Isolation of human fibroadipogenic progenitors and satellite cells from frozen muscle biopsies.

Skeletal muscle contains multiple cell types that work together to maintain tissue homeostasis. Among these, satellite cells (SC) and fibroadipogenic progenitors cells (FAPs) are the two main stem cell pools. Studies of these cells using animal models have shown the importance of interactions between these cells in repair of healthy muscle, and degeneration of dystrophic muscle. Due to the unavailability of fresh patient muscle biopsies, similar analysis of interactions between human FAPs and SCs is limited especially among the muscular dystrophy patients. To address this issue here we describe a method that allows the use of frozen human skeletal muscle biopsies to simultaneously isolate and grow SCs and FAPs from healthy or dystrophic patients. We show that while the purified SCs differentiate into mature myotubes, purified FAPs can differentiate into adipocytes or fibroblasts demonstrating their multipotency. We find that these FAPs can be immortalized and the immortalized FAPs (iFAPs) retain their multipotency. These approaches open the door for carrying out personalized analysis of patient FAPs and interactions with the SCs that lead to muscle loss.

High Numbers of Circulating CD57+ NK Cells Associate with Resistance to HER2-Specific Therapeutic Antibodies in HER2+ Primary Breast Cancer.

Natural killer (NK) cells can orchestrate effective antitumor immunity. The presence of tumor-infiltrating NK cells in diagnostic biopsies predicts pathologic complete response (pCR) to HER2-specific therapeutic antibodies in patients with primary breast cancer. Here, we analyzed whether diversity in circulating NK cells might influence tumor infiltration and HER2-specific therapeutic antibody efficacy. We found that numbers of circulating CD57+ NK cells inversely correlated with pCR to HER2-specific antibody treatment in patients with primary breast cancer independently of age, traditional clinicopathologic factors, and CD16A 158F/V genotype. This association was uncoupled from the expression of other NK-cell receptors, the presence of adaptive NK cells, or changes in major T-cell subsets, reminiscent of cytomegalovirus-induced immunomodulation. NK-cell activation against trastuzumab-coated HER2+ breast cancer cells was comparable in patients with high and low proportions of CD57+ NK cells. However, circulating CD57+ NK cells displayed decreased CXCR3 expression and CD16A-induced IL2-dependent proliferation in vitro Presence of CD57+ NK cells was reduced in breast tumor-associated infiltrates as compared with paired peripheral blood samples, suggesting deficient homing, proliferation, and/or survival of NK cells in the tumor niche. Indeed, numbers of circulating CD57+ were inversely related to tumor-infiltrating NK-cell numbers. Our data reveal that NK-cell differentiation influences their antitumor potential and that CD57+ NK cells may be a biomarker useful for tailoring HER2 antibody-based therapeutic strategies in breast cancer.

Early plant growth and biochemical responses induced by Azospirillum brasilense Sp245 lipopolysaccharides in wheat (Triticum aestivum L.) seedlings are attenuated by procyanidin B2.

This study analyzes the effects of procyanidin B2 on early wheat plant growth and plant biochemical responses promoted by lipopolysaccharides (LPS) derived from the rhizobacteria Azospirillum brasilense Sp245. Measurements of leaf, root length, fresh weight, and dry weight showed in vitro plant growth stimulation 4 days after treatment with A. brasilense as well as LPS. Superoxide anion (O2·-) and hydrogen peroxide (H2O2) levels increased in seedling roots treated with LPS (100 µg mL-1). The chlorophyll content in leaf decreased while the starch content increased 24 h after treatment in seedling roots. The LPS treatment induced a high increase in total peroxidase (POX) (EC 1.11.1.7) activity and ionically bound cell wall POX content in roots, when compared to respective controls. Early plant growth and biochemical responses observed in wheat seedlings treated with LPS were inhibited by the addition of procyanidin B2 (5 µg mL-1), a B type proanthocyanidin (PAC), plant-derived polyphenolic compound with binding properties of LPS. All results suggest first that the ionically bound cell wall POX enzymes could be a molecular target of A. brasilense LPS, and second that the recognition or association of LPS by plant cells is required to activate plant responses. This last event could play a critical role during plant growth regulation by A. brasilense LPS.

Comparison of outcome of tibial plafond fractures managed by hybrid external fixation versus two-stage management with final plate fixation.

BACKGROUND: Tibial platfond fractures are usually associated with massive swelling of the foot and ankle, as well as with open wounds. This swelling may cause significant decrease of the blood flow, so the state of the soft tissue is determinant for the surgical indication and the type of implant. This retrospective study compares the union times in cases of tibial plafond fractures managed with a hybrid external fixation as a definitive procedure versus those managed with a two stage strategy with final plate fixation. MATERIALS AND METHODS: A retrospective study in a polytrauma referral hospital was performed between 2005 and 2011. Patients with a tibial plafond fracture, managed with a hybrid external fixation as a definitive procedure or managed with a two stage strategy with the final plate fixation were included in the study. Postoperative radiographs were evaluated by two senior surgeons. Fracture healing was defined as callus bridging of one cortex, seen on both lateral and anteroposterior X-ray. The clinical outcome was evaluated by means of 11 points Numerical Rating Scale for pain and The American Orthopedic Foot and Ankle Society ankle score, assessed at the last followup visit. Thirteen patients had been managed with a hybrid external fixation and 18 with a two-stage strategy with the final plate fixation. There were 14 males and 17 females with a mean age of 48 years (range 19-82 years). The mean followup was 24 months (range 24-70 months). RESULTS: The mean time from surgery to weight bearing was 7 ± 6.36 days for the hybrid fixation group and 57.43 ± 15.46 days for the plate fixation group (P < 0.0001); and the mean time from fracture to radiological union was 133.82 ± 37.83) and 152.8 ± 72.33 days respectively (P = 0.560). CONCLUSION: Besides the differences between groups regarding the baseline characteristics of patients, the results of this study suggest that in cases of tibial plafond fractures, the management with a hybrid external fixation as a definitive procedure might involve a faster union than a two-stage management with final plate fixation.

