Interleukin-1 is essential for systemic inflammatory bone loss.

OBJECTIVES: Chronic inflammation is a major risk factor for systemic bone loss leading to osteoporotic fracture and substantial morbidity and mortality. Inflammatory cytokines, particularly tumour necrosis factor (TNF) and interleukin-1 (IL1), are thought to play a key role in the pathogenesis of inflammation-induced bone loss, but their exact roles are yet to be determined. METHODS: To determine whether TNF directly triggers bone loss or requires IL1, human TNFalpha mice (hTNFtg) were crossed with mice lacking IL1alpha and IL1beta (IL1(-/-)hTNFtg). Systemic bone architecture was evaluated using CT scanning, static and dynamic bone histomorphometry and serum markers of bone metabolism. RESULTS: hTNFtg mice developed severe bone loss accompanied by a severe distortion of bone microarchitecture. Bone trabeculae were thinner and decreased in numbers, resulting in increased trabecular separation. Histomorphometric analyses revealed strongly increased bone resorption in hTNFtg mice compared with wild-type mice. In contrast, IL1(-/-)hTNFtg mice were fully protected from systemic bone loss despite still developing inflammation in their joints. Lack of IL1 completely reversed increased osteoclast formation and bone resorption in hTNFtg mice and the increased levels of RANKL in these mice. Structural parameters and osteoclast and osteoblast numbers were indistinguishable from wild-type mice. CONCLUSIONS: These data indicate that IL1 is essential for TNF-mediated bone loss. Despite TNF-mediated inflammatory arthritis, systemic bone is fully protected by the absence of IL1, which suggests that IL1 is an essential mediator of inflammatory osteopenia.

Selective p38MAPK isoform expression and activation in antineutrophil cytoplasmatic antibody-associated crescentic glomerulonephritis: role of p38MAPKalpha.

OBJECTIVE: Crescentic glomerulonephritis (crGN) is a frequent and life-threatening manifestation of antineutrophil cytoplasmatic antibody-associated vasculitis. Up-regulation of proinflammatory cytokines contributes to renal damage by activation of p38 mitogen-activated protein kinases (MAPKs). However, it is unclear which of the four p38MAPK isoforms are expressed, activated and hence of major importance in crGN. METHODS: Kidney biopsies of patients with antineutrophil cytoplasmatic antibody-positive crGN and control samples were investigated for the expression and phosphorylation of p38MAPK isoforms and downstream target kinase MAPKAP2 by immunohistochemistry. Expression and functional activation of p38MAPK isoforms by TNF was also assessed in a human podocyte cell line by reverse transcription-polymerase chain reaction, immunoblotting and kinase array. RESULTS: Strong expression of p38MAPKalpha, beta and gamma isoforms was found in glomerular podocytes and crescents. Infiltrating leucocytes showed predominant p38MAPKalpha expression. Activation of p38MAPK and its downstream mediator MAPKAP2 was found in crGN confined to glomerular podocytes, crescents and inflammatory infiltrates. Interestingly, corticosteroid treatment before kidney biopsy diminished p38MAPK activation in crGN. Activated p38MAPK co-localised with alpha, beta and gamma isoforms in podocytes and crescents, while leucocytes showed mainly p38MAPKalpha activation. In a human podocyte cell line mRNA and protein of all four p38MAPK isoforms was expressed but only p38MAPKalpha was activated upon challenge with TNF. CONCLUSIONS: This study shows selective p38MAPK isoform expression and activation in crGN. Podocytes and podocyte-induce crescent formation is the main source of p38MAPK activation in crGN. TNF is a potent and selective activator of the alpha-isoform in podocytes, which therefore appears as a main contributor to proinflammatory signalling in the glomerulum of crGN.

Characterization of the nuclear import of human MutLalpha.

