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Embryonic Stem Cells/Induced Pluripotent Stem Cells
Long-Term Safety and Function of RPE from Human Embryonic Stem Cells in Preclinical Models of Macular Degeneration†‡
Version of Record online: 11 JUN 2009
DOI: 10.1002/stem.149
Copyright © 2009 AlphaMed Press
Volume 27, Issue 9, pages 2126–2135, September 2009
Additional Information
How to Cite
Lu, B., Malcuit, C., Wang, S., Girman, S., Francis, P., Lemieux, L., Lanza, R. and Lund, R. (2009), Long-Term Safety and Function of RPE from Human Embryonic Stem Cells in Preclinical Models of Macular Degeneration. STEM CELLS, 27: 2126–2135. doi: 10.1002/stem.149
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Author contributions: B.L.: conception and design, collection and/or assembly of data, data analysis and interpretation, manuscript writing; C.M.: conception and design, collection and/or assembly of data, data analysis and interpretation, manuscript writing; S.W.: conception and design, collection and/or assembly of data, data analysis and interpretation, manuscript writing and final approval of manuscript; S.G.: collection and/or assembly of data, data analysis and interpretation; P.F.: conception; L.L.: conception and design, collection and/or assembly of data, data analysis and interpretation; R. Lanza: conception and design, collection and/or assembly of data, data analysis and interpretation, manuscript writing and final approval of manuscript; R. Lund: conception and design, collection and/or assembly of data, data analysis and interpretation, manuscript writing and final approval of manuscript. B.L., C.M., and S.W. contributed equally to this article.
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First published online in STEM CELLS EXPRESS June 11, 2009; available online without subscription through the open access option.
Publication History
- Issue online: 8 SEP 2009
- Version of Record online: 11 JUN 2009
- Accepted manuscript online: 11 JUN 2009 12:00AM EST
- Manuscript Accepted: 3 JUN 2009
- Manuscript Received: 20 MAR 2009
- Embryonic stem cells;
- Retinal pigment epithelium;
- Macular degeneration;
- Clinical trials
- Top of page
- Abstract
- Acknowledgements
- Supporting Information
Assessments of safety and efficacy are crucial before human ESC (hESC) therapies can move into the clinic. Two important early potential hESC applications are the use of retinal pigment epithelium (RPE) for the treatment of age-related macular degeneration and Stargardt disease, an untreatable form of macular dystrophy that leads to early-onset blindness. Here we show long-term functional rescue using hESC-derived RPE in both the RCS rat and Elov14 mouse, which are animal models of retinal degeneration and Stargardt, respectively. Good Manufacturing Practice-compliant hESC-RPE survived subretinal transplantation in RCS rats for prolonged periods (>220 days). The cells sustained visual function and photoreceptor integrity in a dose-dependent fashion without teratoma formation or untoward pathological reactions. Near-normal functional measurements were recorded at >60 days survival in RCS rats. To further address safety concerns, a Good Laboratory Practice-compliant study was carried out in the NIH III immune-deficient mouse model. Long-term data (spanning the life of the animals) showed no gross or microscopic evidence of teratoma/tumor formation after subretinal hESC-RPE transplantation. These results suggest that hESCs could serve as a potentially safe and inexhaustible source of RPE for the efficacious treatment of a range of retinal degenerative diseases. STEM CELLS 2009;27:2126–2135
- Top of page
- Abstract
- Acknowledgements
- Supporting Information
Since their discovery over a decade ago [1], human ESCs (hESCs) have been considered a promising source of replacement cells for clinical studies. However, numerous problems continue to plague clinical translation, including the risk of teratoma formation and the need for powerful drugs to overcome the problem of immune rejection. Until somatic cell nuclear transfer or induced pluripotent stem cell technology is further developed, diseases affecting the eye—a relative immunoprivileged site [2]—are likely to be among the first hESC-derived therapies. In the retina, compromised retinal pigment epithelial cell (RPE) function can lead to deteriorated vision and photoreceptor loss in both age-related macular degeneration and some forms of retinitis pigmentosa [3–7]. Although treatment options are presently extremely limited, there is evidence in animals that implantation of RPE cells can sustain photoreceptors and control deterioration of visual function [8–13].
