Relationships between fetal body weight of Wistar rats at term and the extent of skeletal ossification

We investigated the relationship between fetal body weight at term (pregnancy day 21) and the extent of ossification of sternum, metacarpus, metatarsus, phalanges (proximal, medial and distal) of fore- and hindlimbs and cervical and coccygeal vertebrae in Wistar rats. The relationships between fetal body weight and sex, intrauterine position, uterine horn, horn size, and litter size were determined using historical control data (7594 fetuses; 769 litters) of untreated rats. Relationships between body weight and degree of ossification were examined in a subset of 1484 historical control fetuses (154 litters) which were subsequently cleared and stained with alizarin red S. Fetal weight was independent of horn size, uterine horn side (left or right) or intrauterine position. Males were heavier than females and fetal weight decreased with increasing litter size. Evaluation of the skeleton showed that ossification of sternum, metacarpus and metatarsus was extensively complete and independent of fetal weight on pregnancy day 21. In contrast, the extent of ossification of fore- and hindlimb phalanges and of cervical and sacrococcygeal vertebrae was dependent on fetal body weight. The strongest correlation between body weight and degree of ossification was found for hindlimb, medial and proximal phalanges. Our data therefore suggest that, in full-term rat fetuses (day 21), reduced ossification of sternum, metacarpus and metatarsus results from a localized impairment of bone calcification (i.e., a malformation or variation) rather than from general growth retardation and that ossification of hindlimb (medial and proximal) phalanges is a good indicator of treatment-induced fetal growth retardation.

Calcaneal apophysitis: a quantitative radiographic evaluation of the secondary ossification center

Background: Calcaneal apophysitis in children is a self-limited condition that may interfere with walking and physical performance in sports, thus causing concern to the patient and parents. There is still controversy about the significance of the radiographic changes in children with heel pain, since the report of Sever in 1912. One of the reasons is that normal children may display a considerable variation in the radiographic aspects of the secondary ossification center of the calcaneus at different ages. Methods: In this investigation, the developmental aspects of primary and secondary ossification centers of the calcaneus were studied in radiographs obtained from healthy boys and from boys with calcaneal apophysitis. The normal population comprised 392 children and adolescents ranging in age from 6 to 15 years. There were 69 individuals with calcaneal apophysitis ranging in age from 8 to 14 years. Lateral standard radiographs were obtained of both heels, and a copper step wedge was used as a calibration to determine bone density. The following parameters were analyzed on the plain films: time of appearance, fusion and number of fragments of the secondary nucleus, area and bone densitometry of the primary and secondary ossification centers of the calcaneus. Results: In the normal population, the ossification of the secondary nucleus began at 7 years of age, and at 15 years of age, the nucleus was fused in all individuals. In the apophysitis group, the secondary ossification center was present and not fused in all individuals. Both secondary nuclei increased in size with age with no difference between the two groups. Regarding bone density, both the primary and secondary nuclei were less dense in the apophysitis group than their counterparts in the normal population. The most significant difference between the two populations referred to the degree of fragmentation, which was greater in the apophysitis group. Conclusion: Our data showed that the sclerotic aspect of the secondary nucleus of the calcaneus is a normal feature and, therefore, should not be used to establish the diagnosis of Sever's disease. The most consistent difference between the normal and apophysitis group was related to the more fragmented aspect of the secondary nucleus in the latter individuals, which may suggest a mechanical etiology for that condition.

Bayesian Networks for the Age Classification of Living Individuals: a Study on Transition Analysis

Over the past few decades, age estimation of living persons has represented a challenging task for many forensic services worldwide. In general, the process for age estimation includes the observation of the degree of maturity reached by some physical attributes, such as dentition or several ossification centers. The estimated chronological age or the probability that an individual belongs to a meaningful class of ages is then obtained from the observed degree of maturity by means of various statistical methods. Among these methods, those developed in a Bayesian framework offer to users the possibility of coherently dealing with the uncertainty associated with age estimation and of assessing in a transparent and logical way the probability that an examined individual is younger or older than a given age threshold. Recently, a Bayesian network for age estimation has been presented in scientific literature; this kind of probabilistic graphical tool may facilitate the use of the probabilistic approach. Probabilities of interest in the network are assigned by means of transition analysis, a statistical parametric model, which links the chronological age and the degree of maturity by means of specific regression models, such as logit or probit models. Since different regression models can be employed in transition analysis, the aim of this paper is to study the influence of the model in the classification of individuals. The analysis was performed using a dataset related to the ossifications status of the medial clavicular epiphysis and results support that the classification of individuals is not dependent on the choice of the regression model.

