functioning. SAMPLING AND METHODS: Twenty-nine subjects who met the ICD-10 criteria for schizophrenia or a schizo-affective disorder were included. Beliefs about voices and coping responses as measured by the Beliefs about Voices Questionnaire were compared with social functioning as assessed with the Life Skills Profile (LSP). RESULTS: The belief that voices are benevolent was associated with poor communication. Engagement with voices was correlated with the non-turbulence and the compliance factors of the LSP. Patients who held the belief that their voices were benevolent functioned significantly more poorly on the communication factor of the LSP than patients who interpreted their voices as malevolent. DISCUSSION: The results indicate that a positive relationship with voices may affect social functioning. However, the size of the sample is small and patients with benevolent voices are overrepresented. Nonetheless, these results have implications for the use of cognitive therapy for psychotic symptoms.

Completed suicides among the Inuit of northern Quebec, 1982-1996: a case-control study.

BACKGROUND: The rate of completed suicide among Inuit in Canada has been alarmingly high in recent years, and the suicide rate among Inuit in northern Quebec has increased since 1982. Our objectives were to describe the characteristics of Inuit people who died by suicide in Nunavik between 1982 and 1996, and to identify the antecedents and correlates of completed suicide. METHODS: We carried out a case-control study of 71 people who died by suicide between 1982 and 1996 and 71 population-based living control subjects matched for sex, community of residence and age within 1 year. Comprehensive medical charts were reviewed for data on sociodemographic characteristics, medical and psychiatric history, childhood separations and family history, and use of health care services. RESULTS: Most of the case subjects were single males aged 15 to 24 years. The two principal means of suicide were hanging (in 39 cases [54.9%]) and gunshot (in 21 cases [29.6%]). About 33% had been in contact with medical personnel in the month before their death. The case subjects were significantly more likely than the control subjects to have received a lifetime psychiatric diagnosis (one or more of depression, personality disorder or conduct disorder) (odds ratio [OR] 4.3 [95% confidence interval (CI) 1.2-15.2]) and to have had a history of psychiatric symptoms, disorder (including solvent sniffing) or treatment (OR 3.5 [95% CI 1.4-8.7]). The case subjects had experienced more severe types of nonpsychiatric illnesses and injuries than the control subjects (p = 0.04). The case subjects had more lifetime contacts with health care services than the control subjects (p = 0.01) and were more likely than the control subjects to have had contact with health care services in the year before death of the case subject (p = 0.03), even when psychiatric diagnoses were controlled for in conditional regression analysis (OR 1.02 [95% CI 1.01-1.04] and 5.0 [95% CI 1.07-23.7] respectively). INTERPRETATION: Since case subjects had frequent contact with health care services, frontline medical personnel may be in a position to identify people at risk for suicide.

Attachment disorganization and dissociative symptoms in clinically treated adolescents.

OBJECTIVE: To examine the association of unresolved and unclassifiable attachment with dissociative symptomatology in a sample of 133 adolescents in psychiatric treatment. METHOD: The study compared 69 adolescents who were unresolved and unclassifiable with 64 adolescents who were not unresolved and unclassifiable. Attachment organization was assessed using the Adult Attachment Interview (AAI). Dissociative symptomatology was assessed using a scale derived from the Youth Self Report (YSR) behaviour checklist. RESULTS: A continuing unresolved and unclassifiable response to attachment-related trauma was correlated with dissociative symptomatology for both male and female adolescents. CONCLUSIONS: Cognitive disorganization may be an important variable mediating between the effects of earlier traumatic caregiving experiences and later dissociative symptoms.

Gram-positive and Gram-negative bacteria as carrier systems for DNA vaccines.

Vaccination by intradermal or intramuscular injection of eukaryotic antigen expression vectors (so-called DNA vaccines) elicits strong cellular and humoral immune responses. A novel approach employs attenuated mutant strains of Gram-positive and Gram-negative intracellular bacteria as carriers for the delivery of DNA vaccines. This strategy allows the administration of the DNA vaccines via mucosal surfaces and a direct delivery of the plasmid DNA to professional antigen presenting cells (APC), such as macrophages and dendritic cells (DC). In this work, we have found that several Gram-negative bacteria are capable of delivering plasmid vectors to human DC. In addition, we tested the suitability of the Gram-positive bacterium Listeria monocytogenes as a vaccine carrier for the immunization of fish.

Recombinant attenuated bacteria for the delivery of subunit vaccines.