However, despite preliminary data generated using RPE generated from hESCs [9], the transposition of scientific protocols from basic research to clinical application is precluded by the need to adhere to guidelines set forth by the U.S. Food and Drug Administration, collectively referred to as current Good Manufacturing Practices (cGMPs) and current Good Tissue Practices (cGTPs). In the context of clinical manufacturing of a cell therapy product, such as hESC-derived RPE, cGTPs govern donor consent, traceability, and infectious disease screening, whereas the cGMPs are relevant to the facility, processes, testing, and practices to produce a consistently safe and effective product for human use.
Here we augmented previously published RPE-derivation protocols [9], implementing appropriate controls around the cell manufacturing and documentation process to achieve full compliance with the cGMPs/cGTPs. hESC-RPE cell lines were prepared for clinical application and tested for safety and efficacy in several different in vitro and animal models. Three RPE batches from two hESC lines were tested in the RCS rat, an animal in which vision deteriorates over several months because of an RPE functional defect [14, 15]. We focused on longevity of effect, dosing effect, and evidence of untoward pathology. Based on these functional data, an hESC-RPE batch was chosen for a GLP safety study using the NIH III mouse model, an immune-deficient animal model with eye pigmentation but devoid of natural killer (NK) cells and mature T and B cells. Importantly, we also explored whether vision can be rescued in the Elovl4 mouse model for Stargardt's disease (STGD) [16]. STGD is a form of early-onset macular dystrophy that leads to blindness despite an intact Bruch's membrane, which is critical for RPE cell attachment and potential clinical efficacy [17].
- Top of page
- Abstract
- Acknowledgements
- Supporting Information
Animals and Experimental Designs
Pigmented dystrophic RCS rats (n = 79) and ELOVL4 mice (n = 28) were used in the main experiments. NIH III immuno-nude mice (n = 45) were used for safety study. For RCS rats, animals were divided into five groups according to the doses they received. They were 5 × 103 (5,000)/eye (n = 21), 2 × 104 (20,000)/eye (n = 21), and 5 × 104 (50,000)/eye (n = 21). Animals from all dosage groups received cells with low medium and high pigmentation. All above the dosage group animals were received cells with low, medium and high pigmentation (Table 1). For further comparison, two groups were added: one group of animals (n = 8) received 7.5 × 104 (75,000)/eye cells and another group (n = 8) received 1 × 105 (100,000)/eye cells with medium pigmentation. For ELOVL4 mice, the eyes received 5 × 104 (50,000)/eye cells with medium pigmentation. All animals in the main experiments were maintained on oral cyclosporine A administered in the drinking water (210 mg/l, resulting blood concentration of ∼300 μg/l [12]) from 1 day before transplantation until they were sacrificed. An intraperitoneal injection of dexamethasone was given for 2 weeks (1.6 mg/kg/day) after surgery in cell and control injected rats and for 2 weeks alone in untreated animals. All animals were maintained under a 12-hour light/dark cycle. The animals were housed and handled with the authorization and supervision of the Institutional Animal Care Committee at the Oregon Health and Science University.
Experiments were carried out in accordance with the guidelines laid down by the NIH regarding the care and use of animal for experimental procedures and ARVO Statement for the Use of Animals in Ophthalmic and Vision Research.
Cell Preparation
Culture of hESCs and Differentiation into Mature RPE Cells.
