RESUMEN
Phenalenyl is a radical nanographene with a triangular shape hosting an unpaired electron with spin S = 1/2. The open-shell nature of the phenalenyl is expected to be retained in covalently bonded networks. As a first step, we report synthesis of the phenalenyl dimer by combining in-solution synthesis and on-surface activation and its characterization on Au(111) and on a NaCl decoupling layer by means of inelastic electron tunneling spectroscopy (IETS). IETS shows inelastic steps that are identified as singlet-triplet excitation arising from interphenalenyl exchange. Spin excitation energies with and without the NaCl decoupling layer are 48 and 41 meV, respectively, indicating significant renormalization due to exchange with Au(111) electrons. Furthermore, third-neighbor hopping-induced interphenalenyl hybridization is fundamental to explaining the position-dependent bias asymmetry of the inelastic steps and activation of kinetic interphenalenyl exchange. Our results pave the way for bottom-up synthesis of S = 1/2 spin-lattices with large exchange interactions.
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Antineutrophil cytoplasm antibody (ANCA)-associated vasculitis (AAV) comprises a group of multisystem disorders involving severe, systemic, small-vessel vasculitis with short- and long term serious and life-threating complications. Despite the simplification of treatment, fundamental aspects concerning assessment of its efficacy and its adaptation to encountered complications or to the relapsing/remitting/subclinical disease course remain still unknown. The pathogenesis of AAV is complex and unique, and despite the progress achieved in the last years, much has not to be learnt. Foremost, there is still no accurate marker enabling us to monitoring disease and guide therapy. Therefore, the disease management relays often on clinical judgment and follows a" trial and error approach". In the recent years, an increasing number of new molecules s have been explored and used for this purpose including genomics, B- and T-cell subpopulations, complement system factors, cytokines, metabolomics, biospectroscopy and components of our microbiome. The aim of this review is to discuss both the role of known historical and clinically established biomarkers of AAV, as well as to highlight potential new ones, which could be used for timely diagnosis and monitoring of this devastating disease, with the goal to improve the effectiveness and ameliorate the complications of its demanding therapy.
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Vasculitis Asociada a Anticuerpos Citoplasmáticos Antineutrófilos , Metabolómica , Humanos , Vasculitis Asociada a Anticuerpos Citoplasmáticos Antineutrófilos/diagnóstico , GenómicaRESUMEN
BACKGROUND: Minimally invasive left ventricular assist device (LVAD) implantation may reduce peri-/postoperative complications and risks associated with resternotomies. In this study, we describe our first results using a minimally invasive LVAD implantation technique (lateral thoracotomy [LT] group). These results were compared with LVAD implantations done via full median sternotomy (STX group). METHODS: HVAD (HeartWare, Framingham, Massachusetts, United States) implantations in 70 patients (LT group n = 22, 52 ± 15 years old; STX group n = 48, 59 ± 11 years old) were retrospectively analyzed. Minimally invasive access via left thoracotomy was feasible in 22 patients. Peri- and postoperative analyses of survival and adverse events were performed. RESULTS: No survival differences were observed between the LT and STX group (p = 0.43). LT patients without temporary right ventricular assist device (tRVAD) showed a significantly better survival rate compared to LT patients with concomitant tRVAD implantation (p = 0.02), which could not be demonstrated in the STX group (p = 0.11). Two LT and four STX patients were successfully bridged to heart transplantation and three STX patients were successfully weaned with subsequent LVAD explantations. LVAD-related infections (n = 4 LT group vs n = 20 STX group, p = 0.04) were less likely in the LT group. No wound dehiscence occurred in the LT group, whereas five were observed in the STX group (p = 0.17). The amount of perioperative blood transfusions (within the first 7 postoperative days) did not differ in both study groups (p = 0.48). CONCLUSION: The minimally invasive approach is a viable alternative with the possibility to reduce complications and should be particularly considered for bridge-to-transplant patients.