Description du développement épiphysaire du tarse et du grasset équin à l’IRM et au CT : un pas vers la compréhension de l’OCD

L’ostéochondrite disséquante (OCD) est un défaut focal du processus d’ossification endochondrale en des sites spécifiques au niveau épiphysaire. Elle est caractérisée par la présence de fragments ostéochondraux pouvant se détacher de la surface articulaire. Cette maladie a un impact majeur sur les performances athlétiques des chevaux. Les deux hypothèses principales présentement véhiculées quant à sa pathogénie sont une nécrose ischémique du cartilage de croissance et une altération du métabolisme de la matrice de collagène de type II au sein du cartilage de croissance. Malgré de nombreuses années de recherche sur le sujet, plusieurs aspects de cette maladie demeurent inconnus. L’objectif de cette étude était de décrire le développement épiphysaire équin au niveau du membre pelvien à l’aide de l’imagerie médicale afin de déterminer si des variations du processus de maturation à certains sites pouvaient être un facteur prédisposant au développement de lésions d’OCD. Des membres pelviens de fœtus et de jeunes poulains ont été étudiés post-mortem. L’épiphyse du fémur distal, tibia distal et du talus ont été examinées par tomodensitométrie (CT) et résonnance magnétique 1.5 Tesla (IRM) dans le but de documenter le degré et le patron d’ossification, la régularité du front d’ossification, de même que le pourcentage du diamètre épiphysaire demeurant occupé par le complexe de cartilage articulaire-épiphysaire, et ce au niveau de certains sites prédéterminés. Les centres secondaires d’ossification (SOCs) ont été détectés pour la première fois à 7 mois de gestation (MOG) au niveau de l’épiphyse fémorale distale et à 8 MOG au niveau de l’épiphyse tibiale distale et du talus. À 8-9 MOG la lèvre latérale de la trochlée fémorale, la malléole médiale du tibia (MM) et la partie crâniale de la crête intermédiaire du tibia distal (DIRT(Cr)), tous des sites prédisposés à la maladie, avaient le plus haut pourcentage de cartilage de tous les sites évalués. Post-partum, le pourcentage de cartilage de la MM et de la DIRT(Cr) sont demeurés importants. Le CT et l’IRM ont su illustrer le développement épiphysaire équin et soutenir d’avantage le fait qu’un cartilage plus épais à certains sites articulaires pourrait avoir un rôle dans le développement de lésions d’OCD.

Morfologia das 3ª e 4ª vértebras cervicais representativa do surto de crescimento puberal

OBJETIVOS: determinar a morfologia das 3ª e 4ª vértebras cervicais representativa dos estágios de pré-pico, pico e pós-pico de velocidade de crescimento estatural, definidos previamente pelos centros de ossificação do primeiro dedo em radiografias carpais ou do dedo polegar. MÉTODOS: foram utilizadas 120 telerradiografias em norma lateral de 106 pacientes selecionados da clínica de Ortodontia Preventiva e Interceptiva da Faculdade de Odontologia de Araçatuba-UNESP e Profis/HRAC-USP que apresentavam as correspondentes radiografias carpais ou do dedo polegar. As telerradiografias foram divididas em três grupos de 40 radiografias, de acordo com os estágios maturacionais pré-pico, pico e pós-pico, definidos pela imagem do primeiro dedo. A morfologia dos corpos das 3ª e 4ª vértebras cervicais foi determinada por dois examinadores devidamente calibrados em dois tempos diferentes. CONCLUSÕES: concluiu-se que o formato retangular horizontal com borda inferior reta foi representativo do estágio de pré-pico, independentemente da vértebra analisada. Já o formato retangular horizontal com borda inferior curva, especialmente se encontrado em C4, ou o formato quadrado com borda inferior reta caracterizaram o pico de velocidade de crescimento. O formato quadrado ou, principalmente, o retangular vertical com borda inferior curva determinaram o estágio de pós-pico de velocidade de crescimento da adolescência.