All cell manufacturing procedures were carried out in ISO Class 5 biosafety cabinets in a fully validated, state-of-the-art ISO Class 7 clean room facility under strict environmental control monitoring systems and a routine microbial testing regimen. Single-blastomere hESC lines MA01 and MA09 were maintained as previously reported [18]. hESCs were dissociated from the primary mouse embryonic fibroblast layer by treatment with 0.05% trypsin-EDTA (Invitrogen, Carlsbad, CA, http://www.invitrogen.com) and were seeded in 6-well low-attachment plates to allow EB formation in a chemically defined minimal essential medium (MEM)-based medium (MDBK-GM; Sigma, St. Louis, http://www.sigmaaldrich.com) containing B-27 supplement (Invitrogen) for approximately 7 days and plated on gelatin-coated (0.1%; Stem Cell Technologies, Vancouver, BC, Canada, http://www.stemcell.com) dishes until RPE colonies were visible. RPE was purified by 3-hour exposure to 4 mg/ml type IV collagenase (Invitrogen) and manually isolated with a glass pipette. Purified RPE was seeded onto gelatin-coated tissue culture plates and expanded in EGM-2 medium (Lonza Biologics, Basel, Switzerland, http://www.lonzabiologics.com) until desired density was achieved, at which point cultures were reverted to MEM-based medium (MDBK-MM; Sigma) and cultured until the appropriate phenotype was achieved. RPE was dissociated from culture using a 1:1 mixture of 0.25% trypsin-EDTA and Hanks-based cell dissociation buffer (Invitrogen) and was cryopreserved in 90% fetal bovine serum (HyClone, Logan, UT, http://www.hyclone.com) and 10% dimethylsulfoxide (Sigma).
Quantitative, Real-Time, Reverse Transcription-Polymerase Chain Reaction.
RNA was extracted from the cells using TRIzol reagent (Invitrogen) according to the manufacturer's protocol. Eluted RNA was quantitated by spectrophotometry, and 10 μg was subjected to DNase digestion, followed by a reverse transcription reaction using a QuantiTech reverse transcription kit (Qiagen, Hilden, Germany, http://www1.qiagen.com) with a mixture of oligo-dT and random hexamers primers. Fifty nanograms per well of cDNA was used as templates in quantitative polymerase chain reactions (qPCRs) with oligonucleotides specific for hESC and retinal genes (supporting information Table 1). All qPCR reactions were performed in triplicate, with the resultant values being combined into an average threshold cycle (CT). The efficiency of qPCR was calculated from the slope of a relative standard curve using GAPDH primers. Relative quantization was determined using a Stratagene MX3005P QPCR system measuring real-time SYBR Green (PerfeCTa SuperMix, low ROX; Quanta Biosciences, Gaithersburg, MD, http://www.quantabio.com) fluorescence and calculated by the ΔΔCT method as described recently. Fold differences are calculated using the ΔΔCT in the formula 2 – ΔΔCt. Expression profiles for the mRNA transcripts are shown as fold differences in comparison to mRNA levels in hESCs.
Microarray Gene Expression Profiling.
Global gene expression analysis was performed using the human Affymetrix HG-U133 Plus 2.0 microarray platform (Santa Clara, CA, http://www.affymetrix.com) on both of the single blastomere-derived hESC lines MA01 and MA09 and the resulting RPE cells derived from each. Additionally, fetal RPE (donated by Dr. Marco Zarbin), ARPE-19 (American Type Culture Collection, Manassas, VA, http://www.atcc.org), and retinoblastoma (American Type Culture Collection) cell lines were analyzed as controls. See supporting information data for statistical analysis methods.
Western Blot Analysis.
Immunoblot analysis was carried out using standard SDS-PAGE methods using the Bio-Rad Mini-Protean and Mini-Transblot Cell (Hercules, CA, http://www.bio-rad.com). The protein bands were visualized using Western Lightning Chemiluminescence Reagent (PerkinElmer Life and Analytical Sciences, Boston, http://www.perkinelmer.com) and a Kodak 4000MM digital imaging station (Rochester, NY, http://www.kodak.com). Antibodies used were purchased from Abcam (DPPA4 and TDGF1; Cambridge, U.K., http://www.abcam.com), Chemicon (b-actin, CHX-10, Otx2, REX1, RPE65, and all horseradish peroxidase-conjugated secondary antibodies; Temecula, CA, http://www.chemicon.com), Covance (Pax6; Princeton, NJ, http://www.covance.com), Novus Biologicals (Bestrophin, CRALBP, and Pax2; Littleton, CO, http://www.novusbio.com), and Santa Cruz Biotechnology (MitF, Nanog, Oct4, PEDF, and Tyr; Santa Cruz, CA, http://www.scbt.com).
