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Both exogenous and endogenous interleukin-10 affects the maturation of bone-marrow-derived dendritic cells in vitro and strongly influences T-cell priming in vivo.

In order to avoid autoimmunity and excessive tissue destruction, the action of certain immunoinhibitory substances are very important for negative regulation of the immune system. Interleukin-10 (IL-10) is an important immunoregulatory cytokine which is thought to negatively affect both T cells and antigen-presenting cells in vivo. Adoptive transfer of IL-10-treated bone-marrow-derived dendritic cells (BMDCs) may be one therapeutic avenue to inhibit autoimmunity. In this study we present a comprehensive analysis of the effects of IL-10 on murine BMDC. We demonstrate that IL-10 can prevent BMDC maturation, as measured by both cytokine production and T-cell priming capacity in vitro. Furthermore, we show that IL-10 can inhibit DC maturation induced by strong stimulatory signals such as lipopolysaccharide or a mixture of cytokines (interferon-gamma, tumour necrosis factor-alpha, IL-4). Interestingly, maturation of both T helper 1- and T helper 2-inducing DCs, characterized by the induction of high levels of interferon-gamma and IL-4-production by responding T cells, respectively, was inhibited by IL-10 in vitro. Finally, our data suggest that both endogenous and exogenous IL-10 affect the T-cell stimulatory capacity of BMDCs after injection of in vitro-treated BMDCs into naïve mice. These data both support existing data as well as point towards a new understanding of the many aspects of IL-10-mediated immunosuppression.

Animals↗

Attitudes of professionals, students and the general public to HIV/AIDS and people with HIV/AIDS: a review of the research.

This paper provides a review relating to the attitudes of health care professionals, students and the general public to HIV/AIDS and people with HIV/AIDS. This is done by describing the literature on attitudes to HIV and AIDS in three fields of research: medicine/nursing, psychology and the social sciences. In addition, we look at the countries in which research has been conducted in these issues and at the variety of tools and methodological approaches used and at the participants studied. The review shows that, during the 1990s, the research interest in issues related to HIV/AIDS attitudes has rapidly increased. Most of the work on HIV and AIDS has been done in the United States, with the main focus on empirical research using questionnaires. The attention has mostly been on students and their attitudes to AIDS/HIV and sexual behaviour, but increasing interest has also been shown in the attitudes of health care personnel and students. As it seems that attitudes have been highly resistant to change, more consideration needs to be given to finding appropriate ways of educating the general public, students and health care professionals. A comprehensive analysis is also needed of the different tools used in measuring attitudes and in international comparisons of the attitudes of adolescents, health care professionals and students towards AIDS and AIDS patients.

Attitude to Health↗

Video modelling to educate patients.

BACKGROUND: Changes in health care delivery in the United States of America due to economic pressures have required nurses to develop innovative instructional materials for educating patients and families. Educational materials such as videotapes, specifically designed to provide information and promote active participation in treatment decisions, can be effective tools for empowering patients. A comprehensive analysis in 1988 concluded that the concept of 'video modelling' or 'behavioural modelling' offered the greatest benefit of videotaped presentations. AIM OF THE STUDY: The purpose of this integrative literature review was to examine the concept of video modelling and its applications in clinical practice. METHODS: A computer search of the electronic databases of Medline and CINHAL between 1990 and 1999 produced a total of 40 research studies on video instruction for patients. Based on criteria for inclusion, 18 research studies involving video modelling were reviewed and three major uses were identified: (1) assisting decision making regarding treatment options; (2) reducing pre-procedural anxiety and improving coping skills; and (3) teaching self-care practices. RESULTS: The studies reviewed included a variety of research designs, clinical settings, and patient populations. Despite these differences, several benefits to the use of video modelling were found. Patients who viewed videotapes regarding treatment options had a greater understanding of the risks and benefits of those choices and were more apt to be active participants in decision making. Collective results of the studies focusing on stress and coping revealed that preparatory videotapes using video modelling could have a positive effect on reducing anxiety and physiological arousal during stressful procedures. With self-care practices, several of the studies found that there was an increase in desired behaviours in people whose educational programmes included video modelling. CONCLUSIONS: The use of video modelling has potential benefits for clinical practice in facilitating knowledge acquisition, reducing preparatory anxiety, and improving self-care. Nurses must become more actively involved in evaluating various teaching approaches used with patients to enhance practice and outcomes.

