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The host range of retroviral vectors including lentiviral vectors can be expanded or altered by a process known as pseudotyping. Pseudotyped lentiviral vectors consist of vector particles bearing glycoproteins (GPs) derived from other enveloped viruses. Such particles possess the tropism of the virus from which the GP was derived. For example, to exploit the natural neural tropism of rabies virus, vectors designed to target the central nervous system have been pseudotyped using rabies virus-derived GPs. Among the first and still most widely used GPs for pseudotyping lentiviral vectors is the vesicular stomatitis virus GP (VSV-G), due to the very broad tropism and stability of the resulting pseudotypes. Pseudotypes involving VSV-G have become effectively the standard for evaluating the efficiency of other pseudotypes. This review samples a few of the more prominent examples from the ever-expanding list of published lentiviral pseudotypes, noting comparisons made with pseudotypes involving VSV-G in terms of titer, viral particle stability, toxicity, and host-cell specificity. Particular attention is paid to publications of successfully targeting a specific organ or cell types.
One mechanism for expanding the cellular tropism of enveloped viruses is through the formation of phenotypically mixed particles or pseudotypes, a process that commonly occurs during viral assembly in cells infected with two or more viruses [Zavada, 1982]. Human immunodeficiency virus type 1 (HIV-1) has long been known to form pseudotypes by the incorporation of heterologous GPs through phenotypic mixing, allowing an extension of the host range of such virions beyond cells that express the CD4 receptor and an appropriate co-receptor. Several studies have demonstrated that wild type HIV-1 produced in cells infected with xenotropic murine leukemia virus (MLV) [Canivet, 1990; Lusso, 1990], amphotropic MLV [Chesebro, 1990; Spector, 1990], or herpes simplex virus [Zhu, 1990] gave rise to phenotypically mixed virions with an expanded host range, suggesting that pseudotyped virions had been produced. Phenotypic mixing of viral GPs has also been shown to occur between HIV-1 and vesicular stomatitis virus in coinfected cell cultures [Zhu, 1990]. These early observations were key to the subsequent design of HIV-1-based lentiviral vectors bearing heterologous GPs.
Page et al. [Page, 1990] were the first to design and test HIV-1-based vector particles harboring a heterologous GP. In this study, the HIV-1 genome was rendered replication defective by replacing the GP-encoding gp160 sequence with a guanine-phosphoribosyl transferase (gpt) gene driven by the Simian virus 40 (SV40) early promoter. Transient cotransfection of monkey COS-7 cells with this GP-deleted vector and an expression plasmid encoding an amphotropic MLV GP resulted in packaging of the HIV-gpt vector genome into infectious virions. Upon infection of susceptible cells, the gpt gene was transmitted and expressed, allowing transduced cells to be selected in media containing mycophenolic acid [Page, 1990]. Landau et al. [Landau, 1991] subsequently demonstrated that expression of other GPs including those of the ecotropic MLV and human T-cell leukemia virus type I (HTLV-I) in cells transfected with the HIV-gpt vector resulted in the production of vector pseudotypes capable of infecting murine and human cells with titers reaching 105 colony forming units per ml following mycophenolic acid selection.
These early observations were confirmed and extended in 1996 by results reported independently by three groups showing that VSV-G was also efficiently incorporated into HIV-1 virions [Akkina, 1996; Naldini, 1996; Reiser, 1996]. Lentiviral vectors pseudotyped with VSV-G presented significant advantages in that VSV-G appears to interact with a ubiquitous cellular “receptor” on cells, giving the vector a broad host-cell range. The receptor for VSV has long been thought to be a ubiquitous membrane lipid, phosphatidylserine [Schlegel, 1983]. However, recent experimental evidence has indicated that phosphatidylserine is not the cell surface receptor for VSV, although it may be involved in a postbinding step of virus entry [Coil, 2004]. VSV-G pseudotyping also confers high vector particle stability, allowing for concentration by ultracentrifugation [Burns, 1993; Bartz, 1996]. Perhaps the most significant shortcoming for VSV-G is that it is toxic to cells if expressed constitutively. The use of tetracycline-regulatable promoters has alleviated this problem and facilitated the generation of stable cell lines expressing VSV-G [Ory, 1996]. Another potential drawback for in vivo applications is that VSV-G pseudotypes are inactivated by human serum complement [DePolo, 2000]. This problem can be bypassed by using polyethylene glycol-modified (PEGylated) VSV-G pseudotypes. Covalent modification of VSV-G pseudotyped HIV-1 vector particles using monomethoxypoly(ethylene) reduced the rate of vector inactivation by a factor of 1,000 and significantly enhanced transduction efficiency in the bone marrow and spleen after systemic administration of the virus [Croyle, 2004].
To date, VSV-G remains the most popular GP for pseudotyping HIV-1-based vectors as well as other lentiviral vectors including vectors based on HIV-2 [Poeschla, 1998a] and on simian immunodeficiency virus (SIV) [Nakajima, 2000; Negre, 2000; Schnell, 2000]. Vectors based on nonprimate lentiviruses include feline immunodeficiency virus (FIV) [Poeschla, 1998b], equine infectious anemia virus (EIAV) [Mitrophanous, 1999], bovine immunodeficiency virus (BIV) [Berkowitz, 2001a], Jembrana disease virus (JDV) [Metharom, 2000], visna virus (VV) [Berkowitz, 2001b], and caprine arthritis encephalitis virus (CAEV) [Mselli-Lakhal, 2000] (Table 1).
There is an ever-growing list of alternative GPs for pseudotyping lentiviral vectors, each with specific advantages and disadvantages. While space limitations preclude an exhaustive description of all pseudotypes, this review will survey some of the more prominent examples in the recent literature. Other emerging pseudotypes are referred to in Table 1. The widespread use of VSV-G to pseudotype lentiviral vectors has made this GP in effect the standard against which the usefulness of other viral GPs to form pseudotypes are compared. Comparison of pseudotype efficiency inevitably means comparing vector titers on cell lines in vitro. This brings up two important issues: (i) Techniques to determine lentiviral vector titers, and (ii) Cell lines chosen for titration of such vectors. There are different measures used to determine lentiviral vector titers. Some of them are based on the number of vector particles present in a virus stock while others are derived from the number of proviral copies in transduced target cells. Virus particle numbers can be determined using real-time PCR based on strong-stop cDNA present in virions [Scherr, 2001]. Alternatively, the amount of virus proteins present in virus cores such as p24 Gag are determined by ELISA to arrive at relative particle titers [Logan, 2004]. Functional titration assays are based on vector-encoded reporter gene expression. For example, vectors encoding green fluorescent protein (GFP) have been titrated using FACS analysis [Zhang, 2002]. For vectors that do not contain a reporter gene, proviral DNA copy numbers determined by real-time PCR using DNA extracted from transduced cells have been used to determine titers [Sastry, 2002]. As each of these methods produces different results, a reader of the literature must pay close attention to which titration method was used. Also significant for titration is the cell line used as receptors for a given GP may vary from cell line to cell line, possibly producing a falsely depressed titer. Frequently, a given cell line (such as HeLa or 293 cells) is chosen because it is considered generally permissive to infection rather than representing the natural tropism of the virus from which the GP was derived. Additionally, results produced with a given cell line or type of cell in vitro do not always translate into comparable results in vivo due to factors such as complement-mediated inactivation of vectors. Careful attention must also be paid to the given virus strain from which the GP was derived as this may also be a source of confusion and conflicting results. For example, considerable differences in the brain transduction patterns were demonstrated between two related lyssavirus-derived GPs. Transduction of the mouse striatum by HIV-1 vectors pseudotyped with the GP derived from a Mokola virus originally isolated from a cat in Zimbabwe (MokZIM) [Bourhy, 1989] was inefficient [Desmaris, 2001], while HIV-1 vectors bearing a Mokola virus GP obtained from an Ethiopian Mokola virus isolate (MokETH) [Mebatsion, 1995] revealed robust transduction [Watson, 2002].
The following is an overview of prominent examples of lentivirus pseudotypes that have appeared in the literature. Additional examples that are not discussed in the text are referred to in Tables Tables11 and and22.
The capacity of lentiviral vectors to form pseudotypes with lyssavirus GPs was investigated early on with a view toward designing neurotropic lentiviral vectors. The genus Lyssavirus is part of the Rhabdoviridae family and includes the classical rabies viruses and rabies-related viruses including Mokola virus [Badrane, 2001]. Rabies virus (RV), the prototype of the genus Lyssavirus, is an enveloped negative-strand RNA virus encoding five structural proteins including the transmembrane GP. The transmembrane GPs of lyssaviruses appear to exhibit a similar overall structure; however, considerable antigenic differences were noted between RV and rabies-related viruses by using anti-GP monoclonal antibodies. Rabies is a disease of the CNS and neuroinvasiveness is the major defining characteristic of a classical RV infection.
Mochizuki et al. [Mochizuki, 1998] were the first to describe robust pseudotyping of HIV-1 lentiviral vectors using lyssavirus-derived GPs including those of the rabies virus strain RabSADB19 [Conzelmann, 1990] and the Mokola virus MokETH strain [Mebatsion, 1995]. The Lyssavirus pseudotypes were compared to VSV-G pseudotypes by analyzing unconcentrated virus stocks on human osteosarcoma (HOS) cells and on Rat-2 fibroblasts. The lyssavirus GPs produced identical titers, which were 5 times lower than those of VSV-G pseudotypes on HOS cells and 4 times lower on Rat-2 cells. Shortly thereafter, EIAV vectors were successfully pseudotyped with a RV GP [Mitrophanous, 1999] (the source of the rabies GP used for pseudotyping was not revealed).
