The pathways by which chondrocytes of articular cartilage sense their mechanical environment are unclear. Compelling structural evidence suggests that chondrocyte primary cilia are mechanosensory organelles. This study used a 3D agarose culture model to examine the effect of compressive strain on chondrocyte cilia. Chondrocyte/agarose constructs were subjected to cyclic compression (0-15%; 1 Hz) for 0.5-48 h. Additional constructs were compressed for 48 h and allowed to recover for 72 h in uncompressed free-swelling conditions. Incidence and length of cilia labelled with anti-acetylated alpha-tubulin were examined using confocal microscopy. In free-swelling chondrocytes, these parameters increased progressively, but showed a significant decrease following 24 or 48 h compression. A 72 h recovery partially reversed this effect. The reduced cilia incidence and length were not due to increased cell division. We therefore propose that control of primary cilia length is an adaptive signalling mechanism in response to varying levels and duration of mechanical loads during joint activity.

A survival-promoting peptide has been purified from medium conditioned by Y79 human retinoblastoma cells and a mouse hippocampal cell line (HN 33.1) exposed to H 2 O 2 . A 30 residue synthetic peptide was made on the basis of N-terminal sequences obtained during purification, and it was found to exhibit gel mobility and staining properties similar to the purified molecules. The peptide maintains cells and their processes in vitro for the HN 33.1 cell line treated with H 2 O 2 , and in vivo for cortical neurons after lesions of the cerebral cortex. It has weak homology with a fragment of a putative bacterial antigen and, like that molecule, binds IgG. The peptide also contains a motif reminiscent of a critical sequence in the catalytic region of calcineurin-type phosphatases; surprisingly, like several members of this family, the peptide catalyzes the hydrolysis of para-nitrophenylphosphate in the presence of Mn 2ϩ . Application of the peptide to one side of bilateral cerebral cortex lesions centered on area 2 in rats results in an increase in IgG immunoreactivity in the vicinity of the lesions 7 d after surgery. Microglia immunopositive for IgG and ED-1 are, however, dramatically reduced around the lesions in the treated hemisphere. Furthermore, pyramidal neurons that would normally shrink, die, or disintegrate were maintained, as determined by MAP2 immunocytochemistry and Nissl staining. These survival effects were often found in both hemispheres. The results suggest that this peptide operates by diffusion to regulate the immune response and thereby rescue neurons that would usually degenerate after cortical lesions. The phosphatase activity of this molecule also suggests the potential for direct neuron survivalpromoting effects.

CONTEXT: Utilization of primary care settings offers a promising approach to enhance parenting practices that are critical for promoting early childhood development. Determining the impact of existing primary care interventions on key parenting behaviors will aid providers and policy makers as they seek strategies to improve early child outcomes. OBJECTIVE:To evaluate the efficacy of primary care-based interventions on parenting practices that promote early child development among children younger than 36 months.DATA SOURCES: PubMed, Excerpta Medica dataBASE, PsycINFO, and Cumulative Index to Nursing and Allied Health Literature databases were searched electronically.STUDY SELECTION: English-language articles that were quasi-randomized or randomized controlled trials, included parents of children <36 months of age, and reported outcomes related to parenting behaviors that promote early child development. DATA EXTRACTION:Two reviewers independently extracted data regarding participants, interventions, and outcomes. Quantitative meta-analyses were conducted with random effects for study and fitted with restricted maximum likelihood methods. RESULTS:The review included 13 studies reporting parenting outcomes in 2 categories: participation in cognitively stimulating activities and positive parent-child interactions. We found a statistically significant positive effect of primary care-delivered interventions and parent-child interactions (summary standardized mean difference 0.29, 95% confidence interval [CI] 0.06-0.52, P < .0001) and participation in cognitively stimulating activities (summary standardized mean difference 0.34, 95% CI 0.03-0.54; summary odds ratio 0.13, 95% CI 0.01-0.25, P < .0001). LIMITATIONS:Limitations included heterogeneity in measures used, outcomes, and timing of assessments.CONCLUSIONS: Primary care-based interventions modestly affect positive parenting behaviors important for early childhood development. Randomized controlled trials with comparable outcome measures using standardized assessments are needed to assess further beneficial impacts.

Mast cells are well known for their role in allergic disease and have recently been implicated in inflammatory disorders, including autoimmune arthritis, multiple sclerosis, and atherosclerosis. Although aberrant mast cell activation is the focus of many studies, much less is known about normal mast cell homeostasis. Because loss of the normal constraints on mast cell activation, proliferation, and survival may be central to disease etiology, understanding these issues warrants attention. This review summarizes the knowledge of mast cell homeostasis controlled by IgE and the regulatory cytokines IL-4, IL-10, and TGF-beta1. Because each of these proteins plays an important role in immune responses tied to mast cell-associated disease, this group represents a potential set of factors altered in atopic or autoimmune patients. It is interesting to note, for example, that polymorphisms within each of these factors or their receptors are linked to allergic disease. By first understanding how cytokines and IgE regulate mast cell function and survival, we may then predict how these factors may function in disease onset and progression.