Transplantation Protocol.
Before cell transplantation, cells were thawed and washed in balance salt solution (BSS) and suspended in BSS. Three cell lines designated low, medium, and high pigment were given in different dose groups. These are summarized in Table 1. Using techniques described previously [19], a suspension of cells was delivered into the subretinal space of one eye through a small scleral incision, suspended in 2 μl of BSS medium (Invitrogen) using a fine glass pipette (internal diameter, 75-150 μm) attached by tubing to a 25-μl Hamilton syringe (Hamilton, Reno, NV, http://www.hamiltoncompany.com). The cornea was punctured to reduce intraocular pressure and to limit the efflux of cells. A sham-surgery group was treated the same way, except the carrying medium alone was injected. Pigmented dystrophic RCS rats received unilateral subretinal injections of the cell lines (n = 79 eyes) at P21; control rats received sham alone (n = 35 eyes) or were untreated (n = 29 eyes). Elovl4 mice at P28 received cells (n = 12 eyes), sham alone (n = 8 eyes), or were untreated (n = 8 eyes). Immediately after injection, the fundus was examined for retinal damage or signs of vascular distress. Any animal showing such problems was removed from the study and excluded from the final animal counts.
Spatial Visual Acuity.
Animals were tested for spatial visual acuity using an optometry testing apparatus (CerebralMechanics, Lethbridge, Canada, http://www.cerebralmechanics.com) comprising four computer monitors arranged in a square, which projected a virtual three-dimensional space of a rotating cylinder lined with a vertical sine wave grating. Unrestrained animals were placed on a platform in the center of the square, where they tracked the grating with reflexive head movements. The spatial frequency of the grating was clamped at the viewing position by recentering the "cylinder" on the animal's head. The acuity threshold was quantified by increasing the spatial frequency of the grating using a psychophysics staircase progression until the following response was lost, thus defining the acuity. Rats were tested from P60 to P240 at monthly intervals. Elovl4 mice were also tested in this apparatus at 3, 5, 7, and 11 weeks after surgery.
Luminance Threshold.
This was studied to provide a different measure of function from the spatial acuity and was achieved by recording single and multiunit activity close to the surface of the superior colliculus (SC) using glass-coated tungsten electrodes (resistance: 0.5 MΩ; bandpass 500 Hz to 5 KHz) with previously described procedures [20]. Recordings were made only in rats, selected on the basis of good and representative optomotor results: mice were not examined with this test. The brightness of a 5° spot was varied using neutral density filters (minimum steps of 0.1 log unit) over a baseline level of 5.2 log units until a response double the background activity was obtained: this was defined as the threshold level for that point on the visual field. A total of 15-20 positions were recorded from each SC. All animals were recorded at approximately P100, and some were studied again at a second time point at approximately P190. Data are expressed as a graph of percentage of SC area with a luminance threshold below defined levels and as raw results.
At the end of functional tests, all animals were killed with an overdose of sodium pentobarbital (Sigma) and perfused with phosphate-buffered saline. The eyes were removed, immersed in 2% paraformaldehyde for 1 hour, infiltrated with sucrose, embedded in optical cutting temperature, and cut into 10-μm horizontal sections on a cryostat. Four sections (50 μm apart) were collected per slide, providing five series of every fourth section collected. One was stained with cresyl violet for assessing the injection site and integrity of retinal lamination. The remaining slides were used for antibody staining, following previous protocols [21], and were examined by regular and confocal microscopy.
Safety Study.