Adaptation, Psychological↗

Examining out-of-pocket expenditure on health care in Nouna, Burkina Faso: implications for health policy.

OBJECTIVE: To examine household out-of-pocket expenditure on health care, particularly malaria treatment, in rural Burkina Faso. METHOD: Comprehensive analysis of out-of-pocket expenditure on health care through a descriptive analysis and a second, multivariate analysis using the Tobit model with emphasis on malaria, based on 800 urban and rural households in Nouna health district. RESULTS: Households will spend less on malaria, either in or outside the health facility, if given the choice to do so, because they feel confident to self-treat malaria. Seeking health care from a qualified health worker incurs more out-of-pocket expenditure than self-treatment and traditional healers, and if necessary, households sell off assets to offset the expenditure. More than 80% of household out-of-pocket expenditure is allocated to drugs. CONCLUSION: This has policy implications for malaria control and the Roll Back Malaria Initiative. Communities need to be educated on the risks of malaria complications and the potential risk of inappropriate diagnosis and treatment. Drug or health services pricing policy needs to create an incentive to use the health services. In the fight against malaria, building alliances between households, traditional healers and health workers is essential.

Burkina Faso↗

Presynaptic calcium-channel currents in normal and csp mutant Drosophila peptidergic terminals.

The study of regulated vesicle exocytosis, which underlies neurotransmitter and neuropeptide release, has benefited from a convergence of several independent approaches. These include the use of genetically tractable organisms and model preparations that allow a direct characterization of presynaptic ionic currents. Aiming for a comprehensive analysis of release, we had already developed a Drosophila preparation in which electrophysiological recordings from peptidergic terminals are feasible. Here, we report on the characterization of the Ca2+-channel currents present in these terminals. With Ba2+ as the charge carrier, the presynaptic membrane expresses a current type with high-activation threshold and little inactivation. This current is blocked by verapamil and diltiazem at micromolar concentrations, it is relatively insensitive to nifedipine and completely resistant to non-L-type Ca2+-channel antagonists. As a comparison, we also analysed the pharmacology of high-threshold Ba+2 currents on muscle fibres. A high-activation threshold Ca2+-channel current is also present in muscle fibres, albeit with a distinct pharmacological profile. Thus, peptidergic terminals and muscle fibres exhibit different subtypes of voltage-gated Ca2+ channels. The putative role of cysteine string protein (CSP) as a neuronal Ca2+-channel modulator was tested by examining the peptidergic presynaptic current in csp null mutants. We show that CSP is expressed in peptidergic boutons and abolished in the mutant. Direct recordings, under conditions that inhibit calcium influx into glutamatergic terminals, show that Ca2+-currents in peptidergic csp terminals are entirely normal. This result indicates that CSP is not a generic Ca2+-channel modulator and it might perform different functions in fast versus slow forms of release.

Animals↗

Normal structure and age-related changes of the equine retina.

Investigations of the pathophysiology of ocular diseases require a detailed knowledge of the microanatomy of the eye. The available information is still inadequate for the equine retina despite the importance of eye diseases in equine medicine. Here we provide a comprehensive analysis of the histologic features of the horse eye as a reference for future studies. Thirty normal eyes of 15 healthy horses were examined immediately after slaughter. The retina of the horse differs considerably in the degree and quantity of neurons and glial elements as well as in vascular patterns compared to the retina of other domestic animals. Morphometric analysis revealed that the thickness of the retina varies between 80 microm at the ora serrata and 250 microm medial to the optic disc. Approximately 90% of the equine retina is comparatively thin (< 130 microm). This is a physiologic response to the distance that oxygen can diffuse in avascular retina. Ganglion cells form a single layer in all parts of the retina. The majority of ganglion cells are very large Nissl-positive cells. Small Nissl-negative ganglion cells are less abundant. A high ganglion cell density is found only in the central area. Vascularization is virtually absent from the retina with the exception of a narrow strip around the disc of the optic nerve, as revealed by lectin histochemistry. Light microscopy of the eyes of older horses repeatedly revealed cystoid degenerations in the retina adjacent to the pars plana of the ciliary body, as well as a destruction of the regular layering of the peripheral region of the retina.