DNA mismatch repair (MMR) is essential for the maintenance of replication fidelity. Its major task is to recognize mismatches as well as insertion/deletion loops of newly synthesized DNA strands. Although different players of human MMR have been identified, the regulation of essential steps of MMR is poorly understood. Because MMR is initiated in the nucleus, nuclear import might be a mechanism to regulate MMR. Nuclear targeting is accomplished by conserved signal sequences called nuclear localization signals (NLS), which represent clusters of positively charged amino acids (aa). hMLH1 contains two clusters of positively charged amino acids, which are candidate NLS sequences (aa 469-472 and 496-499), while hPMS2 contains one (aa 574-580). To study the effect of these clusters on nuclear import, NLS mutants of hMLH1 and hPMS2 were generated and expressed in 293T cells. The subcellular localization of the mutant constructs was monitored by confocal laser microscopy. We demonstrated that missense mutations of two signal sequences, one in hMLH1 and one in hPMS2, lead to impaired nuclear import, which was especially prominent for mutants of the hMLH1 residues K471 and R472; and hPMS2 residues K577 and R578.

Indicators of quality of in-patient psychiatric treatment: the patients' view.

OBJECTIVES: The object of this study was to find out from psychiatric in-patients which aspects of care and treatment they considered important and how satisfied they were with these. DESIGN: One hundred and ninety-four in-patients were asked to rate the importance of, and their satisfaction with, 22 different aspects of in-patient care and treatment. The questionnaire, developed for the purpose of the study after a pilot phase including professional care givers and patients, contained 92 items and was returned by 52% of all discharged patients from the Psychiatric university hospital during a 3-month period. RESULTS: Patients made a clear distinction between aspects of treatment they considered important and aspects they were satisfied with. Ranked of highest importance were various therapeutic relationships, and respect for their rights and privileges. They were satisfied with their relationships with clinical staff but dissatisfied with medication. CONCLUSION: A well differentiated assessment of importance and satisfaction has implications for the evaluation of the quality of psychiatric care, for specific methods of treatment, and for the improvement of in-patient psychiatric care.

Delay of growth and development in children with bronchial asthma, atopic dermatitis and allergic rhinitis.

The elevated incidence of short stature (body height < (-)x - 2s), skeletal retardation and delayed puberty in children with bronchial asthma or atopic dermatitis is generally attributed to the severity of the disorder. However, a series of findings indicate a causal influence of the atopy and the existence of atopic skeletal retardation per se.The observation that children with atopic disorders, whether bronchial asthma, atopic dermatitis or allergic rhinitis, exhibit a rate of short stature that is twice to five times higher than normal indicates atopic and thus genetically determined influences. The elevated prevalence of short stature associated with allergic rhinitis is especially significant, as this disorder cannot be included among the severe chronic disorders. The fact that skeletal retardation is more prevalent in boys than in girls by a ratio of about 2:1 and that a significantly more marked retardation of bone maturation is found in atopic in comparisons with non-atopic asthmatics also lend support to this postulation. The clinical relevance of atopic growth retardation is also supported by the close interaction of pathophysiological basal mechanisms of bone metabolism and the atopy status. Thus the local growth factor prostaglandin E(2) (PGE(2)), which is important for bone metabolism, is also a messenger substance for the immediate and late allergic reaction. The platelet-activating factor (PAF), as one of the strongest mediators in the pathogenesis of allergic disorders, influences the PGE(2) synthesis in the osteoblasts. These relationships show that atopy-dependent imbalances in the complex system of local and systemic growth factors can certainly lead to disturbance of skeletal maturation which may delay growth and development in atopic children. In order to verify these assumptions it is necessary to research the interaction of local growth factors (particularly the roles of PGE(2), PAF and IGF I) in the skeletons of children of short stature suffering from atopic disorders. This should also include the possible effects on the overall hormonal factors influencing bone maturation. Atopy should be included in the differential diagnosis programme to clarify growth and development disturbances.

[Investigation of growth prognosis in asthmatic children]. / Untersuchungen zur Wachstumsprognose asthmakranker Kinder.