Once successful pseudotyping of lentiviral vectors with lyssavirus-derived GPs had been demonstrated, interest was focused on assessing the neurotropism of such pseudotypes. Desmaris et al. [Desmaris, 2001] initially tested the infectivity of HIV-1 vector pseudotypes bearing a Mokola virus GP (MokZIM strain) or VSV-G. The in vivo tropism of the two pseudotypes was tested by injecting vector particles encoding β-galactosidase (β-gal) into the striatum. The two pseudotypes produced comparable numbers of β-gal expressing cells, and co-labeling for cell-type markers indicated that both vectors had primarily transduced neurons. Watson et al. [Watson, 2002] described a similar analysis involving HIV-1 vectors pseudotyped with the MokETH GP. Intrastriatal injection of such pseudotypes in the mouse also resulted in extensive transgene expression along the corpus callosum and external capsule. Transduction by pseudotypes bearing the MokETH GP in both the striatum and the white matter was considerably more extensive than that observed with pseudotypes involving VSV-G. Mokola pseudotypes however were not transported along projections from the striatum to the substantia nigra [Watson, 2002]. To assess whether lentiviral vector particles pseudotyped with lyssavirus GPs gain access to the CNS by retrograde transport, pseudotyped HIV-1 virions were either injected into the nasal cavity or injected into limb muscles of rats. Both VSV-G and MokZIM pseudotypes produced β-gal-expressing neurons in the olfactory bulb after nasal injection, but neither produced β-gal-positive motor neurons in the spine [Desmaris, 2001]. The investigators concluded that VSV-G and MokZIM pseudotypes were of comparable efficiency in retrograde axonal transport, but that neither was taken up at the neuromuscular junction. In a related study, EIAV-based vectors pseudotyped with the GPs derived from the RV RabERA strain [Sacramento, 1992] or from VSV-G were injected into the striatum to compare the spread of the virus as judged from the number and distribution of transgene-expressing cells [Mazarakis, 2001]. After injection of VSV-G pseudotypes, some 30,000 β-gal-positive cells were found in the striatum. At a comparable titer, RabERA pseudotypes produced 32,000 β-gal-positive cells in the striatum plus 14,800 cells in the thalamus along with 3000 cells in the substantia nigra, indicating both retrograde and anterograde transport. The GPs of the rabies RabCVS [Sacramento, 1992] and RabERA strains produced similar transgene expression patterns in distal brain areas after striatal injection of EIAV vectors [Wong, 2004]. However, a change from arginine to glutamine at amino acid 333 of the RabERA GP resulted in weaker transduction of the caudate putamen. Interestingly, there was no retrograde transport of vectors bearing the mutated RabERA GP to areas such as the thalamus. This suggests that this single mutation in the viral GP was sufficient to abolish the virus' retrograde transport characteristics [Wong, 2004]. Instraspinal injections of EIAV vectors pseudotyped with the RabERA GP have likewise demonstrated spread of transgene expression along nerve tracts. This was not seen after injection of VSV-G pseudotypes [Mazarakis, 2001].
Of potential clinical importance, intramuscular injection of EIAV vectors pseudotyped with RV GPs resulted in transgene expression in spinal motor neurons. Retrograde transport of lentiviral vectors from peripheral muscle to spinal motor neurons has been used to demonstrate a therapeutic effect in an animal model of familial amyotrophic lateral sclerosis (ALS) [Azzouz, 2004b]. This model is based on SOD1G93A mice which have a mutation in the SOD1 gene and experience severe motor neuron degeneration comparable to that seen in human ALS. EIAV-based vectors encoding vascular endothelial growth factor (VEGF) were pseudotyped with the RabERA GP and injected into various muscle groups of 3 week-old SOD1G93A mice. Animals that were injected with a control vector were severely impaired by 100 days of age, while comparable impairment in animals that received the VEGF vector was delayed until a month later. EIAV/VEGF vector treatment also extended lifespan of these animals by 30%. Clinical markers such as rotarod performance were also improved with EIAV/VEGF vector treatment. In a related study, EAIV vectors expressing either a marker gene or the human survival motor neuron (SMN) cDNA and pseudotyped with the RabERA GP were injected into various muscles of spinal muscle atrophy (SMA) mice. Retrograde transport of the virus to spinal motor neurons was confirmed by marker gene expression. Compared to untreated controls, SMN-vector treatment restored SMN protein levels in motor neurons, reduced motor neuron death, and increased the life expectancy by an average of 3 and 5 days (20% and 38%) [Azzouz, 2004a]. These examples demonstrate the capacity of lentivirus vectors pseudotyped with RV GPs to deliver therapeutic genes to otherwise inaccessible neurons by exploiting the lyssavirus' ability to induce retrograde transport along axons.
MLV-based retroviral vectors pseudotyped with the lymphocytic choriomeningitis virus (LCMV) GP have been shown to efficiently transduce cells from different tissues relevant to gene therapy including fibroblasts, epithelial cells, hematopoietic cells, as well as hepatoma, neuroblastoma and glioma cell lines [Miletic, 1999]. Successful transduction involving pseudotypes containing the LCMV-WE GP [Romanowski, 1985] was observed in cell lines derived from various species of human, mouse, hamster, or dog origin. LCMV is a noncytopathic arenavirus, which infects a broad range of different cell types. The LCMV GPs are synthesized as a 74-kDa precursor protein, GP-C, and then cleaved into a 35-kDa transmembrane protein, GP-2, and an external 44-kDa protein, GP-1. Alpha-dystroglycan, a cellular receptor for LCMV [Cao, 1998; Smelt, 2001], is a dystrophin-associated GP that connects the cytoskeleton with the extracellular matrix, and is widely expressed in most tissues.
Pseudotyping of HIV-1-based lentiviral vectors with the GP derived from the LCMV Armstrong 53b strain (LCMV-Arm53b, [Salvato, 1988]) was first reported in 2001 [Christodoulopoulos, 2001]. The titers obtained with such pseudotypes on 293T cells were at least two logs below those observed with vectors pseudotyped with the amphotropic MLV GP or VSV-G. Interestingly, LCMV-pseudotyped lentiviral vectors containing the GP of the LCMV-WE strain revealed vector titers similar to those obtained with pseudotypes containing the amphotropic MLV GP or VSV-G [Beyer, 2002].
A number of applications involving lentiviral vectors pseudotyped with LCMV GPs have been described. The following is an overview of these applications. Loss of insulin-secreting pancreatic islet cells leads to insulin-dependent diabetes. As diabetes is a significant public health problem, genetic manipulation of islet cells aimed at preventing their destruction is important for both clinical applications and understanding islet cell biology. Kobinger et al. [Kobinger, 2004] evaluated the ability of a HIV-1-based lentiviral vector pseudotyped with various viral GPs to target human islets ex vivo, with the goal of improving transduction efficiency while minimizing toxicity. LCMV-pseudotyped vectors transduced insulin-secreting beta cells with the highest efficiency. Moreover, toxicity associated with transduction of islets was found to be lower with LCMV-WE-pseudotyped vectors than with VSV-G-pseudotyped vectors, the second most efficient pseudotype for islet transduction.
A reduced toxic response to LCMV pseudotypes compared to VSV-G pseudotypes was also found in the liver following portal vein injection of HIV-1-based vectors encoding human blood-clotting factor VIII (hFVIII) [Park, 2003]. Transduction of hemophilia A mice with hFVIII expressing lentiviral vectors resulted in transient correction of the bleeding diathesis phenotype. Moreover, the use of vectors pseudotyped with the LCMV-WE GP resulted in similar circulating levels of hFVIII. Similar doses of LCMV-pseudotyped lentiviral vectors resulted in minimal systemic or hepatic injury compared to VSV-G pseudotypes. Taken together, these studies demonstrate the importance of using alternative pseudotypes, such as LCMV, to minimize adverse vector effects in vitro and in vivo.
There is increasing interest in developing gene therapy strategies for malignant gliomas, which are the most frequent primary brain tumors, but have a poor prognosis with current treatments. Work reported by Beyer et al. [Beyer, 2002] showed that LCMV-WE pseudotypes were able to transduce human glioma and neuroblastoma cell lines, as well human primary glioma cells derived from resected human gliomas. In a related study, Miletic et al. [Miletic, 2004] assessed the transduction efficiencies of HIV-1 vectors pseudotyped with the LCMV-WE GP or VSV-G in malignant glioma cells and normal brain cells both in vitro and in vivo. In vitro, LCMV-WE pseudotypes transduced almost exclusively astrocytes, whereas VSV-G pseudotypes transduced neurons as well as astrocytes. In an in vivo glial tumor model based on rats carrying intracerebral grafts of rat 9L gliosarcoma cells, LCMV-WE pseudotypes revealed efficient transduction of solid glioma parts and specific transduction of infiltrating tumor cells. In contrast, VSV-G-pseudotyped lentiviral vectors transduced only a few tumor cells in solid tumor parts and transduced mostly neurons surrounding the tumors. Similar results were reported by Steffens et al. [Steffens, 2004]. These authors compared eight different lentivirus pseudotypes regarding their efficiencies to transduce neuronal and glial tumor cell lines. Pseudotypes involving the GPs of human foamy virus, RV (RabSADB19 strain), Mokola (MokETH strain) or amphotropic MLV displayed the highest transduction efficiency in neuroblastoma cells, whereas pseudotypes involving the LCMV-Arm53b GP resulted in the highest transduction efficiency in gliomas.
In a recent report, an FIV vector expressing β-gal and pseudotyped with the LCMV-WE GP was injected into postnatal mouse brain striatum to determine neural cell-type transduction [Stein, 2005]. β-gal-expressing cells included astrocytes in the striatum and in the subventricular zone (SVZ), neuroblasts along the rostral migratory stream, and neurons in the olfactory bulb. This pattern was suggestive of transduction of neural stem/progenitor cells that reside in the SVZ and continually generate olfactory bulb neurons. Thus, FIV/LCMV-WE pseudotypes are potentially useful for targeting neural stem/progenitor cells in vivo. Taken together, these results demonstrate that targeted transduction in the CNS can be achieved using specific pseudotypes.
