Variable expressivity of osteogenesis imperfecta in a Brazilian family due to p.G1079S mutation in the COL1A1 gene

Osteogenesis imperfecta (OI) is a Mendelian disease with genetic heterogeneity characterized by bone fragility, recurrent fractures, blue sclerae, and short stature, caused mostly by mutations in COL1A1 or COL1A2 genes, which encode the pro-alpha 1(I) and pro-alpha 2(I) chains of type I collagen, respectively. A Brazilian family that showed variable expression of autosomal dominant OI was identified and characterized. Scanning for mutations was carried out using SSCP and DNA sequence analysis. The missense mutation c.3235G>A was identified within exon 45 of the COL1A1 gene in a 16-year-old girl diagnosed as having OI type I; it resulted in substitution of a glycine residue (G) by a serine (S) at codon 1079 (p.G1079S). The proband's mother had the disease signs, but without bone fractures, as did five of nine uncles and aunts of the patient. All of them carried the mutation, which was excluded in four healthy brothers of the patient's mother. This is the first description in a Brazilian family with OI showing variable expression; only one among seven carriers for the c.3235G>A mutation developed bone fractures, the most striking clinical feature of this disease. This finding has a significant implication for prenatal diagnosis in OI disease.

Do patients with osteogenesis imperfecta need individualized nutritional support?

Objective: Information regarding nutrition and body composition in patients diagnosed with osteogenesis imperfecta (OI) is scarce. In the present study, nutritional status, bone mineral density, and biochemical parameters of subjects with Of were evaluated. Methods: Patients with type I OI (n = 13) and type III OI (n = 13) and healthy controls (n = 8) were selected. Nutritional status and bone mineral density were assessed by a 3-d food diary and dual-energy X-ray absorptiometry at the lumbar spine, respectively. Body mass index, serum albumin, calcium, creatinine, cross-linked C-telopeptide, parathyroid hormone, and 25-hydroxivitamin D-3 were also evaluated. Results: Patients with OI had lower bone mineral density (P < 0.05 versus controls). Patients with type III OI had the highest body mass index (P < 0.05 versus patients with type I OI and controls) and the lowest lean body mass (P < 0.05 versus patients with type I OI and controls). In patients with OI, the number of fractures was positively correlated with body mass index (r = 0.581, P = 0.002) and the percentage of body fat (r = 0.451, P = 0.027) and negatively correlated to lean body mass (r = -0.523, P = 0.009). Even when taking dietary supplements, 58% and 12% of subjects with OI did not achieve the calcium and vitamin D recommendations, respectively. Conclusions: Body composition is a risk factor for bone fractures in subjects with OI. Individualized nutritional support is recommended not only to improve body composition but also to potentiate pharmacologic and physical therapies. (C) 2012 Elsevier Inc. All rights reserved.

New insights in osteogenic differentiation revealed by mass spectrometric assessment of phosphorylated substrates in murine skin mesenchymal cells

Abstract Background Bone fractures and loss represent significant costs for the public health system and often affect the patients quality of life, therefore, understanding the molecular basis for bone regeneration is essential. Cytokines, such as IL-6, IL-10 and TNFα, secreted by inflammatory cells at the lesion site, at the very beginning of the repair process, act as chemotactic factors for mesenchymal stem cells, which proliferate and differentiate into osteoblasts through the autocrine and paracrine action of bone morphogenetic proteins (BMPs), mainly BMP-2. Although it is known that BMP-2 binds to ActRI/BMPR and activates the SMAD 1/5/8 downstream effectors, little is known about the intracellular mechanisms participating in osteoblastic differentiation. We assessed differences in the phosphorylation status of different cellular proteins upon BMP-2 osteogenic induction of isolated murine skin mesenchymal stem cells using Triplex Stable Isotope Dimethyl Labeling coupled with LC/MS. Results From 150 μg of starting material, 2,264 proteins were identified and quantified at five different time points, 235 of which are differentially phosphorylated. Kinase motif analysis showed that several substrates display phosphorylation sites for Casein Kinase, p38, CDK and JNK. Gene ontology analysis showed an increase in biological processes related with signaling and differentiation at early time points after BMP2 induction. Moreover, proteins involved in cytoskeleton rearrangement, Wnt and Ras pathways were found to be differentially phosphorylated during all timepoints studied. Conclusions Taken together, these data, allow new insights on the intracellular substrates which are phosphorylated early on during differentiation to BMP2-driven osteoblastic differentiation of skin-derived mesenchymal stem cells.