The mast cell (MC) inflammatory response is now linked not only to atopy, but also to arthritis, multiple sclerosis, heart disease, and resistance to bacterial infection. In the current study, we demonstrate that the signal transducer and activator of transcription 5 (Stat5) is rapidly activated by IgE cross-linkage, and that its expression is critical to the MC response. Stat5-deficient (Stat5KO) MC demonstrated a significant decrease in IgE-mediated degranulation, leukotriene B4 production, cytokine secretion, and survival signals. The defect in cytokine production may be caused by decreased cytokine mRNA stability. Stat5KO MC-induced cytokine mRNAs normally following IgE cross-linkage, but these mRNAs were not sustained over time and were degraded at twice the rate observed in WT cells. Interestingly, the RNA destabilizing protein tristetraprolin was induced following IgE cross-linkage in Stat5KO but not wild-type cells. Moreover, reducing tristetraprolin expression via short hairpin RNA transfection significantly increased IL-13 production in Stat5KO MC. Our work demonstrates that Stat5 is a critical factor in IgE-induced MC activation, acting in part via posttranscriptional control of cytokine mRNA stability. These data have a direct impact on MC-associated inflammatory and autoimmune diseases.

One of the keys to ensuring high expectations for all students is the requirement for inclusive measures of educational accountability. Recognizing this need, Congress enacted Title II, National Education Reform Leadership, Standards, and Assessments, calling for the development of state assessment systems that fully include all students, as a major component of the 1994 Goals 2000: Educate America Act. At present, Kentucky is the only state in the nation that fully includes all students within a statewide educational assessment and accountability system. Students with moderate and severe cognitive disabilities participate in Kentucky's assessment system via the Alternate Portfolio. This article describes the development of Kentucky's alternate assessment, including the content and scoring standards for the Alternate Portfolio. Specific examples of entries at each grade level (4th, 8th, and 12th) are given as well. Initial implementation data, including reliability, validity, and instructional impact measures, are presented. Finally, critical issues in the development of alternate statewide assessments are discussed, with recommendations for future research efforts in this area.

Our recent studies have identified dynamic protein ensembles containing forkhead box protein 3 (FOXP3) that provide insight into the molecular complexity of suppressor T-cell activities, and it is our goal to determine how these ensembles regulate FOXP3's transcriptional activity in vivo. In this review, we summarize our current understanding of how FOXP3 expression is induced and how FOXP3 functions in vivo as a transcriptional regulator by assembling a multisubunit complex involved in histone modification as well as chromatin remodeling.

Tumor necrosis factor alpha (TNF-␣) is an important mediator of inflammation, apoptosis, and the development of secondary lymphoid structures. Multiple polymorphic microsatellites have been identified in and around the gene, and there are also multiple single-base pair biallelic polymorphisms in the introns and promoter. The TNF-␣ ؊308 promoter polymorphism is a G-to-A transition which has been statistically associated with various autoimmune disorders. Some studies have found that it may directly mediate the increased transcription of TNF-␣ in some circumstances. This study characterizes proteins interacting at the polymorphic promoter site. Affinity purification of binding proteins and confirmatory chromatin immunoprecipitation assays were used to identify the proteins. Electrophoretic mobility shift analyses and surface plasmon resonance were used to define binding characteristics. Proteins interacting at this site include GCF2/LRRFIP1 and Ets-1. GCF2/LRRFIP1 appears to act as a repressor and occupies the ؊308 site in cells that do not make TNF-␣. Cells competent to produce TNF-␣ have Ets-1 bound to the ؊308 promoter site. Active transcription is accompanied by NF-B and c-Jun binding to the proximal promoter. Thus, dynamic changes on the TNF-␣ promoter, particularly at the ؊308 site, accompany the transition from repressed to active transcription. GCF2/LRRFIP1 is the first TNF-␣ repressor identified.Tumor necrosis factor (TNF-␣) is a proinflammatory cytokine which plays a role in humoral immunity, chemokine expression, the regulation of adhesion molecule expression, and apoptosis. The expression of TNF-␣ is regulated at many levels, including transcription, message turnover, protein production, and protein release (15,21,25,36). The transcription of TNF-␣ is complex, with tissue-specific expression and stimulusinducible expression, which in turn can be tissue specific. Some of the TNF-␣ promoter motifs which have been implicated in transcription include binding sites for NF-B, NFAT, ATF-2, Ets-1, C/EBP␤, cis-acting replication element, AP-1, and AP-2 (13, 14, 24, 26-28, 35, 37, 39, 40, 48, 49, 58, 59). Several promoter polymorphisms have been identified, and a polymorphism at Ϫ308 has been implicated in the regulation of TNF-␣ transcription (24,29,52,56). (This work uses the traditional numbering system for the promoter polymorphisms [55].)The TNF-␣ promoter polymorphism at Ϫ308 involves a biallelic single-base pair transition from G to A. The polymorphic sequence, designated as TNF-␣ Ϫ308A, has gene frequencies of approximately 0.12 to 0.17 in Caucasians and 0.08 in African Americans (9,10,42,45,4). Studies have found associations of TNF-␣ Ϫ308A with systemic lupus erythematosus, sarcoidosis, alopecia areata, rheumatoid arthritis, and dermatitis herpetiformis (9,17,18,34,42,43,45,54). Other studies have found no association of TNF-␣ Ϫ308A with rheumatoid arthritis, ankylosing spondylitis, and Felty's syndrome (5, 51, 53). Certain infectious diseases, such as cerebral malaria and mucocutaneous leishmania, als...