Colgajo nautilus y colgajo en muleta taurina para defectos quirúrgicos faciales periorificiales / Nautilus Flap and Bullfighter Crutch Flap for the Repair of Periorificial Surgical Defects of the Face

Antecedentes La reconstrucción de defectos quirúrgicos en regiones anatómicas próximas a bordes palpebrales, fosas nasales o boca es un reto debido su tendencia a ser deformadas por la tensión del cierre directo o los colgajos cutáneos. El empleo de nuevas técnicas reconstructivas que eviten la retracción de estas zonas sensibles puede suponer un avance importante. Pacientes y métodos Se emplean 2 nuevos diseños, el colgajo nautilus y el colgajo en muleta taurina, para la reconstrucción de defectos quirúrgicos periorificiales, realizando una recolección retrospectiva de los mismos en las zonas periparpebral, perivestibular nasal y peribucal. El colgajo nautilus se empleó en 4 pacientes con defectos periparpebrales y 2 peribucales. El colgajo en muleta taurina se utilizó para la reconstrucción de 14 defectos en ala nasal. Resultados En los 20 pacientes se obtuvieron resultados muy satisfactorios tanto a nivel estético como funcional, sin la aparición de ectropión, colapso del vestíbulo nasal o asimetría de borde labial. No se observó necrosis en ningún caso. Conclusiones Proponemos el colgajo nautilus y el colgajo en muleta taurina como una excelente opción reconstructiva para defectos quirúrgicos localizados en zonas periorificiales (AU)

Background Reconstruction of surgical defects located close to eyelid edges, nostrils, or the mouth is challenging, as tension generated by direct closure or skin flaps in these sensitive regions tends to cause distortion. New repair techniques that prevent retraction may significantly improve outcomes. Patients and methods Retrospective study of the use of 2 novel flap designs—the nautilus flap and the bullfighter crutch flap—to repair surgical defects in the peripalpebral, perivestibular, nasal, and perioral areas. The nautilus flap was used to repair 4 peripalpebral defects and 2 perioral defects, and the bullfighter crutch flap to repair 14 nasal ala defects.Results Cosmetic and functional outcomes were very satisfactory in all 20 patients, with no cases of ectropion, nasal vestibule collapse, or labial asymmetry. Necrosis did not occur in any of the cases. Conclusions The nautilus and bullfighter crutch flaps appear to be excellent choices for reconstructing surgical defects in periorificial areas (AU)

[Translated article] Nautilus Flap and Bullfighter Crutch Flap for the Repair of Periorificial Surgical Defects of the Face / Colgajo nautilus y colgajo en muleta taurina para defectos quirúrgicos faciales periorificiales

Background Reconstruction of surgical defects located close to eyelid edges, nostrils, or the mouth is challenging, as tension generated by direct closure or skin flaps in these sensitive regions tends to cause distortion. New repair techniques that prevent retraction may significantly improve outcomes. Patients and methods Retrospective study of the use of 2 novel flap designs—the nautilus flap and the bullfighter crutch flap—to repair surgical defects in the peripalpebral, perivestibular, nasal, and perioral areas. The nautilus flap was used to repair 4 peripalpebral defects and 2 perioral defects, and the bullfighter crutch flap to repair 14 nasal ala defects.Results Cosmetic and functional outcomes were very satisfactory in all 20 patients, with no cases of ectropion, nasal vestibule collapse, or labial asymmetry. Necrosis did not occur in any of the cases. Conclusions The nautilus and bullfighter crutch flaps appear to be excellent choices for reconstructing surgical defects in periorificial areas (AU)

Antecedentes La reconstrucción de defectos quirúrgicos en regiones anatómicas próximas a bordes palpebrales, fosas nasales o boca es un reto debido su tendencia a ser deformadas por la tensión del cierre directo o los colgajos cutáneos. El empleo de nuevas técnicas reconstructivas que eviten la retracción de estas zonas sensibles puede suponer un avance importante. Pacientes y métodos Se emplean 2 nuevos diseños, el colgajo nautilus y el colgajo en muleta taurina, para la reconstrucción de defectos quirúrgicos periorificiales, realizando una recolección retrospectiva de los mismos en las zonas periparpebral, perivestibular nasal y peribucal. El colgajo nautilus se empleó en 4 pacientes con defectos periparpebrales y 2 peribucales. El colgajo en muleta taurina se utilizó para la reconstrucción de 14 defectos en ala nasal. Resultados En los 20 pacientes se obtuvieron resultados muy satisfactorios tanto a nivel estético como funcional, sin la aparición de ectropión, colapso del vestíbulo nasal o asimetría de borde labial. No se observó necrosis en ningún caso. Conclusiones Proponemos el colgajo nautilus y el colgajo en muleta taurina como una excelente opción reconstructiva para defectos quirúrgicos localizados en zonas periorificiales (AU)