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Insuficiencia Cardíaca/terapia , Corazón Auxiliar , Implantación de Prótesis/instrumentación , Implantación de Prótesis/métodos , Esternotomía , Toracotomía/métodos , Función Ventricular Izquierda , Adulto , Anciano , Femenino , Alemania , Insuficiencia Cardíaca/diagnóstico , Insuficiencia Cardíaca/mortalidad , Insuficiencia Cardíaca/fisiopatología , Humanos , Masculino , Persona de Mediana Edad , Procedimientos Quirúrgicos Mínimamente Invasivos , Complicaciones Posoperatorias/etiología , Diseño de Prótesis , Implantación de Prótesis/efectos adversos , Implantación de Prótesis/mortalidad , Recuperación de la Función , Estudios Retrospectivos , Esternotomía/efectos adversos , Esternotomía/mortalidad , Toracotomía/efectos adversos , Toracotomía/mortalidad , Factores de Tiempo , Resultado del TratamientoRESUMEN
BACKGROUND: Guidelines summarize medical evidence, they identify the most efficient therapy under study conditions and recommend this therapy for use. The physician now has the challenge to translate a therapy that is efficient under laboratory conditions to a patient who is an individual person. To accomplish this task the physician has to make sure that (I) the ideal typical therapy is applicable and effective in this individual patient taking the special features into consideration, that (II) therapy is compliant with the norm including guidelines, laws and ethical requirements (conformity) and that (III) the therapy meets the patient's needs. OBJECTIVE: How can physicians together with the patients translate the medical evidence into an individually optimized therapy? MATERIAL AND METHODS: At the German Aortic Center in Hamburg we use ISWOT as an instrument to identify such individually optimized therapy. With ISWOT, we present an instrument with which we have developed an (I) efficient, (II) conform and (III) needs-oriented therapeutic strategy for individual patients. RESULTS: I-SWOT cross-tabulates strengths (S) and weaknesses (W) related to therapy with opportunities (O) and threats (T) related to individual patients. This ISWOT matrix identifies four fundamental types of strategy, which comprise "SO" maximizing strengths and opportunities, "WT" minimizing weaknesses and threats, "WO" minimizing weaknesses and maximizing opportunities and "ST" maximizing strengths and minimizing threats. We discuss the case of a patient with asymptomatic thoracoabdominal aneurysm to show how ISWOT is used to identify an individually optimized therapy strategy.
RESUMEN
Cell-based in vitro resorption assays are an important tool to simulate the in vivo biodegradation of resorbable bone graft materials and to predict their clinical performance. The present study analyses the activity of osteoclast-specific enzymes as potential surrogate measures for classical pit assay, which is not applicable on irregular structured materials. Osteoclasts derived from human peripheral blood mononuclear cells were cultivated on different surfaces: calcium phosphate bone cements (CPC), dentin discs, osteoblast-derived extracellular matrix (ECM) and tissue culture polystyrene as control. Pit formation on the resorbable materials was investigated and correlated with the activity of tartrate resistant acid phosphatase (TRAP), carbonic anhydrase II (CAII) and cathepsin K (CTSK). Furthermore, the relation between intra- and extracellular enzyme activities was examined for TRAP and CTSK during resorption of the different materials. Resorbed area of CPC correlated with intracellular TRAP activity and intracellular CAII activity. Highest resorption was detected at around pH 7.2. Resorbed area on dentin correlated with the extracellular CTSK activity and extracellular TRAP activity and was maximal at around pH 6.8. Osteoclasts cultivated on cell-derived mineralised ECM showed a good correlation between both extracellular TRAP and CTSK activity and the release of calcium ions. Based on these data a different regulation of TRAP and CTSK secretion is hypothesised for the resorption of inorganic calcium phosphate compared to the resorption of collagenous mineralised matrix.
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Bioensayo/métodos , Resorción Ósea/enzimología , Osteoclastos/enzimología , Cementos para Huesos/farmacología , Matriz Ósea/efectos de los fármacos , Matriz Ósea/metabolismo , Resorción Ósea/patología , Fosfatos de Calcio/farmacología , Diferenciación Celular/efectos de los fármacos , Dentina/metabolismo , Matriz Extracelular/efectos de los fármacos , Matriz Extracelular/metabolismo , Humanos , Osteoblastos/efectos de los fármacos , Osteoblastos/enzimología , Osteoblastos/patología , Osteoclastos/efectos de los fármacos , Osteoclastos/patología , Osteoclastos/ultraestructura , Poliestirenos/farmacología , Coloración y Etiquetado , Fosfatasa Ácida Tartratorresistente/metabolismoRESUMEN
Studies on tissue-engineering approaches for the regeneration of traumatized cartilage focus increasingly on multipotent human mesenchymal stem cells (hMSCs) as an alternative to autologous chondrocytes. The present study applied porous scaffolds made of collagen from the jellyfish Rhopilema esculentum for the in vitro chondrogenic differentiation of hMSCs. Culture conditions in those scaffolds differ from conditions in high-density pellet cultures, making a re-examination of these data necessary. We systematically investigated the influence of seeding density, basic culture media [Dulbecco's modified Eagle's medium (DMEM), α-minimum essential medium (α-MEM)] with varying glucose content and supplementation with fetal calf serum (FCS) or bovine serum albumin (BSA) on the chondrogenic differentiation of hMSCs. Gene expression analyses of selected markers for chondrogenic differentiation and hypertrophic development were conducted. Furthermore, the production of cartilage extracellular matrix (ECM) was analysed by quantification of sulphated glycosaminoglycan and collagen type II contents. The strongest upregulation of chondrogenic markers, along with the highest ECM deposition was observed in scaffolds seeded with 2.4 × 106 cells/cm3 after cultivation in high-glucose DMEM and 0.125% BSA. Lower seeding densities compared to high-density pellet cultures were sufficient to induce in vitro chondrogenic differentiation of hMSCs in collagen scaffolds, which reduces the amount of cells required for the seeding of scaffolds and thus the monolayer expansion period. Furthermore, examination of the impact of FCS and α-MEM on chondrogenic MSC differentiation is an important prerequisite for the development of an osteochondral medium for simultaneous osteogenic and chondrogenic differentiation in biphasic scaffolds for osteochondral tissue regeneration. Copyright © 2015 John Wiley & Sons, Ltd.