Comparative analysis between three methods of bone estimating age in individuals with down syndrome by mode of the hand and wrist ray

The wrist and hand region has been the most commonly used for estimating age and osseous development due to the great number of ossification centers. The aim was to determine which method, Tanner & Whitehouse's (TW3), Greulich & Pyle's (GP) or Eklof & Ringertz's, more closely relates to the chronological age in subjects with Down syndrome with chronological ages between 61 and 180 months, using wrist and hand radiographs. The sample consisted of 85 radiographs, 52 of males and 33 of females. Eklof & Ringertz's method was computerized (Radiomemory). Greulich & Pyle's atlas was used and compared with the wrist and hand radiographs. For the TW3 method, 13 ossification centers were evaluated; for each one of them, there are seven or eight development stages to which scores are assigned; these scores are then added and the results are transformed into osseous age values. No statistically significant differences were observed between the male and female genders for methods TW3 and GP, contrasting with the observed differences for the Eklof & Ringertz method. Correlation (r2) between osseous and chronological ages was 0.8262 for TW3 and 0.7965 for GP, while for the method of Eklof & Ringertz, it was 0.7656 for females and 0.8353 for males. The author concluded that the osseous age assessment method that better related to the chronological age was the TW3, followed by Greulich & Pyle's and Eklof & Ringertz's.

Morfologia das 3ª e 4ª vértebras cervicais representativa do surto de crescimento da adolescência

Pós-graduação em Odontologia - FOA

Análise comparativa entre três métodos de estimativa da idade óssea em indivíduos com Síndrome de Down, por meio de radiografias de mão e punho

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Conditional expression of Sry, and Wnt4 by gene-targeting: Studies in sexual and skeletal development

Sry and Wnt4 cDNAs were individually introduced into the ubiquitously-expressed Rosa26 ( R26) locus by gene targeting in embryonic stem (ES) cells to create a conditional gene expression system in mice. In the targeted alleles, expression of these cDNAs should be blocked by a neomycin resistance selection cassette that is flanked by loxP sites. Transgene expression should be activated after the blocking cassette is deleted by Cre recombinase. ^ To test this conditional expression system, I have bred R26-stop- Sry and R26-stop-Wnt4 heterozygotes with a MisRII-Cre mouse line that expresses Cre in the gonads of both sexes. Analysis of these two types of bigenic heterozygotes indicated that their gonads developed normally like those of wild types. However, one XX R26-Sry/R26-Sry; MisR2-Cre/+ showed epididymis-like structures resembling those of males. In contrast, only normal phenotypes were observed in XY R26-Wnt4/R26-Wnt4; MisR2-Cre /+ mice. To interpret these results, I have tested for Cre recombinase activity by Southern blot and transcription of the Sry and Wnt4 transgenes by RT-PCR. Results showed that bigenic mutants had insufficient activation of the transgenes in their gonads at E12.5 and E13.5. Therefore, the failure to observe mutant phenotypes may have resulted from low activity of MisR2-Cre recombination at the appropriate time. ^ Col2a1-Cre transgenic mice express Cre in differentiating chondrocytes. R26-Wnt4; Col2a1-Cre bigenic heterozygous mice were found to exhibit a dramatic alteration in growth presumably caused by Wnt4 overexpression during chondrogenesis. R26-Wnt4; Col2a1-Cre mice exhibited dwarfism beginning approximately 10 days after birth. In addition, they also had craniofacial abnormalities, and had delayed ossification of the lumbar vertebrate and pelvic bones. Histological analysis of the growth plates of R26-Wnt4; Col2a1-Cre mice revealed less structural organization and a delay in onset of the primary and secondary ossification centers. Molecular studies confirmed that overexpression of Wnt4 causes decreased proliferation and early maturation of chondrocytes. In addition, R26-Wnt4; Col2a1-Cre mice had decreased expression of vascular endothelial growth factor (VEGF), suggesting that defects in vascularization may contribute to the dwarf phenotype. Finally, 9-month-old R26-Wnt4; Col2a1-Cre mice had significantly more fat cells in the marrow cavities of their metaphysis long bones, implying that long-term overexpression of Wnt4may cause bone marrow pathologies. In conclusion, Wnt4 was activated by Col2a1-Cre recombinase and was overexpressed in the growth plate, resulting in aberrant proliferation and differentiation of chondrocytes, and ultimately leads to dwarfism in mice. ^