The objective of the present study was the ascertainment of correct data related to growth prognosis in asthmatic children. For this purpose, 210 young asthmatics including 153 males and 57 females ranging in age from 1.7 to 18.9 years were evaluated. Data were specified for the parameters: chronological age (CA); bone age (KA); height age (LA); height (KH); severity of illness; target adult height (GZG) and predicted adult height (PEG). Among this group, 5.9 % of the boys and 3.5 % of the girls, were found to be of small stature (KH < - 2 s). This corresponds to about 2.5 and 1.5 times the average in a normal distribution. Children with the highest severity of illness showed the strongest negative deviation for the KH-SDS values (boys - 0.59 plus minus 1.1, girls - 0.97 plus minus 0.8). This proved to be statistically significant for females. Development of CA, KA and LA continued unremarkably until the age of 4 years. From age 5, partially a significant growth retardation of KA and LA could be observed in both sexes. To age 16, KA values and, after that LA values, were more strongly affected. Comparison of the GZG and PEG average values within the degree of severity groups made it clear that the PEG values for both sexes, independent of the severity of illness, were without exception, smaller than the GZG. The differences of both parameters proved to be statistically significant for the group with the most severe symptoms of the boys (180.8 plus minus 6.5 cm to 175.8 plus minus 6.2 cm, p < 0.01) and for the entire groups of both sexes (boys 180.2 plus minus 5.3 cm to 178.6 plus minus 7.3 cm, p < 0.05, girls 167.7 plus minus 4.4 cm to 165.8 plus minus 6.8 cm, p < 0.05). However, a separate comparison of GZG and PEG average values, respectively, among the severity groups showed no significant differences. The results argue for a growth-inhibiting influence from bronchial asthma. The cause has to be severity of illness but direct effects of the atopy on skeletal development is also taken in consideration. Confirmation of these findings requires investigation of a larger group of asthmatics and clarification of the pathophysiological processes in the growing skeletons of atopic children.

Apoptosis regulators and their role in tumorigenesis.

It has become clear that, together with deregulated growth, inhibition of programmed cell death (PCD) plays a pivotal role in tumorigenesis. In this review, we present an overview of the genes and mechanisms involved in PCD. We then summarize the evidence that impaired PCD is a prerequisite for tumorigenesis, as indicated by the fact that more and more neoplastic mutations appear to act by interfering with PCD. This has made the idea of restoration of corrupted 'death programs' an intriguing new area for potential cancer therapy.

Group choice: the ideal free distribution of human social behavior.

Group choice refers to the distribution of group members between two choice alternatives over time. The ideal free distribution (IFD), an optimal foraging model from behavioral ecology, predicts that the ratio of foragers at two resource sites should equal the ratio of obtained resources, a prediction that is formally analogous to the matching law of individual choice, except that group choice is a social phenomenon. Two experiments investigated the usefulness of IFD analyses of human group choice and individual-based explanations that might account for the group-level events. Instead of nonhuman animals foraging at two sites for resources, a group of humans chose blue and red cards to receive points that could earn cash prizes. The groups chose blue and red cards in ratios in positive relation to the ratios of points associated with the cards. When group choice ratios and point ratios were plotted on logarithmic coordinates and fitted with regression lines, the slopes (i.e., sensitivity measures) approached 1.0 but tended to fall short of it (i.e., undermatching), with little bias and little unaccounted for variance. These experiments demonstrate that an IFD analysis of group choice is possible and useful, and suggest that group choice may be explained by the individual members' tendency to optimize reinforcement.

Molar versus as a paradigm clash.

The molar view of behavior arose in response to the demonstrated inadequacy of explanations based on contiguity. Although Dinsmoor's (2001) modifications to two-factor theory render it irrefutable, a more basic criticism arises when we see that the molar and molecular views differ paradigmatically. The molar view has proven more productive.

An improved procedure for the generation of recombinant single-chain Fv antibody fragments reacting with human CD13 on intact cells.

A procedure was developed to generate recombinant single chain Fv (scFv) antibody fragments reacting with the extracellular domain of human cell surface antigen CD13 (hCD13; aminopeptidase N) on intact cells. Membrane fractions prepared from a stably transfected hCD13-positive murine NIH/3T3 cell line were used to immunize BALB/c mice, with the intention that hCD13 would be the major immunogenic molecule recognized by the immune system. Spleen RNA from the immunized mice served to generate a combinatorial scFv phage display library. The library was adsorbed against non-transfected NIH/3T3 or Sf21 insect cells to eliminate nonrelevant binders. The supernatant was then used for panning with either hCD13-transfected Sf21 insect cells or a hCD13-expressing human leukemia-derived cell line. Therefore, the key concepts of the procedure were the presentation of hCD13 as the sole human antigen on murine NIH/3T3 cells and a screening strategy where hCD13 was the major common antigen of the material used for immunization and panning. Two different hCD13-reactive phages were isolated and the soluble scFvs were expressed in E. coli and purified. The two scFvs, anti-hCD13-1 and anti-hCD13-3, differed at four amino acid positions in their V(H) regions and both had high affinities for hCD13 as determined by surface plasmon resonance (K(D)=7 and 33x10(-10) M, respectively). Both efficiently recognized hCD13 on intact cells. Therefore, the procedure allowed the production of high affinity scFvs reacting with a desired antigen in its native conformation without requiring extensive purification of the antigen and should be useful for the preparation of scFvs against other conformation-sensitive cell-surface antigens.