Cells were prepared and transplanted using the same methodology described above for the RCS rat study. A minimum of six NIH III mice per group were injected with either hESCs or hESC-derived RPE from the MA09 single blastomere cell line in three time-based cohort groups (n = 36). The animals were killed by CO2 inhalation followed by exsanguination at 1, 3, and 9+ months based on cohort. Three negative control animals were also put in the study for each cohort (n = 9). Life study assessments included routine clinical assessments and body weight analysis, plus pre-sacrifice clinical chemistry (data not shown). Post mortem, eyes were removed and immersed in cold 4% paraformaldehyde, for up to 1 week. The tissue was embedded in paraffin and sectioned. Select slides were stained with hematoxylin and eosin. Slides were examined microscopically to assess retinal lamination and tumor formation.
- Top of page
- Abstract
- Acknowledgements
- Supporting Information
Differentiation and Characterization of hESC-Derived RPE
Human RPE cells were generated using a cGMP-compliant cellular manufacturing process. Three different batches of RPE were created from each blastomere-derived hESC line based on morphological assessment of pigmentation (Fig. 1), an important indicator of RPE maturation.
With the current process in place, each production run generated approximately 50 × 106 hRPE cells from a single frozen ampule of 1 × 106 hESCs. This represents the lower limit to our manufacturing capabilities, yet is still sufficient to dose approximately 500 rats or 50-100 human subjects. Additionally, our process (outlined in the Materials and Methods section) is completely suitable to available scale-up technologies such as bioreactor culture or large-scale fluid handling systems.
To characterize the developmental stages during RPE differentiation, several assays were used to identify the expression levels of genes key to each stage of development. qPCR was developed to provide a quantitative and relative measurement of the abundance of cell type-specific mRNA transcripts associated with the RPE differentiation process. A panel of genes associated with hESC pluripotency (Oct-4, Nanog, Rex-1, TDGF1, Sox2, DPPA2, and DPPA4), neuroectoderm intermediates (Pax6 and Chx10), and RPE (RPE-65, Bestrophin, CRALBP, PEDF, MitF, Otx-2, Tyr, and Pax2) was established and assayed for each by qPCR. With regard to quality control of cellular manufacturing, the marked decrease in all stem-related genes and concomitant increase in all retinal-associated genes, at a level of 10- to 100-fold, was deemed acceptable release criteria (in conjunction with the additional testing described below).
Figure 2 shows the gene expression profile of the transcripts during differentiation to mature RPE, including samples from hESCs (d0), embryoid bodies (EBs, d7), plated EBs (d14), mixed population of newly formed RPE and less differentiated cells (mixed, d28), purified early RPE (eRPE, d35), and fully matured pigmented RPE (mRPE, d56). A progressive decrease in the expression level of hESC-specific genes (Fig. 2A) was accompanied by an increase in the level of neuroectoderm and RPE-specific genes. Lightly pigmented RPE (Fig. 1) expressed 1,000-fold lower quantities of Oct-4, Nanog, Sox2, and DPPA4; <10,000-fold less TDGF1; and 50-fold less Rex-1 and DPPA2 than hESC. The cells also expressed 10- to 100-fold greater quantities of RPE65, CRALBP, PEDF, Bestrophin, Pax6, and MitF and expressed >100,000,000-fold Tyr, a downstream target of MitF/Otx2 in RPE. The finding that this cell population expresses genes such as Pax6 and CHX10 is not entirely surprising, because this stage represents an "immature" population of hRPE derived from embryonic cells, and as such, may continue to express markers associated with developing cells of the neuretina and/or neurectoderm.
The phenotypic changes that RPE undergoes during the in vitro maturation process were characterized by qPCR (Fig. 1A–1C). Figure 2A shows that RPE with a higher degree of pigmentation and polygonal cell borders (corresponding to Fig. 1C) maintains higher expression of RPE-specific genes. Notably, both pigmentation and the high level of RPE-specific gene expression are correlated with the emergence of Pax2 expression and a sharp increase in MitF, Otx2, and Tyr expression. This finding is consistent with previous reports that MitF expression, and in turn Tyr, is achieved in RPE through synergy of Pax2 and Pax6 during embryonic development [22].