Alphaviruses exhibit a wide cellular tropism that includes important gene therapy targets such as antigen-presenting cells, neurons, and muscle cells. Alphaviruses belong to the family Togaviridae and are found in insects, reptiles, birds, and mammals. Although it remains to be determined as to what molecules act as cellular receptors for these viruses, it has been suggested that several receptors or several receptor-coreceptor combinations are involved in virus entry. The main part of the viral GP complex consists of heterodimers of the E1 and E2 GPs that are further organized into trimers. A total of 80 of these trimeric spikes are distributed on the outer surface of the virion. The E2 GP most likely mediates interactions with target cell receptors, whereas E1 is thought to mediate fusion. Several reports describing successful pseudotyping of lentiviral vectors involving the Ross River virus (RRV), Semliki Forest virus (SFV) and Sindbis virus GPs have appeared in the literature.
Kang et al. [Kang, 2002] were the first to describe lentivirus pseudotypes involving alphavirus GPs. These authors examined the in vivo gene transfer efficiency and tissue or cell tropism of a FIV-based lentiviral vector pseudotyped with the GPs derived from RRV. RRV GPs were efficiently incorporated into FIV virions, generating preparations of FIV vectors, which after concentration attained high titers. After systemic administration, RRV-pseudotyped FIV vectors (RRV/FIV) predominantly transduced the liver of recipient mice. Transduction efficiency in the liver with the RRV/FIV was about 20-fold higher than that achieved with a VSV-G pseudotype. Moreover, in comparison to VSV-G, the RRV GPs caused less cytotoxicity. Although hepatocytes were the main liver cell type transduced, nonhepatocytes (mainly Kupffer cells) were also transduced. After injection into the brain, RRV/FIV preferentially transduced neuroglial cells (astrocytes and oligodendrocytes). In contrast to the VSV-G protein that targets predominantly neurons, <10% of the brain cells transduced with the RRV pseudotyped vector were neurons.
Kahl et al. [Kahl, 2004] subsequently showed that the GPs derived from RRV and SFV can pseudotype HIV-1-derived lentivirus vectors and that both RRV and SFV GPs considerably expanded the host range of these vectors. Also, such pseudotypes could be efficiently concentrated by ultracentrifugation. A systematic analysis comparing the alphaviral GPs to VSV-G revealed that lentivirus vectors incorporated RRV GPs with an efficiency comparable to that of VSV-G. Both pseudotypes had comparable physical titers as judged by quantitative RT-PCR amplification of vector RNA. However, infectious titers with the RRV pseudotype were lower than with VSV-G as determined by FACS analysis of GFP expression in transduced cells. However, incorporation of SFV GPs into lentivirus vector was less efficient, leading to significantly decreased physical and infectious titers.
In a recent report by Kahl et al. [Kahl, 2005] the transduction of hematopoietic cells using RRV-pseudotyped lentiviral was investigated. RRV-mediated transduction of human CD34+ cord blood cells and progenitors was very inefficient, even at multiplicities of infection of 100. Inefficient transduction was also observed in a variety of hematopoietic cell lines. However, exposing the monocytic cell line THP-1 to 12-O-Tetradecanoylphorbol-13-acetate (TPA), a very potent activator of macrophage differentiation, induced a more than 10-fold increase in transduction. The uniquely restricted host range of RRV-pseudotyped vectors in hematopoietic cells may aid in the design of novel cell-selective transduction strategies.
Sustained production of lentiviral vectors pseudotyped with RRV and SFV from stable packaging cell lines was reported by Strang et al. [Strang, 2005]. Cell clones producing high-titer RRV pseudotypes were stable for at least 5 months while clones producing SFV pseudotypes produced relatively low titers. RRV pseudotypes were found to display a number of characteristics that are attractive, including resistance to inactivation by heat-labile components in fresh human sera and thermostability at 37°C. Upon concentration by ultracentrifugation, vector stocks with titers up to 6 × 107 transducing units (TU)/ml were obtained.
One strategy for cell-specific targeting has been the insertion of cell-specific ligands including single chain antibodies into the viral GP, or to bridge the virus and target cell using a ligand or antibody bridge. Morizono et al. [Morizono, 2001] having shown that the GP of Sindbis virus is able to pseudotype oncoretroviruses and lentiviruses, reported an oncoretroviral and lentiviral gene-targeting system based on antibody-mediated specific binding of a modified chimeric Sindbis virus GP. The Fc-binding domain (ZZ domain) of protein A was inserted into the Sindbis E2 protein, to produce the ZZ SINDBIS GP. They also showed that monoclonal antibodies directed to cell-surface antigens can be used to redirect the target specificity of these vectors when pseudotyped with the modified Sindbis virus GP. Of particular note, the vectors maintained high viral titers, which could be further increased by simple ultracentrifugation. Intravenous injection of such pseudotypes into mice resulted in high levels of transduction in liver and spleen cells. An exciting recent report from the same lab [Morizono, 2005] documented successful targeting in a living animal through intravenous injection of a lentiviral vector pseudotyped with a modified chimeric Sindbis virus GP. The modified GP (termed m168) contained a ZZ domain and carried a 4 aa deletion in E3 and two separate point mutations within E2. m168 pseudotypes were found to display a high targeting specificity and, unlike other pseudotypes, had low nonspecific infectivity in liver and spleen. A mouse cancer model of metastatic melanoma was used to test intravenous targeting with m168. Human P-GP was ectopically expressed on the surface of melanoma cells and targeted by the m168 pseudotyped lentiviral vector conjugated with antibody specific for P-glycoprotein. m168 pseudotypes successfully targeted metastatic melanoma cells growing in the lung after systemic administration by tail vein injection. Further development of this targeting technology may result in applications not only for cancers but also for genetic, infectious and immune diseases.
In 2000 Chan et al. [Chan, 2000] described the successful pseudotyping of HIV-1 vectors using Marburg virus and Ebola Zaire (EboZ) virus-derived virion GPs. The folate receptor-alpha was soon thereafter identified as a cofactor for cellular entry by Marburg and Ebola viruses [Chan, 2001]. The Ebola and Marburg filoviruses are highly pathogenic viruses that induce hemorrhagic fevers in humans and nonhuman primates. Of the four identified strains of Ebola virus, Zaire, Ivory Coast, Sudan, and Reston, the Zaire strain induces the highest death rates in humans, while the Reston strain has not caused fatal disease in humans. Mononuclear phagocytic cells form the primary targets for filovirus replication, while endothelial cells serve as secondary targets. The GP-encoding region of Ebola virus gives rise to two products, a nonstructural secreted form of the GP, sGP, encoded by the predominant transcript (80%), and the virion GP, the result of RNA editing during formation of the message (20%).
The in vivo tropism of lentiviral vectors pseudotyped with EboZ as compared to VSV-G was tested by in utero intramuscular and intrahepatic injections [MacKenzie, 2002]. Injected animals were harvested at time points between 5 days and 9 months following injection and detailed histologic assessments were performed. The efficiency and distribution of transduction after in utero administration was highly dependent upon the route of administration and the vector pseudotype used. Biodistribution studies showed widespread distribution of vector sequences in multiple tissues, albeit at very low levels, and transduced cells were found in significant numbers only in liver, heart, and muscle. Overall, vectors pseudotyped with VSV-G were the most efficient in transducing hepatocytes, whereas vectors pseudotyped with the MokETH and EboZ GPs were more efficient in transducing myocytes. Transduction of cardiomyocytes was observed after both intramuscular and intrahepatic injection of all three vectors. EboZ-pseudotyped vectors injected into muscle in utero were found to transduce muscle satellite or stem cells. Examination of transgene expression in regenerated muscle after injury showed that the newly generated tissue had originated from transduced satellite cells [MacKenzie, 2005]. Direct brain injection of EboZ/HIV pseudotypes in adult mice failed to transduce cells in the brain [Watson, 2002] and EboZ pseudotypes were also unable to transduce oligodendrocyte cultures. This may be due to the lack of the proper receptor since folate receptor-alpha expression was found to be low in the CNS.
The application of gene delivery strategies as a treatment for cystic fibrosis is limited by poor gene transfer efficiency with vectors applied to the apical surface of airway epithelia. To overcome this limitation, Kobinger et al. [Kobinger, 2001] carried out a systematic analysis of lentivirus vector pseudotypes to stably transduce airway epithelia in vivo. To do this, HIV-1-based vectors pseudotyped with the amphotropic MLV, Mokola (MokETH), HA (influenza-hemagglutinin), EboZ and Ebola-Reston (EboR) GPs were applied apically and basolaterally to cultures of airway epithelial cells. EboR and HA pseudotypes were completely ineffective, and only EboZ pseudotypes were able to efficiently transduce after apical application. In the same study, human tracheal explants were incubated with EboZ and VSV-G pseudotypes. VSV-G was largely ineffective while the EboZ pseudotype efficiently transduced airway epithelium. EboZ pseudotypes were similarly more effective than VSV-G pseudotypes in an in vivo test in mice receiving vector pseudotypes intratracheally. Sinn et al. [Sinn, 2003] found that polarized human airway epithelia expressed abundant folate receptor alpha on their apical surface. In an attempt to target these apical receptors, FIV-based vectors were pseudotyped using GPs derived from the Marburg and EboZ viruses. Deletion of a heavily O-glycosylated extracellular domain of the EboZ GP including amino acids 309 to 489 resulted in the EboZΔO GP variant. The titer of concentrated FIV vector stocks pseudotyped with EboZΔO was increased some 74-fold. Both EboZ/FIV and EboZΔO/FIV transduced airway epithelia from the apical surface at greater efficiency than from the basolateral surface. In contrast, VSV-G/FIV transduced the basolateral surface more efficiently than the apical one. In addition, the EboZ GP contains a mucin-rich region that was demonstrated to reduce expression of cellular adhesion molecules and induce cytotoxic effects to endothelial cells in vitro and in vivo. Medina et al. [Medina, 2003] developed several mutant EboZ GPs that contained deletions corresponding to the mucin-rich domain. Lentiviral vectors pseudotyped with these mutant GPs possessed higher titers compared to vectors pseudotyped with the unmodified EboZ GP. Also, they resulted in more efficient gene transfer in vivo.