Using attenuated intracellular bacteria as carriers, we have developed two different approaches for the delivery of subunit vaccines encoding heterologous antigens. The first system is based on the direct secretion of the heterologous antigens in Gram-negative bacteria via the hemolysin secretion system of Escherichia coli into either phagosome or cytosol of infected cells. The second approach is based on the transport of eukaryotic antigen expression vectors by intracellular bacteria like Listeria and Salmonella into the host cell and here, preferably, into the cytosolic compartment. After release of the plasmid DNA from the bacteria, the plasmid-encoded antigens can be expressed directly by the host cell. Finally, we combined both types of subunit vaccines in one live vector - we equipped Salmonella strains with a phagosomal escape function by utilization of the hemolysin secretion system and used this recombinant vaccine strain for the delivery of a eukaryotic antigen expression vector into the cytosol of macrophages.

The Adolescent Unresolved Attachment Questionnaire: the assessment of perceptions of parental abdication of caregiving behavior.

This article reports on the Adolescent Unresolved Attachment Questionnaire (AUAQ), a brief questionnaire that assesses the caregiving experiences of unresolved adolescents (as recipients of caregiving). The AUAQ was developed and validated in a large normative sample (n = 691) and a sample of 133 adolescents in psychiatric treatment. It is a self-report questionnaire consisting of 3 scales with Likert-type responses ranging from strongly disagree to strongly agree. The Aloneness/Failed Protection Scale assesses the adolescent's perception of the care provided by the attachment figure. The Fear Scale taps the fear generated by the adolescent's appraisal of failed attachment figure care. The Anger/Dysregulation Scale assesses negative affective responses to the perceived lack of care from the attachment figure. All scales demonstrated satisfactory internal reliability and agreement between scores for adolescents (n = 91) from the normative sample who completed the AUAQ twice. Adolescents in the clinical sample also completed the Adult Attachment Interview (AAI; C. George, N. Kaplan, & M. Main, 1984/1985/1996); the AUAQ demonstrated high convergent validity with the AAI.

Bacterial systems for the delivery of eukaryotic antigen expression vectors.

Attenuated bacterial strains allow the administration of recombinant vaccines via the mucosal surfaces. Whereas attenuated bacteria are generally engineered to express heterologous antigens, a novel approach employs intracellular bacteria for the delivery of eukaryotic antigen expression vectors (so-called DNA vaccines). This strategy allows a direct delivery of DNA to professional antigen-presenting cells (APC), such as macrophages and dendritic cells (DC), through bacterial infection. The bacteria used for DNA vaccine delivery either enter the host cell cytosol after phagocytosis by the APC, for example, Shigella and Listeria, or they remain in the phagosomal compartment, such as Salmonella. Both intracellular localizations of the bacterial carriers seem to be suitable for successful delivery of DNA vaccine vectors.

Delivery of protein antigens and DNA by virulence-attenuated strains of Salmonella typhimurium and Listeria monocytogenes.

Two different plasmid-vector systems were developed which allow the efficient production and presentation of protein antigens in antigen-presenting cells (APC) by means of virulence-attenuated bacteria. The first antigen-delivery system is based on the secretion machinery of the Escherichia coli hemolysin (HlyA-type I secretion system), which transports proteins, possessing the specific HlyA secretion signal (HlyA(s)) at the C-terminus, across both membranes of gram-negative bacteria. This system functions in all gram-negative bacteria that possess the TolC-analogous protein in the outer membrane. This outer membrane protein is necessary for the stable anchoring of the type I secretion apparatus in the cell envelope. Suitable HlyA(s)-fused antigens are secreted with high efficiency by E. coli and by virulence-attenuated strains of Salmonella, Shigella, Vibrio cholerae and Yersinia enterocolitica. The other vector system expresses the heterologous antigen under the control of an eukaryotic promoter in a similar fashion as in plasmids commonly used for vaccination with naked DNA. This plasmid DNA is introduced into APCs with the help of virulence-attenuated self-destructing Listeria monocytogenes mutants. After synthesis of the heterologous protein, epitopes of the antigen are presented by the APC together with MHC class I molecules. This system functions in macrophages and dendritic cells in vitro and can also be used in a modified form in animal models.

Identification of immunogenic antigens of Helicobacter pylori via the Escherichia coli hemolysin secretion system(1).