Nautilus Flap and Bullfighter Crutch Flap for the Repair of Periorificial Surgical Defects of the Face. / Colgajo nautilus y colgajo en muleta taurina para defectos quirúrgicos faciales periorificiales.

BACKGROUND: Reconstruction of surgical defects located close to eyelid edges, nostrils, or the mouth is challenging, as tension generated by direct closure or skin flaps in these sensitive regions tends to cause distortion. New repair techniques that prevent retraction may significantly improve outcomes. PATIENTS AND METHODS: Retrospective study of the use of 2 novel flap designs-the nautilus flap and the bullfighter crutch flap-to repair surgical defects in the peripalpebral, perivestibular, nasal, and perioral areas. The nautilus flap was used to repair 4 peripalpebral defects and 2 perioral defects, and the bullfighter crutch flap to repair 14 nasal ala defects. RESULTS: Cosmetic and functional outcomes were very satisfactory in all 20 patients, with no cases of ectropion, nasal vestibule collapse, or labial asymmetry. Necrosis did not occur in any of the cases. CONCLUSIONS: The nautilus and bullfighter crutch flaps appear to be excellent choices for reconstructing surgical defects in periorificial areas.

[Translated article] Nautilus Flap and Bullfighter Crutch Flap for the Repair of Periorificial Surgical Defects of the Face.

BACKGROUND: Reconstruction of surgical defects located close to eyelid edges, nostrils, or the mouth is challenging, as tension generated by direct closure or skin flaps in these sensitive regions tends to cause distortion. New repair techniques that prevent retraction may significantly improve outcomes. PATIENTS AND METHODS: Retrospective study of the use of 2 novel flap designs-the nautilus flap and the bullfighter crutch flap-to repair surgical defects in the peripalpebral, perivestibular, nasal, and perioral areas. The nautilus flap was used to repair 4 peripalpebral defects and 2 perioral defects, and the bullfighter crutch flap to repair 14 nasal ala defects. RESULTS: Cosmetic and functional outcomes were very satisfactory in all 20 patients, with no cases of ectropion, nasal vestibule collapse, or labial asymmetry. Necrosis did not occur in any of the cases. CONCLUSIONS: The nautilus and bullfighter crutch flaps appear to be excellent choices for reconstructing surgical defects in periorificial areas.

Estesioneuroblastoma o neuroblastoma olfatorio / Esthesioneuroblastoma or olfactory neuroblastoma

Estesioneuroblastoma o también llamado neuroblastoma olfatorio es un tumor maligno infrecuente derivado del neuroepitelio olfatorio. La historia clínica de un paciente con un neuroblastoma olfatorio es muy inespecífica. Durante la última década, los métodos endoscópicos se han aplicado gradualmente para el tratamiento de tumores de la base del cráneo. El tratamiento puramente endoscópico endonasal del estesioneuroblastoma ha mostrado excelentes resultados de supervivencia con disminución de las complicaciones. Presentamos un paciente afectado por un neuroblastoma olfatorio tratado solo con cirugía endoscópica endonasal.

Esthesioneuroblastoma or olfactory neuroblastoma is an uncommon malignant tumor arising from the olfactory neuroepithelium. The clinical symptoms of a patient with olfactory neuroblastoma are very inespecific. During the past decade, endoscopic approaches have been gradually applied in treating skull base tumors. The purely endoscopic endonasal surgery for esthesioneuroblastoma showed successful survival results with remarkably decreased complications. We reported a patient who suffered olfactory neuroblastoma treated with only endoscopic endonasal surgery.

Paving the way of systems biology and precision medicine in allergic diseases: the MeDALL success story: Mechanisms of the Development of ALLergy; EU FP7-CP-IP; Project No: 261357; 2010-2015.