Gp130 and ras mediated signaling in human plasma cell line INA-6: a cytokine-regulated tumor model for plasmacytoma.

INTRODUCTION: Cytokines of the gp130-family, particularly interleukin(IL)-6, play a crucial role in the propagation of malignant plasma cells. MATERIALS AND METHODS: The role of IL-6 and other gp130-cytokines was studied in the human plasma cell line INA-6 in vitro and in INA-6 xenografts. The proliferative response to gp130-cytokines was evaluated and activated components of gp130-signaling pathways were identified by Western blotting and DNA binding studies. Specifically, expression of IL-6 and receptors for IL-6 and leukemia inhibitory factor were analysed by RT-PCR and ELISA. RESULTS: The plasma cell line INA-6 was cultured for several years remaining strictly dependent on exogenous IL-6. Other gp130-cytokines had no significant effect on INA-6 cell proliferation in vitro. Due to an activating mutation in the N-ras gene, mitogen-activated protein kinases (MAPK) were constitutively phosphorylated. In contrast, signal transducer and activator of transcription(STAT)-3 activation was dependent on stimulation with IL-6. Blocking of either one of these pathways resulted in a significant decrease of INA-6 cell proliferation. Remarkably, INA-6 xenografts did not require exogeneous IL-6 for proliferation in vivo. Instead, an autocrine IL-6 loop and, in certain tumor sublines, responsiveness to additional gp130-cytokines was induced during in vivo growth. CONCLUSION: Activation of the gp130 signal transducer is mandatory for INA-6 cell growth in vitro and in vivo. Both the MAPK and the Jak/STAT pathway are operative in malignant plasma cells and either one is essential for plasma cell growth. The INA-6 cell line provides a preclinical model to study growth regulation of human plasmacytoma cells and to evaluate novel therapeutic strategies.

A novel protein (Fbf-1) that binds to CD95/APO-1/FAS and shows sequence similarity to trichohyalin and plectin.

The Fas/Apo-1/CD95 cell surface receptor belongs to the TNF receptor family of cell death inducing molecules. A number of cytosolic adapter proteins that mediate signal transduction of CD95 have been characterized, but some features of the molecular mechanisms of CD95-induced cell death remain elusive. We describe here a novel protein that can interact with the cytosolic domain of the murine CD95 receptor in a yeast two-hybrid assay. This novel protein was termed Fbf-1 for Fas binding factor and bears no sequence similarity to the known CD95 adapter proteins. Fbf-1 is 1173 aa long and has a theoretical molecular weight of around 130 kDa. The protein is expressed in a wide variety of tissues and is localized in the cytoplasm. Fbf-1 is a very hydrophilic protein, highly conserved between mouse and human and bears a carboxyterminal leucine heptad repeat reminiscent of leucine zipper protein interaction domains. In addition, it shows sequence similarity to trichohyalin and plectin pointing to a function as a structural protein.

Choice in a variable environment: every reinforcer counts.

Six pigeons were trained in sessions composed of seven components, each arranged with a different concurrent-schedule reinforcer ratio. These components occurred in an irregular order with equal frequency, separated by 10-s blackouts. No signals differentiated the different reinforcer ratios. Conditions lasted 50 sessions, and data were collected from the last 35 sessions. In Part 1, the arranged overall reinforcer rate was 2.22 reinforcers per minute. Over conditions, number of reinforcers per component was varied from 4 to 12. In Part 2, the overall reinforcer rate was six per minute, with both 4 and 12 reinforcers per component. Within components, log response-allocation ratios adjusted rapidly as more reinforcers were delivered in the component, and the slope of the choice relation (sensitivity) leveled off at moderately high levels after only about eight reinforcers. When the carryover from previous components was taken into account, the number of reinforcers in the components appeared to have no systematic effect on the speed at which behavior changed after a component started. Consequently, sensitivity values at each reinforcer delivery were superimposable. However, adjustment to changing reinforcer ratios was faster, and reached greater sensitivity values, when overall reinforcer rate was higher. Within a component, each successive reinforcer from the same alternative ("confirming") had a smaller effect than the one before, but single reinforcers from the other alternative ("disconfirming") always had a large effect. Choice in the prior component carried over into the next component, and its effects could be discerned even after five or six reinforcement and nonreinforcement is suggested.