We describe a new procedure allowing the generation and detection of immunogenic antigens from Helicobacter pylori via the hemolysin secretion apparatus of Escherichia coli. The gene (or gene fragment) encoding the H. pylori protein (or protein domain) is inserted in-frame into a residual portion of the hemolysin gene (hlyA), encoding the HlyA secretion signal (HlyA(s)). These fusion proteins are secreted efficiently by E. coli. This new approach allows the identification of immunodominant antigens by using sera derived from H. pylori-infected patients suffering from different gastroduodenal pathologies. Three immunodominant antigens bearing the ureB (urease B-subunit), flaA (flagellin A-subunit), and an unknown ORF (HP0888) encoding an E. coli FecE analogous protein fused to hlyA(s) were identified and characterized.

Novel bacterial systems for the delivery of recombinant protein or DNA.

On the basis of attenuated intracellular bacteria, we have developed two delivery systems for either heterologous proteins or DNA vaccine vectors. The first system utilizes attenuated strains of Gram-negative bacteria which are engineered to secrete heterologous antigens via the alpha-hemolysin secretion system of Escherichia coli. The second system is based on attenuated suicide strains of Listeria monocytogenes, which are used for the direct delivery of eukaryotic antigen expression vectors into professional antigen presenting cells (APC) like macrophages in vitro as well as in vivo.

The relationship between health-related quality of life and frequency of spells in patients with syncope.

Chronic syncope has a wide range of symptom burden, and anecdotal data suggest substantial but variable physical and psychosocial morbidity. We hypothesized that health-related quality of life (HRQL) is impaired in syncope patients and the degree of impairment is proportional to syncope frequency. The EuroQol EQ-5D was completed by 136 patients (79 female and 57 male) with mean age 40 (SD = 17) prior to assessment. HRQL was substantially impaired in syncope patients compared to population norms in all five dimensions of health measured by the EQ-5D. In patients with six or more lifetime syncopal spells there was a significant (P < 0.001) negative relationship between the frequency of spells and overall perception of health, which was not evident in those who had a history of less than six lifetime spells. These relationships were maintained after controlling for comorbid conditions.

Salmonella vaccines secreting measles virus epitopes induce protective immune responses against measles virus encephalitis.

In the present study we describe a live vaccine against measles virus (MV) infection on the basis of attenuated Salmonella typhimurium aroA secreting MV antigens via the Escherichia coli alpha-hemolysin secretion system. Two well-characterized MV epitopes, a B-cell epitope of the MV fusion protein (amino acids 404-414) and a T-cell epitope of the MV nucleocapsid protein (amino acids 79-99) were fused as single or repeating units to the C-terminal secretion signal of the E. coli hemolysin and expressed in secreted form by the attenuated S. typhimurium aroA SL7207. Immunization of MV-susceptible C3H mice revealed that S. typhimurium SL7207 secreting these antigens provoked a humoral and a cellular MV-specific immune response, respectively. Mice vaccinated orally with a combination of both recombinant S. typhimurium strains showed partial protection against a lethal MV encephalitis after intracerebral challenge with a rodent-adapted, neurotropic MV strain.

Relationship between attachment-felt security and history of suicidal behaviours in clinical adolescents.

OBJECTIVE: This study was designed to test the hypothesis that adolescents who perceive their attachment figures as unavailable (low felt security) would be overrepresented in the case group of adolescents with a history of suicidal behaviours. METHOD: One hundred and eighty-seven adolescents in psychiatric treatment participated in this retrospective case-comparison study of attachment-felt security and history of suicidal behaviours. All participants completed the following measures: Adolescent Attachment Questionnaire, Perceived Social Support From Friends Scale, Rosenberg Self-Esteem Scale, Beck Hopelessness Scale, the depression syndrome scale of the Youth Self Report, and Adam's Suicidal Ideation and Behavior protocol. RESULTS: The comparison group comprised 101 adolescents who had never experienced suicidal ideation or behaviour; the case group included 86 adolescents with a history of suicidal behaviour. We found that perceived unavailability and high levels of depressive symptomatology were predictive of suicidal behaviours. We also found a strong association between being older and having high levels of angry distress in adolescents with a history of suicidal behaviours. CONCLUSION: The advantage of including an assessment of parent-adolescent attachment with clinical adolescents is noted.

Anxious attachment and severity of depressive symptomatology in women.

The present research evaluated a conceptual model that links anxious attachment to depressive symptomatology in women. Four hundred and twenty women completed the following measures: Socio-demographic questionnaire, the Centre for Epidemiological Studies Depression Scale (CES-D), the Reciprocal Attachment Questionnaire, the Social Support Questionnaire, the Rosenberg Self-Esteem Scale and the Global Assessment of Recent Stress Scale. Results indicate that feared loss of the attachment figure, intense proximity seeking and lack of use of the attachment figure were predictive of depressive symptomatology. We also found that lower levels of self-esteem and higher levels of recent stress were predictive of depressive symptomatology. The implications of these findings for the understanding of symptoms of depression in women is discussed.

