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.

Choice in free-ranging wild pigeons.

A flock of free-ranging wild pigeons were trained to peck at buttons which, when operated, allowed brief access to grain. Although only one bird at a time could have access to the buttons, the pecks of the group were treated as an aggregate. When they chose between two buttons, each of which could occasionally produce grain, the ratios of pecks at the buttons approximately equaled the ratios of the grain presentations obtained from them. This accords with a relation well substantiated in the laboratory, the matching law. It suggests that the matching law may apply to the behavior of higher organisms in natural environments.

Preliminary results from the viking orbiter imaging experiment.

During its first 30 orbits around Mars, the Viking orbiter took approximately 1000 photographic frames of the surface of Mars with resolutions that ranged from 100 meters to a little more than 1 kilometer. Most were of potential landing sites in Chryse Planitia and Cydonia and near Capri Chasma. Contiguous high-resolution coverage in these areas has led to an increased understanding of surface processes, particularly cratering, fluvial, and mass-wasting phenomena. Most of the surfaces examined appear relatively old, channel features abound, and a variety of features suggestive of permafrost have been identified. The ejecta patterns around large craters imply that fluid flow of ejecta occurred after ballistic deposition. Variable features in the photographed area appear to have changed little since observed 5 years ago from Mariner 9. A variety of atmospheric phenomena were observed, including diffuse morning hazes, both stationary and moving discrete white clouds, and wave clouds covering extensive areas.

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.

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.

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.

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.

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.

Time-based and count-based measurement of preference.

Rats' pressing on two levers was reinforced according to two independent variable-interval schedules that were varied during the experiment. Since the levers were connected directly to the programming equipment, bypassing the standard pulseformers, reinforcement could occur while a lever was held down. Although the time a lever was pressed might, therefore, have varied independently of number of presses, these two measures covaried substantially, because the average duration of the presses remained roughly constant. This rough invariance may have resulted from the rats' tendency to make bursts of brief presses (i.e., to jiggle the levers), even though the contingencies encouraged holding. When duration did vary, presses on the two levers tended to vary together. As a result, relative time spent pressing corresponded closely to relative number of presses. Both of these measures conformed well to the matching law. Absolute behavioral frequency at a lever, measured either way, varied directly with proportion of reinforcement for that lever, in accordance with the generalized version of the matching law. Number of presses seemed, on balance, to be a slightly more reliable measure than pressing time. The substantial interchangeability may prove more significant than the slight disparity, however, because it supports the notion that all behavior can be measured on a common scale of time.

Time allocation in human vigilance.

Three human subjects detected unpredictable signals by pressing either of two telegraph keys. The relative frequencies with which detections occurred for the two alternatives were varied. The procedure included a changeover delay and response cost for letting go of a key. All subjects matched the relative time spent holding each key to the relative number of detections for that key, in conformity with the matching law. One subject's performance, which at first deviated from the relation, came into conformity with it when response cost was increased. Another subject's performance approximated matching more closely when the changeover delay was increased. The results confirm and extend the notions that choice consists in time allocation and that all behavior can be measured on the common scale of time.

On two types of deviation from the matching law: bias and undermatching.

DATA ON CHOICE GENERALLY CONFORM CLOSELY TO AN EQUATION OF THE FORM: log(B(1)/B(2))=a log(r(1)/r(2)+log k, where B(1) and B(2) are the frequencies of responding at Alternatives 1 and 2, r(1) and r(2) are the obtained reinforcement from Alternatives 1 and 2, and a and k are empirical constants. When a and k equal one, this equation is equivalent to the matching relation: B(1)/B(2)=r(1)/r(2). Two types of deviation from matching can occur with this formulation: a and k not equal to one. In some experiments, a systematically falls short of one. This deviation is undermatching. The reasons for undermatching are obscure at present. Some evidence suggests, however, that factors favoring discrimination also favor matching. Matching (a=1) may represent the norm in choice when discrimination is maximal. When k differs from one, its magnitude indicates the degree of bias in choice. The generalized matching law predicts that bias should take this form (adding a constant proportion of responding to the favored alternative). Data from a variety of experiments indicate that it generally does.

Chained concurrent schedules: reinforcement as situation transition.