Aging↗

Modulation of keratinocyte gene expression and differentiation by PPAR-selective ligands and tetradecylthioacetic acid.

Peroxisome proliferator-activated receptors (PPARs) are pleiotropic regulators of growth and differentiation of many cell types. We have performed a comprehensive analysis of the expression of PPARs, transcriptional cofactors, and marker genes during differentiation of normal human keratinocytes using a combination of reverse transcriptase polymerase chain reaction, Northern and Western blotting, and immunohistochemistry. PPARdelta was the predominant PPAR subtype in human keratinocytes and highly expressed in basal cells and suprabasal cells. Induction of PPARalpha and PPARgamma expression was linked to differentiation, and accordingly, expression of PPARalpha and PPARgamma was in essence confined to suprabasal cells. Differentiation was not accompanied by significant changes in the expression of the coactivators CREB-binding protein, p300, steroid receptor coactivator 1, or the corepressors nuclear receptor corepressor and silence mediator for retinoid and thyroid hormone receptors. We critically evaluated the effects of selective PPAR ligands and a synthetic fatty acid analog, tetradecylthioacetic acid. Tetradecylthioacetic acid activated all human PPAR subtypes in the ranking order PPARdelta >> PPARalpha > PPARgamma. All selective PPAR ligands marginally induced transglutaminase-1 expression with the PPARdelta-selective ligand L165041 being the most potent. The PPARalpha- and PPARgamma-selective ligands Wy14643 and BRL49653 had negligible effect on involucrin expression, whereas a dose-dependent induction was observed with L165041. Simultaneous addition of L165041 and BRL49653 synergistically induced strong involucrin expression. Additionally, L165041 potently induced CD36 mRNA expression. Administration of tetradecylthioacetic acid resulted in a dramatic decrease in proliferation and a robust upregulation of the expression of involucrin and transglutaminase. Our results indicate that tetradecylthioacetic acid may affect keratinocyte gene expression and differentiation via PPAR-dependent and PPAR-independent pathways, and that the latter play an important role.

Adult↗

Integrative biology and the developing limb bud.

The identification or selective construction of mutations within genes has allowed researchers to explore the downstream effects of gene disruption. Although these approaches have been successful, a limitation in our assessment of the consequences of conditional changes, and thereby our understanding of roles or function of genes, limits the degree to which we examine the effects of our manipulations. It is also clear that linear associations are incorrect models for describing development, and newer methods now give us an opportunity to practice an integrative biology. In our attempts to explore the consequences of Hoxa13 disruption in mice and humans, it has become clear that a better understanding of the consequences of gene alteration may be achievable by taking a broader approach with a long-term view. Fundamental questions regarding Hox gene function in vertebrates, including those related to the number of target genes; the degree of overlap of target gene regulation among paralogs; the magnitude of modulation exerted; and the identity of genes that are activated versus repressed need to be explored if a more thorough mechanistic understanding is to be achieved. To begin to address these questions, we undertook a comprehensive analysis of the expression of genes within developing limb buds of mice, and here we present some of our preliminary results. Our efforts will further (1) the exploration of the broader genetic relationships of expressed genes, (2) the determination of parallels or variations in target usage for a given gene in different tissues and between different organisms, (3) the evaluation of limb patterning mechanisms in other animal model systems, and (4) the exploration of gene expression hierarchies regulated by HOX proteins in developmental systems.

Animals↗

Reporting the potential benefits of new technologies for cervical cancer screening.

The AHCPR released its evidence-based report, "Evaluation of Cervical Cytology" in early 1999. This report represents the most comprehensive analysis available to date of Pap smears and new technologies designed to improve cervical cancer screening. Both the ACOG and the AHCPR have released simplified summaries of the results of the evidence report that may lead to misunderstandings of the potential clinical impact of these new technologies. This report reviews the 4 major statements in these summaries and discusses how they are either incorrect based on the full AHCPR report or may be misinterpreted because their ramifications are not fully discussed. New screening technology has the potential to finally bring the sensitivity of a new Pap test to an acceptable level. The increased sensitivity afforded by these new technologies can reduce negative outcomes at reasonable cost-effectiveness ratios and at an equivalent or superior specificity compared to the conventional Pap smear.