Studio della patogenesi di HIV-1 nel compartimento osseo: effetti della terapia antiretrovirale convenzionale e nuovi approcci all'eradicazione

L’obiettivo della tesi è studiare il virus HIV-1 in relazione alle alterazioni sistemiche, riscontrate nel paziente HIV-infetto, in particolare alterazioni a carico del sistema scheletrico, indotte dal virus o dall’azione dei farmaci utilizzati nella terapia antiretrovirale (HAART). L’incidenza dell’osteoporosi nei pazienti HIV-positivi è drammaticamente elevata rispetto alla popolazione sana. Studi clinici hanno evidenziato come alcuni farmaci, ad esempio inibitori della proteasi virale, portino alla compromissione dell’omeostasi ossea, con aumento del rischio fratturativo. Il nostro studio prevede un follow-up di 12 mesi dall’inizio della HAART in una coorte di pazienti naïve, monitorando diversi markers ossei. I risultati ottenuti mostrano un incremento dei markers metabolici del turnover osseo, confermando l’impatto della HAART sull’omeostasi ossea. Successivamente abbiamo focalizzato la nostra attenzione sugli osteoblasti, il citotipo che regola la sintesi di nuova matrice ossea. Gli esperimenti condotti sulla linea HOBIT mettono in evidenza come il trattamento, in particolare con inibitori della proteasi, porti ad apoptosi nel caso in cui vi sia una concentrazione di farmaco maggiore di quella fisiologica. Tuttavia, anche concentrazioni fisiologiche di farmaci possono regolare negativamente alcuni marker ossei, come ALP e osteocalcina. Infine esiste la problematica dell’eradicazione di HIV-1 dai reservoirs virali. La HAART riesce a controllare i livelli viremici, ciononostante diversi studi propongono alcuni citotipi come potenziali reservoir di infezione, vanificando l’effetto della terapia. Abbiamo, perciò, sviluppato un nuovo approccio molecolare all’eradicazione: sfruttare l’enzima virale integrasi per riconoscere in modo selettivo le sequenze LTR virali per colpire il virus integrato. Fondendo integrasi e l’endonucleasi FokI, abbiamo generato diversi cloni. Questi sono stati transfettati stabilmente in cellule Jurkat, suscettibili all’infezione. Una volta infettate, abbiamo ottenuto una significativa riduzione dei markers di infezione. Successivamente la transfezione nella linea linfoblastica 8E5/LAV, che porta integrata nel genoma una copia di HIV, ha dato risultati molto incoraggianti, come la forte riduzione del DNA virale integrato.

Getting in touch-3D printing in Forensic Imaging

With the increasing use of medical imaging in forensics, as well as the technological advances in rapid prototyping, we suggest combining these techniques to generate displays of forensic findings. We used computed tomography (CT), CT angiography, magnetic resonance imaging (MRI) and surface scanning with photogrammetry in conjunction with segmentation techniques to generate 3D polygon meshes. Based on these data sets, a 3D printer created colored models of the anatomical structures. Using this technique, we could create models of bone fractures, vessels, cardiac infarctions, ruptured organs as well as bitemark wounds. The final models are anatomically accurate, fully colored representations of bones, vessels and soft tissue, and they demonstrate radiologically visible pathologies. The models are more easily understood by laypersons than volume rendering or 2D reconstructions. Therefore, they are suitable for presentations in courtrooms and for educational purposes.

