José M. López

Collagenase 3 is a target of Cbfa1, a transcription factor of the runt gene family involved in bone formation.

ABSTRACT Collagenase 3 (MMP-13) is a recently identified member of the matrix metalloproteinase (MMP) gene family that is expressed at high levels in diverse human carcinomas and in articular cartilage from arthritic patients. In addition to its expression in pathological conditions, collagenase 3 has been detected in osteoblasts and hypertrophic chondrocytes during fetal ossification. In this work, we have evaluated the possibility that Cbfa1 (core binding factor 1), a transcription factor playing a major role in the expression of osteoblastic specific genes, is involved in the expression of collagenase 3 during bone formation. We have functionally characterized a Cbfa motif present in the promoter region of collagenase 3 gene and demonstrated, by cotransfection experiments and gel mobility shift assays, that this element is involved in the inducibility of the collagenase 3 promoter by Cbfa1 in osteoblastic and chondrocytic cells. Furthermore, overexpression of Cbfa1 in osteoblastic cells unable to produce collagenase 3 leads to the expression of this gene after stimulation with transforming growth factor β. Finally, we show that mutant mice deficient in Cbfa1, lacking mature osteoblasts but containing hypertrophic chondrocytes which are also a major source of collagenase 3, do not express this protease during fetal development. These results provide in vivo evidence that collagenase 3 is a target of the transcriptional activator Cbfa1 in these cells. On the basis of these transcriptional regulation studies, together with the potent proteolytic activity of collagenase 3 on diverse collagenous and noncollagenous bone and cartilage components, we proposed that this enzyme may play a key role in the process of bone formation and remodeling.

Stochastic analysis of periodic real-time systems

This paper describes a stochastic analysis method for general periodic real-time systems. The proposed method accurately computes the response time distribution of each task in the system, thus making it possible to determine the deadline miss probability of individual tasks, even for systems with maximum utilization factor greater than one. The method uniformly covers both fixed-priority scheduling (such as rate monotonic) as well as dynamic-priority scheduling (such as earliest deadline first) and can handle arbitrary relative deadlines and execution time distributions. The accuracy of the method is proven by comparing the results from the analysis with those obtained from simulations, as well as other methodologies in the literature.

Worst-case utilization bound for EDF scheduling on real-time multiprocessor systems

Presents the utilization bound for earliest deadline first (EDF) scheduling on homogeneous multiprocessor systems with partitioning strategies. Assuming that tasks are pre-emptively scheduled on each processor according to the EDF algorithm, and allocated according to the first-fit (FF) heuristic, we prove that the worst-case achievable utilization is 0.5(n+1), where n is the number of processors. This bound is valid for arbitrary utilization factors. Moreover, if all the tasks have utilization factors under a value /spl alpha/, the previous bound is raised, and the new utilization bound considering /spl alpha/ is calculated. In addition, we prove that no uniprocessor scheduling algorithm/allocation algorithm pair can provide a higher worst-case achievable utilization than that of EDF-FF. Finally, simulation provides the average-case achievable utilization for EDF-FF.

Utilization Bounds for EDF Scheduling on Real-Time Multiprocessor Systems

The utilization bound for earliest deadline first (EDF) scheduling is extended from uniprocessors to homogeneous multiprocessor systems with partitioning strategies. First results are provided for a basic task model, which includes periodic and independent tasks with deadlines equal to periods. Since the multiprocessor utilization bounds depend on the allocation algorithm, different allocation algorithms have been considered, ranging from simple heuristics to optimal allocation algorithms. As multiprocessor utilization bounds for EDF scheduling depend strongly on task sizes, all these bounds have been obtained as a function of a parameter which takes task sizes into account. Theoretically, the utilization bounds for multiprocessor EDF scheduling can be considered a partial solution to the bin-packing problem, which is known to be NP-complete. The basic task model is extended to include resource sharing, release jitter, deadlines less than periods, aperiodic tasks, non-preemptive sections, context switches, and mode changes.

A regulatory cascade involving retinoic acid, Cbfa1, and matrix metalloproteinases is coupled to the development of a process of perichondrial invasion and osteogenic differentiation during bone formation

Tissue-remodeling processes are largely mediated by members of the matrix metalloproteinase (MMP) family of endopeptidases whose expression is strictly controlled both spatially and temporally. In this article, we have examined the molecular mechanisms that could contribute to modulate the expression of MMPs like collagenase-3 and MT1-MMP during bone formation. We have found that all-trans retinoic acid (RA), which usually downregulates MMPs, strongly induces collagenase-3 expression in cultures of embryonic metatarsal cartilage rudiments and in chondrocytic cells. This effect is dose and time dependent, requires the de novo synthesis of proteins, and is mediated by RAR-RXR heterodimers. Analysis of the signal transduction mechanisms underlying the upregulating effect of RA on collagenase-3 expression demonstrated that this factor acts through a signaling pathway involving p38 mitogen-activated protein kinase. RA treatment of chondrocytic cells also induces the production of MT1-MMP, a membrane-bound metalloproteinase essential for skeletal formation, which participates in a proteolytic cascade with collagenase-3. The production of these MMPs is concomitant with the development of an RA-induced differentiation program characterized by formation of a mineralized bone matrix, downregulation of chondrocyte markers like type II collagen, and upregulation of osteoblastic markers such as osteocalcin. These effects are attenuated in metatarsal rudiments in which RA induces the invasion of perichondrial osteogenic cells from the perichondrium into the cartilage rudiment. RA treatment also resulted in the upregulation of Cbfa1, a transcription factor responsible for collagenase-3 and osteocalcin induction in osteoblastic cells. The dynamics of Cbfa1, MMPs, and osteocalcin expression is consistent with the fact that these genes could be part of a regulatory cascade initiated by RA and leading to the induction of Cbfa1, which in turn would upregulate the expression of some of their target genes like collagenase-3 and osteocalcin.