Journal Article↗

Cellular gene expression survey of PseudoRabies Virus (PRV) infected Human Embryonic Kidney cells (HEK-293).

Pseudorabies virus (PRV) is an alpha herpesvirus that causes Aujezsky disease in the pig. To characterize the impact of PRV infection on cellular expression, we used microarrays consisting of 9850 oligonucleotides corresponding to human genes and examined the expression levels of mRNA isolated 0.5, 3, 6, and 9 h post infection (hpi) from cultures of infected HEK-293 cells. Very few changes were observed during the first 3 h of infection but significant modifications in the cell expression of more than 1000 genes were clearly apparent by 6 hpi. More than 2400 genes were either up- or down-regulated during the 9 h experiment. These results were then analyzed using gene ontology and the MAPP and MAPPFinder software. This comprehensive analysis clearly shows that the down-regulated genes were mainly involved in macromolecular synthesis (DNA, RNA and proteins) and the cell cycle. The up-regulated genes primarily concerned the regulation of DNA transcription, developmental processes (central nervous system development, neurogenesis, angiogenesis), cell adhesion and potassium transport. This study is the first qualitative analysis of a gene expression survey in a human cell line following PRV infection. It demonstrates global changes in the cell expression profile, and identifies the main biological processes that are altered during virus replication.

Animals↗

Red blood cell fatty acid profile of chronic renal failure patients receiving maintenance haemodialysis treatment.

The fatty acid profile of chronic renal failure (CRF) patients on maintenance haemodialysis treatment (MHT) is abnormal and results point towards an essential fatty acid (EFA) deficiency. However, controversies still exist as to which of the essential fatty acids (EFAs) or EFA-products are decreased. In this study, the results of a comprehensive analysis of the fatty acids, performed on the red blood cells of 14 CRF patients on MHT, are presented. The red blood cell membrane fatty acids determined in this study include a range of saturated fatty acids (SFAs), mono-unsaturated fatty acids (MUFAs) and poly-unsaturated fatty acids (PUFAs). Results confirmed the suggested presence of an essential fatty acid deficiency in CRF patients on MHT. It showed the total content of the n-6 fatty acids (31.66 +/- 3.21 vs 34.67 +/- 2.05), as well as the total content of the PUFAs (37.22 +/- 4.08 vs 40.93 +/- 2.35), to be significantly decreased. The total MUFA content was, in contrast, significantly increased (16.87 +/- 0.91 vs 15.49 +/- 1.18). The EFA deficiency profile seen in this study points towards that of a chronic inflammatory condition. This is borne out by the fact that all three precursors of the eicosanoids--the mediators of various inflammatory and immune responses--were reduced in the presence of an increase in MUFAs as well as SFAs. The possibility of the fatty acid profile of CRF patients on MHT being that of a chronic inflammatory condition is supported by the fact that continuous complement-dependent and complement-independent immune activation, due to bio-incompatibility between blood cells and the dialysis membranes, is known to occur during the dialysis process.

Adolescent↗

[Deletion of chromosome 10q--a marker for metastasis of head-neck carcinomas?].

BACKGROUND: Comparative genomic hybridization (CGH) was applied to squamous cell carcinomas of the head and neck to define genetic alterations that are associated with the metastatic phenotype. METHODS: CGH is a molecular cytogenetic method allowing the comprehensive analysis of a tumor genome for chromosomal imbalances. In total, 23 primary squamous cell carcinomas without evidence of metastasis formation and 20 lymph node metastases were investigated. RESULTS: Prevalent changes observed in more than 50% of the primary tumors included deletions on chromosomes 3p, 4p/q, 5q, 6q, 9p, 11q, 13q, and 18q, and DNA overrepresentations on chromosomes 1p, 3q, 5p, 8q, 9q, 11q13, 16p, 17q, 19p, 20q, and 22q. To evaluate the differences between both groups we used a histogram representation, calculation of a difference histogram, and statistical analysis. The analysis revealed that the lymph node metastases were frequently characterized by deletions on chromosomes 10, 11, and 14. In particular, DNA loss of the chromosomal bands 5p12, 10p11.2-12, 10q21, 10q22-23, 10q24-26, 11p13-14, 11q24-25, and 14q22-24 were significantly associated with metastases formation. The statistical analysis indicated that particularly the deletions on chromosome 10q were highly significant markers for the incidence of lymph node metastases. CONCLUSION: Our data indicate that tumor phenotypes are determined by patterns of chromosomal alterations, and that 10q deletions may predict the metastatic phenotype in head and neck squamous cell carcinomas.

