RESUMEN

People who inject drugs (PWID) and prisoners are considered key populations at risk for human immunodeficiency virus (HIV) and/or Hepatitis C Virus (HCV). In 2016, the Joint United Nations Program on HIV/AIDS (UNAIDS) was implemented to eliminate HIV and AIDS by 2030 and the World Health Organization (WHO) presented the first strategy to eliminate viral hepatitis by 2030 as well. Following the objectives of the WHO and the United Nations, the German Federal Ministry of Health (BMG) presented the first integrated overall strategy for HIV and HCV in 2017. This article discusses the situation of PWID and prisoners in Germany with regard to HIV and HCV five years after the adoption of this strategy, on the basis of available data and against the background of the most recent practice in the field. In order to meet the elimination goals by 2030, Germany will have to improve the situation of PWID and prisoners substantially, mainly through the implementation of evidence-based harm reduction measures as well as the promotion of diagnosis and treatment in prisons and in freedom.

Asunto(s)

Consumidores de Drogas , Infecciones por VIH , Hepatitis C , Prisioneros , Abuso de Sustancias por Vía Intravenosa , Humanos , Hepacivirus , VIH , Prisiones , Abuso de Sustancias por Vía Intravenosa/epidemiología , Hepatitis C/epidemiología , Hepatitis C/prevención & control , Hepatitis C/tratamiento farmacológico , Alemania/epidemiología , Organización Mundial de la Salud , Infecciones por VIH/epidemiología , Infecciones por VIH/prevención & control

RESUMEN

Colchicine is commonly prescribed for treatment of inflammatory conditions but has a narrow therapeutic window and dangerous toxicity profile. Here we describe a case of survival after massive unintentional colchicine overdose treated with plasmapheresis and renal replacement therapy. A 37 year old male with history of pericarditis presented to the Emergency Department with a chief complaint of nausea, vomiting, and diarrhea after unintentionally ingesting 36 mg of colchicine 17 h prior to arrival. An initial colchicine concentration resulted at 5.1 ng/mL (30 h post-ingestion) and peaked at 12 ng/mL (40 h post-ingestion). He was treated with continuous kidney replacement therapy (CKRT) beginning on his first day of hospitalization and with plasmapheresis on hospital days two through four. The patient's course was complicated by multiorgan failure including coagulopathy, respiratory failure, neuropathy, renal failure, pancytopenia, and heart failure. He was discharged to inpatient rehabilitation on hospital day 24. On clinical follow up four months after discharge the patient was found to have no significant persistent morbidity related to colchicine overdose.

RESUMEN

Lentiviral vectors pseudotyped with the baculovirus envelope protein GP64 transduce primary cultures of human airway epithelia (HAE) at their apical surface. Our goal in this study was to harness a directed evolution approach to develop a novel envelope glycoprotein with increased transduction properties for HAE. Using error-prone PCR, a library of GP64 mutants was generated and used to prepare a diverse pool of lentiviral virions pseudotyped with GP64 variants. The library was serially passaged on HAE and three GP64 mutations were recovered. Single-, double- and the triple-combination mutant envelope glycoproteins were compared with wild-type GP64 for their ability to transduce HAE. Our results suggest that lentiviral vectors pseudotyped with evolved GP64 transduced HAE with greater efficiency than wild-type GP64. This effect was not observed in primary cultures of porcine airway epithelial cells, suggesting that the directed evolution protocol was species specific. In summary, our studies indicate that serial passage of a GP64 mutant library yielded specific variants with improved HAE cell tropism, yielding tools with the potential to improve the success of gene therapy for airway diseases.

Asunto(s)

Técnicas de Transferencia de Gen , Mucosa Respiratoria/metabolismo , Proteínas del Envoltorio Viral/genética , Animales , Baculoviridae/genética , Células Cultivadas , Terapia Genética/métodos , Vectores Genéticos/genética , Humanos , Lentivirus/genética , Ratones , Ratones Endogámicos BALB C , Mutación , Mucosa Respiratoria/citología , Proteínas del Envoltorio Viral/metabolismo