A variety of different envelope GPs derived from gammaretroviruses were shown to pseudotype lentivirus vectors. The GPs included those derived from the ecotropic MLV [Landau, 1991; Reiser, 1996], amphotropic 4070A MLV [Reiser, 1996], 10A1 MLV [Stitz, 2000; Christodoulopoulos, 2001], xenotropic NZB MLV and that of the polytropic mink cell focus-forming virus [Christodoulopoulos, 2001]. Other GPs included that of the gibbon ape leukemia virus (GALV) [Stitz, 2000; Christodoulopoulos, 2001] and the RD114 GP [Hanawa, 2002; Sandrin, 2002; Zhang, 2004]. The titers of lentiviral vectors pseudotyped with these GPs were usually lower than those observed with VSV-G. However these pseudotypes were stable and retained full infectivity during concentration by ultrafiltration or ultracentrifugation [Reiser, 1996; Reiser, 2000; Hanawa, 2002; Sena-Esteves, 2004].
Hanawa et al. [Hanawa, 2002] showed that HIV-1 vectors pseudotyped with the RD114 and amphotropic MLV GPs were more efficient than VSV-G pseudotypes at transducing human cord blood-derived CD34+ cells and clonogenic progenitors. In addition, amphotropic MLV pseudotypes transduced cytokine-mobilized, human peripheral blood CD34+ cells capable of establishing hematopoiesis in immunodeficient mice more efficiently than VSV-G pseudotypes. Work reported by Muhlebach et al. [Muhlebach, 2003] revealed that MLV vectors pseudotyped with the MLV 10A1 and the GALV GPs resulted in efficient transduction of preactivated human primary T lymphocytes while pseudotypes bearing the amphotropic MLV, RD114 and VSV-G GPs were less efficient. In contrast, HIV-1 vectors pseudotyped with the same GPs transduced preactivated T lymphocytes with similar efficiencies. This may reflect differences in the half-lives of lentiviral vector pseudotypes compared to MLV pseudotypes containing the same GPs.
In a study involving pseudotyped SIVmac251-based vectors, Sandrin et al. [Sandrin, 2002] showed that GALV and RD114 pseudotypes displayed a much reduced infectivity relative to vectors pseudotyped with the VSV-G, LCMV and amphotropic 4070A MLV GPs. Interestingly, SIV vectors pseudotypes bearing chimeric GPs with the extracellular and transmembrane domains of GALV or RD114 fused to the cytoplasmic tail of the amphotropic 4070A MLV GP, augmented the transduction efficiency of such pseudotypes in human and macaque primary blood lymphocytes and CD34+ cells. This may have been caused by changes in the intracellular trafficking and assembly of the GPs on viral cores due to changes of the cytoplasmic tails [Sandrin, 2004]. Similar results were reported by Zhang et al. [Zhang, 2004] who showed that transduction efficiencies in human mesenchymal stem cells (MSCs) with HIV-1 vectors pseudotyped with the unmodified RD114 GP were 1 to 2 orders of magnitude below those observed with VSV-G pseudotypes. However, HIV-1 vectors pseudotyped with a chimeric RD114 GP harboring an amphotropic 4070A MLV cytoplasmic tail revealed about 15-fold higher titers relative to the unmodified RD114 GP.
Baculoviruses, which are used as biopesticides, primarily infect members of the Lepidopteran family. It was found that recombinant baculoviruses are able to deliver transgenes into mammalian cells provided that they are controlled by a promoter that is active in mammalian cells [Ghosh, 2002]. GP64 is the major baculovirus GP and interest has recently been directed towards using the GP64 GP to pseudotype lentiviral vectors [Kumar, 2003]. GP64-pseudotyped HIV-1 vectors tolerated ultracentrifugation well and efficiently transduced various cell types although GP64 pseudotypes showed a more restricted tropism than VSV-G pseudotypes, with an especially poor ability to transduce hematopoietic cell types including dendritic cells [Schauber, 2004]. More notably, GP64 expression was not cytotoxic to host cells. Thus, derivation of a permanent cell line constitutively expressing GP64 at high levels over many passages was successful [Kumar, 2003]. Lentiviral vectors pseudotyped with the baculovirus GP64 were found to be inactivated by human complement [Schauber, 2004]. In animal sera, the vectors were mostly resistant to inactivation by rodent complement, whereas canine complement caused a moderate reduction in titer [Schauber-Plewa, 2005]. Human complement-resistant pseudotyped HIV-1 vector particles were subsequently produced through incorporation of complement regulatory proteins. Co-display of decay accelerating factor (DAF)/CD55 or of a GP64-DAF fusion protein stabilized GP64-bearing vector particles against inactivation by human and nonhuman primate serum [Guibinga, 2005; Schauber-Plewa, 2005].
To facilitate large-scale reproducible lentiviral vector production, a number of groups have established lentivirus packaging cell lines. Several of the proteins required to assemble lentiviral vector particles including VSV-G [Burns, 1993], HIV-1 Vpr [Bartz, 1996], Tat [Li, 1995], Rev [Miyazaki, 1995], and protease [Konvalinka, 1995] are detrimental to mammalian cells when overexpressed. Thus, for stable production of HIV-1 vectors pseudotyped with VSV-G, 293-based cell lines allowing conditional production of virus-encoded proteins and VSV-G using tetracycline regulated promoters were generated that yielded titers up to 106 TU/ml [Kafri, 1999; Klages, 2000; Farson, 2001; Xu, 2001]. Inducible cell lines for the production of SIV vectors pseudotyped with VSV-G were also established [Kuate, 2002]. This was achieved with expression plasmids containing a codon-optimized gag-pol coding region of SIV and the VSV-G coding region under the control of a ponasterone-inducible promoter. Vector titers at around 105 TU/ml were obtained.
To rapidly and reproducibly generate high-titer HIV-1 vector pseudotypes, Ikeda et al. [Ikeda, 2003] generated stable cell lines expressing GPs of gammaretroviruses including the amphotropic 4070A MLV GP, the RD114 GP with an HIV protease site introduced at the R-peptide cleavage site (RDpro), or the GALV glycoprotein with an MLV cytoplasmic tail. In concert with a codon-optimized HIV-1 gagpol coding region, these producer cells could make lentiviral pseudotyped particles with titers up to 107 TU/ml (20 TU/cell/day) for at least three months in culture. Compared to vectors pseudotyped with VSV-G, the gammaretrovirus pseudotypes were relatively stable at 37°C and were resistant to inactivation by freeze/thaw cycling or incubation with human sera [Strang, 2004]. More recently, Strang et al. [Strang, 2005] established stable producer cell lines for HIV-1 vectors pseudotyped with the alphavirus RRV and SFV E2E1 GPs. Stable clones produced high-titer RRV-pseudotyped HIV-1 vectors for at least 5 months. Clones producing SFV pseudotypes, however, revealed relatively low titers.
Typical titers of lentiviral vectors pseudotyped with VSV-G obtained by transient transfection range from 106 to 107 TU/ml. Increased titers can be achieved by physical concentration including ultracentrifugation and ultrafiltration of virus-containing cell culture supernatants. Using a protocol initially designed for the generation of high-titer MLV vector stocks [Burns, 1993], pseudotyped lentiviral particles containing VSV-G have proven to be stable and able to withstand concentration by ultracentrifugation without significant loss in titer [Bartz, 1996]. This greatly facilitated the generation of highly concentrated vector stocks for in vivo applications. Lentiviral vectors harboring a wide variety of different GPs have since been shown to lend themselves to concentration by ultracentrifugation [Reiser, 2000; Kobinger, 2001; Sena-Esteves, 2004].
Concentration protocols based on ultrafiltration were also established for HIV-1-based vectors. Facile ultrafiltration procedures were developed early on [Reiser, 1996] to concentrate HIV-1 pseudotypes. Good recoveries were observed with HIV-1 particles containing VSV-G or the amphotropic MLV GP. Improved procedures for concentration by ultrafiltration of other pseudotypes were subsequently reported [Reiser, 2000; Cui, 2002; Hanawa, 2002; Coleman, 2003; Sena-Esteves, 2004].
Vector production for large-scale in vivo applications that require high-titer stocks is challenging due to the lack of simple procedures capable of rapidly processing large volumes of cell culture supernatant. The traditional ultracentrifugation-based approaches are limited in terms of their capacity to handle large volumes, thus making this procedure extremely tedious. Centrifugation at low speed overnight has been used to process volumes in excess of one liter [VandenDriessche, 2002]. However, this low speed approach is time consuming. Thus, there is an emerging need for quick, reproducible and less laborious procedures that rapidly reduce the volume of the cell culture supernatant to be processed. Pham et al. [Pham, 2001] have described a gentle and simple precipitation method to rapidly concentrate pseudotyped lentiviral vectors involving the co-precipitation of pH-adjusted viral supernatants with calcium phosphate, low speed centrifugation, and dialysis. Volumes were decreased from 300 ml to 10 ml, with 50-100% recovery of infectious virus. Zhang et al. [Zhang, 2001] have presented an alternative precipitation method based on poly-L-lysine. Viral supernatants were mixed with poly-L-lysine, incubated for 30 min at 4°C, and then centrifuged at 10,000 g for 2 h. Recovery of infectious virus ranged from 26-32% after processing up to 3 liters of cell culture supernatant.
One problem with the methods outlined above is that cell-derived components are concentrated along with the vector particles leading to potential immune and inflammation responses [Baekelandt, 2003]. Thus, ultracentrifugation/ultrafiltration and precipitation approaches generate rather impure virus preparations and additional steps including chromatography-based approaches are needed in order to purify the virus of contaminating host cell components. Methods based on anion exchange chromatography of HIV-1 vectors pseudotyped with VSV-G have been established [Scherr, 2002; Yamada, 2003]. Schauber et al. [Schauber, 2004] described a similar procedure for HIV-1 vectors pseudotyped with the baculovirus GP64 GP. Yields and purity of the virus stocks resulting from these procedure were not reported, but these approaches may lead to vector stocks of improved purity, increased infectivity and reduced toxicity.
VSV-G-pseudotyped lentiviral vectors recently entered clinical application. The production, purification and testing protocols were adapted to GMP standards, and eventually gained FDA clearance [Lu, 2004; Manilla, 2005].
We acknowledge the support of the US National Institutes of Health (NS044832).