Pigeons' pecks at two white response keys (initial-link situation) occasionally turned both keys red (terminal-link situation). When the two keys were red, pecks occasionally produced food, after which the keys were again white. In both situations, a changeover delay prevented the response-produced outcome from immediately following a change of responding from either key to the other. In the initial-link situation, the ratio of pecks at the keys closely paralleled the ratio of transitions into the terminal-link situation produced by the pecks, conforming to the well-known matching relation. In the terminal-link situation, the peck ratios deviated from the matching relation toward indifference. Overall response rate and rate of changeover were generally higher in the terminal-link situation than in the initial-link situation. The finding of matching in the initial-link situation supports a definition of reinforcement as situation transition. The differences in performance between the two situations, viewed in the light of other recent findings, suggest that the effects of a changeover delay depend on the overall reinforcing value of the choice alternatives.

Performances on ratio and interval schedules of reinforcement: Data and theory.

TWO DIFFERENCES BETWEEN RATIO AND INTERVAL PERFORMANCE ARE WELL KNOWN: (a) Higher rates occur on ratio schedules, and (b) ratio schedules are unable to maintain responding at low rates of reinforcement (ratio "strain"). A third phenomenon, a downturn in response rate at the highest rates of reinforcement, is well documented for ratio schedules and is predicted for interval schedules. Pigeons were exposed to multiple variable-ratio variable-interval schedules in which the intervals generated in the variable-ratio component were programmed in the variable-interval component, thereby "yoking" or approximately matching reinforcement in the two components. The full range of ratio performances was studied, from strained to continuous reinforcement. In addition to the expected phenomena, a new phenomenon was observed: an upturn in variable-interval response rate in the midrange of rates of reinforcement that brought response rates on the two schedules to equality before the downturn at the highest rates of reinforcement. When the average response rate was corrected by eliminating pausing after reinforcement, the downturn in response rate vanished, leaving a strictly monotonic performance curve. This apparent functional independence of the postreinforcement pause and the qualitative shift in response implied by the upturn in variable-interval response rate suggest that theoretical accounts will require thinking of behavior as partitioned among at least three categories, and probably four: postreinforcement activity, other unprogrammed activity, ratio-typical operant behavior, and interval-typical operant behavior.

The correlation-based law of effect.

It is commonly understood that the interactions between an organism and its environment constitute a feedback system. This implies that instrumental behavior should be viewed as a continuous exchange between the organism and the environment. It follows that orderly relations between behavior and environment should emerge at the level of aggregate flow in time, rather than momentary events. These notions require a simple, but fundamental, change in the law of effect: from a law based on contiguity of events to a law based on correlation between events. Much recent research and argument favors such a change. If the correlation-based law of effect is accepted, it favors measures and units of analysis that transcend momentary events, extending through time. One can measure all consequences on a common scale, called value. One can define a unit of analysis called the behavioral situation, which circumscribes a set of values. These concepts allow redefinition of reinforcement and punishment, and clarification of their relation to discriminative stimuli.

Time allocation and negative reinforcement.

Pigeons' standing on one or the other side of a chamber was reinforced with timeout from electric shock on two concurrent variable-interval schedules. For two pigeons, the ratio of time spent on the left to time spent on the right approximately matched the ratio of timeouts obtained on the left to timeouts obtained on the right. The data of two other birds deviated from this relation, although in opposite directions. Overall, the results suggest that reduction in rate of electric shock plays a role in behavioral allocation analogous to that played by rate of positive reinforcement. It appears possible to describe aversive control and positive control within the same conceptual framework-that provided by the matching relation.

In search of the feedback function for variable-interval schedules.

Finding a theoretically sound feedback function for variable-interval schedules remains an important unsolved problem. It is important because interval schedules model a significant feature of the world: the dependence of reinforcement on factors beyond the organism's control. The problem remains unsolved because no feedback function yet proposed satisfies all the theoretical and empirical requirements. Previous suggestions that succeed in fitting data fail theoretically because they violate a newly recognized theoretical requirement: The slope of the function must approach or equal 1.0 at the origin. A function is presented that satisfies all requirements but lacks any theoretical justification. This function and two suggested by Prelec and Herrnstein (1978) and Nevin and Baum (1980) are evaluated against several sets of data. All three fitted the data well. The success of the two theoretically incorrect functions raises an empirical puzzle: Low rates of reinforcement are coupled with response rates that seem anomalously high. It remains to be discovered what this reflects about the temporal patterning of operant behavior at low reinforcement rates. A theoretically and empirically correct function derived from basic assumptions about operant behavior also remains to be discovered.

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.