Statistical issues encountered in the comparison of health-related quality of life in diseased patients to published general population norms: problems and solutions.

The objectives of this study were (1) to illustrate the statistical problems encountered when comparing health-related quality of life (HRQL) measured by the Medical Outcome Study Short Form-36 (SF-36) in a diseased group to general population norms, and (2) to define age- and gender-standardized dichotomous indicator variables for each health concept and show that these indicator variables facilitate comparisons between the diseased sample and the general population. Our "diseased" group consisted of 136 sequentially consenting patients referred to the syncope clinic for assessment and treatment. Participants completed the SF-36 questionnaire before undergoing diagnostic testing. General population norms for the SF-36 are available from the responses of 2474 participants in the National Survey of Functional Health Status, conducted in 1990 in the United States. Comparison of the SF-36 in a diseased sample with general population norms is difficult, owing to skewed and unusual distributions in both groups. In addition, making comparisons within age and gender strata is difficult if the within strata sample size is small. We propose a dichotomous indicator variable for each health concept that classifies an individual as having impaired health if he or she scored lower than the 25th percentile for the appropriate age and gender general population strata. By definition, the prevalence of impaired health in the general population is 25% for all eight health concepts. Comparison between the eight health-concept variables is easy because the population norm is the same for each of them. These indicator variables are age and gender adjusted, so that even if the sample did not have the age and gender distribution as the general population, comparisons can still be made with the value of 25.

A comparison of two measures to assess adult attachment.

The purpose of this study was to determine the relationship between two different approaches to the assessment of adult attachment. The approaches were C. Hazan and P. R. Shaver's (1987) categorization of three attachment styles and the dimensional system of the Reciprocal Attachment Questionnaire (RAQ; M. West & A. Sheldon-Keller, 1992). Participants were 196 (72 men and 124 women) respondents to a community survey. Multiple regression results showed that the three attachment groups differed significantly on three dimensions of the RAQ: feared loss of the attachment figure, perceived availability of the attachment figure, and use of the attachment figure. In addition, men and women differed significantly on use of the attachment figure, independent of their attachment style. The results indicate that the two approaches to the assessment of adult attachment are interrelated in meaningful ways.

Construction of chromosomally encoded secreted hemolysin fusion proteins by use of mini-TnhlyAs transposon.

We used the minitransposon TnhlyAs [Gentschev, I., Maier, G., Kranig, A. and Goebel, W. (1996) Mol. Gen. Genet. 252, 266-274] for random insertion of the secretion signal (HlyAs) of Escherichia coli hemolysin (HlyA) into chromosomal genes. Four mini-TnhlyAs derivatives bearing the gltA (citrate synthase), deoC (2 deoxyribose-5 phosphate aldolase), tig (trigger factor) genes and an unknown ORF fused to hlyAs were identified and characterized. Our data suggest that TnhlyAs-generated hemolysin fusion proteins are secreted efficiently by the HlyB/HlyD/TolC hemolysin secretion machinery and that this can be useful for studies of gene expression or function.

Anxious attachment and self-reported depressive symptomatology in women.

OBJECTIVE: Lack of intimacy has been identified as an important provoking agent that increases the risk of depressive symptoms in women. This study precisely characterized lack of intimacy by assessing a woman's attachment style and investigated the specificity of association between depressive symptoms and an anxious attachment pattern. METHOD: Four hundred and twenty women participated in this cross-sectional study of depressive symptomatology and anxious attachment. All participants completed the following measures: a sociodemographic questionnaire, the Centre for Epidemiological Studies Depression Scale (CES-D), the Reciprocal Attachment Questionnaire, the Social Support Questionnaire, the Rosenberg Self-Esteem Scale, and the Global Assessment of Recent Stress Scale. RESULTS: A score of 16 or above on the CES-D, which indicates the presence of depressive symptoms, was used to divide the sample into 2 groups: a depressed group (N = 129) and a nondepressed group (N = 291). We found that women in the depressive symptomatology group were more likely than women in the nondepressive symptomatology group to exhibit anxious attachment and adverse social and cognitive characteristics. Lower levels of self-esteem and higher levels of recent stress were also predictive of depressive symptomatology. Feared loss of the attachment figure and a lack of use of the attachment figure were independent predictors of depressive symptomatology in the same model. CONCLUSION: The feared loss of security associated with an attachment figure seems to be related to an increased likelihood of depressive symptoms.