text:
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Wish Upon A Paper Plane
A second shot at first love. Lee Sooyeon is 29 and has never been in a relationship. Consoled by everyone that she’ll find the one someday, she believes her failed first love has doomed her love life. One day, a guardian angel takes her back in time. Will she manage to change the future or will everything crumble?

domain: literature


text:
There are thousands of Colorado football fans who believe they already know who their starting quarterback will be when the 2014 season kicks off in six months.
There is one man wearing black and gold on the practice fields this spring believing he has a chance to surprise them.
Sophomore Jordan Gehrke spent his first season in the program last fall No. 3 on the depth chart behind Sefo Liufua and Connor Wood. Liufua took over the starting job at midseason after Wood’s hot start faded into inconsistency.
Liufua threw for nearly 1,800 yards and led the Buffs to a pair of wins, including a Pac-12 Conference victory over California. After watching Liufau throw for 12 touchdowns and complete 60 percent of his passes as a true freshman while keeping the Buffs competitive in more games than they were in 2012, most believe Liufau has little standing in his way from at least beginning the 2014 season as the Buffs’ starter. Wood’s decision to quit football before his junior season only adds to that belief.
Gehrke believes he can push Liufau for the job.
“I think it’s just executing, honestly,” Gehrke said Friday after the Buffs’ first practice of spring drills in the bubble on the lower practice fields. “At this level, everyone is talented. So it just comes down to who is more efficient. I just take it one play at a time and just win every play I guess.”
Coach Mike MacIntyre said he didn’t sleep much Thursday night because he was excited about the first practice. He was rewarded with a good first practice by his quarterbacks, including newcomer Trent Sessions, who joined the team as a walk-on after serving as a member of the training staff last season.
“I think Sefo was throwing the ball today better than he did at any time last year,” MacIntyre said.
Coaches like Gehrke’s athletic ability, but he is several inches shorter than Liufau and is susceptible to passes being batted down on occasion. His ability to make plays on the run is an area where he clearly has an advantage over Liufau.
Coaches began telling Gehrke last August that if he ever wants to be able to maximize that ability on game days, he will have to take better care of the ball and carry it the proper way in a protected fashion when he does choose to run. He too often carried it one-handed, flailing at his side in August camp, which led to fumbles.
“All day today he’s putting it away, he’s protecting it,” MacIntyre said. “All those types of things you can tell they’re listening to what we’re saying.”
Gehrke admitted Friday he didn’t come to CU last summer in the right frame of mind to truly compete for a job. He said he was “star struck” by some of his teammates and didn’t play with the level of confidence needed to excel.
He passed for 2,388 yards and 22 touchdowns two years ago at Scottsdale Community College. He said he has no doubt he can play at this level after spending a season in the program.
“You’ve got to compete,” he said. “Every single practice matters. I think when I came in, I was just star struck, not really knowing what to expect. I got that out of the way and I’ve got three more years left and I’m going to compete every single down.”

domain: news


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86 coupe GT crash
This past thursday my 86 GT was involved in a pretty good crash. My
wife was driving in Boulder, in the left turn lane, sitting in the
waiting for traffic to clear. Across from her was a big chebby suburban,
awaiting to also turn left. She waited and waited, then light changed to
red and she attempted to complete her turn. As she started moving a
phone comany van comes through the intersection doing 35mph,
or possibly more. The left corner of the van appears to have made
contact with the inside edge of the passenger headlight. The van
popped the GT pretty good, and spun my wife 180 degrees and then
she had steer the car away from hitting the car which was in the turn
lane immediately behind her. Her chest hit the steering wheel pretty
hard and she got a brush burn on her collar bone from the seat belt.
She was pretty dazed. I got the call, turned off the grill, loaded the
and took her to the emergency room for come chest x rays. There was
no internal chest injury, just the muscle tissue is very sore from the
impact. She is still sore, but getting better.
As for the accident report, she says the light was red, the van driver
it was yellow when he entered the intersection and no tickets were
As for the coupe, it performed its job and protected my wife and she
away. The front of the car is pretty trashed. The impact broke off the
compressor, sheered off the oil filter and the a/c compressor ended up
down the street. The valence, grill, passenger headlight are destroyed.
drivers headlight is cracked. The impact created a V in the valence &
strutcure about 8 in deep. The hood release latch got moved about 6
towards the drivers side. The hood remained latched. It looks like the
got shoved into the radiator, but I'm not totally sure. The windshield
break.Both fenders got crunched. The passenger door is fine, the drivers
opens but has some interference with the drivers fender. The safety
of the car did its job, if you look from the A pillar on back the car is
if you look at the car from the A pillar forward, its trashed. My
is that the car is totalled. I am planning on trying to keep the car and
There are many parts which can find a new home on the ur-q and there are
many coupe specific parts which people should be interested in, i.e.
/h&r suspension w/5k miles? 14 in msw 5 spoke wheels w/ re71s? And other
goodies on the shelf which i've been storing, yes Bruce B., a set has
name on it. I also need to pull the Nakamichi/Proton stereo system, no
yard deserves a stereo system like that.
As for a replacement, my wife and I have been discussing a 5000/200
TQ wagon/avante. No real hurry right now, as the 83 ur-q will take the
driver duty, but any leads on 1986 - 1990 quattro wagons appreciated.
I've been on the q-list since 93, and lightly follow the 5000/200 turbo
talk. I've dug
out my original 1987 5000 and 1989 200 brouchures to familiarize myself
the cars. But I do need a primer of sorts. If anyone has a 5000/200
FAQ, I would appreciate a copy.
A convert to the green stuff?
86 GT, graphite metallic, bent
83 ur-q, green, daily driver w/89 200 MC engine installed


text:
One Of The Top Ten Haunted Hotels In The Country Is Right Here In Texas
It’s officially October, which means it’s time to commence the Halloween festivities. Texas is absolutely full of spooky attractions, but one rises above all the rest. Beaumont’s Haunted Hotel has been voted the scariest place of its kind in the entire state, and for good reason. You’ll be frightened out of your mind in the best way possible and leave with the spirit of the holiday engulfing you — if you ever get to check out, that is.
The Haunted Hotel is located at 447 Orleans St, Beaumont, Texas 77701.
Have you ever been here? Is it as scary as the hype claims it to be?

domain: geography


text:
UNIVERSITY OF COPENHAGEN
Office: 26.1.22 (1st floor)
University of Copenhagen
Department of Economics
Øster Farimagsgade 5, bygn. 26
DK-1353 København K
Phone: +45 35 32 32 58
Email: shb "at" econ.ku.dk
SIMON HALPHEN BOSERUP
Department of Economics, University of Copenhagen
o "Stability and persistence of intergenerational wealth formation: Evidence from Danish wealth records of three generations"
(with Wojciech Kopczuk and Claus Thustrup Kreiner). October 2014. (pdf)
(Previously a chapter in my PhD thesis entitled "Intergenerational Wealth Mobility: Evidence from Danish Wealth Records of Three Generations")
o "Tax evasion, information reporting, and the regressive bias prediction"
(with Jori Veng Pinje). August 2013. (pdf).
WORK IN PROGRESS
o "Intergenerational wealth formation over the life-cycle: Evidence from Danish wealth records 1984-2013"
(with Wojciech Kopczuk and Claus Thustrup Kreiner).
o "Born with a silver spoon? Danish evidence on wealth inequality in childhood"
(Previously entitled "Born with a silver spoon: Danish evidence on intergenerational wealth formation from cradle to adulthood")
o "The Role of Bequests in Shaping Wealth Inequality: Evidence from Danish Wealth Records"
"Essays on Tax Evasion and Enforcement and Intergenerational Wealth Mobility". PhD dissertation. March 2013. (pdf)
o Danish evidence on wealth inequality in childhood in VOX, CEPR's Policy Portal, November 4 2016.
o Bequests and wealth inequality: Evidence from Denmark in VOX, CEPR's Policy Portal, March 11 2016.
Public finance, taxation, tax evasion, intergenerational mobility
Assistant professor, University of Copenhagen, 2017-present.
Postdoctoral research fellow, University of Copenhagen, 2013-2017.
Visiting student researcher, University of California, Berkeley, 2011.
PhD in Economics, University of Copenhagen, 2008-2013 (Supervisor: Prof. Claus Thustrup Kreiner (link)).
MSc in Economics, University of Copenhagen, 2010.
BSc in Economics, University of Copenhagen, 2007.
Research assistant, CEPOS, 2005-2006
HONORS AND AWARDS
Postdoctoral research grant, 2013 (Danish Council for Independent Research, Social Sciences, DFF - 1329-00046, visit project page).
Euroclear Scholarship 2010 for studies at UC Berkeley, spring 2011.
Nominated for McKinsey Prize (best seminar paper), spring 2008, at Econonomics, Univ. of Copenhagen.
Public Finance, spring 2015-2016.
Econometrics A, fall 2009.
Quantitative Methods 2, spring 2009.
Theoretical Statistics 2, fall 2005.

domain: finance


Random thoughts of a Birmingham-based political junkie and party hack, Labour member and activist. And a Councillor for the City of Birmingham, representing Acocks Green ward.
RECENT ACTIVITYNo to the building of Halfway Houses opposite East Park School edited their Description and News.
Lol the PC brigade.
Post a Comment


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Chapel Sessions Music presents Andrea Valeri www.andreavaleri.com
We are delighted to welcome back to the UK, the world renowned Italian guitarist Andrea Valeri
“A Guitar Genius, one of the best Guitar players in the world” – John Doan
“He’s Like a brother to me, One of the best Guitarists around” – Francois Sciortino
“The shining light of the guitar world” – Michael Fix
“His potential is one of the world’s Best” – Eric Spitzer-Marlyn
“I met him in a Festival in Italy, And I was completely blown away by this new fingerpicking sensation” -Thomas Leeb
These are just some of the accolades his fellow guitarists from around the world have bestowed on Andrea.
Andrea Valeri was born in Pontedera February 27, 1991: from Australia to New Zealand, from Russia to Canada, from Europe to the African continent, Andrea continues to reap a worldwide success since the age of 16 years.
Defined as the “Prince of the Acoustic guitar”, and a new ambassador of acoustic music, Andrea, despite his young age is now a veteran of many of the biggest stages around the globe.
He has never been more in demand, and this year he includes 7 dates in Japan so we are honoured that he has once again committed to his loyal English fans to perform at Chapel Sessions.
In addition to his busy solo touring schedule he has also recently embarked on a very exciting new project where he has teamed up with some of Europe’s and The World’s best musicians, bringing his music to a star studded Eurochestra to give birth to a truly unique Musical Experience! Andrea will be able to explain in person where this project is going next?!
Guitarists who have inspired Andrea define him as a “Star of the guitar world.”
Check out Andrea’s music on his website or visit some of his many youtube videos
Doors open 7pm, concert starts 7.30pm, Real Ale/Wine bar & Tea/Coffee available from 6.30pm in Church Hall.