Systemic response of the GH/IGF-I axis in timely versus delayed fracture healing

The GH-IGF axis has profound effects on the local and systemic regulation of bone metabolism and may be important for quality of fracture healing. To test the hypothesis that deficiency of the GH/IGF axis may play a role in the pathogenesis of fracture non-union we investigated whether alterations of serum concentrations of the GH-IGF axis could be related to failed fracture healing compared to timely fracture healing in trauma patients. Serum probes were prospectively collected from 186 patients with surgical treatment of long bone fractures up to 6 months after surgery. Samples from 14 patients with atrophic type of non-union have been compared to 14 matched patients with normal bone healing. Postoperative time courses of serum concentrations have been analyzed using commercially available chemiluminescence sandwich assays (GH), fully automated assay systems (IGF-I, IGFBP-3) or sandwich immunometric assays (ALS). Comparison between both collectives revealed significantly lower serum concentrations of GH dependent ALS during early (1st week after surgery) and of both IGFBP-3 and ALS during late stages of fracture healing (6 and 8 weeks after surgery) in non-union patients, coinciding clinically with failed fracture healing. Tendentially lower serum levels of IGF-I in the non-union group over the entire investigation period were statistically not significant. We have been able to show time courses of serum concentrations of the GH/IGF-I axis during normal and failed fracture healing in humans. An impairment of the GH/IGF-I axis might be involved in the biochemical mechanisms determining delayed or failed fracture healing.

Haptics in forensics: the possibilities and advantages in using the haptic device for reconstruction approaches in forensic science

Non-invasive documentation methods such as surface scanning and radiological imaging are gaining in importance in the forensic field. These three-dimensional technologies provide digital 3D data, which are processed and handled in the computer. However, the sense of touch gets lost using the virtual approach. The haptic device enables the use of the sense of touch to handle and feel digital 3D data. The multifunctional application of a haptic device for forensic approaches is evaluated and illustrated in three different cases: the representation of bone fractures of the lower extremities, by traffic accidents, in a non-invasive manner; the comparison of bone injuries with the presumed injury-inflicting instrument; and in a gunshot case, the identification of the gun by the muzzle imprint, and the reconstruction of the holding position of the gun. The 3D models of the bones are generated from the Computed Tomography (CT) images. The 3D models of the exterior injuries, the injury-inflicting tools and the bone injuries, where a higher resolution is necessary, are created by the optical surface scan. The haptic device is used in combination with the software FreeForm Modelling Plus for touching the surface of the 3D models to feel the minute injuries and the surface of tools, to reposition displaced bone parts and to compare an injury-causing instrument with an injury. The repositioning of 3D models in a reconstruction is easier, faster and more precisely executed by means of using the sense of touch and with the user-friendly movement in the 3D space. For representation purposes, the fracture lines of bones are coloured. This work demonstrates that the haptic device is a suitable and efficient application in forensic science. The haptic device offers a new way in the handling of digital data in the virtual 3D space.

Letrozole compared with tamoxifen for elderly patients with endocrine-responsive early breast cancer: the BIG 1-98 trial

PURPOSE: To explore potential differences in efficacy, treatment completion, and adverse events (AEs) in elderly women receiving adjuvant tamoxifen or letrozole for five years in the Breast International Group (BIG) 1-98 trial. METHODS: This report includes the 4,922 patients allocated to 5 years of letrozole or tamoxifen in the BIG 1-98 trial. The median follow-up was 40.4 months. Subpopulation Treatment Effect Pattern Plot (STEPP) analysis was used to examine the patterns of differences in disease-free survival and incidences of AEs according to age. In addition, three categoric age groups were defined: "younger postmenopausal" patients were younger than 65 years (n = 3,127), "older" patients were 65 to 74 years old (n = 1,500), and "elderly" patients were 75 years of age or older (n = 295). RESULTS: Efficacy results for subpopulations defined by age were similar to the overall trial results: Letrozole significantly improved disease-free survival (DFS), the primary end point, compared with tamoxifen. Elderly patients were less likely to complete trial treatment, but at rates that were similar in the two treatment groups. The incidence of bone fractures, observed more often in the letrozole group, did not differ by age. In elderly patients, letrozole had a significantly higher incidence of any grade 3 to 5 protocol-specified non-fracture AE compared with tamoxifen (P = .002), but differences were not significant for thromboembolic or cardiac AEs. CONCLUSION: Adjuvant treatment with letrozole had superior efficacy (DFS) compared with tamoxifen in all age groups. On the basis of a small number of patients older than 75 years (6%), age per se should not unduly affect the choice of adjuvant endocrine therapy.

