Electroporación irreversible: ampliando las fronteras de la ablación / Irreversible electroporation: Beyond the limits of tumor ablation

La electroporación irreversible o IRE (irreversible electroporation) es una técnica de ablación tumoral no térmica basada en la aplicación de pulsos eléctricos de alto voltaje entre pares de agujas insertadas alrededor de un tumor. La corriente generada favorece la creación de nanoporos en la membrana plasmática, desencadenando la apoptosis. Por ello, la IRE puede utilizarse de manera segura en localizaciones cercanas a estructuras vasculares delicadas, contraindicadas para el resto de técnicas termoablativas. Actualmente la IRE se emplea con éxito para la ablación de tumores en páncreas, riñón e hígado y, de manera muy extendida, como opción terapéutica focal para el cáncer de próstata. La necesidad de un manejo anestésico específico y la colocación precisa y en paralelo de múltiples agujas implican un alto nivel de complejidad, siendo necesaria una gran experiencia del equipo intervencionista. No obstante, se trata de una técnica muy prometedora con una gran capacidad inmunológica sistémica que puede provocar un efecto a distancia del tumor tratado (efecto abscopal).(AU)

Irreversible electroporation (IRE) is a non-thermal tumor ablation technique. High-voltage electrical pulses are applied between pairs of electrodes inserted around and/or inside a tumor. The generated electric current induces the creation of nanopores in the cell membrane, triggering apoptosis. As a result, IRE can be safely used in areas near delicate vascular structures where other thermal ablation methods are contraindicated. Currently, IRE has demonstrated to be a successful ablation technique for pancreatic, renal, and liver tumors and is widely used as a focal therapeutic option for prostate cancer. The need for specific anesthetic management and accurate parallel placement of multiple electrodes entails a high level of complexity and great expertise from the interventional team is required. Nevertheless, IRE is a very promising technique with a remarkable systemic immunological capability and may impact on distant metastases (abscopal effect).(AU)

Irreversible electroporation: Beyond the limits of tumor ablation.

Irreversible Electroporation (IRE) is a non-thermal tumor ablation technique. High-voltage electrical pulses are applied between pairs of electrodes inserted around and/or inside a tumor. The generated electric current induces the creation of nanopores in the cell membrane, triggering apoptosis. As a result, IRE can be safely used in areas near delicate vascular structures where other thermal ablation methods are contraindicated. Currently, IRE has demonstrated to be a successful ablation technique for pancreatic, renal, and liver tumors and is widely used as a focal therapeutic option for prostate cancer. The need for specific anesthetic management and accurate parallel placement of multiple electrodes entails a high level of complexity and great expertise from the interventional team is required. Nevertheless, IRE is a very promising technique with a remarkable systemic immunological capability and may impact on distant metastases (abscopal effect).

Evaluation of the biocompatibility of a PVA/SA scaffold with a human gingival fibroblast (HGF) by using electrochemical impedance spectroscopy.

The biocompatibility of human gingival fibroblasts (HGF) was evaluated in different concentrations of poly(vinyl alcohol) and sodium alginate (PVA/SA) nanofibres (3.5 wt% 4 wt% and 5 wt%). The PVA/SA nanofibres were deposited on the surface of an electrode microchip by using the electrospinning technique. Electrochemical impedance spectroscopy (EIS) was applied to measure the dielectric properties of each system. In order to provide a detailed analysis as well as a right physical interpretation of the EIS results, the data was fitted with an electric equivalent circuit based on the EIS and the microscopic assessments. The results registered three different time constants (TCs) of the PVA/SA scaffold which indicated different layers at different depths of the scaffold. The TCs changed their dielectric properties depending on the PVA/SA concentration. The 4 wt% system showed the highest biocompatibility properties, given that its resistance and electrochemical capacitance show the formation of a mature-stage cell interaction of HGF. The EIS data offers an exhaustive analysis of the biological activity of the cell response in real time to determine its biocompatibility features. Fluorescence analysis demonstrated a heterogeneous growth of the HGF on the PVA/SA scaffold surface.