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Cartílago , Condrogénesis , Colágeno/química , Células Madre Mesenquimatosas/metabolismo , Escifozoos/química , Andamios del Tejido/química , Animales , Técnicas de Cultivo de Célula/métodos , Medios de Cultivo/química , Humanos , Células Madre Mesenquimatosas/citología , Ingeniería de Tejidos/métodosRESUMEN
UNLABELLED: Strontium ions were discovered to exert a dual effect on bone turnover, namely an inhibition of cell-driven bone resorption and a simultaneous stimulation of new bone tissue formation. A variety of strontium containing calcium phosphate bone cements (SrCPC) have been developed to benefit from both effects to locally support the healing of osteoporotic bone defects. While the stimulating effect of strontium modification on bone forming cells has been demonstrated in a number of studies, this study focuses on the inhibition and/or reduction of osteoclastogenesis and osteoclastic resorption by a strontium substituted calcium phosphate bone cement (SrCPC). Human peripheral blood mononuclear cells (PBMC) were differentiated into osteoclasts in the presence of different Sr(2+)-concentrations as well as on the surface of SrCPC disks. Osteoclastogenesis of PBMC was shown to be merely unaffected by medium Sr(2+)-concentrations comparable to those released from SrCPC in vitro (0.05-0.15mM). However, an altering effect of 0.1mM strontium on the cytoskeleton of osteoclast-like cells was shown. In direct contact to SrCPC disks, these cells exhibited typical morphological features and osteoclast markers on both RNA and protein level were formed. However, calcium phosphate resorption was significantly decreased on strontium-containing cements in comparison to a strontium-free control. This was accompanied by an intracellular accumulation of strontium that increased with substrate strontium content as demonstrated by Time of Flight Secondary Ion Mass Spectrometry (ToF-SIMS). This study illustrates that SrCPC do not inhibit osteoclastogenesis but significantly attenuate osteoclastic substrate resorption in vitro. STATEMENT OF SIGNIFICANCE: Strontium ions have been shown to promote bone formation and inhibit bone resorption. Therefore strontium is successfully used in the treatment of osteoporosis and also inspired the development of strontium-containing strontium/calcium phosphate bone cements (SrCPC). Studies have shown the positive effects of SrCPC on bone formation, however, the inhibiting effect of strontium on bone resorption in the context of such cements has not been shown so far. We found that the formation of bone-resorbing osteoclasts is not inhibited, but that their resorption activity is decreased in contact to SrCPC. The former is important since those cells play an important role in the bone cell signaling. The latter is a key requirement in osteoporosis therapy, which addresses excess bone resorption.
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Apatitas/farmacología , Cementos para Huesos/farmacología , Resorción Ósea/patología , Fosfatos de Calcio/farmacología , Osteoclastos/patología , Osteogénesis/efectos de los fármacos , Estroncio/farmacología , Adulto , Calcio/metabolismo , Células Cultivadas , ADN/metabolismo , Regulación de la Expresión Génica/efectos de los fármacos , Humanos , Espacio Intracelular/metabolismo , Microscopía Fluorescente , Osteoclastos/efectos de los fármacos , Osteoclastos/metabolismo , Osteogénesis/genéticaRESUMEN
Marine, hybrid constructs of porous scaffolds from fibrillized jellyfish collagen and alginate hydrogel are mimicking both of the main tissue components of cartilage, thus being a promising approach for chondrogenic differentiation of human mesenchymal stem cells (hMSC). Investigating their potential for articular cartilage repair, the present study examined scaffolds being either infiltrated with an alginate-cell-suspension (ACS) or seeded with hMSC and embedded in alginate after cell adhesion (EAS). Hybrid constructs with 2×10(5) and 4.5×10(5)hMSC/scaffold were compared to hMSC encapsulated in pure alginate discs, both chondrogenically stimulated for 21days. Typical round, chondrocyte-like morphology was observed in pure alginate gels and ACS scaffolds, while cells in EAS were elongated and tightly attached to the collagen pores. Col 2 gene expression was comparable in all scaffold types examined. However, the Col 2/Col 1 ratio was higher for pure alginate discs and ACS scaffolds compared to EAS. In contrast, cells in EAS scaffolds displayed higher gene expression of Sox 9, Col 11 and ACAN compared to ACS and pure alginate. Secretion of sulfated glycosaminoglycans (sGAG) was comparable for ACS and EAS scaffolds. In conclusion hybrid constructs of jellyfish collagen and alginate support hMSC chondrogenic differentiation and provide more stable and constructs compared to pure hydrogels.