In vitro biomechanical testing of a micro external skeletal fixator

OBJECTIVE To biomechanically test the properties of three different Universal Micro External Fixator (UMEX™) configurations with regard to their use in very small animals (<5kg) and compare the UMEX system to the widely used IMEX External Skeletal Fixation (SK™) system in terms of stiffness, space needed for pin placement and weight. METHODS Three different UMEX configurations (type Ia, type Ib, and type II modified) and one SK configuration type Ia were used to stabilize Delrin plastic rods in a 1 cm fracture gap model. These constructs were tested in axial compression, craniocaudal bending, mediolateral bending, and torsion. Testing was conducted within the elastic range and mean stiffness in each mode was determined from the slope of the linear portion of the load-deformation curve. A Kruskal Wallis one-way analysis of variance on ranks test was utilized to assess differences between constructs (p <0.05). RESULTS The UMEX type II modified configuration was significantly stiffer than the other UMEX configurations and the SK type Ia, except in craniocaudal bending, where the SK type Ia configuration was stiffer than all UMEX constructs. The UMEX type Ia configuration was significantly the weakest of those frames. The UMEX constructs were lighter and smaller than the SK, thus facilitating closer pin placement. CONCLUSIONS Results supported previous reports concerning the superiority of more complex constructs regarding stiffness. The UMEX system appears to be a valid alternative for the treatment of long-bone fractures in very small animals.

Periodontal healing after application of enamel matrix derivative in surgical supra/infrabony periodontal defects in rats with streptozotocin-induced diabetes

BACKGROUND AND OBJECTIVE Epidemiologic and clinical studies have indicated that diabetes is a risk factor for periodontal disease progression and healing. The aim of the present study was to evaluate short-term healing after enamel matrix derivative (EMD) application in combined supra/infrabony periodontal defects in diabetic rats. MATERIAL AND METHODS Thirty male Wistar rats were initially divided into two groups, one with streptozotocin-induced diabetes and another one with healthy (non-diabetic) animals. Bony defects were surgically created on the mesial root of the first maxillary molars. After root surface planing and EDTA conditioning, EMD was applied to the roots at one side of the maxillae, while those on the contralateral sides were left untreated. Animals were killed 3 wk after surgery, and block sections were prepared for histologic and histomorphometric analysis. RESULTS There was statistically significant more gingival recession in diabetic animals than in non-diabetic animals. The length of the junctional epithelium was significantly shorter in the EMD-treated sites in both diabetic and normoglycemic rats. Sulcus depth and length of supracrestal soft connective tissue showed no statistically significant differences between groups. In all animals, new bone formation was observed. Although new bone occurred more frequently in healthy animals, the extent of new bone was not significantly different between groups. In none of the teeth, a layer of new cementum was detectable. EMD had no influence on bone or cementum regeneration. Adverse reactions such as excessive inflammation due to bacterial root colonization, ankylosis and bone fractures were exclusively observed in diabetic animals, irrespective of EMD treatment. CONCLUSION Within the limits of the present study, it can be concluded that periodontal healing was impaired in streptozotocin-induced diabetic rats. EMD had no beneficial effects on new bone and cementum formation during short-term healing in this defect model and could not ameliorate the adverse effects in the systemically compromised animals.

