VEGF production by osteoarthritic chondrocytes cultured in micromass and stimulated by IL-17 and TNF-alpha

We compared the secretion of vascular endothelial growth factor (VEGF) by chondrocytes isolated from cartilage of patients with osteoarthritis (OA) and maintained in monolayer or in three-dimensional culture. Chondrocytes, immediately after isolation, or after one passage in monolayer culture (subculture), were seeded in monolayer or pelleted in micromasses. Cells cultured differently were immunoassayed for the secretion of VEGF in basal conditions or after stimulation with IL-17, TNF-alpha or B IL-17+TNF-alpha. Chondrocytes cultured in micromasses secreted lower levels of VEGF in comparison with those in monolayer culture. In micromass cultures TNF-alpha was more stimulating than IL-17 and their combined effect was additional. On the basis of alkaline phosphatase/cathespin B activities, primary cultures in micromass, characterized by low VEGF secretion, represented the best condition for maximal cell differentiation. Our results suggest that the culture of chondrocytes in micromasses represents a good condition to study the physiology of these cells to a maximum degree when micromasses are obtained with freshly isolated chondrocytes.     

Advances in adult asthma 2006: its risk factors, course, and management

This Advances article updates our understanding of risk factors for asthma and its course and management. Studies relevant to clinical practice are discussed, with special attention to their clinical research methods.     

Influence of substratum surface chemistry/energy and topography on the human fetal osteoblastic cell line hFOB 1.19: Phenotypic and genotypic responses observed in vitro

Time-dependent phenotypic response of a model osteoblast cell line (hFOB 1.19, ATCC, and CRL-11372) to substrata with varying surface chemistry and topography is reviewed within the context of extant cell-adhesion theory. Cell-attachment and proliferation kinetics are compared using morphology as a leading indicator of cell phenotype. Expression of (alpha2, alpha3, alpha4, alpha5, alphav, beta1, and beta3) integrins, vinculin, as well as secretion of osteopontin (OP) and type I collagen (Col I) supplement this visual assessment of hFOB growth. It is concluded that significant cell-adhesion events-contact, attachment, spreading, and proliferation-are similar on all surfaces, independent of substratum surface chemistry/energy. However, this sequence of events is significantly delayed and attenuated on hydrophobic (poorly water-wettable) surfaces exhibiting characteristically low-attachment efficiency and long induction periods before cells engage in an exponential-growth phase. Results suggest that a 'time-cell-substratum-compatibility-superposition principle' is at work wherein similar bioadhesive outcomes can be ultimately achieved on all surface types with varying hydrophilicity, but the time required to arrive at this outcome increases with decreasing cell-substratum-compatibility. Genomic and proteomic tools offer unprecedented opportunity to directly measure changes in the cellular machinery that lead to observed cell responses to different materials. But for the purpose of measuring structure-property relationships that can guide biomaterial development, genomic/proteomic tools should be applied early in the adhesion/spreading process before cells have an opportunity to significantly remodel the cell-substratum interface, effectively erasing cause and effect relationships between cell-substratum-compatibility and substratum properties. IMPACT STATEMENT: This review quantifies relationships among cell phenotype, substratum surface chemistry/energy, topography, and cell-substratum contact time for the model osteoblast cell line hFOB 1.19, revealing that genomic/proteomic tools are most useful in the pursuit of understanding cell adhesion if applied early in the adhesion/spreading process.     

Multilevel models for repeated measures research designs in psychophysiology: an introduction to growth curve modeling

Psychophysiologists often use repeated measures analysis of variance (RMANOVA) and multivariate analysis of variance (MANOVA) to analyze data collected in repeated measures research designs. ANOVA and MANOVA are nomothetic approaches that focus on group means. Newer multilevel modeling techniques are more informative than ANOVA because they characterize both group-level (nomothetic) and individual-level (idiographic) effects, yielding a more complete understanding of the phenomena under study. This article was written as an introduction to growth curve modeling for applied researchers. A growth model is defined that can be used in place of RMANOVAs and MANOVAs for single-group and mixed repeated measures designs. The model is expanded to test and control for the effects of baseline levels of physiological activity on stimulus-specific responses. Practical, conceptual, and statistical advantages of growth curve modeling are discussed.     

Role of precise spike timing in coding of dynamic vibrissa stimuli in somatosensory thalamus

Rats discriminate texture by whisking their vibrissae across the surfaces of objects. This process induces corresponding vibrissa vibrations, which must be accurately represented by neurons in the somatosensory pathway. In this study, we investigated the neural code for vibrissa motion in the ventroposterior medial (VPm) nucleus of the thalamus by single-unit recording. We found that neurons conveyed a great deal of information (up to 77.9 bits/s) about vibrissa dynamics. The key was precise spike timing, which typically varied by less than a millisecond from trial to trial. The neural code was sparse, the average spike being remarkably informative (5.8 bits/spike). This implies that as few as four VPm spikes, coding independent information, might reliably differentiate between 10⁶ textures. To probe the mechanism of information transmission, we compared the role of time-varying firing rate to that of temporally correlated spike patterns in two ways: 93.9% of the information encoded by a neuron could be accounted for by a hypothetical neuron with the same time-dependent firing rate but no correlations between spikes; moreover, 93.4% of the information in the spike trains could be decoded even if temporal correlations were ignored. Taken together, these results suggest that the essence of the VPm code for vibrissa motion is firing rate modulation on a submillisecond timescale. The significance of such a code may be that it enables a small number of neurons, firing only few spikes, to convey distinctions between very many different textures to the barrel cortex.     

Marshmallows used as saliva stimulant do not affect cortisol concentrations: finally a palatable alternative for toddler saliva collection

Two studies were conducted to validate marshmallows as a saliva stimulant for use with toddlers. First, cortisol concentrations from 14 subjects (ages 6-46 years) were compared using three saliva collection methods: (1) plain cotton dental roll, (2) dental roll with one mini-marshmallow, and (3) expectorating into a collection tube using no cotton or stimulant. EIA was used for analyses. There were no significant differences among cortisol concentrations. Second, saliva collection compliance rate was compared for 21-month-olds (n = 51) using either flavored drink crystal- (compliance rate = 16.7%) or marshmallow-flavored (compliance rate = 60%) dental rolls for saliva collection (chi(2) (1) = 4.02, p = .045). These studies indicate that marshmallow is a viable option for saliva stimulation to determine toddler cortisol concentrations using EIA.