Proteomic Validation of Selected Transcripts in hESC-Derived RPE
To verify that genes of interest were expressed at the protein level, all targets of the initial transcriptional profile panel were assayed by Western analysis. As an internal control, hESC-derived RPE was compared with the well-studied ARPE-19 cell line by both qPCR and Western analysis. Figure 2A shows that, although hESC-RPE expresses similar levels of RPE-specific transcripts to ARPE-19, the hESC-RPE seems to express more abundant levels of these proteins (RPE65, PEDF, Pax2, and Bestrophin). Additionally, proteins expressed by hESCs are all downregulated in the final differentiated cell product. This disappearance of stem-related proteins (by immunoblot) and concomitant emergence of retinal-associated proteins is considered necessary to determine final product identity and is essential for lot release under our current production and quality scheme.
Bioinformatic Analysis of Global Gene Expression in hESC-RPE
The biological relevance of the morphological changes observed in vitro were assessed by gene expression profiling and subsequent informatic analysis of both hESC lines, each with three different morphologies: a control and several "reference" cell lines on the human Affymetrix HG-U133 Plus 2.0 microarray platform. Figure 3 shows a principal component analysis (PCA) scatter plot, indicating the contribution to variance that the two major variables, cell type and cell line (x- and y-axis, respectively), yield on global gene expression. A linear progression was observed from the undifferentiated (hESCs) state through the three levels of RPE pigmentation. Interestingly, the depigmented RPE cells (light green markers and Fig. 1A) cluster closer to both ARPE-19 (orange marker) and fetal RPE (black marker); the latter display strikingly similar morphological characteristics to this batch of cells in vitro (data not shown). The more heavily pigmented batches of RPE cells (red and purple markers) appear to cluster farther from hESCs (green markers) and retinoblastoma cells (RB, blue marker) than any other cell type tested. Whereas the pigmented batches of hRPE from MA01 and MA09 do not overlap by PCA, they are within a similar order of magnitude to each other to that of fetal RPE and ARPE-19. Whereas the majority of genes associated with the RPE state are similar in expression between these two groups when evaluated by qPCR (data presented above and not shown), a more detailed study into which gene expression values—by microarray—are generating this result is underway, Regardless, taken together, these data suggest that the more heavily pigmented hESC-RPE cells are the most differentiated, and from a safety standpoint, the most genetically divergent from cells possessing "stemness" or expressing cancer-related genes.
Pathogen Testing and Stability of hRPE
The most important criterion to consider in release of a product for clinical application is product safety. To ensure that our hRPE product was free of contamination during the extensive culture and differentiation process, we carried out the following testing according to U.S. Food and Drug Administration and International Conference on Harmonization guidelines for applicable microbial and viral agents on our master cell bank: United States Pharmacopeia membrane filtration sterility, fluorochrome-based mycoplasma, transmission electron microscopy for viral particles, in vitro tissue culture safety testing for adventitious agents, in vivo inapparent virus detection, PCR-based reverse transcriptase detection, HIV-1, HIV-2, HBV, HCV, CMV, HTLV-1 and -2, parvovirus B19, Epstein-Barr virus, and herpesvirus 6. Additionally, the master cell bank was cytogenetically analyzed by G-banding karyotype analysis. Results confirmed that these cell lines are karyotypically stable with no presence of infectious pathogens (outsourced data not shown).
Dosing Studies in RCS Rats
The effect of different doses on efficacy was titrated using the optomotor response as an indicator. The results at P90 (70 days after transplantation) are summarized in Figure 4A. Improved rescue of spatial acuity occurred from 5,000 to 50,000, after which even doubling the dose of cells to 100,000 had no significant effect on efficacy. Best performers among the cell-injected group gave a figure of 0.536 cycles/degree (c/d) compared with 0.6 c/d in normal rats, which is approximately 90% of normal value. There was no significant difference between sham and untreated groups, which performed significantly worse than the cell-injected group (p < .01).