Cistinuria: la importancia del sedimento / Cystinuria: the importance of sediment

La cistinuria es causada por el exceso de un aminoácido llamado cistina (dímero del aminoácido cisteína) en la orina. Fue descrita por primera vez a principios del siglo XIX. Es una enfermedad metabólica congénita con un patrón de herencia autosómico recesivo, se caracteriza por un defecto en el transporte que afecta a determinados aminoácidos dibásicos: cistina, ornitina, lisina y arginina (COLA) en su reabsorción en el túbulo renal y tracto gastrointestinal; como resultado solo la cistina, debido a su gran insolubilidad en orinas ácidas por aumento de la excreción y sobresaturación en la orina, favorece la formación de cristales o precipitados formando cálculos. Presentamos un caso de cistinuria en un adulto que fue diagnosticado por los cristales encontrados en el sedimento urinario, después de hacer diagnóstico diferencial con los cristales de colesterol anhidro, con los que pueden confundirse

Cystinuria is caused by the excess of an amino acid called cystine (amino acid dimer cysteine), in the urine. It was first described at the beginning of the 19th century. It is a congenital metabolic disease with an autosomal recessive inheritance pattern. It is characterised by a defect in transport, which affects certain dibasic amino acids, cystine, ornithine, lysine, and arginine in its reabsorption into the renal tubule and gastrointestinal tract. As a result, only cystine (due to its great insolubility in acid urine owing to increased excretion and supersaturation in urine), promotes the formation of crystals or precipitates that form stones. The case is presented of an adult with cystinuria, who was diagnosed by the crystals found in the urinary sediment, after making a differential diagnosis with the crystals of anhydrous cholesterol, with which they can be confused

Polycaprolactone- and polycaprolactone/ceramic-based 3D-bioplotted porous scaffolds for bone regeneration: A comparative study.

One of the critical challenges that scaffolding faces in the organ and tissue regeneration field lies in mimicking the structure, and the chemical and biological properties of natural tissue. A high-level control over the architecture, mechanical properties and composition of the materials in contact with cells is essential to overcome such challenge. Therefore, definition of the method, materials and parameters for the production of scaffolds during the fabrication stage is critical. With the recent emergence of rapid prototyping (RP), it is now possible to create three-dimensional (3D) scaffolds with the essential characteristics for the proliferation and regeneration of tissues, such as porosity, mechanical strength, pore size and pore interconnectivity, and biocompatibility. In this study, we employed 3D bioplotting, a RP technology, to fabricate scaffolds made from (i) pure polycaprolactone (PCL) and (ii) a composite based on PCL and ceramic micro-powder. The ceramics used for the composite were bovine bone filling Nukbone® (NKB), and hydroxyapatite (HA) with 5%, 10% or 20% wt. CONTENT: The scaffolds were fabricated in a cellular lattice structure (i.e. meshing mode) using a 0/90° lay down pattern with a continuous contour filament in order to achieve interconnected porous reticular structures. We varied the temperature, as well as injection speed and pressure during the bioplotting process to achieve scaffolds with pore size ranging between 200 and 400µm and adequate mechanical stability. The resulting scaffolds had an average pore size of 323µm and an average porosity of 32%. Characterization through ATR-FTIR revealed the presence of the characteristic bands of hydroxyapatite in the PCL matrix, and presented an increase of the intensity of the phosphate and carbonyl bands as the ceramic content increased. The bioplotted 3D scaffolds have a Young's modulus (E) in the range between 0.121 and 0.171GPa, which is compatible with the modulus of natural bone. PCL/NKB scaffolds, particularly 10NKBP (10% NKB wt.) exhibited the highest proliferation optical density, demonstrating an evident osteoconductive effect when cultured in Dulbecco's Modified Eagle Medium (DMEM). Scanning electron microscopy (SEM) confirmed osteoblast anchorage to all composite scaffolds, but a low adhesion to the all-PCL scaffold, as well as cell proliferation. The results from this study demonstrate the potential of PCL/NKB 3D bioplotted scaffolds as viable platforms to enable osseous tissue formation, which can be used in several tissue engineering applications, including improvement of bone tissue regeneration.