Roles of p38 mitogen-activated protein kinase pathway in the regulation of SOX9 activity and chondrogenesis

Sox9 is a master transcription factor in chondrocyte differentiation. Several lines of evidence suggest that the p38 mitogen-activated protein kinase (MAPK) pathway is involved in chondrocyte differentiation. In the present study, we examined the roles of p38 in the regulation of SOX9 activity and chondrogenesis. ^ COS7 cells were transfected with a SOX9 expression vector and 4x48-p89, a luciferase construction harboring four tandem copies of a SOX9-dependent 48-bp enhancer in Col2a1. Coexpression of MKK6EE, a constitutively active mutant of MKK6, a MAPKK that specifically activates p38, further increased the activity of the SOX9-dependent 48-bp enhancer about 5-fold, and SOX9 protein levels were not increased under these conditions. This increase in enhancer activity was not observed in a mutant enhancer construct harboring mutations that abolish SOX9 binding. These data strongly suggested that activation of the p38 pathway results in increased activity of SOX9. In addition, the increase of the activity of the SOX9-dependent 48-bp enhancer by MKK6EE was also observed in primary chondrocytes, and this increase was abolished by coexpression of a p38 phosphatase, MKP5, and p38 specific inhibitors. Furthermore, treatment of primary chondrocytes with p38 inhibitors decreased the expression of Col2a1, a downstream target of Sox9, without affecting Sox9 RNA levels, further supporting the hypothesis that p38 plays a role in regulating Sox9 activity in chondrocytes. ^ To further study the role of the p38 MAPK pathway in chondrogenesis, we generated transgenic mice that express MKK6EE in chondrocytes under the control of the Col2a1 promoter/intron regulatory sequences. These mice showed a dwarf phenotype characterized by reduced chondrocyte proliferation and a delay in the formation of primary and secondary ossification centers. Histological analysis using in situ hybridization showed reduced expression of Indian hedgehog, PTH/PTHrP receptor, cyclin D1 and increased expression of p21. In addition, consistent with the notion that Sox9 activity was increased in these mice, transgenic mice that express MKK6EE in chondrocytes showed phenotypes similar to those of mice that overexpress SOX9 in chondrocytes. Therefore, our study provides in vivo evidence for the role of p38 in chondrocyte differentiation and suggests that Sox9 is a downstream target of the p38 MAPK pathway. ^

Variables Associated With The Performance Of Centers For Dental Specialties In Brazil.

The aim of this study was to evaluate the performance of the Centers for Dental Specialties (CDS) in the country and associations with sociodemographic indicators of the municipalities, structural variables of services and primary health care organization in the years 2004-2009. The study used secondary data from procedures performed in the CDS to the specialties of periodontics, endodontics, surgery and primary care. Bivariate analysis by χ2 test was used to test the association between the dependent variable (performance of the CDS) with the independents. Then, Poisson regression analysis was performed. With regard to the overall achievement of targets, it was observed that the majority of CDS (69.25%) performance was considered poor/regular. The independent factors associated with poor/regular performance of CDS were: municipalities belonging to the Northeast, South and Southeast regions, with lower Human Development Index (HDI), lower population density, and reduced time to deployment. HDI and population density are important for the performance of the CDS in Brazil. Similarly, the peculiarities related to less populated areas as well as regional location and time of service implementation CDS should be taken into account in the planning of these services.

Oral health in Brazil - Part II: Dental Specialty Centers (CEOs)

The concepts of health promotion, self-care and community participation emerged during the 1970s and, since then, their application has grown rapidly in the developed world, showing evidence of effectiveness. In spite of this, a major part of the population in the developing countries still has no access to specialized dental care such as endodontic treatment, dental care for patients with special needs, minor oral surgery, periodontal treatment and oral diagnosis. This review focuses on a program of the Brazilian Federal Government named CEOs (Dental Specialty Centers), which is an attempt to solve the dental care deficit of a population that is suffering from oral diseases and whose oral health care needs have not been addressed by the regular programs offered by the SUS (Unified National Health System). Literature published from 2000 to the present day, using electronic searches by Medline, Scielo, Google and hand-searching was considered. The descriptors used were Brazil, Oral health, Health policy, Health programs, and Dental Specialty Centers. There are currently 640 CEOs in Brazil, distributed in 545 municipal districts, carrying out dental procedures with major complexity. Based on this data, it was possible to conclude that public actions on oral health must involve both preventive and curative procedures aiming to minimize the oral health distortions still prevailing in developing countries like Brazil.