MeDALL (Mechanisms of the Development of ALLergy; EU FP7-CP-IP; Project No: 261357; 2010-2015) has proposed an innovative approach to develop early indicators for the prediction, diagnosis, prevention and targets for therapy. MeDALL has linked epidemiological, clinical and basic research using a stepwise, large-scale and integrative approach: MeDALL data of precisely phenotyped children followed in 14 birth cohorts spread across Europe were combined with systems biology (omics, IgE measurement using microarrays) and environmental data. Multimorbidity in the same child is more common than expected by chance alone, suggesting that these diseases share causal mechanisms irrespective of IgE sensitization. IgE sensitization should be considered differently in monosensitized and polysensitized individuals. Allergic multimorbidities and IgE polysensitization are often associated with the persistence or severity of allergic diseases. Environmental exposures are relevant for the development of allergy-related diseases. To complement the population-based studies in children, MeDALL included mechanistic experimental animal studies and in vitro studies in humans. The integration of multimorbidities and polysensitization has resulted in a new classification framework of allergic diseases that could help to improve the understanding of genetic and epigenetic mechanisms of allergy as well as to better manage allergic diseases. Ethics and gender were considered. MeDALL has deployed translational activities within the EU agenda.

Phenotyping asthma, rhinitis and eczema in MeDALL population-based birth cohorts: an allergic comorbidity cluster.

BACKGROUND: Asthma, rhinitis and eczema often co-occur in children, but their interrelationships at the population level have been poorly addressed. We assessed co-occurrence of childhood asthma, rhinitis and eczema using unsupervised statistical techniques. METHODS: We included 17 209 children at 4 years and 14 585 at 8 years from seven European population-based birth cohorts (MeDALL project). At each age period, children were grouped, using partitioning cluster analysis, according to the distribution of 23 variables covering symptoms 'ever' and 'in the last 12 months', doctor diagnosis, age of onset and treatments of asthma, rhinitis and eczema; immunoglobulin E sensitization; weight; and height. We tested the sensitivity of our estimates to subject and variable selections, and to different statistical approaches, including latent class analysis and self-organizing maps. RESULTS: Two groups were identified as the optimal way to cluster the data at both age periods and in all sensitivity analyses. The first (reference) group at 4 and 8 years (including 70% and 79% of children, respectively) was characterized by a low prevalence of symptoms and sensitization, whereas the second (symptomatic) group exhibited more frequent symptoms and sensitization. Ninety-nine percentage of children with comorbidities (co-occurrence of asthma, rhinitis and/or eczema) were included in the symptomatic group at both ages. The children's characteristics in both groups were consistent in all sensitivity analyses. CONCLUSION: At 4 and 8 years, at the population level, asthma, rhinitis and eczema can be classified together as an allergic comorbidity cluster. Future research including time-repeated assessments and biological data will help understanding the interrelationships between these diseases.

Are allergic multimorbidities and IgE polysensitization associated with the persistence or re-occurrence of foetal type 2 signalling? The MeDALL hypothesis.

Allergic diseases [asthma, rhinitis and atopic dermatitis (AD)] are complex. They are associated with allergen-specific IgE and nonallergic mechanisms that may coexist in the same patient. In addition, these diseases tend to cluster and patients present concomitant or consecutive diseases (multimorbidity). IgE sensitization should be considered as a quantitative trait. Important clinical and immunological differences exist between mono- and polysensitized subjects. Multimorbidities of allergic diseases share common causal mechanisms that are only partly IgE-mediated. Persistence of allergic diseases over time is associated with multimorbidity and/or IgE polysensitization. The importance of the family history of allergy may decrease with age. This review puts forward the hypothesis that allergic multimorbidities and IgE polysensitization are associated and related to the persistence or re-occurrence of foetal type 2 signalling. Asthma, rhinitis and AD are manifestations of a common systemic immune imbalance (mesodermal origin) with specific patterns of remodelling (ectodermal or endodermal origin). This study proposes a new classification of IgE-mediated allergic diseases that allows the definition of novel phenotypes to (i) better understand genetic and epigenetic mechanisms, (ii) better stratify allergic preschool children for prognosis and (iii) propose novel strategies of treatment and prevention.

Inhibition of VEGF expression through blockade of Hif1α and STAT3 signalling mediates the anti-angiogenic effect of melatonin in HepG2 liver cancer cells.