Adhesión y proliferación de las células troncales mesenquimales de médula ósea en andamios fibrilares de poli (ácido L-láctico) (PLA) / Adhesion and proliferation of bone marrow mesenchymal stem cells onto poli-L lactic acid (PLA) fiber scaffolds

Resumen: Los andamios fibrilares han recibido un enorme interés como futuros biomateriales con potencial aplicación en el campo de la biomedicina regenerativa. En este sentido, hemos optimizado los parámetros para la síntesis de diferentes concentraciones (6, 7, y 10 %) de andamios de ácido poli-láctico (PLA) por la técnica de hilado por propulsión de gas (AJS). Dichos andamios fueron caracterizados por Microscopía Electrónica de Barrido (SEM) y por espectrometría Infrarroja con Transformada de Fourier (FTIR). Nuestros resultados mostraron que los andamios son fibrilares con diámetros en escalas nanométricas. Asimismo; se estudió la biocompatibilidad celular in vitro al realizar ensayos de adhesión, proliferación y de interacción célula-material al cultivar células troncales mesenquimales derivadas de médula ósea. Nuestros datos indican que las membranas fibrilares de PLA aumentan la respuesta celular, no son citotóxicas al compararse con las películas delgadas de PLA. Por lo tanto; el método de síntesis propuesto tiene potencial para la fabricación de membranas hiladas con una facilidad de procesamiento y podría ser un prometedor biomaterial económico con futuras aplicaciones en la regeneración de tejidos.

Abstract: Fiber scaffolds have received increasing interest as promising biomaterials for potential application in the field of tissue regeneration. In this sense, we optimized the parameters for the synthesis of different concentrations (6, 7, and 10 %) of poly-lactic acid (PLA) scaffolds by air jet spinning technology (AJS). The PLA scaffolds were characterized by Scanning Electron Microscopy (SEM) and Fourier Transform Infrared Spectroscopy (FTIR) analysis. Our results by SEM micrographs showed that scaffolds have a fibrilar morphology with nanoscale diameter of fibers. Biocompatibility assay was observed through an in vitro experiment based on cell attachment, MTT and cell-material interaction assay when culturing bone marrow-derived mesenchymal stem cells onto the PLA spun membrane scaffolds. Our data indicate that fiber membrane of PLA scaffold increase the cellular response, are not cytotoxic when compared to thin films of PLA. Thus; the proposed synthesis method has potential for easy processing of spun fibrilar scaffolds with good biocompatibility and could be a promising economical biomaterial with future potential applications in tissue regeneration.

Properties of carbon nanotube-dispersed Sr-hydroxyapatite injectable material for bone defects.

This study concerns the synthesis of gel materials based on carbon nanotubes dispersed strontium-modified hydroxyapatite (Sr-HA) at different compositions obtained by sol-gel technology and their influence on human-bone-marrow-derived mesenchymal stem cells. Furthermore, an evaluation of the influence of nanotubes and Strontium on physico-chemical, morphological, rheological and biological properties of hydroxyapatite gel was also performed. Morphological analysis (scanning electron microscopy) shows a homogeneous distribution of modified nanotubes in the ceramic matrix improving the bioactive properties of materials. The biological investigations proved that Sr-HA/carbon nanotube gel containing 0-20 mol (%) of Sr showed no toxic effect and promote the expression of early and late markers of osteogenic differentiation in cell culture performed in basal medium without osteogenic factors. Finally, the SrHA/carbon nanotube gels could have a good potential application as filler in bone repair and regeneration and may be used in the osteoporotic disease treatment.

Lesión medular traumática en la infancia y adolescencia en Asturias / Traumatic spinal cord injury in children and adolescents in Asturias

Introducción. La lesión medular traumática es rara en la infancia, pero con graves complicaciones. Objetivo. Conocer la incidencia, etiología y características sociodemográficas de los pacientes pediátricos y adolescentes con lesión medular traumática en Asturias. Pacientes y métodos. Censo de pacientes entre 0-17 años codificados como ‘lesión medular traumática con o sin fractura vertebral’ en Asturias entre 1934 y 2013. Se recogen variables sociodemográficas, hospitalarias y clínicas. Resultados. Se registran 72 pacientes, la mayoría varones (79,2%), con una edad media de 16 años (rango: 5-17 años). La incidencia en 2011 fue de 7,3 por millón. Predominaron las lesiones completas (68,1%) y la localización dorsal (44,4%). Hubo lesión ósea en el 95,8%, y en la mitad de ellos, afectación de dos o más vértebras. La etiología predominante fueron los accidentes de tráfico (52,1%), sobre todo de vehículos de cuatro ruedas (63,4%), con una mayoría de varones con una edad media de 17 años y una localización dorsal (46,3%). En segundo y tercer lugar se situaron la etiología laboral y la deportiva, con el mismo número de casos. En la laboral predominaron los varones y el nivel torácico (77,8%), y el mecanismo fundamental fue el golpe directo. En la deportiva, la mayoría fueron varones, y la zambullida fue la causa más importante, de predominio cervical. Conclusiones. Son necesarios estudios centrados en este grupo poblacional que permitan determinar sus características diferenciadoras para establecer programas de cuidados adaptados a sus necesidades, así como las medidas preventivas más oportunas para reducir las posibilidades de una lesión con tan graves repercusiones (AU)