RESUMEN

BACKGROUND: Dissociative drugs are used for chemical restraint in monkeys. The aim was to evaluate muscle relaxation, recovery, and ophthalmic and hemodynamic parameters in 24 capuchin monkeys subjected to four dissociative anesthesia protocols. METHODS: Animals were anesthetized with tiletamine-zolazepam (TZ), ketamine-xylazine (KX), ketamine-midazolam (KM), or ketamine-dexmedetomidine (KD). Muscle relaxation, digital reflex, lacrimal production, intraocular pressure (IOP), heart and respiratory rates, oxygen saturation (SpO2 ), rectal temperature, non-invasive arterial blood pressure, palpebral and pupillary reflexes, and eyeball positioning were evaluated every 5 minutes for 20 minutes. RESULTS: Muscle relaxation was highest in KM and KD. At 5-minute post-injection, IOP was higher in TZ than in all other groups. There was a significant difference between groups and times in heart and respiratory rates and temperature. There were no significant differences in SpO2, arterial blood pressure, and lacrimal production between groups. CONCLUSIONS: The established parameters may help in clinical and ophthalmic examinations of primates.

Asunto(s)

Anestesia/veterinaria , Anestésicos Disociativos/farmacología , Presión Sanguínea/efectos de los fármacos , Cebus/fisiología , Presión Intraocular/efectos de los fármacos , Lágrimas/efectos de los fármacos , Anestesia General/veterinaria , Anestésicos Combinados/farmacología , Animales , Temperatura Corporal , Femenino , Frecuencia Cardíaca/efectos de los fármacos , Respiración/efectos de los fármacos

RESUMEN

Adeno-associated virus (AAV) vectors have achieved clinical efficacy in treating several diseases. However, enhanced vectors are required to extend these landmark successes to other indications and protein engineering approaches may provide the necessary vector improvements to address such unmet medical needs. To generate new capsid variants with potentially enhanced infectious properties and to gain insights into AAV's evolutionary history, we computationally designed and experimentally constructed a putative ancestral AAV library. Combinatorial variations at 32 amino acid sites were introduced to account for uncertainty in their identities. We then analyzed the evolutionary flexibility of these residues, the majority of which have not been previously studied, by subjecting the library to iterative selection on a representative cell line panel. The resulting variants exhibited transduction efficiencies comparable to the most efficient extant serotypes and, in general, ancestral libraries were broadly infectious across the cell line panel, indicating that they favored promiscuity over specificity. Interestingly, putative ancestral AAVs were more thermostable than modern serotypes and did not use sialic acids, galactose or heparan sulfate proteoglycans for cellular entry. Finally, variants mediated 19- to 31-fold higher gene expression in the muscle compared with AAV1, a clinically used serotype for muscle delivery, highlighting their promise for gene therapy.

Asunto(s)

Dependovirus/genética , Biblioteca de Genes , Animales , Cápside/metabolismo , Línea Celular , Línea Celular Tumoral , Femenino , Terapia Genética/métodos , Vectores Genéticos/genética , Células HEK293 , Humanos , Ratones , Ratones Endogámicos BALB C , Filogenia , Análisis de Secuencia de ADN , Análisis de Secuencia de Proteína

RESUMEN

Transmembrane receptors are the predominant conduit through which cells sense and transduce extracellular information into intracellular biochemical signals. Current methods to control and study receptor function, however, suffer from poor resolution in space and time and often employ receptor overexpression, which can introduce experimental artefacts. We report a genetically encoded approach, termed Clustering Indirectly using Cryptochrome 2 (CLICR), for spatiotemporal control over endogenous transmembrane receptor activation, enabled through the optical regulation of target receptor clustering and downstream signalling using noncovalent interactions with engineered Arabidopsis Cryptochrome 2 (Cry2). CLICR offers a modular platform to enable photocontrol of the clustering of diverse transmembrane receptors including fibroblast growth factor receptor (FGFR), platelet-derived growth factor receptor (PDGFR) and integrins in multiple cell types including neural stem cells. Furthermore, light-inducible manipulation of endogenous receptor tyrosine kinase (RTK) activity can modulate cell polarity and establish phototaxis in fibroblasts. The resulting spatiotemporal control over cellular signalling represents a powerful new optogenetic framework for investigating and controlling cell function and fate.

