Abstract Background Delay discounting (DD), a decline in the subjective value of reward with increasing delay until its receipt, is an established behavioral model of impulsive choice, a key ...component of a broader impulsivity construct. Greater DD, i.e., a tendency to choose smaller immediate over larger delayed rewards, has been implicated as a potential intermediate phenotype (endophenotype) for addictive disorders and comorbid externalizing psychopathology, particularly in adolescence. However, genetic and environmental origins of DD remain unclear. Accordingly, the goal of the present study was to assess heritability of DD, an important aspect of its utility as an endophenotype. Methods A commonly used computerized procedure involving choice between varying amounts of money available immediately and a standard amount of $100 presented at variable delays was administered to a population-based sample of twins aged 16 and 18 ( n = 560, including 134 monozygotic and 142 dizygotic pairs). DD was quantified using area under the discounting curve and the k coefficient estimated by fitting a hyperbolic model to individual data. Heritability was assessed using linear structural equation modeling of twin data. Results The genetic analysis revealed significant heritability of both DD measures (area under the discounting curve: 46% and 62%; k: 35% and 55% at age 16 and 18, respectively). Conclusions The present study provides evidence for heritability of both model-based and model-free DD measures and suggests that DD is a promising intermediate phenotype for genetic dissection of impulsivity and externalizing spectrum disorders.
The transcriptional profiles of mouse embryonic, neural, and hematopoietic stem cells were compared to define a genetic program for stem cells. A total of 216 genes are enriched in all three types of ...stem cells, and several of these genes are clustered in the genome. When compared to differentiated cell types, stem cells express a significantly higher number of genes (represented by expressed sequence tags) whose functions are unknown. Embryonic and neural stem cells have many similarities at the transcriptional level. These results provide a foundation for a more detailed understanding of stem cell biology.
Failure of Bone Marrow to Reconstitute Lung Epithelium Kotton, Darrell N; Fabian, Attila J; Mulligan, Richard C
American journal of respiratory cell and molecular biology,
10/2005, Letnik:
33, Številka:
4
Journal Article
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A new paradigm of epithelial tissue reconstitution has been suggested whereby circulating cells derived from bone marrow contribute to a variety of epithelial cell types. With regard to the lung, ...several recent reports have used immunofluorescence microscopy to demonstrate engraftment of bone marrow-derived cells as type II pneumocytes, the endogenous progenitors of the lung alveolus. We show here that immunofluorescence microscopy, as has been used in previous reports, cannot reliably identify rare engrafted cells in lung tissue sections after transplantation of bone marrow cells or purified hematopoietic stem cells tracked with ubiquitous labels. We have employed a lineage-specific reporter system based on transgenic mice that express the GFP reporter gene only in lung epithelial cells (surfactant protein C-GFP) to assay for engrafted cells by flow cytometry, histology, and molecular methods. Using this approach to evaluate transplant recipients, including those subjected to bleomycin-induced lung injury, we demonstrate that when autofluorescence, dead cells, and contaminating blood cells are excluded from analysis, there is no detectable reconstitution of lung alveolar epithelial cells by unfractionated bone marrow cells or purified hematopoietic stem cells.
Single-cell transplantation analysis revealed that the cells that had the strongest dye efflux activity (“Tip”-SP cells) and had the phenotype CD34− c-Kit+ Sca-1+ Lin− (CD34− KSL cells) exhibited ...very strong proliferation and multilineage differentiation capacity. Ninety-six percent of the lethally irradiated mice that received a single “Tip”-SP CD34− KSL cell showed significant donor cell engraftment for long term. These findings support the hypothesis that “Tip”-SP CD34− KSL cells represent the most primitive hematopoietic stem cells that are capable of migrating into the primary site and surviving and/or proliferating with nearly absolute efficiency. This led us to propose high marrow-seeding efficiency as a specific characteristic of primitive HSCs, in addition to their self-renewal and multipotent capacity.
One of the major obstacles in engineering thick, complex tissues such as muscle is the need to vascularize the tissue in vitro. Vascularization in vitro could maintain cell viability during tissue ...growth, induce structural organization and promote vascularization upon implantation. Here we describe the induction of endothelial vessel networks in engineered skeletal muscle tissue constructs using a three-dimensional multiculture system consisting of myoblasts, embryonic fibroblasts and endothelial cells coseeded on highly porous, biodegradable polymer scaffolds. Analysis of the conditions for induction and stabilization of the vessels in vitro showed that addition of embryonic fibroblasts increased the levels of vascular endothelial growth factor expression in the construct and promoted formation and stabilization of the endothelial vessels. We studied the survival and vascularization of the engineered muscle implants in vivo in three different models. Prevascularization improved the vascularization, blood perfusion and survival of the muscle tissue constructs after transplantation.