Introduction. Traumatic spinal cord injury (TSCI) is rare in childhood, but it has serious complications. Aim. To assess incidence, causes and socio-demographic aspects of TSCI among children and adolescents in Asturias. Patients and methods. It was made a census of children among aged 0-17 years codified as ‘TSCI with or without vertebral fracture’ in Asturias between 1934 and 2013. Socio-demographic, hospital and clinical variables were picked up. Results. In total 72 patients were registered, most of them males (79.2%), with an average age of 16 (range: 5-17). The estimated incidence in 2011 was found to be 7.3/million children/year for children aged 0-17 years. Complete TSCIs (68.1%) and dorsal localization (44.4%) were found to be more common. There was bone injury in 95.8% with two or more vertebras affected in a half of the cases. The main cause was traffic accidents (52.1%) mainly the four wheels vehicles (63.4%), most of them males with an average age of 17 and dorsal localization (46.3%). On second and third place were work-related injuries and sports-related injuries, with the same number of cases. In work related injuries, males were the majority, with dorsal level (77.8%) and the direct blow was the fundamental mechanism. In sports there were males also the dominant, being diving the most important cause, with cervical level. Conclusions. It is necessary to carry out studies focused on this population group in order to indentify the own characteristics, to establish care and preventive programs, adapted to their necessities (AU)

[Traumatic spinal cord injury in children and adolescents in Asturias]. / Lesion medular traumatica en la infancia y adolescencia en Asturias.

INTRODUCTION: Traumatic spinal cord injury (TSCI) is rare in childhood, but it has serious complications. AIM: To assess incidence, causes and socio-demographic aspects of TSCI among children and adolescents in Asturias. PATIENTS AND METHODS: It was made a census of children among aged 0-17 years codified as 'TSCI with or without vertebral fracture' in Asturias between 1934 and 2013. Socio-demographic, hospital and clinical variables were picked up. RESULTS: In total 72 patients were registered, most of them males (79.2%), with an average age of 16 (range: 5-17). The estimated incidence in 2011 was found to be 7.3/million children/year for children aged 0-17 years. Complete TSCIs (68.1%) and dorsal localization (44.4%) were found to be more common. There was bone injury in 95.8% with two or more vertebras affected in a half of the cases. The main cause was traffic accidents (52.1%) mainly the four wheels vehicles (63.4%), most of them males with an average age of 17 and dorsal localization (46.3%). On second and third place were work-related injuries and sports-related injuries, with the same number of cases. In work related injuries, males were the majority, with dorsal level (77.8%) and the direct blow was the fundamental mechanism. In sports there were males also the dominant, being diving the most important cause, with cervical level. CONCLUSIONS: It is necessary to carry out studies focused on this population group in order to indentify the own characteristics, to establish care and preventive programs, adapted to their necessities.

TITLE: Lesion medular traumatica en la infancia y adolescencia en Asturias.Introduccion. La lesion medular traumatica es rara en la infancia, pero con graves complicaciones. Objetivo. Conocer la incidencia, etiologia y caracteristicas sociodemograficas de los pacientes pediatricos y adolescentes con lesion medular traumatica en Asturias. Pacientes y metodos. Censo de pacientes entre 0-17 años codificados como 'lesion medular traumatica con o sin fractura vertebral' en Asturias entre 1934 y 2013. Se recogen variables sociodemograficas, hospitalarias y clinicas. Resultados. Se registran 72 pacientes, la mayoria varones (79,2%), con una edad media de 16 años (rango: 5-17 años). La incidencia en 2011 fue de 7,3 por millon. Predominaron las lesiones completas (68,1%) y la localizacion dorsal (44,4%). Hubo lesion osea en el 95,8%, y en la mitad de ellos, afectacion de dos o mas vertebras. La etiologia predominante fueron los accidentes de trafico (52,1%), sobre todo de vehiculos de cuatro ruedas (63,4%), con una mayoria de varones con una edad media de 17 años y una localizacion dorsal (46,3%). En segundo y tercer lugar se situaron la etiologia laboral y la deportiva, con el mismo numero de casos. En la laboral predominaron los varones y el nivel toracico (77,8%), y el mecanismo fundamental fue el golpe directo. En la deportiva, la mayoria fueron varones, y la zambullida fue la causa mas importante, de predominio cervical. Conclusiones. Son necesarios estudios centrados en este grupo poblacional que permitan determinar sus caracteristicas diferenciadoras para establecer programas de cuidados adaptados a sus necesidades, asi como las medidas preventivas mas oportunas para reducir las posibilidades de una lesion con tan graves repercusiones.