Asunto(s)

Proteínas de Arabidopsis/metabolismo , Membrana Celular/metabolismo , Criptocromos/metabolismo , Optogenética/métodos , Células 3T3 , Animales , Western Blotting , Polaridad Celular , Fibroblastos , Células HEK293 , Humanos , Inmunoprecipitación , Luz , Ratones , Microscopía Confocal , Proteínas Tirosina Quinasas Receptoras/metabolismo , Transducción de Señal

RESUMEN

Gene delivery vectors based on adeno-associated viruses (AAV) have exhibited promise in both preclinical disease models and human clinical trials for numerous disease targets, including the retinal degenerative disorders Leber's congenital amaurosis and choroideremia. One general challenge for AAV is that preexisting immunity, as well as subsequent development of immunity following vector administration, can severely inhibit systemic AAV vector gene delivery. However, the role of neutralizing antibodies (NABs) in AAV transduction of tissues considered to be immune privileged, such as the eye, is unclear in large animals. Intravitreal AAV administration allows for broad retinal delivery, but is more susceptible to interactions with the immune system than subretinal administration. To assess the effects of systemic anti-AAV antibody levels on intravitreal gene delivery, we quantified the anti-AAV antibodies present in sera from non-human primates before and after intravitreal injections with various AAV capsids. Analysis showed that intravitreal administration resulted in an increase in anti-AAV antibodies regardless of the capsid serotype, transgene or dosage of virus injected. For monkeys injected with wild-type AAV2 and/or an AAV2 mutant, the variable that most significantly affected the production of anti-AAV2 antibodies was the amount of virus delivered. In addition, post-injection antibody titers were highest against the serotype administered, but the antibodies were also cross-reactive against other AAV serotypes. Furthermore, NAB levels in serum correlated with those in vitreal fluid, demonstrating both that this route of administration exposes AAV capsid epitopes to the adaptive immune system and that serum measurements are predictive of vitreous fluid NAB titers. Moreover, the presence of preexisting NAB titers in the serum of monkeys correlated strongly (R=0.76) with weak, decaying or no transgene expression following intravitreal administration of AAV. Investigating anti-AAV antibody development will aid in understanding the interactions between gene therapy vectors and the immune system during ocular administration and can form a basis for future clinical studies applying intravitreal gene delivery.

Asunto(s)

Anticuerpos Neutralizantes/fisiología , Anticuerpos Antivirales/fisiología , Dependovirus/inmunología , Degeneración Retiniana/terapia , Animales , Dependovirus/genética , Terapia Genética , Vectores Genéticos , Células HEK293 , Humanos , Inyecciones Intravítreas , Macaca mulatta , Transducción Genética

RESUMEN

X-linked retinoschisis, a disease characterized by splitting of the retina, is caused by mutations in the retinoschisin gene, which encodes a putative secreted cell adhesion protein. Currently, there is no effective treatment for retinoschisis, though viral vector-mediated gene replacement therapies offer promise. We used intravitreal delivery of three different AAV vectors to target delivery of the RS1 gene to Müller glia, photoreceptors or multiple cell types throughout the retina. Müller glia radially span the entire retina, are accessible from the vitreous, and remain intact throughout progression of the disease. However, photoreceptors, not glia, normally secrete retinoschisin. We compared the efficacy of rescue mediated by retinoschisin secretion from these specific subtypes of retinal cells in the Rs1h-/- mouse model of retinoschisis. Our results indicate that all three vectors deliver the RS1 gene, and that several cell types can secrete retinoschisin, leading to transport of the protein across the retina. The greatest long-term rescue was observed when photoreceptors produce retinoschisin. Similar rescue was observed with photoreceptor-specific or generalized expression, although photoreceptor secretion may contribute to rescue in the latter case. These results collectively point to the importance of cell targeting and appropriate vector choice in the success of retinal gene therapies.

Asunto(s)

Proteínas del Ojo/genética , Terapia Genética/métodos , Retina/citología , Envejecimiento , Animales , Moléculas de Adhesión Celular/genética , Modelos Animales de Enfermedad , Electrorretinografía , Vectores Genéticos/genética , Proteínas Fluorescentes Verdes/genética , Proteínas Fluorescentes Verdes/metabolismo , Humanos , Ratones Endogámicos C57BL , Ratones Mutantes , Técnicas de Cultivo de Órganos , Células Fotorreceptoras de Vertebrados/fisiología , Retina/fisiología , Retinosquisis/genética , Retinosquisis/terapia