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Alginatos/química , Diferenciación Celular , Condrogénesis , Colágeno/química , Células Madre Mesenquimatosas/metabolismo , Escifozoos/química , Animales , Células Cultivadas , Ácido Glucurónico/química , Ácidos Hexurónicos/química , Humanos , Células Madre Mesenquimatosas/citologíaRESUMEN
Scaffolds for bone regeneration are mostly prepared with an isotropic, sponge-like structure mimicking the architecture of trabecular bone. We have developed an anisotropic bioceramic with parallel aligned pores resembling the honeycomb arrangement of Haversian canals of cortical bone and investigated its potential as a scaffold for tissue engineering. Parallel channel-like pores were generated by ionotropic gelation of an alginate-hydroxyapatite (HA) slurry, followed by ceramic processing. Organic components were thermally removed at 650 °C, whereas the pore system was preserved in the obtained HA bioceramic in the processing stage of a bisque. Even without further sintering at higher temperatures, the anisotropic HA bisque (AHAB) became mechanically stable with a compressive strength (4.3 MPa) comparable to that of native trabecular bone. Owing to the low-temperature treatment, a nanocrystalline microstructure with high porosity (82%) and surface area (24.9 m(2)/g) was achieved that kept the material dissolvable in acidic conditions, similar to osteoclastic degradation of bone. Human mesenchymal stem cells (hMSCs) adhered, proliferated and differentiated into osteoblasts when osteogenically induced, indicating the cytocompatibility of the bisque scaffold. Furthermore, we demonstrated fusion of human monocytes to osteoclast-like cells in vitro on this substrate, similar to the natural pathway. Biocompatibility was demonstrated in vivo by implantation of the bisque ceramic into cortical rabbit femur defects, followed by histological analysis, where new bone formation inside the channel-like pores and generation of an osteon-like tissue morphology was observed.
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Sustitutos de Huesos , Durapatita , Fémur/metabolismo , Nanopartículas/química , Andamios del Tejido/química , Animales , Anisotropía , Sustitutos de Huesos/química , Sustitutos de Huesos/farmacología , Diferenciación Celular/efectos de los fármacos , Durapatita/química , Durapatita/farmacología , Femenino , Fémur/química , Fémur/patología , Humanos , Masculino , Células Madre Mesenquimatosas/metabolismo , Células Madre Mesenquimatosas/patología , Osteoblastos/metabolismo , Osteoblastos/patología , Osteogénesis/efectos de los fármacos , Porosidad , ConejosRESUMEN
BACKGROUND: Skin lesions due to Fusarium spp. occur either secondarily following hematological spread in systemic infection or represent primary cutaneous infections following traumatic inoculation. CASE REPORT: A 34-year-old woman with insulin-dependent diabetes mellitus presented with a most likely posttraumatic leg ulcer present for 4 weeks. The ulcer showed superficial necrosis with cellular debris, neutrophils, and leukocytoclasia. Septate hyphae were detected both in the necrotic area and between the collagen fibers on initial H & E stained sections. Using PAS and Grocott-Gomori silver staining, the dichotomous branching hyphae were clearly visible. Unfortunately, cultural detection of the fungi was impossible. After extraction and purification of the fungal DNA from formalin-fixed and paraffin embedded (FFPE) tissue sections, the amplification of the ITS region of rDNA was done. Using sequencing and comparison with reference sequences of a gene bank, Fusarium oxysporum was identified. THERAPY: Therapy was performed by surgical excision of the entire ulcer followed by topical antiseptic treatment and wound conditioning. No systemic antifungal treatment was given. The lesion healed without any problems. DISCUSSION: Cutaneous fusarium infections are rare but emerging opportunistic infections. Histological examination represents the quickest diagnostic method for detection of the fungal infection. An alternative approach represents the species identification based on molecular techniques.