Effects on growth and osteogenic differentiation of mesenchymal stem cells by the strontium-added sol-gel hydroxyapatite gel materials.

In the present study, strontium-modified hydroxyapatite gels (Sr-HA) at different concentrations were prepared using sol-gel approach and their effect on human-bone-marrow-derived mesenchymal stem cells, were evaluated. The effect of Strontium on physico-chemical and morphological properties of hydroxyapatite gel were evaluated. Morphological analyses (SEM and TEM) demonstrate that an increasing in the amount of Sr ions doped into HA made the agglomerated particles smaller. The substitution of large Sr2+ for small Ca2+ lead to denser atomic packing of the system causing retardation of crystals growth. The biological results demonstrated that hydroxyapatite gel containing from 0 to 20 mol% of Sr presented no cytotoxicity and promote the expression of osteogenesis related genes including an early marker for osteogenic differentiation ALP; a non-collagen protein OPN and a late marker for osteogenic differentiation OCN. Finally, the Sr-HA gels could have a great potential application as filler in bone repair and regeneration and used in especially in the osteoporotic disease.

Effect of citric acid crosslinking cellulose-based hydrogels on osteogenic differentiation.

Understanding the relationships between material surface properties and cellular responses is essential to designing optimal material surfaces for implantation and tissue engineering. In this study, cellulose hydrogels were crosslinked using a non-toxic and natural component namely citric acid. The chemical treatment induces COOH functional groups that improve the hydrophilicity, roughness, and materials rheological properties. The physiochemical, morphological, and mechanical analyses were performed to analyze the material surface before and after crosslinking. This approach would help determine if the effect of chemical treatment on cellulose hydrogel improves the hydrophilicity, roughness, and rheological properties of the scaffold. In this study, it was demonstrated that the biological responses of human mesenchymal stem cell with regard to cell adhesion, proliferation, and differentiation were influenced in vitro by changing the surface chemistry and roughness.

Bioactivation of calcium deficient hydroxyapatite with foamed gelatin gel. A new injectable self-setting bone analogue.

An alternative approach to bone repair for less invasive surgical techniques, involves the development of biomaterials directly injectable into the injury sites and able to replicate a spatially organized platform with features of bone tissue. Here, the preparation and characterization of an innovative injectable bone analogue made of calcium deficient hydroxyapatite and foamed gelatin is presented. The biopolymer features and the cement self-setting reaction were investigated by rheological analysis. The porous architecture, the evolution of surface morphology and the grains dimension were analyzed with electron microscopy (SEM/ESEM/TEM). The physico-chemical properties were characterized by X-ray diffraction and FTIR analysis. Moreover, an injection test was carried out to prove the positive effect of gelatin on the flow ensuing that cement is fully injectable. The cement mechanical properties are adequate to function as temporary substrate for bone tissue regeneration. Furthermore, MG63 cells and bone marrow-derived human mesenchymal stem cells (hMSCs) were able to migrate and proliferate inside the pores, and hMSCs differentiated to the osteoblastic phenotype. The results are paving the way for an injectable bone substitute with properties that mimic natural bone tissue allowing the successful use as bone filler for craniofacial and orthopedic reconstructions in regenerative medicine.

Disfunción sacroilíaca y dolor lumbar relacionado con el embarazo: ¿son efectivas las terapias médico-manuales? / Sacroiliac malfunction and lumbar pain related with pregnancy: Are medical manual therapies effective?