RESUMEN

Vesicular stomatitis virus G glycoprotein (VSV-G) is the most widely used envelope protein for retroviral and lentiviral vector pseudotyping; however, serum inactivation of VSV-G pseudotyped vectors is a significant challenge for in vivo gene delivery. To address this problem, we conducted directed evolution of VSV-G to increase its resistance to human serum neutralization. After six selection cycles, numerous common mutations were present. On the basis of their location within VSV-G, we analyzed whether substitutions in several surface exposed residues could endow viral vectors with higher resistance to serum. S162T, T230N and T368A mutations enhanced serum resistance, and additionally K66T, T368A and E380K substitutions increased the thermostability of VSV-G pseudotyped retroviral vectors, an advantageous byproduct of the selection strategy. Analysis of a number of combined mutants revealed that VSV-G harboring T230N+T368A or K66T+S162T+T230N+T368A mutations exhibited both higher in vitro resistance to human serum and higher thermostability, as well as enhanced resistance to rabbit and mouse serum. Finally, lentiviral vectors pseudotyped with these variants were more resistant to human serum in a murine model. These serum-resistant and thermostable VSV-G variants may aid the application of retroviral and lentiviral vectors to gene therapy.

Asunto(s)

Evolución Molecular Dirigida , Técnicas de Transferencia de Gen , Terapia Genética , Glicoproteínas de Membrana/genética , Proteínas del Envoltorio Viral/genética , Animales , Vectores Genéticos , Humanos , Lentivirus/genética , Glicoproteínas de Membrana/metabolismo , Ratones , Mutación , Retroviridae/genética , Suero/química , Suero/virología , Proteínas del Envoltorio Viral/metabolismo

RESUMEN

Gene therapy vectors based on adeno-associated virus (AAV) are currently in clinical trials for numerous disease targets, such as muscular dystrophy, hemophilia, Parkinson's disease, Leber's congenital amaurosis and macular degeneration. Despite its considerable promise and emerging clinical success, several challenges impede the broader implementation of AAV gene therapy, including the prevalence of neutralizing antibodies in the human population, low transduction of a number of therapeutically relevant cell and tissue types, an inability to overcome physical and cellular barriers in vivo and a relatively limited carrying capacity. These challenges arise as the demands we place on AAV vectors are often different from or even at odds with the properties nature bestowed on their parent viruses. Viral-directed evolution-the iterative generation of large, diverse libraries of viral mutants and selection for variants with specific properties of interest-offers an approach to address these problems. Here we outline progress in creating novel classes of AAV variant libraries and highlight the successful isolation of variants with novel and advantageous in vitro and in vivo gene delivery properties.

Asunto(s)

Dependovirus/genética , Evolución Molecular Dirigida , Técnicas de Transferencia de Gen , Vectores Genéticos , Animales , Dependovirus/fisiología , Modelos Animales de Enfermedad , Humanos , Selección Genética , Células Madre

RESUMEN

Delivery of therapeutic genes to a large region of the retina with minimal damage from intraocular surgery is a central goal of treatment for retinal degenerations. Recent studies have shown that AAV9 can reach the central nervous system (CNS) and retina when administered systemically to neonates, which is a promising strategy for some retinal diseases. We investigated whether the retinal transduction efficiency of systemically delivered AAV9 could be improved by mutating capsid surface tyrosines, previously shown to increase the infectivity of several AAV vectors. Specifically, we evaluated retinal transduction following neonatal intravascular administration of AAV9 vectors containing tyrosine to phenylalanine mutations at two highly conserved sites. Our results show that a novel, double tyrosine mutant of AAV9 significantly enhanced gene delivery to the CNS and retina, and that gene expression can be restricted to rod photoreceptor cells by incorporating a rhodopsin promoter. This approach provides a new methodology for the development of retinal gene therapies or creation of animal models of neurodegenerative disease.

Asunto(s)

Sistema Nervioso Central , Dependovirus/genética , Terapia Genética , Retina/patología , Degeneración Retiniana/terapia , Animales , Modelos Animales de Enfermedad , Regulación del Desarrollo de la Expresión Génica , Vectores Genéticos/administración & dosificación , Proteínas Fluorescentes Verdes , Humanos , Ratones , Ratones Endogámicos C57BL , Mutación , Regiones Promotoras Genéticas , Retina/citología , Retina/crecimiento & desarrollo , Degeneración Retiniana/genética , Células Fotorreceptoras Retinianas Bastones/metabolismo , Rodopsina/genética