BACKGROUND: Hepatocellular carcinoma (HCC) growth relies on angiogenesis via vascular endothelial growth factor (VEGF) release. Hypoxia within tumour environment leads to intracellular stabilisation of hypoxia inducible factor 1 alpha (Hif1α) and signal transducer and activator of transcription (STAT3). Melatonin induces apoptosis in HCC, and shows anti-angiogenic features in several tumours. In this study, we used human HepG2 liver cancer cells as an in vitro model to investigate the anti-angiogenic effects of melatonin. METHODS: HepG2 cells were treated with melatonin under normoxic or CoCl2-induced hypoxia. Gene expression was analysed by RT-qPCR and western blot. Melatonin-induced anti-angiogenic activity was confirmed by in vivo human umbilical vein endothelial cells (HUVECs) tube formation assay. Secreted VEGF was measured by ELISA. Immunofluorescence was performed to analyse Hif1α cellular localisation. Physical interaction between Hif1α and its co-activators was analysed by immunoprecipitation and chromatin immunoprecipitation (ChIP). RESULTS: Melatonin at a pharmacological concentration (1 mM) decreases cellular and secreted VEGF levels, and prevents HUVECs tube formation under hypoxia, associated with a reduction in Hif1α protein expression, nuclear localisation, and transcriptional activity. While hypoxia increases phospho-STAT3, Hif1α, and CBP/p300 recruitment as a transcriptional complex within the VEGF promoter, melatonin 1 mM decreases their physical interaction. Melatonin and the selective STAT3 inhibitor Stattic show a synergic effect on Hif1α, STAT3, and VEGF expression. CONCLUSION: Melatonin exerts an anti-angiogenic activity in HepG2 cells by interfering with the transcriptional activation of VEGF, via Hif1α and STAT3. Our results provide evidence to consider this indole as a powerful anti-angiogenic agent for HCC treatment.

Severe chronic allergic (and related) diseases: a uniform approach--a MeDALL--GA2LEN--ARIA position paper.

Concepts of disease severity, activity, control and responsiveness to treatment are linked but different. Severity refers to the loss of function of the organs induced by the disease process or to the occurrence of severe acute exacerbations. Severity may vary over time and needs regular follow-up. Control is the degree to which therapy goals are currently met. These concepts have evolved over time for asthma in guidelines, task forces or consensus meetings. The aim of this paper is to generalize the approach of the uniform definition of severe asthma presented to WHO for chronic allergic and associated diseases (rhinitis, chronic rhinosinusitis, chronic urticaria and atopic dermatitis) in order to have a uniform definition of severity, control and risk, usable in most situations. It is based on the appropriate diagnosis, availability and accessibility of treatments, treatment responsiveness and associated factors such as comorbidities and risk factors. This uniform definition will allow a better definition of the phenotypes of severe allergic (and related) diseases for clinical practice, research (including epidemiology), public health purposes, education and the discovery of novel therapies.

Oleic acid content is responsible for the reduction in blood pressure induced by olive oil.

Numerous studies have shown that high olive oil intake reduces blood pressure (BP). These positive effects of olive oil have frequently been ascribed to its minor components, such as alpha-tocopherol, polyphenols, and other phenolic compounds that are not present in other oils. However, in this study we demonstrate that the hypotensive effect of olive oil is caused by its high oleic acid (OA) content (approximately 70-80%). We propose that olive oil intake increases OA levels in membranes, which regulates membrane lipid structure (H(II) phase propensity) in such a way as to control G protein-mediated signaling, causing a reduction in BP. This effect is in part caused by its regulatory action on G protein-associated cascades that regulate adenylyl cyclase and phospholipase C. In turn, the OA analogues, elaidic and stearic acids, had no hypotensive activity, indicating that the molecular mechanisms that link membrane lipid structure and BP regulation are very specific. Similarly, soybean oil (with low OA content) did not reduce BP. This study demonstrates that olive oil induces its hypotensive effects through the action of OA.