LiLa-Klassifikation für Frakturen der langen Röhrenknochen im Wachstumsalter

Background. There are two child-specific fracture classification systems for long bone fractures: the AO classification of pediatric long-bone fractures (PCCF) and the LiLa classification of pediatric fractures of long bones (LiLa classification). Both are still not widely established in comparison to the adult AO classification for long bone fractures. Methods. During a period of 12 months all long bone fractures in children were documented and classified according to the LiLa classification by experts and non-experts. Intraobserver and interobserver reliability were calculated according to Cohen (kappa). Results. A total of 408 fractures were classified. The intraobserver reliability for location in the skeletal and bone segment showed an almost perfect agreement (K=0.91-0.95) and also the morphology (joint/shaft fracture) (K=0.87-0.93). Due to different judgment of the fracture displacement in the second classification round, the intraobserver reliability of the whole classification revealed moderate agreement (K=0.53-0.58). Interobserver reliability showed moderate agreement (K=0.55) often due to the low quality of the X-rays. Further differences occurred due to difficulties in assigning the precise transition from metaphysis to diaphysis. Conclusions. The LiLa classification is suitable and in most cases user-friendly for classifying long bone fractures in children. Reliability is higher than in established fracture specific classifications and comparable to the AO classification of pediatric long bone fractures. Some mistakes were due to a low quality of the X-rays and some due to difficulties to classify the fractures themselves. Improvements include a more precise definition of the metaphysis and the kind of displacement. Overall the LiLa classification should still be considered as an alternative for classifying pediatric long bone fractures.

Application, tolerance and safety of fondaparinux therapy in a German hospital: a prospective single-centre experience.

INTRODUCTION The pentasaccharide fondaparinux is widely approved for prophylaxis and treatment of thromboembolic diseases and therapy of acute coronary syndrome. It is also used off-label in patients with acute, suspected or antecedent heparin-induced thrombocytopenia (HIT). The aim of this prospective observational cohort study was to document fondaparinux' prescription practice, tolerance and therapy safety in a representative mixed German single-centre patient cohort. PATIENTS AND METHODS Between 09/2008 - 04/2009, 231 consecutive patients treated with fondaparinux were enrolled. Medical data were obtained from patient's records. The patients were clinically screened for thrombosis (Wells score), sequelae of HIT (4T's score), and bleeding complications (ISTH-criteria) and subjected to further assessment (i.e. sonography, HIT-diagnostics), if necessary. The mortality rate was assessed 30 days after therapy start. RESULTS Overall, 153/231 patients had a prophylactic, 74/231 patients a therapeutic, and 4/231 patients a successive prophylactic/therapeutic indication. In 11/231 patients fondaparinux was used due to suspected/antecedent HIT, in 5/231 patients due to a previous cutaneous delayed-type hypersensitivity to heparins. Other indications were rare. Three new/progressive thromboses were detected. No cases of HIT, major bleedings, or fatalities occurred. CONCLUSIONS Fondaparinux was well tolerated and was safe in prophylaxis and therapy; prescriptions mostly followed the current approval guidelines and were rarely related to HIT-associated indications (<5% of prescriptions), which is in contrast to previous study results in the U.S. (>94% of prescriptions were HIT-associated). A trend towards an individualised fondaparinux use based on the compound's inherent properties and the patients' risk profiles, i.e., antecedent HIT, bone fractures, heparin allergy, was observed.