RESUMEN

The etiology of depression is still poorly understood, but two major causative hypotheses have been put forth: the monoamine deficiency and the stress hypotheses of depression. We evaluate these hypotheses using animal models of endogenous depression and chronic stress. The endogenously depressed rat and its control strain were developed by bidirectional selective breeding from the Wistar-Kyoto (WKY) rat, an accepted model of major depressive disorder (MDD). The WKY More Immobile (WMI) substrain shows high immobility/despair-like behavior in the forced swim test (FST), while the control substrain, WKY Less Immobile (WLI), shows no depressive behavior in the FST. Chronic stress responses were investigated by using Brown Norway, Fischer 344, Lewis and WKY, genetically and behaviorally distinct strains of rats. Animals were either not stressed (NS) or exposed to chronic restraint stress (CRS). Genome-wide microarray analyses identified differentially expressed genes in hippocampi and amygdalae of the endogenous depression and the chronic stress models. No significant difference was observed in the expression of monoaminergic transmission-related genes in either model. Furthermore, very few genes showed overlapping changes in the WMI vs WLI and CRS vs NS comparisons, strongly suggesting divergence between endogenous depressive behavior- and chronic stress-related molecular mechanisms. Taken together, these results posit that although chronic stress may induce depressive behavior, its molecular underpinnings differ from those of endogenous depression in animals and possibly in humans, suggesting the need for different treatments. The identification of novel endogenous depression-related and chronic stress response genes suggests that unexplored molecular mechanisms could be targeted for the development of novel therapeutic agents.

Asunto(s)

Amígdala del Cerebelo/fisiopatología , Trastorno Depresivo/patología , Regulación de la Expresión Génica/fisiología , Hipocampo/fisiopatología , Estrés Psicológico/patología , Corticoesteroides/sangre , Glándulas Suprarrenales/patología , Animales , Peso Corporal , Trastorno Depresivo/sangre , Trastorno Depresivo/genética , Modelos Animales de Enfermedad , Reacción Cataléptica de Congelación/fisiología , Perfilación de la Expresión Génica , Masculino , Análisis de Secuencia por Matrices de Oligonucleótidos , Tamaño de los Órganos , Radioinmunoensayo , Ratas , Ratas Endogámicas F344 , Ratas Endogámicas WKY , Estrés Psicológico/sangre , Natación/psicología

RESUMEN

Nogo-A, an axonal growth inhibitory protein known to be mostly present in CNS myelin, was upregulated in retinal ganglion cells (RGCs) after optic nerve injury in adult mice. Nogo-A increased concomitantly with the endoplasmic reticulum stress (ER stress) marker C/EBP homologous protein (CHOP), but CHOP immunostaining and the apoptosis marker annexin V did not co-localize with Nogo-A in individual RGC cell bodies, suggesting that injury-induced Nogo-A upregulation is not involved in axotomy-induced cell death. Silencing Nogo-A with an adeno-associated virus serotype 2 containing a short hairpin RNA (AAV2.shRNA-Nogo-A) or Nogo-A gene ablation in knock-out (KO) animals had little effect on the lesion-induced cell stress or death. On the other hand, Nogo-A overexpression mediated by AAV2.Nogo-A exacerbated RGC cell death after injury. Strikingly, however, injury-induced sprouting of the cut axons and the expression of growth-associated molecules were markedly reduced by AAV2.shRNA-Nogo-A. The axonal growth in the optic nerve activated by the intraocular injection of the inflammatory molecule Pam3Cys tended to be lower in Nogo-A KO mice than in WT mice. Nogo-A overexpression in RGCs in vivo or in the neuronal cell line F11 in vitro promoted regeneration, demonstrating a positive, cell-autonomous role for neuronal Nogo-A in the modulation of axonal regeneration.

Asunto(s)

Apoptosis/efectos de los fármacos , Estrés del Retículo Endoplásmico , Proteínas de la Mielina/metabolismo , Neuronas/metabolismo , Retina/citología , Regulación hacia Arriba , Animales , Anexina A5/metabolismo , Axotomía , Células Cultivadas , Dependovirus/genética , Lipoproteínas/farmacología , Masculino , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Proteínas de la Mielina/antagonistas & inhibidores , Proteínas de la Mielina/genética , Neuritas/fisiología , Proteínas Nogo , Interferencia de ARN , ARN Interferente Pequeño/metabolismo , Regeneración/efectos de los fármacos , Retina/metabolismo , Células Ganglionares de la Retina/citología , Células Ganglionares de la Retina/metabolismo , Factor de Transcripción CHOP/genética , Factor de Transcripción CHOP/metabolismo