KANSAS CITY, Mo. — Community activists gathered Tuesday morning near the area where a 22-month-old boy was shot and killed when someone fired into a vehicle less than 24 hours earlier.
Karl Oakman, a deputy chief with the Kansas City, Missouri, Police Department, said Tyron Payton will have "the full power of KCPD" to find the person responsible for the shooting.
"This little boy did not deserve this," Oakman said at the news conference, where nearly a dozen activists gathered outside a Kansas City fire station. " ... We have failed when a situation like this happens in this city that I love and this city that each of us love. We have failed, we have to do better, and we have to make a difference."
Oakman and others encouraged anyone who witnessed the shooting or may have information in Tyron's death to call KCPD or the Greater Kansas City Crime Stoppers Tips Hotline.
Community activists are holding a news conference encouraging community members to come forward with information regarding the shooting death of 1-year-old Tyron Payton. @41actionnews pic.twitter.com/VMcF06QG2S— Emma James (@Emma_JamesTV) September 22, 2020
Bishop Tony Caldwell with the Justice and Dignity Center said there was a $25,000 reward for information leading to an arrest in the deadly shooting.
"When's it gonna stop?" Caldwell said. "Somebody out there saw something. This is about that child's life."
Emanuel Cleaver III, pastor at St. James United Methodist Church, said it was time for Kansas City to "step up."
"We need the community to step up and if you have any information to share that," Cleaver said. "I come from a tradition where we teach that what you do for the least of these, you also do for God. And I think a 2-year-old falls into that category."
On Monday afternoon, Tyron Payton and his parents were shot while inside a vehicle near East 33rd Street and Agnes Avenue. The driver of the vehicle went to a nearby fire station for help after the shooting.
Tyron's parents, who were in the front seat, remained in stable condition late Monday night. A fourth adult in the vehicle was not injured.
Police said that detectives were “making headway” in identifying persons of interest in the shooting but were still asking anyone with information to come forward.
The vehicle, which was towed shortly after the shooting, showed at least eight bullet holes in the driver’s side door.
Zach Fatall with the Kansas City Fire Department was one of the firefighters who responded when the car arrived outside the station. He said he immediately noticed the bullet holes riddled across the car.
While first responders "did everything we could" to save the child, Fatall said it was obvious that he had been badly injured.
Sgt. Jake Becchina, a police spokesman, said Monday that it appeared a person had fired multiple shots into the vehicle at close range.
Speaking Monday, Mayor Quinton Lucas lamented the death of another young child in the community due to gun violence.
In late June, 4-year-old LeGend Taliferro was shot and killed while sleeping in his family’s apartment. His death spurred the launch of Operation LeGend, a federal anti-crime initiative that has since expanded to eight other U.S. cities.
Anyone with information in Monday’s deadly shooting is asked to contact the Greater Kansas City Crime Stoppers Tips Hotline at 816-474-8477 (TIPS).
For jurisdictions that utilize the Greater Kansas City Crime Stoppers Tips Hotline, anonymous tips can be made by calling 816-474-TIPS (8477), submitting the tip online or through the free mobile app at P3Tips.com.
Annual homicide details and data for the Kansas City area are available through the 41 Action News Homicide Tracker, which was launched in 2015.


Lot 50: lever action Winchester Mod. 94 "Klondike", .30-30 Win., #KGR4049, § C
Classic Auction for hunting guns and rifles, accessories, hunting related collectible items and hunting art
November 3, 2016
Wien, Wien, AustriaLive Auction
Description: 50 cm long barrel, gold receiver and mounts, commemorative model engraved celebrating the famous gold rush of 1896, stock of walnut with buttplate 34 cm, inlaid medal in buttstock, 2,9 kg, year of manufacture 1975, condition 1.


text:
Our amazing children are the reason we run camps!
Each of our programs is individually tailored to ensure that each child on camp feels safe, cared for, supported and most of all, happy!
Our unique 1:1 ratio of children to volunteers allows our leaders to provide intensive support, genuine friendship and a whole lot of fun to each child on camp.
Edmund Rice Camps NSW is a not-for-profit organisation that supports children, young people and families experiencing significant and complex disadvantage. We provide a variety of group-based recreational programs, including week-long and weekend camps and follow up day programs. Activities centre on building relationships, affirming peers, developing soft skills, and renewing confidence and self-worth.
Our camps are led by amazing young adult volunteers (17-25 years) who plan and deliver an empowering experience in a safe, fun and supportive environment. Our 1:1 ratio of children to volunteers enables us to provide intensive support and enrichment for all of our participants – each with their own abilities, needs and circumstances.

domain: sociology


text:
Paper planes are something of a metaphor. I see it now. A metaphor of life, of dreams and of the many times gravity couldn’t be defied
Pictures do not lie, until the days of Photoshop, but you cannot Photoshop the experience.
This was the thought that ran through my mind as I swiped through the gallery on Phil’s cracked phone screen Continue reading AWOOF DEY PURGE (tales of Philemon Omoluabi)
A LIFE IN ABEYANCE The torment we face as individuals on the path to greatness vary and differ, one man … Continue reading
Steps 1. Click the link, it takes you to a blog 2. Vote by making a comment on the poem … Continue reading Please vote for me


text:
Extend your holiday and add an additional evening in Sandton, Johannesburg for 65 euro per person (double occupancy).
Single surcharge: 55 euro
Pre-Extension includes :
- Transfer from O.R. Tambo International Airport upon arrival
- 1 nights’ accommodation at hotel with breakfast
PRE-EXTENSION JOHANNESBURG & SOWETO
Take the time to extend your holiday and add an additional evening (Saturday) in Johannesburg with a 4 hour cycling tour in Soweto.
- 1 night’s accommodation at a hotel with breakfast
- Transfer to Soweto and back to the hotel
- 4 Hour Cycle tour in Soweto with lunch (10 am or 2 pm)
Price per person: 175 euro (double occupancy)
Single surcharge: 65 euro
Guided bicycle tour through Soweto
Enjoy the guided bike ride through Soweto’s various neighborhoods where you will experience the life and culture by interacting with people and at the same time learning about Soweto’s rich history.
The 4 hour tour starts at 10am and ends with a lunch at the "outdoor restaurant" after the tour. Alternatively your tour can start with lunch at 2pm and then the cycling.
Note: "Bunny Chow" is a delicious African stew traditionally cooked over an open fire, served with fresh bread.

domain: travel


text:
Usain Bolt defended his gold medal at the Summer Olympics in the men's 200 meters Thursday night, winning in a time of 19.32 seconds. He becomes not only the first ever to win the 200m at back-to-back Games, but also the first to win both the 100m and 200m in consecutive Olympics, firmly cementing his legacy as one of the greatest sprinters the world has ever seen.
Yohan Blake -- who beat Bolt in the 200m at Jamaica's Olympic team trials -- won the silver medal, finishing in 19.44 seconds, his second-fast time ever run. It was also the fastest losing time in history. Warren Weir was third, making it a podium sweep for Jamaica. It's the first time any country other than the United States has swept the 200m.
Wallace Spearmon, the lone American in the race, was fourth, running a 19.90.
It was billed as the Lightning Bolt versus The Beast, and indeed it was, as the 0.12-second margin of victory -- and Bolt's constant glances over to where Blake was in relation to him -- showed. But in the end, it was all Bolt, again.
Bolt separated himself from the field almost immediately and already had a big lead as they made their town for the homestretch. Blake, however, did what maybe no other human being can do -- he closed the gap on Bolt, and made it at least somewhat interesting over the final 30 meters. But, like every other human being, Blake was simply not fast enough, allowing Bolt to be Bolt as he crossed the finish line -- in the photo-finish shot, he already has his finger to his lips.
(Photo by Omega via Getty Images)
Bolt's winning time was 0.02 seconds slower than what he won with in Beijing, but 0.08 faster than the 2011 World Championships.
For more on the Olympics, check out SB Nation's London 2012 Olympics Hub.


May 9, 2020
5 years ago – May 9, 2015
The Roseau High School graduation speakers for 2015 were Alexis Melby, DyAnna Grondahl and Kaitlyn Christianson.
It was seven national titles in a row for Byfuglien Trucking at the USA Hockey National Championships in Florida. Team members wer Steve Arnot, Mike Lundbohm, Kim Newman, Huffer Christianson, Bob Lund, Bryan Grand, Bruce Falk, Ryan Brandt, Rick Torinato, Brush Christianson, Steve Miklos, Peter Hegg, Charlie Brown and Larry Stordahl.
Mayville State University awarded Jacob Eaton of Roseau with the Outstanding Freshman Math Student of the Year Award....
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text:
My grandpa at the beginning of World War 1
I’m sitting here today and thinking about how much I owe to the men and women who have served our country in the military from the earliest days of our existence to the present. I have never looked out my window to see tanks and military vehicles on my street; never heard the sounds of gunfire or bombs falling. I am free to express my opinion on the government, free to follow the religion of my choice. No one excludes me from any building or ceremony because of the colour of my skin or my choice of attire. I owe these freedoms to others who were willing to put their lives on the line, some of whom gave up their lives on the battlefield. They don’t ask much of us…just some respect and two minutes of our time on November 11 at 11 a.m. They ask that we buy and wear a poppy. They ask that we remember them. A small price to pay for our priceless freedom.