Celotno besedilo
Dostopno za:
DOBA, IJS, IZUM, KILJ, NUK, PILJ, PNG, SAZU, SIK, UILJ, UKNU, UL, UM, UPUK
The hematopoietic stem cell (HSC) is a unique cell type found in bone marrow, which has the capacity for both self-renewal and differentiation into all blood lineages. The identification of genes ...expressed specifically in HSCs may help identify gene products vital to the control of self-renewal and/or differentiation, as well as antigens capable of forming the basis for improved methods of stem cell isolation. In previous studies, we identified a number of genes that appeared to be differentially expressed in murine bone marrow–derived HSCs, using microarray technology. We report here that one of those genes, encoding the murine endothelial protein C receptor (EPCR), is expressed at high levels within the bone marrow in HSCs. Bone marrow cells isolated on the basis of EPCR expression alone are highly enriched for hematopoietic reconstitution activity, showing levels of engraftment in vivo comparable to that of stem cells purified using the most effective conventional methods. Moreover, evaluation of cell populations first enriched for stem cell activity by conventional methods and subsequently fractionated on the basis of EPCR expression indicates that stem cell activity is always associated with EPCR-expressing cells. Based on our findings, we believe EPCR represents the first known marker that ‘explicitly’ identifies hematopoietic stem cells within murine bone marrow.
•Discounting of delayed rewards decreases during late adolescence.•Individual differences in delay discounting show strong test–retest stability.
Delay discounting (DD), a decline in subjective value ...of a reward with increasing temporal delay in receipt of that reward, is an established behavioral indicator of impulsivity. Preference for smaller-immediate over larger-delayed rewards has been implicated in the basic neurobehavioral mechanisms of risk for addictive disorders and related externalizing psychopathology. Establishing long-term stability of DD in adolescence is a necessary step towards its validation as an intermediate phenotype, or marker of risk, in neurobiological and genetic studies. Previous studies have demonstrated moderate to high test–retest reliability of DD, however, these studies utilized adult samples and examined relatively short retest intervals. Due to continuing development of brain and behavior, stability of temporal discounting behavior in adolescence may differ from that in adulthood. Here, two cohorts of adolescents aged 16 (n=126) and 18 (n=111) were administered a computerized test of DD and re-tested two years later. DD rate showed a modest but significant decrease with age, suggesting a reduction in overall impulsivity from middle to late adolescence. Significant test–retest correlations were observed in both cohorts (.67 and .76, respectively, p<.001) indicating longitudinal stability of individual differences in decision-making behavior during middle and late adolescence.
This article is part of a Special Issue entitled: insert SI title.
Artemis gene mutations are responsible for the development of a severe combined immunodeficiency radiation-sensitive (RS) SCID characterized by a severe B and T cell deficiency and a normal natural ...killer cell population. To establish the feasibility of a gene therapy approach to the treatment of RS-SCID, we generated a series of lentiviral vectors expressing human Artemis from different promoters and used them to transduce highly purified hematopoietic stem cells (HSCs) from Artemis knockout mice. HSCs transduced by the different viruses were transplanted into either lethally irradiated Rag-l-deficient animals or Artemis knockout mice treated with a nonmyeloablative dose of Busulfan. In both models, transplantation of HSCs transduced by a vector that used a murine phosphoglycerate kinase (PGK) promoter led to a complete functional correction of the immunodeficiency. Corrected animals displayed rescue of mature B cells with normal levels of serum immunoglobulins, together with complete rescue of the T cell compartment as evidenced by the presence of mature T lymphocytes in peripheral blood as well as normal values of thymocytes in thymus. Those B and T cells were capable of activation, as shown both by in vitro stimulation responses and in vivo after immune challenge. Overall, the results indicate that a gene therapy approach for RS-SCID involving the transplantation of genetically modified HSCs is indeed feasible. Furthermore, our studies suggest the possibility that nonmyeloablative conditioning regimens might be effectively used to promote engraftment of genetically modified cells in the case of diseases where standard irradiation-based myeloablative bone marrow transplantation protocols may prove problematic.
γ-Sarcoglycanopathy or limb girdle muscular dystrophy type 2C is an untreatable disease caused by autosomal recessively inherited mutations of the γ-sarcoglycan gene. Nine non-ambulatory patients ...(two males, seven females, mean age 27 years; range 16-38 years) with del525T homozygous mutation of the γ-sarcoglycan gene and no γ-sarcoglycan immunostaining on muscle biopsy were divided into three equal groups to receive three escalating doses of an adeno-associated virus serotype 1 vector expressing the human γ-sarcoglycan gene under the control of the desmin promoter, by local injection into the extensor carpi radialis muscle. The first group received a single injection of 3 × 109 viral genomes in 100 µl, the second group received a single injection of 1.5 × 1010 viral genomes in 100 µl, and the third group received three simultaneous 100-µl injections at the same site, delivering a total dose of 4.5 × 1010 viral genomes. No serious adverse effects occurred during 6 months of follow-up. All nine patients became adeno-associated virus serotype 1 seropositive and one developed a cytotoxic response to the adeno-associated virus serotype 1 capsid. Thirty days later, immunohistochemical analysis of injected-muscle biopsy specimens showed γ-sarcoglycan expression in all three patients who received the highest dose (4.7-10.5% positively stained fibres), while real-time polymerase chain reaction detected γ-sarcoglycan messenger RNA. In one patient, γ-sarcoglycan protein was detected by western blot. For two other patients who received the low and intermediate doses, discrete levels of γ-sarcoglycan expression (<1% positively stained fibres) were also detectable. Expression of γ-sarcoglycan protein can be induced in patients with limb girdle muscular dystrophy type 2C by adeno-associated virus serotype 1 gene transfer, with no serious adverse effects.
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