RESUMEN

Adeno-associated viral (AAV) vectors, which are undergoing broad exploration in clinical trials, have significant promise for therapeutic gene delivery because of their safety and delivery efficiency. Gene delivery technologies capable of mediating localized gene expression may further enhance the potential of AAV in a variety of therapeutic applications by reducing spread outside a target region, which may thereby reduce off-target side effects. We have genetically engineered an AAV variant capable of binding to surfaces with high affinity through a hexa-histidine metal-binding interaction. This immobilized AAV vector system mediates high-efficiency delivery to cells that contact the surface and thus may have promise for localized gene delivery, which may aid numerous applications of AAV delivery to gene therapy.

Asunto(s)

Dependovirus/genética , Terapia Genética/métodos , Vectores Genéticos/genética , Histidina/química , Oligopéptidos/química , Células Cultivadas , Técnicas de Transferencia de Gen , Humanos , Transducción Genética

RESUMEN

Stem cells have the capability to self-renew and maintain their undifferentiated state or to differentiate into one or more specialised cell types. Stem cell expansion and manipulation ex vivo is a promising approach for engineering cell replacement therapies, and endogenous stem cells represent potential drugable targets for tissue repair. Before we can harness stem cells' therapeutic potential, we must first understand the intracellular mechanisms controlling their fate choices. These mechanisms involve complex signal transduction and gene regulation networks that feature, for example, intricate feed-forward loops, feedback loops and cross-talk between multiple signalling pathways. Systems biology applies computational and experimental approaches to investigate the emergent behaviour of collections of molecules and strives to explain how these numerous components interact to regulate molecular, cellular and organismal behaviour. Here we review systems biology, and in particular computational, efforts to understand the intracellular mechanisms of stem cell fate choice. We first discuss deterministic and stochastic models that synthesise molecular knowledge into mathematical formalism, enable simulation of important system behaviours and stimulate further experimentation. In addition, statistical analyses such as Bayesian networks and principal components analysis (PCA)/partial least squares (PLS) regression can distill large datasets into more readily managed networks and principal components that provide insights into the critical aspects and components of regulatory networks. Collectively, integrating modelling with experimentation has strong potential for enabling a deeper understanding of stem cell fate choice and thereby aiding the development of therapies to harness stem cells' therapeutic potential.

Asunto(s)

Algoritmos , Modelos Biológicos , Proteoma/metabolismo , Células Madre/citología , Células Madre/fisiología , Animales , Diferenciación Celular/fisiología , Simulación por Computador , Humanos , Biología de Sistemas

RESUMEN

The field of directed RNA interference (RNAi) has rapidly developed into a highly promising approach for specifically down regulating genes to alleviate disease pathology. This technology is especially well-suited to treating viral infections, and numerous examples now illustrate that a wide range of viruses can be inhibited with RNAi, both in vitro and in vivo. One principle that has arisen from this work is that antiviral RNAi therapies must be tailored to the unique life cycle of each pathogen, including the choice of delivery vehicle, route of administration, gene(s) targeted and regulation and duration of RNAi induction. Although effective strategies will be customized to each virus, all such therapies must overcome similar challenges. Importantly, treatment strategies must compensate for the inevitable fact that viral genome sequences evolve extremely rapidly, and computational and bioinformatics approaches may aid in the development of therapies that resist viral escape. Furthermore, all RNAi strategies involve the delivery of nucleic acids to target cells, and all will therefore benefit from the development of enhanced gene design and delivery technologies. Here, we review the substantial progress that has been made towards identifying effective antiviral RNAi targets and discuss strategies for translating these findings into effective clinical therapies.