Introducción. El dolor de espalda durante el embarazo es una entidad de indudable relevancia, y la disfunción sacroilíaca (DASI) una de sus causas más frecuentes. El objetivo del estudio es presentar una serie de casos con DASI y su evolución tras el tratamiento médico-manual. Material y métodos. Serie de 15 pacientes embarazadas, diagnosticadas de DASI mediante la prueba de flexión de cadera en bipedestación (Kirkaldy-Gillet) y prueba en sedestación (Piedallu), que se trataron con una técnica adaptada de medicina manual. Para su seguimiento se aplicaron el test de discapacidad en dolor lumbar de Oswestry y la escala visual analógica (EVA). Resultados. Se observó una mejoría de 2 puntos en la escala EVA y un descenso de 10 puntos en el test de Oswestry. A los 40 días posparto ninguna de las pacientes refería persistencia de los síntomas. Conclusiones. La DASI es una entidad importante en la embarazada con dolor lumbar irradiado, y su manejo difiere de la clásica lumbalgia inespecífica. Las técnicas de medicina manual han demostrado beneficios en esta enfermedad en términos de dolor y discapacidad, pero son necesarios más ensayos clínicos que avalen su eficacia (AU)

Introduction. Back pain during pregnancy is a condition having unquestionable importance, sacroiliac malfunction being one of the most common causes. This study has aimed to analyze the efficiency of medical manual treatment for pregnant women with irradiated lumbar pain and symptoms consistent with sacroiliac malfunction. Material and methods. A study was conducted on 15 pregnant women with sacroiliac malfunction, diagnosed with the standing hip flexion test (Kirkaldy-Gillet) and the sitting test (Piedallu), who were treated with a technique adapted from manual medicine. For monitoring results, lumbar pain disability Oswestry test and visual analog numerical scale (VAS) were used. Results. A 2-point improvement was observed on the VAS scale and a decrease of 10 points on the Oswestry test. At 40 days after giving birth, none of the women reported sacroliac malfunction symptoms. Conclusions. Sacroiliac malfunction is an important condition in pregnant women with irradiated lumbar pain, and its treatment differs from the classic unspecific lower back pain. Medical manual techniques have demonstrated significant benefits in this condition in terms of pain and disability. However, further clinical trials are required to support its efficiency (AU)

Optimizing PANi doped electroactive substrates as patches for the regeneration of cardiac muscle.

In scaffold aided regeneration of muscular tissue, composite materials are currently utilized as a temporary substrate to stimulate tissue formation by controlled electrochemical signals as well as continuous mechanical stimulation until the regeneration processes are completed. Among them, composites from the blending of conductive (CPs) and biocompatible polymers are powerfully emerging as a successful strategy for the regeneration of myocardium due to their unique conductive and biological recognition properties able to assure a more efficient electroactive stimulation of cells. Here, different composite substrates made of synthesized polyaniline (sPANi) and polycaprolactone (PCL) were investigated as platforms for cardiac tissue regeneration. Preliminary, a comparative analysis of substrates conductivity performed on casted films endowed with synthesized polyaniline (sPANi) short fibres or blended with emeraldine base polyaniline (EBPANi) allows to study the attitude of charge transport, depending on the conducting filler amount, shape and spatial distribution. In particular, conducibility tests indicated that sPANi short fibres provide a more efficient transfer of electric signal due to the spatial organization of electroactive needle-like phases up to form a percolative network. On the basis of this characterization, sPANi/PCL electrospun membranes have been also optimized to mimic either the morphological and functional features of the cardiac muscle ECM. The presence of sPANi does not relevantly affect the fibre architecture as confirmed by SEM/image analysis investigation which shows a broader distribution of fibres with only a slight reduction of the average fibre diameter from 7.1 to 6.4 µm. Meanwhile, biological assays--evaluation of cell survival rate by MTT assay and immunostaining of sarcomeric α-actinin of cardiomyocites-like cells--clearly indicate that conductive signals offered by PANi needles, promote the cardiogenic differentiation of hMSC into cardiomyocite-like cells. These preliminary results concur to promise the development of electroactive biodegradable substrates able to efficiently stimulate the basic cell mechanisms, paving the way towards a new generation of synthetic patches for the support of the regeneration of damaged myocardium.