An exact stochastic analysis of priority-driven periodic real-time systems and its approximations

This paper describes a stochastic analysis framework which computes the response time distribution and the deadline miss probability of individual tasks, even for systems with a maximum utilization greater than one. The framework is uniformly applied to fixed-priority and dynamic-priority systems and can handle, tasks with arbitrary relative deadlines and execution time distributions.

Different bone growth rates are associated with changes in the expression pattern of types II and X collagens and collagenase 3 in proximal growth plates of the rat tibia.

Skeletal growth depends on endochondral ossification in growth plate cartilage, where proliferation of chondrocytes, matrix synthesis, and increases in chondrocyte size all contribute to the final length of a bone. To learn more about the potential role of matrix synthesis/degradation dynamics in the determination of bone growth rate, we investigated the expression of matrix collagens and collagenase 3 in tibial growth plates in three age groups of rats (21, 35, and 80 days after birth), each characterized by specific growth rates. By combining stereological and in situ hybridization techniques, it was found that the expression of matrix collagens and collagenase 3 was specifically turned on or off at specific stages of the chondrocyte‐differentiation cycle, and these changes occurred as a temporal sequence that varied depending of animal growth rate. Furthermore, the expression of these matrix proteins by a growth plate chondrocyte was found to be sped up or slowed down depending of the growth rate. In addition to expression of types II and X collagen, collagenase‐3 expression was found to constitute a constant event in the series of changes in gene expression that takes place during the chondrocyte‐differentiation process. Collagenase‐3 expression was found to show a biphasic pattern: it was intermittently expressed at the proliferative phase and uniformly expressed at the hypertrophic stage. An intimate relationship between morphological and kinetic changes associated with chondrocyte hypertrophy and changes in the expression pattern of matrix collagens and collagenase 3 was observed. Present data prove that the matrix synthesis/degradation dynamics of the growth plate cartilage varied depending on growth rate; these results support the hypothesis that changes in matrix degradation and synthesis are a critical link in the sequence of tightly regulated events that lead to chondrocytic differentiation.

Stochastic analysis of real-time systems under preemptive priority-driven scheduling

Exact stochastic analysis of most real-time systems under preemptive priority-driven scheduling is unaffordable in current practice. Even assuming a periodic and independent task model, the exact calculation of the response time distribution of tasks is not possible except for simple task sets. Furthermore, in practice, tasks introduce complexities such as release jitter, blocking in shared resources, etc., which cannot be handled by the periodic independent task set model.In order to solve these problems, exact analysis must be abandoned for an approximated analysis. However, in the real-time field, approximations must not be optimistic, i.e. the deadline miss ratios predicted by the approximated analysis must be greater than or equal to the exact ones. In order to achieve this goal, the concept of pessimism needs to be mathematically defined in the stochastic context, and the pessimistic properties of the analysis carefully derived.This paper provides a mathematical framework for reasoning about stochastic pessimism, and obtaining mathematical properties of the analysis and its approximations. This framework allows us to prove the safety of several proposed approximations and extensions. We analyze and solve some practical problems in the implementation of the stochastic analysis, such as the problem of the finite precision arithmetic or the truncation of the probability functions. In addition, we extend the basic model in several ways, such as the inclusion of shared resources, release jitter or non-preemptive sections.

Minimum and maximum utilization bounds for multiprocessor rate monotonic scheduling

The utilization bound for real-time rate monotonic (RM) scheduling on uniprocessors is extended to multiprocessors with partitioning-based scheduling. This allows fast schedulability tests to be performed on multiprocessors and quantifies the influence of key parameters, such as the number of processors and task sizes on the schedulability of the system. The multiprocessor utilization bound is a function of the allocation algorithm, so among all the allocation algorithms there exists at least one allocation algorithm providing the minimum multiprocessor utilization bound, and one allocation algorithm providing the maximum multiprocessor utilization bound. We prove that the multiprocessor utilization bound associated with the allocation heuristic worst fit (WF) coincides with that minimum if we use Liu and Laylands bound (LLB) as the uniprocessor schedulability condition. In addition, we present a class of allocation algorithms sharing the same multiprocessor utilization bound which coincides with the aforementioned maximum using LLB. The heuristics first fit decreasing (FFD) and best fit decreasing (BFD) belong to this class. Thus, not even an optimal allocation algorithm can guarantee a higher multiprocessor utilization bound than that of FFD and BFD using LLB. Finally, the pessimism of the multiprocessor utilization bounds is estimated through extensive simulations.

Utilization Bounds for Multiprocessor Rate-Monotonic Scheduling

In this paper, we extend Liu and Laylands utilization bound for fixed priority scheduling on uniprocessors to homogeneous multiprocessor systems under a partitioning strategy. Assuming that tasks are pre-emptively scheduled on each processor according to fixed priorities assigned by the Rate-Monotonic policy, and allocated to processors by the First Fit algorithm, we prove that the utilization bound is (n−1)(21/2−1)+(m−n+1)(21/(m−n+1)−1), where m and n are the number of tasks and processors, respectively. This bound is valid for arbitrary utilization factors. Moreover, if all the tasks have utilization factors under a value α, the previous bound is raised and the new utilization bound considering α is calculated. Finally, simulation provides the average-case behavior.