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We’ve already written about how the Bulgarian compromise proposal for both Article 13 and Article 11 are too broken to fix. Their proposal for Article 3 (Text and Data Mining) does little to alter the major problems standing in the way of a progressive exception for text and data mining.
We’ve continued to follow Article 3 since the European Commission published its proposal on copyright in the Digital Single Market. Even though the Commission’s exception for TDM would be mandatory, we criticised their plan as not going far enough, as it would limit the beneficiaries of the exception only to research organisations, and only for purposes of scientific research.
TDM in the Bulgarian presidency proposal
The Bulgarian proposal is nearly identical to the changes already offered by the earlier Estonian plan. It leaves intact the Commission’s obligatory TDM exception that would apply to research organisations (including cultural heritage institutions) for purposes of scientific research. The Bulgarian proposal similarly introduces an additional and optional exception in Article 3 for temporary reproductions and extractions. This additional exception would apply to beneficiaries other than research organisations, and for uses other than scientific research. But those acts would be limited in that they only would cover temporary reproductions and extractions, and only if the rightsholder does not prohibit it.
In our earlier blog post we wondered whether the existing (and mandatory) exception in the InfoSoc and Database Directives on temporary reproductions arguably already covers the temporary reproductions for text and data mining purposes. In any case, this additional and merely optional exception, for acts that might already be covered under existing law, which can easily be neutralised if rights holders don’t want it, is a weak compromise. It doesn’t address the main concerns we’ve had with Article 3 since the beginning. It also fails to bring much needed harmonization and will instead further the already existing fragmentation of users rights in EU. Continue reading
We just got done criticising how Bulgaria’s weak consolidated presidency compromise handles Article 11 of the proposed Copyright in the Digital Single Market (DSM) directive. The Bulgarian plan doubles down on the Commission’s original bad idea and ignores most of the positive protections offered by some members of parliament.
But we shouldn’t have been surprised when MEP Axel Voss came out with an even worse plan for the press publishers right. This week MEP Voss released his proposal for a compromise on Article 11, and the changes he is proposing are even more radical and more broken than anything we’ve seen thus far.
Expanding the scope to cover facts
First, Voss proposes to expand the scope of beneficiaries of Article 11 from to cover not only press publishers, but also news agencies (who aggressively lobbied for being included in the scope of Article 11 late last year). But in doing so, he introduces the risk of inappropriately granting copyright – like protection to facts and compilations of basic information. This is a dangerous extension of the scope of exclusive rights that would endanger the right of access to information. It is especially appalling since even the founders of the “modern” European copyright system wisely choose to make sure that news of the day and facts cannot be exclusively claimed: The 1986 Berne convention explicitly states that copyright shall “not apply to news of the day or to miscellaneous facts having the character of mere items of press information”.
Perhaps the most extreme change is that press publishers and news agencies would be granted (or saddled with?) an “inalienable right to obtain an fair and proportionate remuneration for such uses.” What does this mean? It means that publishers would be required to demand payment from news aggregators and other users. Continue reading
Last week more than a hundred of copyright reform activists got together in Brussels for the the European Copyright Action Days to make it clear to EU lawmakers that the copyright reform effort that is currently being discussed in the European Parliament and the European Council is not good enough. In a series of events organized by Copyright 4 Creativity, Create.Refresh, Communia and others, activists and other stakeholders discussed the shortcomings of the current reform proposal as well as ideas for a more future-proof overhaul of the outdated EU copyright system.
As part of the Copyright Action Days we organized a a roundtable on the future of education in the European Parliament, our first ever COMMUNIA Salon on the future of copyright in the Museum of Natural Sciences and two workshops for copyright reform activists.
Video documentation by Sebastiaan ter Burg.
Roundtable on the future of education
The roundtable on the the future of education hosted by Dutch MEP Marietje Schaake was a full room event at the European Parliament, with over 40 policymakers and stakeholders attending. We discussed the intersection of educational policy, technology, copyright reform and open licensing policies. Irish school teacher Leanne Lynch talked about the use of technology, social media platforms and digital copyrighted materials in the classroom. Mitja Jermol – UNESCO Chair on Open Technologies for Open Educational Resources and Open Education – talked about how new technologies can support educational goals. Andreia Inamorato dos Santos from EC’s Institute for Prospective Technology Studies presented results of their latest report on open education policies in Europe. Finally, Damjan Harisch from the Slovenian Ministry of Education and Maja Bogataj Jančič, Director of the Slovenian Intellectual Property Institute, presented the position of Slovenian Ministry of Education on the copyright reform During the event, Teresa Nobre also presented our latest research on licences for educational uses. We are happy that we had the opportunity to exchange views on the matter with representatives of publishers and CMOs.
The Future of Technology in Education roundtable, photo by Sebastiaan ter Burg. More photos here.
The COMMUNIA salon in the Museum of Natural Sciences brought together more than 70 activists, academics and policy makers to discuss challenges on the intersection of creativity, value creation and copyright in the online environment. Under the title “Copyright for the future” the discussions attempted to draw up a perspective that looks beyond the current legislative proposal. Continue reading
As we are heading into the final phases of the discussion on article 13 in both the European Parliament and the Council the issue that article 13 has the potential to do substantial damage to the European digital economy is getting more attention from lawmakers. So far the answer to this problem by the proponents of Article 13 is to exempt more and more types of online platforms from the filtering and or licensing requirements established by Article 13. By now the list of services to be excluded contains “Non-for profit online encyclopaedia“, “educational or scientific repositories, where the content is uploaded by the rightholder“, “providers of cloud services for individual use which do not provide access to the public“, “online market places whose main activity is online retail of physical goods” (European Parliament draft), “non-for-profit open source software developing platforms” and “internet access service providers” (Council compromise proposal).
Generally speaking it is a good indication that a policy is bad if there is a need to make a large number of exceptions to prevent it from doing lots of unintended harm. This principle is on full display in the discussion about article 13. As we (and many others) have argued before, article 13 is broken so badly that it cannot be fixed and should be deleted. The key problem with article 13 is that the music industry is employing its old weapon of choice (copyright law) in an attempt to reign in behaviour of a very small group of online platforms that is perceived as problematic by the music industry. By using copyright law as a trigger for the licensing and filtering obligations contained in article 13, the article inevitably effects every other online platform that deals with copyrighted contents (i.e pretty much all online platforms).
It is not surprising that other platforms that operate in completely different markets (like GitHub which has nothing to do with uploading music) have started to realise that article 13 is a threat to their businesses and are demanding to be excluded from the scope of article 13. While excluding such platforms seems like an obvious choice to prevent some of the worst side effects of the provisions contained in article 13, it will not fix the underlying problem: In an age where copyright touches almost every online business model, copyright law is not a suitable regulatory instrument to adjust the bargaining positions of specific industries anymore. In order to make sure that article 13 has no negative side effects it would need to come with a list of exceptions that excludes every single business model that it is not targeted at. Continue reading
The European Union is coming closer to approving a mandatory educational exception that may address some of the limitations copyright law places on everyday educational activities. However, the current proposal for a Directive on Copyright in the Digital Single Market would allow licences that are easily available in the market to take precedence over the educational exception.
Our new report “Educational Licences in Europe“, covering the analysis of 10 agreements in Finland, France, and the United Kingdom, shows that educational licences contain terms and conditions disadvantageous to schools:Continue reading
This spring the ongoing effort to modernize the outdated copyright rules enters into the decisive fase. It is widely expected that both the European Parliament and the EU Member states will their position on the proposed Copyright in the Digital Single Market Directive. Right now things are not looking good: instead of a much needed update of the copyright framework that would enable new uses driven by technological innovation, policy makers in Brussels are working towards new restrictions that would would limit how information and creativity can be shared and enjoyed online.
Against this backdrop we are organising European Copyright Action Days on 19-21 march in Brussels. During these days we want to highlight the broad opposition of civil society, libraries, the users industry and many others concerning the restrictive aspects of the copyright reform proposal. During these days activists will convene in Brussels to discuss with lawmakers and advocate for a more future proof reform and to raise attention for the dangers of the proposed measures. Continue reading
After more than a year of discussions MEP Axel Voss has finally come forward with his ideas about one of the most controversial aspects of the EU copyright reform proposal. On Wednesday he shared his compromise proposals for Article 13 of the proposed copyright in the DSM directive, that deals with filtering measures aimed at online platforms. The “compromises” drafted by MEP Voss make it clear that with regards to article 13 he has chosen to do the bidding of the music industry at the expense of users, open platforms and pretty much the rest of the internet.
Let’s focus on two main aspects of the approach that Voss is backing (for a more comprehensive analysis of his compromise see Julia Reda’s excellent write-up here). What would the new rules mean for users sharing materials via platforms, and what would the new rules mean for online platforms?
Online platforms: License or cease to exist!
In the version supported by MEP Voss, article 13 establishes two different obligations for online platforms that allow user uploads. In a first step, all platforms are required to obtain licenses from rightsholders. Those platforms that hold “significant amounts” of content also need to take “appropriate and proportionate measures to ensure the functioning of these agreements”. In the case that platforms do not have licensing agreements with rightsholders they need to take “appropriate and proportionate measures to prevent the availability on their services of works or other subject-matter”.
These rules would effectively end the current situation in which online platforms are not directly responsible for content that their users upload. The new rules would mean that all online platforms “that store and provide access to the public to copyright protected works or other subject-matter uploaded by their users” (which means pretty much all platforms) will be directly responsible for the content uploaded by their users and must obtain licenses from (unspecified) rightsholders. If they don’t (which is a strange condition given that all platforms must do so) they must implement filtering rules that prevent all copyrighted works from becoming available on their services. In other words, platforms must obtain licenses from rightsholders or they must cease to exist (as it is somewhat hard to make a business case for a platform on which nothing is available). Continue reading
One of the biggest shortcomings of the discussion on copyright is that most of it seems stuck in a fairly outdated creators vs users dichotomy. Copyright laws around the world are generally structured in such a way that they grant exclusive rights to creators and try to balance these with a limited set of rights for users (in the form of exceptions or limitations to copyright). Based on this design it is widely assumed that more (or stronger) exclusive rights benefit creators and that more (or broader) exceptions to copyright benefit users.
This conception is problematic on a number of levels. For one it is clear that creators benefit from user rights that ensure that users have a basic level of access to culture through educational systems and via public institutions such as museums and libraries. On the other hand users benefit from the exclusive rights granted to creators as they incentivise the very production of culture and knowledge that they want to access.
A more fundamental challenge to this general understanding of copyright is posed by the fact that the roles of users and creators are not mutually exclusive, but overlapping. Many creators are also users of copyrighted materials and the other way around. The technological development of the past two decades has contributed to this blurring of the boundaries between creators and users. Digital technologies greatly facilitate both the creative re-use of existing works and the distribution of the resulting new works. This development has resulted in the emergence of the (somewhat nonsensical) category of “user generated content” and concepts like the “prosumer“.
These concepts deal with users becoming creators, and there are relatively straightforward answers to the challenges posed, such as the need to introduce an exception for user generated content in the EU copyright framework that we have been advocating for. But there is another more interesting side of the coin: creators becoming users. While it is true that creators have always appropriated the works of those authors who came before them, these dynamics have been turbocharged by the digital revolution. Creators have entire libraries of content at their fingertips, and the tools to manipulate, incorporate and build on existing works are becoming increasingly sophisticated. These are exciting times to be a creator, but this new reality also brings creators into contact with the limitations to their creative freedom imposed by copyright law. Continue reading
We have been arguing for quite sometime now that handing out the power to define the scope of users rights to right holders – in the form of license agreements that they can (almost unilateral) draft and frame as they wish – is bad. Really bad: licenses fragment the legal framework that mandatory exceptions try to harmonize; licenses contain abusive terms or impose obligations on users that are not foreseen in the laws; and licenses have a huge impact on national budgets.
Unfortunately, this message has not come through to all, or not everyone understands what we are saying, or worse right holders have done a nice job in convincing lawmakers that’s the right way to go.
Allowing licenses to override exceptions is the only treat that publishers want
The current copyright reform carried the promise of being a landmark in the history of the EU copyright law. Lawmakers would finally show they understand that copyright is not superior to any of the other fundamental rights that every constitutional law across Europe grants to their citizens, and would make things right. Sadly, however, the prospects of that being the case for education are now very low.
MEPs passed the last year negotiating the scope of the educational exception. On the one hand, those who side with schools, teachers and students, proposed amendments to eliminate some of the constraints that the educational exception contains. On the other hand, those who side with publishers have been pushing for more restrictions, in order to narrow down the scope of the proposed exception even further.
Not enough MEPs understood that the most problematic aspect of art. 4 is not the scope of the mandatory exception (n.º 1) but the fact that Member States may choose not to apply such mandatory exception if licenses covering those uses are easily available in the market (n.º 2).
It is our understanding that publishers could not care less about the scope of the educational exception, provided that they can rule out the application of said exception with their own license agreements. This is copyright “taking the back seat”, as Professor Niva Elkin-Koren would put it.Continue reading
Today, a group of Portuguese organizations, including an important innovation acceleration hub, software companies, free culture and users rights advocates, and the Portuguese association of librarians, archivists and documentalists, sent an open letter to the Portuguese Government asking to the Government to reconsider its position in relation to art. 13 (the proposal to require online platforms to filter all uploads by their users).
As we have noted before, Portugal is, along with France and Spain, one of the countries that supports the Commission’s plan to force online platforms to install upload filters that would prevent any uses of copyright protected not explicitly approved by rightsholders. Portugal has also been pushing forward amendments proposed by the French Government that would significantly change the way online platforms operate. Under the rules proposed by the French, operating open platforms would only be possible with permission from rights holders.
Portugal can still make it right!
The signatories of the letter acknowledge the negative impact that such proposals would have on the fundamental rights of the Portuguese citizens and on the booming Portuguese ecosystem of startups and entrepreneurs, which is as important to the Portuguese economy as the tourism industry. They, thus, ask to the Portuguese Government to depart from its initial position, which privileges the interests of a small class of commercial copyright holders, and to embrace the future of digital innovation instead.
This open letter is yet another reminder that copyright policy cannot be based on the interests of commercial rightsholders alone and a reminder that it is important to challenge the positions of national governments on this important issue (see this helpful overview by MEP Julia Reda for other governments that need to be reminded that we need copyright rules that embrace the future instead of the past).