Biomarkers, Tumor↗

Single versus repeated-course antenatal corticosteroids: outcomes in singleton and multiple-gestation pregnancies.

The objectives of this study are to compare the neonatal risks and benefits of antenatal single-course versus repeated-course corticosteroids in singleton and multiple-gestation pregnancies. A comprehensive analysis was performed of the inpatient records of all neonates admitted to our center from 1 January 1994 through 31 May 1999. The primary outcome measure was survival without chronic lung disease (CLD). Secondary outcome measures included birth weight; head circumference; interval weight ratios; respiratory disease severity; intraventricular hemorrhage rate and severity; severe retinopathy of prematurity; early infection; and hospital days. All singletons 27-32 completed weeks' gestation, and multiples 26-32 weeks' gestation, whose mothers had received betamethasone before delivery, were included. One hundred and fifteen singleton and 53 multiple-gestation infants (total 168) were stratified by multiplicity, gestational-age (< or =29 or > or =30 weeks), and number of steroid courses. Repeated courses of antenatal betamethasone were not associated with greater survival without CLD, in either singleton- or multiple-gestation infants. In singletons there was no difference in any outcome measure between groups. In multiples, the only difference was greater postnatal weight gain in the lower gestation group. Mean birth head circumference was smaller in repetitively-treated singletons < or =29 weeks. There are no clinically significant neonatal benefits of repeated-course antenatal steroids in singletons > or =27 weeks estimated gestational age (EGA) or multiple-gestation infants > or =26 weeks EGA. Prospective randomized trials of single-course versus repetitive antenatal corticosteroid therapy are warranted.

Adult↗

Toll-like receptor 2 and Toll-like receptor 4 expression in human adrenals.

Toll-like receptors (TLRs) are key elements in the innate immune response, functioning as pattern-recognition receptors for the detection and response to endotoxins and other microbial ligands. Inflammatory cytokines play an important role in the activation of the hypothalamic-pituitary-adrenal HPA axis during inflammation and sepsis. The newly recognized major role of TLR2 and TLR4 and the adrenal stress response during critical illnesses such as inflammation and sepsis demand comprehensive analysis of their interactions. Therefore, we analyzed TLR2 and TLR4 expression in human adrenal glands. Western blot analysis demonstrated the expression of TLR2 and TLR4 in the human adrenocortical cell line NCI-H295. Immunohistochemical analysis of normal human adrenal glands revealed TLR2 and TLR4 expression in the adrenal cortex, but not in the adrenal medulla. Considering the crucial role of the HPA axis and the innate immune response during acute sepsis or septic shock, elucidating the functional interaction of these systems should be of great clinical relevance.

Adrenal Cortex↗

Genetic aspects of hepatocellular carcinogenesis.

Hepatocellular carcinoma (HCC) is linked etiologically to viruses (hepatitis B virus [HBV] and hepatitis C virus [HCV]), chemical carcinogens (i.e., aflatoxins), and other environmental and host factors causing chronic liver injury. Some hepatoblastomas may be linked to inherited gene mutations, but adult hereditary HCC appears to be rare. HCCs display gross genomic alterations, including DNA rearrangements associated with HBV DNA integration, loss of heterozygosity, and, less importantly, chromosomal amplifications and loss of imprinting. Many genes with somatic mutations have now been identified in these tumors. Most frequently involved genes are tumor suppressor genes such as p53, M6P/IGF2R, beta-catenin, p16INK4A, and retinoblastoma genes. Most identified mutations are somatic, but germline mutations of p16INK4A, APC, and BRCA2 have also been reported. Oncogenic activation of several cellular genes such as cyclin D and cyclin A have been described in HCC, but the possible implication of candidate viral oncogenes (i.e., X protein of HBV) is still debated. A comprehensive analysis of all the genetic changes described for HCC demonstrates that at least four different growth regulatory pathways are altered in these tumors. However, each pathway appears to be implicated in a limited fraction of these tumors, suggesting that HCCs are genetically heterogenous neoplasms. This genetic heterogeneity correlates with the heterogeneity of etiologic factors implicated in HCC.