Lesiones apocrinas de la mama / Apocrine lesions of the breast

El cambio apocrino del epitelio mamario es una alteraciónfrecuente habitualmente conocida como metaplasia apocrina. Aunque comúnmente se observa en el epitelio de revestimiento de quistes mamarios, existen otras lesiones, histológicamente más complejas, desde las hiperplasias de diverso grado, la adenosis esclerosante, al carcinoma in situ o invasor, también con citología apocrina. Su particular morfología se acompaña, además, de un perfil de marcadores inmunohistoquímicos distinto al de las lesiones ductales comunes similares: se caracterizan por la expresión de GCDFP, pero, además por la negatividad para receptores de estrógenos, progesterona y bcl-2, y positividad para receptores de andrógenos. El significado biológico de las lesiones apocrinas benignas, en cuanto a riesgo de carcinoma posterior, es controvertido, al igual que su posible carácter preneoplásico

Apocrine change of breast epithelium is a frequent lesion usually known as apocrine metaplasia. It is commonly observed on the lining epithelium of breast cysts, but there are also another histologycally more complex lesions, from hyperplasias or sclerosing adenosis to in situ or infiltrating carcinoma, with apocrine cytology. Besides its special morphology, apocrine metaplasia shows a distinctive immunohistochemical profile, different from similar usual ductal lesions: positivity for GCDFP and androgens receptors and negativity for oestrogen and progesterone receptors and for bcl-2. The risk of subsequent carcinoma in patients with benign apocrine lesions and its possible precancerous condition are controversial subjects

Immunolocalization of a human cementoblastoma-conditioned medium-derived protein.

Little is known about the molecular mechanisms that regulate the cementogenesis process, because specific cementum markers are not yet available. To investigate whether a cementoblastoma-conditioned medium-derived protein (CP) could be useful as a cementum biological marker, we studied its expression and distribution in human periodontal tissues, human periodontal ligament, alveolar bone, and cementoblastoma-derived cells. In human periodontal tissues, immunoreactivity to anti-CP was observed throughout the cementoid phase of acellular and cellular cementum, cementoblasts, cementocytes, cells located in the endosteal spaces of human alveolar bone, and in cells in the periodontal ligament located near the blood vessels. Immunopurified CP promoted cell attachment on human periodontal ligament, alveolar bone-derived cells, and gingival fibroblasts. A monoclonal antibody against bovine cementum attachment protein (CAP) cross-reacted with CP. These findings indicate that CP identifies potential cementoblast progenitor cells, is immunologically related to CAP species, and serves as a biological marker for cementum.

Electron microscopy, micro-analysis, and X-ray diffraction characterization of the mineral-like tissue deposited by human cementum tumor-derived cells.

The nature and characteristics of the mineralized-like tissue deposited by cementoblasts are not well-known due to the difficulties in obtaining and culturing cells representing the cementum phenotype. We hypothesized that a putative cementoblastic cell line derived from a human cementoblastoma could serve as a suitable model to study the physical, chemical, and morphological features of the cementum-like tissue deposited in vitro. The cementoblastoma cell line was studied by transmission electron, high resolution, scanning, and atomic force microscopy and compared with human cellular cementum, human osteoblasts, and human alveolar bone. The analyses of the crystals and mineral-like tissue in the cell line were performed by x-ray diffraction microscopy and energy-dispersive x-ray micro-analysis. TEM examination of cementoblastoma cells revealed the presence of electron-dense intracellular vesicles surrounded by a membrane that contained filaments and needle-like structures. The diffraction patterns obtained from the intracellular material and human cellular cementum were similar, with D-spacings of 3.36 and 2.8, consistent with those of hydroxyapatite (3.440 and 2.814). The composition of the mineral-like tissue had a Ca/P ratio of 1.60 for cementoblastoma cells and 1.97 for human cellular cementum. Na (5.29%) and Cl (1.47%) were present in the composition of cementoblastoma cells. Human cellular cementum additionally contained Mg (4.95%). Osteoblastic cells showed a Ca/P ratio of 1.6280. Na represented 4.52% and Cl 1.22% of its composition. Human alveolar bone had a Ca/P ratio value of 2.01. Na (6.63%), Mg (2.10%), and Cl (0.84%) were also present. All samples examined represented biological-type hydroxyapatite. Based on the compositional and morphological features, these findings indicate that cementoblastoma-derived cells express the human cellular cementum phenotype.

[Bladder transitional cell carcinoma rating, systematization of grading and value of DNA ploidy]. / Contribución a la categorización de los CCT de vejiga, sistemátización de la gradación y utilidad de la ploidía de ADN.