Sobrecarga articular en el astragalo por sobreentrenamiento en la gimnasia artística: tratamiento podológico / No disponible

La media de preparación de un gimnasta hasta alcanzarlos máximos niveles deportivos es de 11-12años, que un año de entrenamiento puede significarentre 300/310 días de trabajo, que un día de entrenamientopuede tener 2 o 3 sesiones y con un totalde 1000/1500 horas al año de entrenamiento año,es de suponer, que tales intensidades y volúmenesde entrenamiento necesitan que la preparación delos gimnastas sea cuidada hasta el más mínimodetalle, incluyendo la seguridad y la protecciónfrente a lesiones. Es en la fase de recepción y en lasextremidades inferiores donde se localiza el mayoríndice de lesiones y se generan fuerzas de impactocon una magnitud comprendida entre 10-18 vecesel peso corporal.Se presenta un caso clínico y el tratamiento ortopodológicocorrespondiente que se aplico en una jovengimnasta con una lesión por sobrecarga articular enla cabeza del astrágalo debido a un sobreentrenamientodeportivo, más acentuado en el pie derecho,producido por un rozamiento cartilaginoso de lasuperficie articular distal de la tibia con la cúpulaastragalina debido a un pie cavo valgo e hiperlaxo(AU)

The average preparation time for a gymnast to reachthe highest competitive levels is between 11 and 12years. In this period a year’s training may meanbetween 300 to 310 days work. Each day of trainingcould include 2 or 3 sessions, a total of between1000 and 1500 hours training a year. Such intensityand volume of training means the preparation ofgymnasts needs to be carefully controlled to thelast detail, including safety and protection againstinjuries. The majority of injuries occur in the receptionphase, and in the lower extremities. Theimpact blows which are generated are understood tobe between 10 and 18 times the body weight.Here presented is a clinical case and the correspondingorthochiropodical treatment which was appliedto a young gymnast with a joint overload injuryin the head of the astragalus due to competitiveovertraining. This overload was heavier in the rightfoot, produced by cartilaginous chaffing of the distalarticular surface of the tibia with the talar dome dueto a hyperlax concave-valgus foot(AU)

Regular physical activity reduces hospital admission and mortality in chronic obstructive pulmonary disease: a population based cohort study.

BACKGROUND: Information about the influence of regular physical activity on the course of chronic obstructive pulmonary disease (COPD) is scarce. A study was undertaken to examine the association between regular physical activity and both hospital admissions for COPD and all-cause and specific mortality in COPD subjects. METHODS: From a population-based sample recruited in Copenhagen in 1981-3 and 1991-4, 2386 individuals with COPD (according to lung function tests) were identified and followed until 2000. Self-reported regular physical activity at baseline was classified into four categories (very low, low, moderate, and high). Dates and causes of hospital admissions and mortality were obtained from Danish registers. Adjusted associations between physical activity and hospital admissions for COPD and mortality were obtained using negative binomial and Cox regression models, respectively. RESULTS: After adjustment for relevant confounders, subjects reporting low, moderate or high physical activity had a lower risk of hospital admission for COPD during the follow up period than those who reported very low physical activity (incidence rate ratio 0.72, 95% confidence interval (CI) 0.53 to 0.97). Low, moderate and high levels of regular physical activity were associated with an adjusted lower risk of all-cause mortality (hazard ratio (HR) 0.76, 95% CI 0.65 to 0.90) and respiratory mortality (HR 0.70, 95% CI 0.48 to 1.02). No effect modification was found for sex, age group, COPD severity, or a background of ischaemic heart disease. CONCLUSIONS: Subjects with COPD who perform some level of regular physical activity have a lower risk of both COPD admissions and mortality. The recommendation that COPD patients be encouraged to maintain or increase their levels of regular physical activity should be considered in future COPD guidelines, since it is likely to result in a relevant public health benefit.

Factores de riesgo cardiovascular, conocimientos y actitudes acerca de la alimentación / Cardiovascular risk factors: knowledge of and attitudes to diet

Objetivos. Conocer la prevalencia de factores de riesgo cardiovascular, evaluar la asociación entre presentar factores de riesgo cardiovascular y el nivel de conocimientos acerca de alimentos y, finalmente, medir la asociación entre presentar factores de riesgo cardiovascular y seguimiento dietético. Método. Estudio transversal de prevalencia, realizado en un área de salud semirrural. La recogida de datos se hizo mediante entrevista personal en el domicilio de 402 personas de entre 18 y 75 años. El modelo de cuestionario se obtuvo básicamente de la Evaluación Nutricional de la Población Catalana, realizada los años 1992-1993. El presente estudio forma parte del más extenso: "Evaluación del estado nutricional del Área de Salud Lleida 2001-2002". Resultados. Se cuantificó un elevado nivel de conocimientos acerca de los alimentos, tanto en su influencia sobre el colesterol como sobre la salud en general, y no se encontraron diferencias importantes entre los individuos que presentan factores de riesgo cardiovascular y los que no. El hecho de presentar un factor de riesgo está ligado a un mayor seguimiento de dieta, pero sólo es mayoritario en el caso de los sujetos diabéticos (71,4%). Los individuos hipercolesterolémicos que reciben tratamiento farmacológico por este motivo disminuyen el seguimiento dietético respecto a los que no lo reciben (el 28,6 frente al 17,6%). Conclusión. El bajo seguimiento dietético que se ha constatado para algunos factores de riesgo no está ligado a conocimientos deficientes, sino a un problema de actitud ante la dieta