Asunto(s)

Terapia Genética/métodos , Interferencia de ARN , ARN Interferente Pequeño , Virosis/terapia , Virus/genética , Ingeniería Genética , Humanos

Asunto(s)

Antiinflamatorios no Esteroideos/uso terapéutico , Antiulcerosos/uso terapéutico , Inhibidores de la Ciclooxigenasa/uso terapéutico , Pautas de la Práctica en Medicina/estadística & datos numéricos , Enfermedades Reumáticas/tratamiento farmacológico , Análisis Costo-Beneficio , Inhibidores de la Ciclooxigenasa/economía , Prescripciones de Medicamentos/estadística & datos numéricos , Humanos , Enfermedades Reumáticas/enzimología

RESUMEN

Gene therapy vehicles must be engineered to overcome numerous barriers that limit delivery efficiency. These barriers arise at every step of the delivery process, including the transit of the vector from injection to a cell surface, receptor binding and uptake, intracellular trafficking, and nuclear entry. The gene transfer properties of the highly promising adeno-associated viral (AAV) vector at each step are determined by its capsid structure. Previous capsid modifications that alter AAV tropism, as well as the existence of multiple AAV serotypes, suggest that the AAV capsid is reasonably plastic. We have taken advantage of this remarkable capsid plasticity to generate a large mutant AAV library (1e6) and select for mutant AAV virons that can overcome several barriers to infection. Specifically, we have selected AAV2 library for infectious particles with altered heparan sulfate (HS) affinity and for the ability to evade an AAV2 immune response. We have generated mutants with lower and higher affinity to heparin, which could prove valuable in controlling the therapeutic zone of an AAV vector in tissues where ECM HS hinders AAV2 diffusion. Furthermore, we have generated vector variants that have resistance to human serum that neutralizes wild type AAV2, yet retain AAV2 gene delivery efficiency. These vectors may enable high gene delivery efficiency even in patients with preexisting immunity, and the locations of point mutations on the capsid surface suggest new regions of functional importance to the virus. These AAV libraries therefore both provide useful variants for gene therapy application and offer a means to dissect AAV biology.

RESUMEN

Physical activity has been inconsistently associated with rectal cancer despite the consistent association between physical activity and colon cancer. In this study, the authors evaluated the association between physical activity and rectal cancer using the same questionnaire used to evaluate the previously reported association with colon cancer. A population-based study of 952 incident cases of cancer in the rectum and rectosigmoid junction and 1,205 age- and sex-matched controls was conducted in Utah and northern California at the Kaiser Permanente Medical Care Program between 1997 and 2002. Vigorous physical activity was associated with reduced risk of rectal cancer in both men and women (odds ratio (OR) = 0.60, 95% confidence interval (CI): 0.44, 0.81 for men; OR = 0.59, 95% CI: 0.40, 0.86 for women). Among men, moderate levels of physical activity also were associated with reduced risk of rectal cancer (OR = 0.70, 95% CI: 0.51, 0.97). Participation in vigorous activity over the past 20 years conferred the greatest protection for both men and women (OR = 0.55, 95% CI: 0.39, 0.78 for men; OR = 0.44, 95% CI: 0.30, 0.67 for women). In summary, physical activity was associated with reduced risk of rectal cancer in these data. The reduced risk was similar to that previously observed for colon cancer.

Asunto(s)

Neoplasias del Colon/etiología , Neoplasias del Colon/prevención & control , Ejercicio Físico , Aptitud Física , Neoplasias del Recto/etiología , Neoplasias del Recto/prevención & control , Adulto , Anciano , Estudios de Casos y Controles , Estudios Epidemiológicos , Femenino , Encuestas Epidemiológicas , Humanos , Estilo de Vida , Masculino , Persona de Mediana Edad , Oportunidad Relativa , Factores de Riesgo

RESUMEN

Fallow deer farming for venison production on pasture and fallow land has gained importance in Germany during recent years. As fallow deer farming constitutes a relatively young farming practice, many questions concerning a welfare-conforming and ecologically sound keeping of fallow deer are still open. Based on the recommendations on the keeping of fallow deer in enclosures for the purpose of venison meat production including by-products from 2 November 1979, a critical comparative review considering the current knowledge from research and practical deer farming experience was conducted. Recommendations on measures for breeding management and administration (statistics on farms and stocks, security and control of enclosures, training and experience of stockperson, practical skills for immobilisation and killing) are proposed. The cultivation goals for the care of landscape by fallow deer farming in protected areas need to be defined precisely. Due to infection risk, mixed herds with domesticated ruminants are not recommended. Potential progress can be foreseen in preventive health measures (control of endoparasites) and by appropriate feeding and water supply, considering feeding and drinking place design, feeding behaviour and water needs. The knowledge on handling and immobilisation methods should also be applied to small herd sizes. More research is needed on transportation of fallow deer.

Asunto(s)

Crianza de Animales Domésticos/métodos , Bienestar del Animal , Ciervos , Animales , Alemania