UNLABELLED: TCCs are an heterogenous group of tumors with an uncertain biologic behaviour, especially intermediate grade (G2). Histologic grade and pathologic stage have revealed only partially useful on predicting the outcome. OBJECTIVE: Search and statement of objective and cuantitative parameters able to define prognostic subgroups to TCCs. MATERIAL AND METHOD: We studied consecutive 106 patients with TCC in order to: 1) Identify every variable with independent predictive value in classifying cases of TCC in a three (1,2,3) or two (high/low) grades systems. 2) Make a correlation between DNA ploidy obtained by image analysis and DNA ploidy and S-phase obtained by flow cytometry. RESULTS: Every variable used to define the histologic grade showed significative correlation with both gradation systems. In multivariate analysis, the presence of superficial cells and the mitotic counts revealed us the most valuable variables in predicting the histologic grade. DNA-ploidy (both obtained by static an flow cytometry) correlated well with grade, stage, growth pattern and necrosis, whereas S-phase did so with grade, mitotic index and DNA ploidy. Image cytometry showed similars results to those of flow cytometry and also was able to detect aneuploidy when an situ carcinoma or dysplasia were present in the adyacent mucosa.

[Value of Markovian nuclear texture to predict progression in surface transitional cell carcinoma of the bladder]. / Utilidad de las texturas nucleares Markovianas en la predicción de la progresión en los CCT superficiales de vejiga.

The aim of this work has been to determine if the DNA and nuclear Markovian textures of tissue sections evaluated by image cytometry correlate with the histologic grade and the progression probability of superficial transitional carcinoma of the bladder. In our study, DNA ploidy is related to histologic grade in that aneuploidy frequency increases with a higher grade of malignancy. Image analyses of superficial TCC tissue sections of the bladder, has allowed us to identify "sub-visual parameters" as well as nuclear textures that according to our results can be useful in clarifying the evolutive behaviour of these tumours. Two Markovian textures that identify entropy (TXI) and mean internal contrast (TXB) allow to discriminate between histologic grades as well as progression or non-progression. In conclusion, image analysis cytometry of paraffin embedded tissue sections of TCC of the bladder supplies densitometric parameters related to grade and provides valuable information for the prediction of progression. Quantification of chromatin pattern description in Feulgen-stained nuclei using the Markovian method can be useful in this context.

Human cementum tumor cells have different features from human osteoblastic cells in vitro.

Cells obtained from human cementoblastoma and alveolar bone were isolated and cultured. Initial and late stages of mineralization were assessed by using atomic force microscopy, scanning electron microscopy and X-ray microanalysis. In cultures of cementoblastoma-derived cells the initial stages of mineralization showed well-defined spherical-shaped structures, while the osteoblastic cells showed plaque-like deposits. These morphological patterns of mineral deposition could serve as nucleation centers for hydroxyapatite crystals. Late stages of mineralization at 28 and 35 d maintained those morphological differences established in initial cultures. The material deposited by cementoblastoma and osteoblastic cells, analyzed by EDX spectra, revealed similar Ca/P ratios for both cell types. These values were similar to those reported for hydroxyapatite in enamel and bone. Alkaline phosphatase specific activity (AlP), of osteoblastic cells at 3, 7 and 11 d, showed an increase of 27.9, 50.9 and 37.0% (p < 0.001), respectively. However, at 15 and 19 d there was an increase of AlP activity of cementoblastoma cells by 39.4 and 34.5% over osteoblastic cells (p < 0.001). Immunostaining of cementoblastoma and osteoblastic cells using a specific mAb against a cementum-derived attachment protein revealed strong immunostaining of cementoblastoma cells which was localized to the cell membrane and fibril-like structures (96.2 +/- 1.3). A few osteoblastic cells also stained weakly with the anti-CAP mAb (6.4 +/- 0.6). Sections of decalcified paraffin embedded cementoblastoma specimens, when immunostained with anti-CAP mAb, showed strong immunostaining of the cells surrounding the regular and irregularly-shaped calcified masses of the tumor. Putative cementocytes also stained positively. Immunostaining with a polyclonal antibody against osteopontin strongly stained the osteoblastic cells (89.0 +/- 3.6). Cementoblastoma cells showed weaker staining (54.2 +/- 2.4). The results suggest that cementoblastoma cells could be a major source of specific cementum proteins. These cells could provide the opportunity to elucidate the regulation of the cementogenesis process.