Estimating HIV-1 incidence using the serologic testing algorithm for recent HIV infections at HIV counseling and testing centers in the city of São Paulo, Brazil

The network of HIV counseling and testing centers in São Paulo, Brazil is a major source of data used to build epidemiological profiles of the client population. We examined HIV-1 incidence from November 2000 to April 2001, comparing epidemiological and socio-behavioral data of recently-infected individuals with those with long-standing infection. A less sensitive ELISA was employed to identify recent infection. The overall incidence of HIV-1 infection was 0.53/100/year (95% CI: 0.31-0.85/100/year): 0.77/100/year for males (95% CI: 0.42-1.27/100/year) and 0.22/100/ year (95% CI: 0.05-0.59/100/year) for females. Overall HIV-1 prevalence was 3.2% (95% CI: 2.8-3.7%), being 4.0% among males (95% CI: 3.3-4.7%) and 2.1% among females (95% CI: 1.6-2.8%). Recent infections accounted for 15% of the total (95% CI: 10.2-20.8%). Recent infection correlated with being younger and male (p = 0.019). Therefore, recent infection was more common among younger males and older females.

Prevalence of Human T-Cell Lymphotropic Virus (HTLV-1/2) in Individuals from Public Health Centers in Mozambique

The prevalence of human T-cell lymphotropic viruses types 1 and 2 (HTLV-1/2) in Mozambique is not known. The present study examined blood samples from 208, 226, and 318 individuals from Northern, Central, and Southern Mozambique, respectively, of all socioeconomic and demographic strata attending public health centers in Mozambique for HTLV-1/2-specific antibodies. Serum samples were assessed for HIV- and HTLV-1/2-specific antibodies by using enzyme immunoassays, and infections with HTLV-1 and -2 were confirmed by using Western blot. An overall HTLV-1/2 prevalence of 2.3% (2.9% in female and 1.1% in male subjects) was observed, and the prevalence of infection increased with age. Regional variation in the prevalence of HIV and HTLV-1/2 was observed; 32.2%, 65.5%, and 44% of individuals tested HIV positive in Northern, Central, and Southern Mozambique, respectively, and 2.4%, 3.9%, and 0.9% tested HTLV-1/2 positive in the same regions. HTLV-1 infection was confirmed in these individuals. No association between HTLV-1 infection and socio-demographic variables or HIV status was detected, although the low number of HTLV-1-positive cases did not allow robust statistical analyses. The results obtained suggest different risk factors and epidemiologic correlates of HIV and HTLV-1 transmission in Mozambique. Furthermore, our results suggested that North and Central Mozambique should be considered endemic regions for HTLV-1 infection. As no cases of HTLV-2 were detected, HTLV-2 appears to have not been introduced into Mozambique.

Optimization of Heat Treatment Profiles Applied to Nanometric-Scale Nb(3)Sn Wires With Cu-Sn Artificial Pinning Centers

Nb(3)Sn is one of the most used superconducting materials for applications in high magnetic fields. The improvement of the critical current densities (J(c)) is important, and must be analyzed together with the optimization of the flux pinning acting in the material. For Nb(3)Sn, it is known that the grain boundaries are the most effective pinning centers. However, the introduction of artificial pinning centers (APCs) with different superconducting properties has been proved to be beneficial for J(c). As these APCs are normally in the nanometric-scale, the conventional heat treatment profiles used for Nb(3)Sn wires cannot be directly applied, leading to excessive grain growth and/or increase of the APCs cross sections. In this work, the heat treatment profiles for Nb(3)Sn superconductor wires with Cu(Sn) artificial pinning centers in nanometric-scale were analyzed in an attempt to improve J(c) . It is described a methodology to optimize the heat treatment profiles in respect to diffusion, reaction and formation of the superconducting phases. Microstructural, transport and magnetic characterization were performed in an attempt to find the pinning mechanisms acting in the samples. It was concluded that the maximum current densities were found when normal phases (due to the introduction of the APCs) are acting as main pinning centers in the global behavior of the Nb(3)Sn superconducting wire.