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Dermatomicosis/diagnóstico , Dermatomicosis/microbiología , Fusariosis/diagnóstico , Fusariosis/microbiología , Fusarium/genética , Úlcera de la Pierna/diagnóstico , Úlcera de la Pierna/microbiología , Adulto , ADN Ribosómico/genética , Dermatomicosis/cirugía , Femenino , Formaldehído , Fusariosis/cirugía , Fusarium/clasificación , Fusarium/aislamiento & purificación , Marcadores Genéticos/genética , Humanos , Úlcera de la Pierna/cirugía , Técnicas de Diagnóstico Molecular , Adhesión en Parafina , Análisis de Secuencia de ADN/métodos , Fijación del Tejido , Resultado del TratamientoRESUMEN
OBJECTIVES: To gain further data on the hormonal control of pregnancy in the donkey and to obtain reference values for hormonal pregnancy testing. MATERIAL AND METHODS: Blood samples were collected at monthly intervals from 23 donkey mares with normal singleton pregnancies. Further samples were obtained from six mares displaying pregnancies with clinical disorders. Progesterone (P4), total estrone (TE), free (E) and conjugated estrone (ES) were determined using radioimmunoassay. RESULTS: Mean duration of pregnancy was 372 ± 16 days. It was longer (p < 0.05) in large (375.9 ± 5.7 days) and standard (385.8 ± 20.7 days) donkeys than in miniature donkeys (357.4 ± 5.7 days) and negatively correlated to the age of the mare (p = 0.043). P4-concentrations varied between 12-35 ng/ml during weeks 2-5 of pregnancy and increased thereafter in eight jennies concomitant with the formation of the secondary corpora lutea (sCL), reaching values of 40-110 ng/ml during weeks 12-17. The decrease observed thereafter resulted in concentrations between 5-16 ng/ml until week 46, followed by a slight increase in most of the mares prior to parturition. Concentrations of TE remained < 1 ng/ml until week 6. They increased thereafter to 600-2700 ng/ml during midpregnancy and displayed a decrease to 1-20 ng/ml during the last 2 weeks of pregnancy. The course of E and ES was correlated (p < 0.0001) and E concentrations were up to 1000 times lower than those of ES. The course of hormone concentrations did not provide any clear indications in relation to the observed clinical disorders. CONCLUSION: The course of P4-concentrations resembles largely the situation in the horse. In contrast to the horse, the course of ES does not show an increase concomitant with the formation of the sCL. Breed-specific effects became apparent regarding pregnancy duration. CLINICAL RELEVANCE: Hormonal pregnancy diagnostic in the jenny could be put on a solid basis with TE values > 5 ng/ml being indicative for pregnancy. At present, monitoring of P4 and estrone during pregnancy does not allow the prediction of clinical disorders.
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Equidae/fisiología , Estrona/análogos & derivados , Estrona/sangre , Preñez/sangre , Progesterona/sangre , Animales , Femenino , EmbarazoRESUMEN
BACKGROUND: Due to the lack of histopathological differentiation the unequivocal identification of fungal pathogens is rarely possible. In order to understand the pathogen spectrum causing cephalic mycosis the use of alternative methods is essential. MATERIAL AND METHODS: In a retrospective study 24 formalin-fixed, paraffin-embedded (FFPE) samples from patients with histologically confirmed cerebral or cephalic mycosis were analyzed with molecular biological methods. RESULTS: In two samples obtained during the patients' lifetime human as well as fungal DNA was detected, making an unambiguous diagnosis possible. For tissue that had been fixed over a longer period, detection of human and fungal DNA was possible merely in 60% and 47 % of the samples, respectively. Most frequently diagnosed were aspergillosis (n = 9), followed by mucormycosis (n = 2) and imported blastomycosis (n = 1). CONCLUSIONS: Using biopsy material a DNA analysis seems promising although only with limited success using brain samples taken at autopsy which have been fixed over a longer period. For unambiguous retrospective diagnostics of pathogens when cephalic mycosis is suspected, the sample extraction for postmortem diagnostics should be performed prior to a long period of formalin fixation.