[Mendelson syndrome in infancy and childhood]. / Das Mendelson-Syndrom im Säuglings- und Kindesalter.

Acute pulmonary failure caused by gastric acid aspiration is designated as Mendelson's syndrome. It is characterized by a trias of symptoms comprising bronchial obstruction, pulmonary oedema, and right ventricular failure. The pathomorphological pulmonary alterations show the typical symptoms of ARDS and allow the differentiation of three phases. The initial phase of injury is characterized by cauterization of the bronchial and alveolar epithelium. It is followed by the exsudative second phase during which alveolar oedema are developing. They impair the pulmonary surfactant synthesis and the formation of hyaline membranes. Fibrosis processes are typical of the proliferative third phase. Every of the mentioned three phases may be classified by their corresponding clinical symptoms. The therapy is entirely symptomatic and follows the intensive medical standards of the ARDS-therapy.

[Effect of the beta-2 mimetic drug terbutaline of growth hormone secretion in prepubertal asthmatic children]. / Zum Einfluss des Beta-2-Mimetikums Terbutalin auf die Wachstumshormonsekretion präpubertärer asthmakranker Kinder.

Asthmatics display a tendency to retarded growth and hyposomia in childhood. The reasons for this are not yet clear, although the atopic disposition seems to occupy a key role. It is a known fact that stimulation of the beta-2 receptors results in inhibiting growth hormone secretion. The purpose of our study was to find out whether the beta-2 mimetic terbutalin, often used in asthma therapy, exercises a negative influence on the spontaneous release of growth hormone in children suffering from asthma. The growth hormone release was studied in 10 prepuberal children suffering from atopic asthma who received intravenous therapeutic doses of terbutalin: testing was done for a total period of 24 hours before and during administration. Terbutalin effected significant inhibition of growth hormone secretion merely during the waking phase (6.2 +/- 1.0 to 3.7 +/- 0.7 ng/ml), but not during the sleep phase (13.1 +/- 1.8 to 12.5 +/- 2.0 ng/ml) and the 24-hour period (11.0 +/- 1.0 to 9.8 +/- 1.5 ng/ml). There was also no significant influence on the average group value for the maximum growth hormone peak (40.8 +/- 9.5 to 42.7 +/- 11.0 ng/ml). These results point to a short-term inhibition of growth hormone secretion, exercised by intravenously administered terbutalin. Terbutalin does not seem to be responsible for any clinically relevant inhibition of growth and development.

Therapy with CD7 monoclonal antibody TH-69 is highly effective for xenografted human T-cell ALL.

Human T-cell acute lymphocytic leukaemia (ALL) was established in athymic nude or severe combined immunodeficient (SCID) mice by injecting CEM or MOLT-16 cells. When nude mice bearing approx. 2 g of tumour were treated with a single injection of CD7 antibody TH-69, 82.6% reached complete remission within 10 d whereas 13.0% showed partial remission. Similarly, in SCID mice with advanced disease a significant prolongation of survival was seen. The therapeutic effects were dependent upon dose and affinity of the antibody. TH-69 is a high-affinity antibody (7.6 x 10(9) M-1) that rapidly induced modulation during treatment. The Fc-portion of the antibody was required for effective tumour cell killing. Complement deposition was found on tumour sections after TH-69 treatment and in part may account for tumour destruction. There was no evidence for antibody-dependent cellular cytotoxicity (ADCC). The kinetics of tumour disappearance suggested the initiation of a programmed cell death (PCD), despite the lack of significant DNA fragmentation. Unmodified high-affinity antibodies to the T-cell antigen CD7 have potential for T-cell ALL therapy.