Luminance thresholds were also measured in a subset of rats selected by their performance on the optomotor response. The area with best sensitivity corresponded to the area of retina in which the cells were introduced, as indicated in Figure 4C–4F. For statistical comparison, however, we presented the data for this part of the study as a percentage of the area of the visual field representation from which thresholds better than designated levels were recorded without regard to position. This gives a simple indicator of overall efficacy, as well as a best response figure, dissociated from spatial considerations. It is clear that the overall sensitivity recorded at 50,000 is superior to 20,000, but as with spatial acuity, it does not change significantly between 50,000 and 75,000. For example, approximately 45% of the SC gave thresholds of 2.2 log units with 50,000 cells/eye and approximately 40% with 75,000 cells/eye. Although the mean response levels at 100,000 were better and gave more long-lasting rescue than did lower doses (Figs. 4 and 5), the high variance precluded significance.
Batch and Longevity of Effect in RCS Rats
Although slight differences in optomotor acuity were seen between the different pigment levels (Table 1), they were not significant. In contrast, there was a significant difference at all time points studied between the cell-injected groups and medium-injected and untreated controls (Fig. 5A). Over time, there was a reduction in acuity response for all the cell groups and dose levels. There was no evidence, however, that any one group showed a preferential rescue or that a higher density of cells might lead to slower deterioration over time.
To examine how luminance responses deteriorated with time, thresholds were recorded at two time points in individual rats. An example is shown in Figure 5B, 5C. As shown, the luminance thresholds show serious deterioration on the untreated side, with more than one half the area being nonresponsive at P187 compared with P98, whereas responsiveness is still sensitive on the cell-injected side, although some reduction in thresholds has occurred (0.7 log units at P98 vs. 1.0 log units at P187).
Efficacy in Elovl4 Mice
Visual acuity in normal mice tested by the same optomotor device was lower than that in rats (0.35 vs. 0.6 c/d). In untreated Elovl4 mice, the visual acuity deteriorated as photoreceptor degeneration progressed from 0.34 c/d at P28 to 0.24 c/d at P105 (Fig. 4B). Subretinal injection of hESC-RPE improved the visual acuity over controls at all time points tested. Cell-injected eyes had a figure of 0.32 ± 0.04 c/d at P63 (5 weeks after surgery) compared with 0.26 ± 0.03 c/d in sham-injected and untreated controls (Fig. 4B). Statistical analysis indicated that the difference between cell-injected and controls was significant (t test, p < .05). Because there is significant variability in the degree of photoreceptor survival across the untreated Elovl4 retina at the terminal time point, photoreceptor rescue was hard to document. Further studies will explore morphological rescue at later time points against background levels when degeneration is more advanced.
Histological Examination of RCS Rats
General Retina Structure.
Retinal sections from cell-injected, sham, untreated, and normal control rats were stained with cresyl violet and examined under light microscopy. At P90, compared with normal control (Fig. 6A), the cell-injected retina had five to six layers of photoreceptors (Fig. 6B), whereas the untreated retina had only a single layer remaining (Fig. 6C). In accordance with the functional results, the 5,000/eye doses had slightly better photoreceptor rescue (Fig. 6D) than sham-operated (primarily with localized photoreceptor rescue around injection site, data not shown), whereas the 20,000/eye produced better photoreceptor rescue (data not shown). The 50,000/eye and greater doses gave consistent photoreceptor rescue, covering a larger area of the retina (Fig. 6E, 6F) with preserved cones. At P150, cell-injected retinas still had an outer nuclear layer two to three cells deep, and the inner retina lamination was not disrupted. In contrast, both untreated and sham-operated retinas showed a typical secondary pathology, including abnormal vascular formation, RPE cells, and inner retinal neurons migrating along abnormal vessels, leading to distortion of retina lamination [19, 20]. At P240, cell-injected retinas still had an outer nuclear layer of one to two cells deep, and the inner retina still showed an orderly lamination. In contrast, advanced degeneration was very evident in control retinas: the inner nuclear layer became very irregular in thickness, ranging from one layer to multiple layers; RPE cells had migrated into the inner retina; and abnormal blood vessels were seen (Fig. 6I).