domain: politics


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Cameron’s Janéa Pol’Anrhy just submitted his new track La Danse du Chien Menchant (The dance of the vicious dog). All I want to see now is a music video for this song with a viscous american pitbull dancing passionately ot this. The song s great combination of traditional sounds with EDM and rock elements.
[soundcloud url=”https://api.soundcloud.com/tracks/270161263″ params=”color=c88937″ width=”100%” height=”166″ iframe=”true” /]
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domain: artistic


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Wanted--An off-farm lease!
I really want to convince my parents to let me lease. I think it will really help me learn about horses. I would like to see a cheap-ish off-farm lease or a farm close to New Tripoli, PA so I would be able to get lessons from MY trainer and also trailer to shows with my lease horse. It would be a hunter horse that is at least schooled at 2' cross-rails. But, if there is not any horses like this in PA or anything. I will exercise horses and such for money to hopefully, lease a hunter I have my eye on. School is out in about 3 weeks so I can start working probably then. :-| Hope someone can help me :wink:.
Before you even start looking into leasing you should make sure you have your parents complete ok. After all they do have control over you until you're 18. lol
Off-farm leases can be very difficult to find as you have seen for your self. Mainly because during an off-farm lease a horse can pick up nasty habits or even be ruined, and no good owner wants that. Most responsible owners want what's best for their horse and want to be able to keep an eye on said horse.
Also, the only cases of off-farm leases I've had experience with required the lessee to use the owner's trainer, vet, and farrier so that the horse would be taken care of to the owner's standard. Plus, I don't know of anyone who would let a trainer they don't know or trust do anything with their horse. I know I wouldn't. I love my boys too much to take a chance with a stranger.
I thought you were able to use you trainer's horse Secret, and that was basically like you owned her? A new horse won't be a miracle solution to any issue you have with Secret. In horse riding 98.5%(if not more) of issues are rider error, and are transfered onto any horse you get on. It's up to the rider to know their weak spots and work to improve them. No one is perfect. I've got multiple titles and championships under my belt, and I know I'm still far from perfection. As riders we must take responsibility for ourselves and work to better our riding. When things go wrong on the horse in 99% of cases the rider has no one to blame but her/herself. You don't get better by blaming your shortcomings on others. I think your best bet would be to stick with Secret or maybe another horse of your trainer's. Off-farm leases are hard to come by and not always the best solution.
^^Thx. Secret is a nice horse but I REALLY wanna' do hunters. Secret is in no place to do that and most of the horses at my barn are the same way. They can trot over cross-rails but I have been riding for 3 years and feel I am ready to lease. I am willing to wait but once I start showing alot more, I will need a horse to condition, train, work with,ect.
Please get permission before advertising that you are looking. If they say no, you won't have wasted the potential lessor's time.
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I think leasing is a great idea! I've gained so much confidence and expirience (not saying I'm expirienced just that compared to how I used to ride? You'd never know that I was the girl who just cantered around uncontrollably not even bothering to look at the lead or use transitions or not even ride for a reason. lol) and it's helped Cheyenne too- she now has one person taking care of her and loving her instead of a whole bunch of pure beginner little kids riding who have no idea what their doing riding her. And it's even helped her owner too. :) Hope you can find a horse :)
When looking to lease you basically have to sell yourself. So posting more about your riding level, specifically what you want to do and/or posting pictures/video of you riding will help people judge whether you would be suitable for their horse.
Ok, sell myself, thanks NittanyEquestrian.
I consider myself a intermediate/advanced rider. I have worked with a few horses in my life. I learned to ride on a naughty horse. Not like bucking and rearing, as I have never been on a bucking or rearing horse but rude and lazy. She wouldn't trot. She would do sharp turns at a trot and the saddle would slip to the side. I fell off once like that. She ripped reins out of my hands and took off when I didn't have stirrups. I have already jumped 18" cross-rails and will pop over them confidently. I can canter a horse easily. I learned to ride on a horse with a very bumpy canter. I can canter basically any horse. I don't like horses with vices like bucking and rearing. I wouldn't want to own one till I'm more experienced. I want to do hunters. Here is a recent video.
Leasing is a great idea when you want to earn more experience but aren't ready to buy (which comes with alot of spending and responsibility!) I've been leasing various horses for the past 4 years! 1 ottb, 1 regular tb, an Arabian, and even a perchX. From each I learned so much! Now just curious.. And I really don't want to stir things up but, if you have been riding 3 years how come the recent video you posted says " second cantering lesson" in the description?
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I won't lie. I went through alot. I rode and was OK then a winter off (no indoor) I had to start again. Then took and was about to canter. I cantered, fell off and totally lost my confidence. I switched to western and went to my first show. I finally realized that if I wanna jump I have to be brave and just canter. So my 2nd lesson cantering at least 50 %.
Also just to add I think you should put yourself as intermediate maybe not advanced. I could be wrong but I see an advanced rider as someone who can ride horses with vices such as bucking and maybe rearing. They also would be able to jump on just about any horse and ride it confidently at all paces and be able to fix any problems as they show up or just before. Just as an example a rider of that level I'm pretty sure would have fixed that the horse in that video was on the wrong lead in the canter both times. Just my 2 cents :D
Good luck. Oh and make sure your parents approve first before getting your hopes up.
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domain: hobby


Oktoberfest Cork has been cancelled - even after 15,000 tickets are sold
It came as a shock to many.
Oktoberfest Beag in Cork has been cancelled, it was announced last night.
It appears that it was cancelled due an objection that was lodged with the Vintners Federation of Ireland.
OKTOBERFEST CORK CANCELLED - 15,000 tickets for one of the city's biggest festivals had already been sold. Organisers say they had been 'dealing with significant resistance from the Vintners Federation of Ireland - Cork division. #NPRedFM email@example.com/ 1850 104 106 pic.twitter.com/jzl6gslu7v
— Neil Prendeville (@NeilRedFM) September 7, 2018
The beer fest took to Facebook to share the news:
"It is with great sadness that we announce the cancellation of the 9th annual Oktoberfest Beag Cork 2018.
"Over the past 24hrs behind the scenes we have been dealing with significant resistance from the Vintners Federation of Ireland - Cork division, where yesterday we were made aware of an objection lodged against the Oktoberfest licence application.
"We exhausted all efforts to resolve the few concerns they had however due to the frustration of the application and lack of support from the Cork Vintners, the deadline passed where we could not proceed with the plans without risking significant losses and disruption for all involved."
It continued by thanking the fans of the festival and those who worked to make it run smoothly:
"To our loyal ticket holders and corporate groups who plan their annual night out with us each year celebrating Oktoberfest - we deeply apologise. We sincerely thank you for all your support over the past number of years. We are all absolutely gutted!"
For more information, you can check out the Facebook post below.