Adult↗

[Stress on the Achilles tendon in Alpine skiing].

In an attempt to gain more systematic knowledge of the biomechanical mechanism involved in the genesis of injuries caused by a fall during skiing, we undertook a comprehensive analysis of more than 100 experimental ski falls simulated under controlled conditions. The well-known relation between speed, release time of the ski binding and injury risk of the lower extremity was confirmed by our experimental results. The EMG investigations revealed the central importance of the musculus triceps surae, and hence of the Achilles tendon, during a fall. The results of our biomechanical model indicate that the present method of setting the ski binding according to the tibial strength, is rather questionable, to say the least.

Achilles Tendon↗

[Concepts for measuring quality of life in patients with chronic sinusitis].

BACKGROUND: In 1997, Benninger developed the Rhinosinusitis Disability Index (RSBI) for patients who suffered from chronic sinusitis. Its content related validity and construct related validity were established, as were its sensitivity and reliability. The aim of our study is to introduce a concept to measure quality of life in these patients according to circumstances prevalent in Germany. METHODS: The questionnaire (Rhinusitis-Beeinträchtigungs-Index, RSBI) contains 30 questions that describe the 5 aspects of quality of life. Moreover we used a standardized data sheet to acquire further information about history and diagnostic results. Using this method it is possible to specificity more precisely the patients current complaints and condition. Evaluation is possible according to complex of questions as well as simple items. The influence of treatment on quality of life can be measured as the sum of the specific scores and is expressed comprehensively in the total score. The concept of the study includes a prospective inquiry as well as before and after surgical intervention. CONCLUSION: A disadvantage of the American RSDI is that the clinical symptoms are not described in a detailed catalogue which allows the correlation of the clinical factors. Therefore we added a detailed questionnaire concerning the clinical symptoms and the individual treatment of the patient (RSBI). That permits comprehensive analysis of quality of life as it relates to different aspects of disease and different strategies in treatment.

Chronic Disease↗

[Genetic screening of head-neck carcinomas using comparative genomic hybridization (CGH)].

BACKGROUND: Comparative Genomic Hybridization (CGH) is a novel cytogenetic method that allows the comprehensive analysis of a tumor genome for DNA gains and losses. METHODS: CGH was performed on genomic DNA extracted from 14 primary head and neck squamous cell carcinomas. Equal amounts of biotin-labeled tumor DNA and digoxigenin-labeled normal reference DNA were hybridized to normal metaphase chromosomes. The tumor DNA was visualized with fluorescein (FITC) and the normal DNA with rhodamine (TRITC) and detected in a fluorescence microscope. The signal intensities of the different fluorochromes were quantitated as gray levels along the single chromosomes. The over-and underrepresented DNA segments were quantified by computation of FITC/TRITC ratio images and average ratio profiles. RESULTS: Consensus deletion regions were most frequently observed on chromosome arms 3p (14 cases), 9p (11), 13q (10), 18q (10), 5q (9), 4q (9), 4p (7), 11q (7), 6q (6), 8p (6), and 11p (6). Copy number increases were identified for chromosomes 3q (11), 16p (9), 17q (9), 19p (8), 19q (7), 22 (7), 1p (6), 8q (6), 9q (6), and 20q (6). Particularly, the 3q isochromosome formation (3p loss/3q amplification), found in 10 cases, was a basic alteration. In addition, 11 tumors showed a 11 q13 amplification. CONCLUSION: CGH analysis allows the identification of recurrent genetic alterations in head and neck squamous cell carcinomas that will be associated with specific tumor phenotypes like metastatic behavior and prognosis.

Adult↗