Objectives. To determine the prevalence of cardiovascular risk factors, evaluate the association between the presence of cardiovascular risk factors and the degree of knowledge about nutrition, and finally to measure the association between the presence of cardiovascular risk factors and dietary control. Method. A cross sectional prevalence study was performed in a semirural health area. Data collection was carried out through personal interviews in the homes of 402 individuals aged between 18 and 75 years old. The questionnaire model was obtained from the "Nutritional Evaluation of the Catalan Population" carried out in 1992-1993. This study is part of the more extensive study: "Evaluation of nutritional status in the Health Area of Lleida 2001-2002". Results. Knowledge about diet and its influence on both cholesterol and health in general was good; no significant differences were found between persons with cardiovascular risk factors and those without. Individuals who knew they had one risk factor showed a greater tendency to control their diets but the percentage of individuals doing so was only high among diabetics (71.4%). Individuals with high cholesterol levels who received cholesterol-lowering drugs showed lower dietary control than those not receiving drug treatment (28.6% versus 17.6%). Conclusion. The poor dietary control found for some risk factors was not associated with a lack of knowledge but rather to a lack of motivation regarding diet

Hydrodynamic liver gene transfer mechanism involves transient sinusoidal blood stasis and massive hepatocyte endocytic vesicles.

The present study contributes to clarify the mechanism underlying the high efficacy of hepatocyte gene transfer mediated by hydrodynamic injection. Gene transfer experiments were performed employing the hAAT gene, and the efficacy and differential identification in mouse plasma of human transgene versus mouse gene was assessed by ELISA and proteomic procedures, respectively. By applying different experimental strategies such as cumulative dose-response efficacy, hemodynamic changes reflected by venous pressures, intravital microscopy, and morphological changes established by transmission electron microscopy, we found that: (a) cumulative multiple doses of transgene by hydrodynamic injection are efficient and well tolerated, resulting in therapeutic plasma levels of hAAT; (b) hydrodynamic injection mediates a transient inversion of intrahepatic blood flow, with circulatory stasis for a few minutes mainly in pericentral vein sinusoids; (c) transmission electron microscopy shows hydrodynamic injection to promote massive megafluid endocytic vesicles among hepatocytes around the central vein but not in hepatocytes around the periportal vein. We suggest that the mechanism of hydrodynamic liver gene transfer involves transient inversion of intrahepatic flow, sinusoidal blood stasis, and massive fluid endocytic vesicles in pericentral vein hepatocytes.

Adenocarcinoma papilar de bajo grado de malignidad de glándulas salivares menores: a propósito de un caso clínico / Low-grade papillary adenocarcinoma of the minor salivary glands: a case report

El adenocarcinoma papilar de bajo grado de malignidad de glándulas salivales menores es un tumor poco frecuente que tiene predilección por la mucosa palatina, de todos los adenocarcinomas es el que tiene un comportamiento clínico más agresivo. Es de crecimiento lento y con tendencia a infiltrar estructuras vecinas, dando en un 25 por ciento de los casos metástasis. El propósito de este artículo es presentar un caso de un adenocarcinoma en mucosa yugal derecha, su forma de presentación y la evolución clínica del mismo (AU)

Stability of PEI-DNA and DOTAP-DNA complexes: effect of alkaline pH, heparin and serum.