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Encefalopatías/microbiología , Encefalopatías/patología , Infecciones Fúngicas del Sistema Nervioso Central/microbiología , Infecciones Fúngicas del Sistema Nervioso Central/patología , Enfermedades de los Senos Paranasales/patología , Adulto , Anciano , Encéfalo/microbiología , Encéfalo/patología , ADN de Hongos/análisis , ADN de Hongos/genética , Femenino , Fijadores , Formaldehído , Hongos/clasificación , Hongos/genética , Hongos/aislamiento & purificación , Humanos , Masculino , Persona de Mediana Edad , Técnicas de Tipificación Micológica , Infecciones Oportunistas/microbiología , Infecciones Oportunistas/patología , Adhesión en Parafina , Enfermedades de los Senos Paranasales/microbiologíaRESUMEN
BACKGROUND AND PURPOSE: Endovascular navigation under MR imaging guidance can be facilitated by a catheter with steerable microcoils on the tip. Not only do microcoils create visible artifacts allowing catheter tracking, but also they create a small magnetic moment permitting remote-controlled catheter tip deflection. A side product of catheter tip electrical currents, however, is the heat that might damage blood vessels. We sought to determine the upper boundary of electrical currents safely usable at 1.5T in a coil-tipped microcatheter system. MATERIALS AND METHODS: Alumina tubes with solenoid copper coils were attached to neurovascular microcatheters with heat shrink-wrap. Catheters were tested in carotid arteries of 8 pigs. The catheters were advanced under x-ray fluoroscopy and MR imaging. Currents from 0 mA to 700 mA were applied to test heating and potential vascular damage. Postmortem histologic analysis was the primary endpoint. RESULTS: Several heat-mitigation strategies demonstrated negligible vascular damage compared with control arteries. Coil currents ≤300 mA resulted in no damage (0/58 samples) compared with 9 (25%) of 36 samples for > 300-mA activations (P = .0001). Tip coil activation ≤1 minute and a proximal carotid guide catheter saline drip > 2 mL/minute also had a nonsignificantly lower likelihood of vascular damage. For catheter tip coil activations ≤300 mA for ≤1 minute in normal carotid flow, 0 of 43 samples had tissue damage. CONCLUSIONS: Activations of copper coils at the tip of microcatheters at low currents in 1.5T MR scanners can be achieved without significant damage to blood vessel walls in a controlled experimental setting. Further optimization of catheter design and procedure protocols is necessary for safe remote control magnetic catheter guidance.
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Quemaduras por Electricidad/etiología , Arterias Carótidas/patología , Arterias Carótidas/cirugía , Traumatismos de las Arterias Carótidas/etiología , Cateterismo/instrumentación , Imagen por Resonancia Magnética Intervencional/efectos adversos , Imagen por Resonancia Magnética Intervencional/instrumentación , Animales , Quemaduras por Electricidad/diagnóstico , Traumatismos de las Arterias Carótidas/patología , Traumatismos de las Arterias Carótidas/prevención & control , Cateterismo/efectos adversos , Diseño de Equipo , Análisis de Falla de Equipo , Seguridad de Equipos , PorcinosRESUMEN
BACKGROUND: Scedosporium and Pseudallescheria species are the second most common lung-colonising fungi in cystic fibrosis (CF) patients. For epidemiological reasons it is important to trace sources of infection, routes of transmission and to determine whether these fungi are transient or permanent colonisers of the respiratory tract. Molecular typing methods like multilocus sequence typing (MLST) help provide this data. METHODS: Clinical isolates of the P. boydii complex (including S. apiospermum and P. boydii) from CF patients in different regions of Germany were studied using MLST. Five gene loci, ACT, CAL, RPB2, BT2 and SOD2, were analysed. RESULTS: The S. apiospermum isolates from 34 patients were assigned to 32 sequence types (STs), and the P. boydii isolates from 14 patients to 8 STs. The results revealed that patients can be colonised by individual strains for years. CONCLUSIONS: The MLST scheme developed for S. apiospermum and P. boydii is a highly effective tool for epidemiologic studies worldwide. The MLST data are accessible at http://mlst.mycologylab.org/.
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Técnicas de Tipificación Bacteriana , Fibrosis Quística/microbiología , Tipificación de Secuencias Multilocus , Técnicas de Tipificación Micológica , Pseudallescheria/clasificación , Scedosporium/clasificación , Adolescente , Adulto , Niño , Preescolar , ADN Bacteriano/genética , ADN de Hongos/genética , Femenino , Frecuencia de los Genes , Variación Genética , Técnicas de Genotipaje , Humanos , Masculino , Reacción en Cadena de la Polimerasa/métodos , Pseudallescheria/aislamiento & purificación , Scedosporium/aislamiento & purificación , Adulto JovenRESUMEN
Conventionally sintered hydroxyapatite-based materials for bone repair show poor resorbability due to the loss of nanocrystallinity. The present study describes a method to establish nanocrystalline hydroxyapatite granules. The material was prepared by ionotropic gelation of an alginate sol containing hydroxyapatite (HA) powder. Subsequent thermal elimination of alginate at 650 °C yielded non-sintered, but unexpectedly stable hydroxyapatite granules. By adding stearic acid as an organic filler to the alginate/HA suspension, the granules exhibited macropores after thermal treatment. A third type of material was achieved by additional coating of the granules with silica particles. Microstructure and specific surface area of the different materials were characterized in comparison to the already established granular calcium phosphate material Cerasorb M(®). Cytocompatibility and potential for bone regeneration of the materials was evaluated by in vitro examinations with osteosarcoma cells and osteoclasts. Osteoblast-like SaOS-2 cells proliferated on all examined materials and showed the typical increase of alkaline phosphatase (ALP) activity during cultivation. Expression of bone-related genes coding for ALP, osteonectin, osteopontin, osteocalcin and bone sialoprotein II on the materials was proven by RT-PCR. Human monocytes were seeded onto the different granules and osteoclastogenesis was examined by activity measurement of tartrate-specific acid phosphatase (TRAP). Gene expression analysis after 23 days of cultivation revealed an increased expression of osteoclast-related genes TRAP, vitronectin receptor and cathepsin K, which was on the same level for all examined materials. These results indicate, that the nanocrystalline granular materials are of clinical interest, especially for bone regeneration.