De novo mutations in PLXND1 and REV3L cause Möbius syndrome.

Möbius syndrome (MBS) is a neurological disorder that is characterized by paralysis of the facial nerves and variable other congenital anomalies. The aetiology of this syndrome has been enigmatic since the initial descriptions by von Graefe in 1880 and by Möbius in 1888, and it has been debated for decades whether MBS has a genetic or a non-genetic aetiology. Here, we report de novo mutations affecting two genes, PLXND1 and REV3L in MBS patients. PLXND1 and REV3L represent totally unrelated pathways involved in hindbrain development: neural migration and DNA translesion synthesis, essential for the replication of endogenously damaged DNA, respectively. Interestingly, analysis of Plxnd1 and Rev3l mutant mice shows that disruption of these separate pathways converge at the facial branchiomotor nucleus, affecting either motoneuron migration or proliferation. The finding that PLXND1 and REV3L mutations are responsible for a proportion of MBS patients suggests that de novo mutations in other genes might account for other MBS patients.

Pseudoartrosis de calcáneo en contexto de pie catastrófico / Pseudarthrose du calcanéum dans le cadre d'une fracture-luxation de pied / Calcaneal pseduarthrosis in catastrophic foot

La pseudoartrosis de calcáneo es excepcional. La incidencia es del 1,3 por ciento de todas las fracturas intraarticulares. Se relaciona con fracturas intraarticulares tratadas conservadoramente, diabetes mellitus, tabaquismo y fracturas abiertas. Presentamos un caso de pseudoartrosis de calcáneo, secuela de fractura de luxación abierta en el contexto de un pie catastrófico(AU)

Calcaneal pseudoarthrosis is rare and its incidence accounts for 1.3 percent of all the intra-articular fractures. It is associated to intra-articular fractures treated with conservative methods, diabetes mellitus, smoking and open fractures. This is a calcaneal pseudoarthrosis case as a sequel of open dislocation fracture in a catastrophic foot injure(AU)

La pseudarthrose du calcanéum est une affection assez rare. Son incidence est 1.3 pourcent de toutes les fractures interarticulaires. Elle est associée aux fractures interarticulaires traitées de façon conservatrice, au diabète sucré, au tabagisme, et aux fractures ouvertes. Un cas de pseudarthrose de calcanéum, séquelle d'une fracture-luxation ouverte du pied après accident de route, est présenté(AU)

The increase of pericyte population in human neuromuscular disorders supports their role in muscle regeneration in vivo.

Pericytes are periendothelial cells that have been involved in many different functions including a possible role as mesodermal stem/progenitor cells. In the present study we demonstrate that alkaline phosphatase (AP) expression is specific for human muscular pericytes and can be used as a marker to identify them in skeletal muscle biopsies. We studied the pericyte population in skeletal muscle biopsies from controls, myopathic and neuropathic patients. We observed a significant increase in the number of pericytes only in myopathies that correlated with the number of NCAM(+) fibres, suggesting that an active muscular degenerative/regenerative process is related to an increase in the pericyte population. AP(+) pericytes sorted from skeletal muscle samples were able to activate the myogenic programme and fuse with both mononucleate satellite cells and mature multinucleated myotubes in vitro, demonstrating that they could participate in muscle regeneration. In accordance, pericytes expressing the myogenic transcription factor MyoD were found in biopsies of myopathic biopsies. All these data support the hypothesis that, apart from satellite cells, pericytes may play an important role in muscle regeneration in adult human muscles in vivo.

Proteomics identification of differentially expressed proteins in the muscle of dysferlin myopathy patients.

The muscular dystrophies are a large and heterogeneous group of neuromuscular disorders that can be classified according to the mode of inheritance, the clinical phenotype and the molecular defect. To better understand the pathological mechanisms of dysferlin myopathy we compared the protein-expression pattern in the muscle biopsies of six patients with this disease with six patients with limb girdle muscular dystrophy 2A, five with facioscapulohumeral dystrophy and six normal control subjects. To investigate differences in the expression levels of skeletal muscle proteins we used 2-DE and MS. Western blot or immunohistochemistry confirmed relevant results. The study showed specific increase expression of proteins involved in fast-to-slow fiber type conversion (ankyrin repeat protein 2), type I predominance (phosphorylated forms of slow troponin T), sarcomere stabilization (actinin-associated LIM protein), protein ubiquitination (TRIM 72) and skeletal muscle differentiation (Rho-GDP-dissociation inhibitor ly-GDI) in dysferlin myopathy. As anticipated, we also found differential expression of proteins common to all the muscular dystrophies studied. This comparative proteomic analysis suggests that in dysferlin myopathy (i) the type I fiber predominance is an active process of fiber type conversion rather than a selective loss of type II fibers and (ii) the dysregulation of proteins involved in muscle differentiation further confirms the role of dysferlin in this process.

Experiencia en el tratamiento de las deformidades axiales de la rodilla por la técnica de Blount en el Hospital San Juan de Dios, Caracas / Experience in the deformities of the knee for the Blount technic in the San Juan de Dios Hospital, Caracas

Se hace un estudio de 54 pacientes a los cuales se les practicó epifisiodesis con grapas según Blount (7 Varos, 47 Valgos). Según los parámetros de evaluación se obtuvieron 34 resultados buenos, 9 regulares y 11 malos. En nuestro medio la espifisiodesis temporal según Blount ha demostrado ser la técnica de elección para la corrección de deformidades axiales de rodilla