Estudo retrospectivo das afecções orais em 754 felinos domésticos (Felis catus) atendidos no Laboratório de Odontologia Comparada da Universidade de São Paulo

Objetivou-se fazer um estudo retrospectivo avaliando quais as afecções da cavidade oral foram mais frequentes nos gatos domésticos atendidos no Laboratório de Odontologia Comparada da Faculdade de Medicina Veterinária e Zootecnia da Universidade de São Paulo, relatando estatisticamente a prevalência das afecções da cavidade oral de gatos, enfatizando se há correlação entre elas e com características como raça, sexo, faixa etária e estado reprodutivo. Os dados analisados dos 754 prontuários foram raça, idade, sexo, estado reprodutivo, diagnóstico, tratamento e, no caso de neoplasia, sua localização e diagnóstico histopatológico. As principais doenças diagnosticadas foram doença periodontal, fratura dentária, gengivoestomatite crônica felina, lesão de reabsorção dentária felina, neoplasia oral e traumatismo do sistema estomatognático (luxação de articulação temporomandibular, fenda palatina, fratura de processo coronoide, fratura de zigomático, disjunção de sínfise, fratura de maxila e mandíbula). A idade dos animais variou de menos de um ano a 20 anos, sendo que, os animais tinham, em média 7,2 anos (desvio padrão = 4,9) e a faixa etária mais frequente foi de um a cinco anos. Os gatos sem raça definida (66,5%), siameses (19,0%) e persas (10,2%) totalizaram 95,7% de todos os felinos atendidos no LOC. A doença periodontal foi a afecção mais frequente e esteve presente em 38,3% da população estudada. A fratura dentária, segunda mais frequente, esteve presente em 27,2% dos animais. Houve associação estatisticamente significativa (p=0,026) entre fratura dentária e faixa etária, já que a proporção de animais entre um e cinco anos de idade com fratura foi maior do que a das outras faixas etárias. A lesão de reabsorção dentária felina (LRDF) esteve presente em 19,6% dos gatos estudados, sendo a terceira afecção mais prevalente dentre as pesquisadas. Esta lesão foi mais frequente em gatos com idade entre 11 e 15 anos e houve associação estatisticamente significativa entre a LRDF e a doença periodontal e entre LRDF e gengivite. A prevalência de gengivoestomatite crônica felina foi de 15,7% entre os felinos pesquisados e a proporção de animais com idades entre seis e dez anos com esta doença foi maior do que em outras faixas etárias. As neoplasias estavam presentes em 9,8% dos gatos, sendo que em 46 dos 72 animais que apresentaram alguma neoplasia tinham mais de dez anos de idade. O carcinoma de células escamosas foi o neoplasma mais comum, correspondendo a 63,2% das neoformações que foram submetidas ao exame histopatológico. As fraturas ósseas do sistema estomatognático corresponderam a 19,3% dos atendimentos, sendo a sínfise mentoniana e o corpo da mandíbula os locais mais comuns de fraturas. Concluiu-se que: existe grande variedade de afecções que acometem a cavidade oral de gatos, sendo a doença periodontal, fratura dentária, lesão de reabsorção dentária, gengivite, gengivoestomatite crônica, neoplasias orais e fraturas dos ossos do sistema estomatognático as mais prevalentes delas; é de extrema importância que as anotações nas fichas de atendimento sejam feitas da maneira mais completa possível, para que informações não sejam perdidas

Safety of antidiabetes medications:an update

Adverse iatrogenic effects of especial relevance for antidiabetes medications include hypoglycemic episodes, major adverse cardiovascular (CV) events, cancer, bone fractures, pancreatic effects, genital/urinary tract infections, and weight gain. Here, recent clinical studies addressing safety profiles of antidiabetes medications are reviewed. On balance, new prospective and population-based studies continue to indicate that the benefits of improved glucose control outweigh the risks associated with antidiabetes medications in most patients with type 2 diabetes.

Avaliação histomorfométrica do efeito do ultrasom pulsado nas falhas ósseas provocadas em fêmures de rato: estudo experimental

SILVA, J. S. P. Avaliação histomorfométrica do efeito do ultrasom pulsado nas falhas ósseas provocadas em fêmures de rato: estudo experimental . 2000. 85 f. Dissertação (Mestrado) – Faculdade de Medicina, Universidade de São Paulo. São Paulo, 2000.