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Regeneración Ósea , Durapatita/química , Durapatita/farmacología , Nanopartículas/química , Nanopartículas/uso terapéutico , Osteoblastos/efectos de los fármacos , Osteoclastos/efectos de los fármacos , Adulto , Regeneración Ósea/efectos de los fármacos , Regeneración Ósea/genética , Sustitutos de Huesos/química , Sustitutos de Huesos/farmacología , Sustitutos de Huesos/uso terapéutico , Adhesión Celular/efectos de los fármacos , Proliferación Celular/efectos de los fármacos , Células Cultivadas , Durapatita/uso terapéutico , Expresión Génica/efectos de los fármacos , Regeneración Tisular Dirigida/instrumentación , Regeneración Tisular Dirigida/métodos , Humanos , Osteoblastos/citología , Osteoblastos/metabolismo , Osteoblastos/fisiología , Osteoclastos/citología , Osteoclastos/metabolismo , Osteoclastos/fisiología , Osteogénesis/efectos de los fármacos , Osteogénesis/genética , Osteogénesis/fisiología , Tamaño de la Partícula , Polvos/química , Polvos/farmacología , Polvos/uso terapéutico , Ingeniería de Tejidos/instrumentación , Ingeniería de Tejidos/métodosRESUMEN
BACKGROUND: Marfan syndrome is a heritable connective tissue disease. Definitive diagnosis is complex, and requires sequencing of a large gene, FBN1. AIM: We aimed to develop a simple model to estimate the pre-test probability of Marfan syndrome. DESIGN: Prospective cross-sectional study. METHODS: We applied diagnostic standards for definitive diagnosis or exclusion of Marfan syndrome in 329 consecutive persons. In 208 persons with random assignment to our derivation group, we performed multivariate logistic regression to assess 14 clinical variables for inclusion in a prediction model with derivation of score points from the estimated coefficients. We created cut-offs to classify low, moderate and high probability of Marfan syndrome. For validation, we applied the model to the remaining 121 persons. RESULTS: We identified seven variables for inclusion in the final model, where we assigned four score points to ectopia lentis, two points to a family history of Marfan syndrome, and one point to previous thoracic aortic surgery, to pectus excavatum, to a wrist and thumb sign, to previous pneumothorax, and to skin striae. In the derivation group 12, 42 and 92% of persons with low (≤1 point), moderate (>1-3.5 points) or high pre-test probability (>3.5 points) had Marfan syndrome, compared to 12, 57 and 91%, respectively, in the validation group. Positive likelihood ratios were 13.96 and 8.54 in the high probability group of the derivation and validation group, respectively. CONCLUSION: A simple prediction model provides evidence for Marfan syndrome. This model can be used to identify patients who require definitive diagnostic work-up.
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Técnicas de Apoyo para la Decisión , Síndrome de Marfan/diagnóstico , Adolescente , Adulto , Anciano , Estudios Transversales , Femenino , Fibrilina-1 , Fibrilinas , Humanos , Masculino , Síndrome de Marfan/genética , Proteínas de Microfilamentos/genética , Persona de Mediana Edad , Mutación/genética , Valor Predictivo de las Pruebas , Estudios Prospectivos , Adulto JovenRESUMEN
Marfan syndrome is considered a clinical diagnosis. Three diagnostic classifications comprising first, Marfan genotype with a causative FBN1 gene mutation; second, Marfan phenotype with clinical criteria of the original Ghent nosology (Ghent-1); and third, phenotype with clinical criteria of its current revision (Ghent-2) in 300 consecutive persons referred for confirmation or exclusion of Marfan syndrome (150 men, 150 women aged 35 ± 13 years) were used. Sequencing of TGBR1/2 genes was performed in 128 persons without FBN1 mutation. Marfan genotype was present in 140, Ghent-1 phenotype in 139, and Ghent-2 phenotype in 124 of 300 study patients. Marfan syndrome was confirmed in 94 and excluded in 129 persons consistently by all classifications, but classifications were discordant in 77 persons. With combined genotype and phenotype information confirmation of Marfan syndrome was finally achieved in 126 persons by Ghent-1 and in 125 persons by Ghent-2 among 140 persons with Marfan genotype, and exclusion was accomplished in 139 persons by Ghent-1 and in 141 persons by Ghent-2 among 160 persons without Marfan genotype. In total, genotype information changed final diagnoses in 22 persons with Ghent-1, and in 32 persons with Ghent-2. It is concluded that genotype information is essential for diagnosis or exclusion of Marfan syndrome.