DNA complexes formed with nonviral vectors such as polyethylenimine (PEI) or 1,2-dioleoyl-3-trimethylammonium-propane (DOTAP) are widely used in gene therapy. These complexes prevent the interaction of DNA with the fluorescent probes usually employed to quantify DNA. We thus studied the procedures for DNA quantification from DNA complexes as well as their stability in the presence of DNase or mouse, rat and human sera. Release of the DNA from its complexes was accomplished by increasing the pH of the medium (from 7.3 to 13.4) or by adding heparin. The stability against degradation was tested in vitro, by incubating the complexes at 37 degrees C in the presence of DNase I and sera from the three species. Both high pH and heparin were able to release DNA from its complexes. Naked DNA formed aggregates with serum proteins that delayed electrophoresis migration, and this effect was reversed in the presence of heparin. However, these aggregates did not protect DNA from digestion by serum DNase, and the DNA digesting ability of serum was: mouse>rat>human. The DNA from the complexes was resistant to degradation by DNase I, although a low proportion of DNA from the complexes was partially digested, as determined by electrophoresis. In contrast, PEI-DNA and DOTAP-DNA complexes were stable in the presence of all sera. Heparin and high pH release DNA from its complexes. The order of DNA degradation is: mouse>rat>human, but DOTAP and PEI avoid degradation of DNA by serum compounds.

Asialofetuin liposome-mediated human alpha1-antitrypsin gene transfer in vivo results in stationary long-term gene expression.

The development of nonviral vectors for in vivo gene delivery to hepatocytes is an interesting topic in view of their safety and tremendous gene therapy potential. Since cationic liposomes and liposome uptake by receptor-mediated mechanisms could offer advantages in the efficacy of liposome-mediated gene transfer, we studied the effect of liposome charge (anionic vs. cationic) and the covalently coupled asialofetuin ligand on the liposome surface in mediating human alpha1-antitrypsin (hAAT) gene transfer to mice in vivo. The changes in liposome charge were made by adding the following lipids to the backbone liposomes: anionic phosphatidylserine, cationic N-[1-(2,3-dioleoyloxy)propyl]-N,N,N-trimethyl-ammonium methylsulfate or a lipopeptide synthesized from dipalmitoylphosphatidylethanolamine and covalently coupled to the cationic nuclear localization signal peptide. Two plasmids containing the hAAT gene were used: pTG7101, containing the complete genomic sequence of the human gene driven by the natural promoter, and p216, containing the human hAAT cDNA under the control of the CMV promoter. The results indicate that both untargeted anionic and cationic liposomes mediate plasma levels of hAAT that decline over time. However, asialofetuin liposomes increase the plasma levels of hAAT and can mediate long-term gene expression (>12 months) with stationary plasma levels of protein. Results from quantitative and qualitative reverse transcriptase polymerase chain reaction match those from protein plasma levels and confirm both the human origin of the message and the liver as source of the protein. The use of asialofetuin liposomes in hepatic gene therapy may both increase and prolong in vivo gene expression of hAAT and other clinically important genes.

Usefulness of the I/D angiotensin-converting enzyme genotype for detecting the risk of left ventricular hypertrophy in pharmacologically treated hypertensive men.

The insertion/deletion polymorphism (I/D) of the angiotensin-converting enzyme (ACE) gene has been associated in some studies with a higher prevalence of left ventricular hypertrophy (LVH), but few of them were performed on pharmacologically treated hypertensive patients. The present study was undertaken to determine whether ACE genotype determination could help in the identification of pharmacologically treated hypertensive patients at a higher risk of LVH. Ninety-six consecutive men with essential hypertension were selected for the study. Left ventricular mass (LVM) was assessed by echocardiography and indexed by body surface area and 82 patients were considered suitable for the study. Three groups of patients were defined on the basis of their I/D ACE genotype: DD (n = 39), ID (n = 33) and II (n = 10). There were no statistically significant differences between the three groups regarding to the severity of hypertension at diagnosis, degree of control of blood pressure or type of antihypertensive drug therapy used. No statistically significant differences were found between the three groups regarding to LVM index (total 124 +/- 31, DD 121 +/- 29, ID 127 +/- 35 and II 122 +/- 18 g/m2), relative wall thickness (total 0.5 +/- 0. 2, DD 0.5 +/- 0.3, ID 0.48 +/- 0.07 and II 0.47 +/- 0.04) or prevalence of LVH (total 34%, DD 31%, ID 39% and II 30% by Cornell criteria and total 39%, DD 33%, ID 45% and II 40% by Framingham criteria). Furthermore, the I and D allele frequency distribution was similar in the whole group of patients, in patients with LVH, and in a control group of healthy volunteers. Our data do not support that the I/D ACE genotype determination helps in identifying treated hypertensive patients at higher risk of LVH. Journal of Human Hypertension (2000) 14, 327-331