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Genotipo , Síndrome de Marfan/diagnóstico , Síndrome de Marfan/genética , Fenotipo , Adolescente , Adulto , Anciano , Femenino , Humanos , Masculino , Persona de Mediana EdadRESUMEN
Mutations in the genes FBN1, TGFBR1, and TGFBR2 can result in heritable connective tissue disorders comprising the Marfan syndrome and the Loeys-Dietz syndrome. Dural ectasia is a characteristic manifestation of both syndromes. However, dural ectasia has not yet been investigated in connective tissue disorders that are unrelated to mutations in the FBN1, TGFBR1 or TGFBR2 genes. Here, we assessed dural ectasia in 33 individuals both with typical manifestations of heritable connective tissue disease and in whom mutations in all three genes had been excluded. We identified 19 individuals with dural ectasia (58%), who exhibited major skeletal manifestations of the Marfan syndrome more frequently than the remaining 14 persons without dural ectasia (p = 0.06). Moreover, only persons with dural ectasia fulfilled clinical criteria of the Marfan syndrome (p = 0.01). Conversely, aortic aneurysm (12 patients; p = 0.8), aortic dissection (five patients; p = 0.1), spontaneous dissection of the carotid arteries (five patients; p = 1), and mitral valve prolapse (13 patients; p = 0.4) were similarly frequent irrespective of dural ectasia. We conclude that dural ectasia is a marker for connective tissue disease which coincides with skeletal rather than with cardiovascular manifestations, and which may involve currently uncharacterized pathogenetic mechanisms and syndromes.
Asunto(s)
Duramadre/anomalías , Síndrome de Marfan/diagnóstico , Proteínas de Microfilamentos/genética , Proteínas Serina-Treonina Quinasas/genética , Receptores de Factores de Crecimiento Transformadores beta/genética , Seno Aórtico/anomalías , Adolescente , Adulto , Niño , Análisis Mutacional de ADN , Diagnóstico Diferencial , Dilatación Patológica/diagnóstico , Dilatación Patológica/genética , Femenino , Fibrilina-1 , Fibrilinas , Pruebas Genéticas , Humanos , Masculino , Persona de Mediana Edad , Mutación , Receptor Tipo I de Factor de Crecimiento Transformador beta , Receptor Tipo II de Factor de Crecimiento Transformador beta , Adulto JovenRESUMEN
Bone remodeling and, therefore, integration of implant materials require the coordinated regulation of osteoblast and osteoclast activity. This is why the in vitro evaluation of biomaterials for bone regeneration should involve not only the analysis of osteoblast differentiation but also the formation and differentiation of osteoclasts. In the present study, we applied a material made of mineralized collagen I that mimics extracellular bone matrix to establish a culture system, which allows the cocultivation of human monocytes and human mesenchymal stem cells (hMSCs), which were differentiated into osteoclast-like cells and osteoblasts, respectively. Both cell types were cultivated on membrane-like structures from mineralized collagen. Transwell inserts were used to spatially separate the cell types but allowed exchange of soluble factors. The osteoclastogenesis and osteogenic differentiation were evaluated by analysis of gene expression, determination of alkaline phosphatase (ALP), and tartrate-resistant acidic phosphatase (TRAP) activity. Furthermore, cell morphology was studied using scanning electron and transmission electron microscopy. Osteogenically induced hMSC showed an increased specific ALP activity as well as increased gene expression of gene coding for alkaline phosphatase (ALPL), when cocultivated with differentiating osteoclasts. Adipogenic differentiation of hMSCs was suppressed by the presence of osteoclasts as indicated by a major decrease in adipocyte cell number and a decrease in gene expression of adipogenic markers. The formation of multinucleated osteoclasts seems to be decreased in the presence of osteogenically induced hMSC as indicated by electron microscopic evaluation and determination of TRAP activity. However, gene expression of osteoclast markers was not decreased in coculture with osteogenically induced hMSC.