Antibody Staining.
The human specific nuclear marker, MAB1281 (Chemicon) was used to identify the donor cells. They formed a layer, one to two cells deep, and integrated into the host RPE layer (Fig. 6G, 6H), as was seen in our previous study [9]. Photoreceptor rescue continued beyond the limits of distribution of donor cells, suggesting that rescue was at least in part caused by a diffusible effect. Cone arrestin antibody showed that cone photoreceptors were preserved with disorganized segments (Fig. 6F) at P90. Donor cells were still evident up to at least P249, the longest time point studied (Fig. 6G, 6H). There was no indication of continued donor cell division (shown by the proliferating cell nuclear antigen marker; data not shown).
Safety Assessment
Studies in NIH III Mice.
The long-term risk of teratoma formation was tested in the NIH III mouse model. The NIH III mouse was chosen for its immune-deficient status; the nude mouse has three mutations rendering it devoid of T cells, NK cells, and mature T-independent B lymphocytes. However, the NIH III mouse retains eye pigmentation, which provides better visualization for subretinal transplantation surgery. The surgical technique was the same as performed in the RCS study. The study compared the hESC-RPE to undifferentiated hESCs (positive control) to determine the teratoma formation potential of the 100,000 RPE cell dose over three time points: 1, 3, and 9 months (the approximate lifespan of the animal; n = 6 per cohort). In contrast to the animals that received undifferentiated hESCs, no teratoma or tumor formation was found in any of the animals injected with the hESC-derived RPE. In addition, basic animal safety assessments were normal compared with controls.
Absence of Tumorigenic Growth in RCS Rats.
In the RCS rat transplant study, none of the 79 cell-injected retinas examined, including the longest time points, showed any evidence of uncontrolled cell proliferation. There was no evidence of teratoma and/or tumor formation.
- Top of page
- Abstract
- Acknowledgements
- Supporting Information
We showed the long-term safety and efficacy of hESC-derived RPE cells produced under manufacturing conditions applicable for use in human clinical trials. In addition to the development of assays with qualified range limits (which constituted the "identity" of the final hRPE product), extensive pathogen testing was carried out to ensure that the manufacturing procedure did not introduce any infectious diseases or adventitious agents into the final product. To confirm the functionality of these GMP-compliant cells, both dose-response and long-term efficacy were evaluated in homologous models of human retinal disease. Because of the proliferative nature of hESCs, evidence of safety under Good Laboratory Practice conditions (21CFR58) is imperative for translating hESC-derived cellular products into the clinic. The extensive characterization detailed above provides assurance of cellular identity, whereas the long-term tumorigenicity study presented here provides strong evidence that the hESC-RPE cells are safe and do not form teratomas and/or tumors during the lifetime of NIH III immune-deficient mice. After introduction to the subretinal space of RCS rats, the hESC-derived cells also survived for more than 8 months without evidence of pathological consequences. Importantly, the hESC-RPE cells also rescued visual functions in a dose-dependent fashion: with increased cell concentrations from 5,000 to 50,000, there was an improvement in functional rescue measured with both visual acuity and luminance threshold response. From 50,000 to 100,000, there was no obvious change in efficacy, which suggests that there is some tolerance in numbers of cells introduced and that twice the optimal dose is still effective. This is particularly important for phase I clinical trials, because extrapolation to humans is difficult to make. Previous rodent work [21] has shown that RPE cells quickly disperse as a single or double layer and that 20,000 cells of an immortalized RPE cell line can occupy at best approximately 20% of the retinal area (12.56 mm2). For the age-related macular degeneration retina, the inner macular is 3 mm in diameter: this would mean that a dose of approximately 40,000 cells would be needed to cover the inner macular area but that a larger cell number would most likely cover a larger area and would not be detrimental.
The significantly improved visual performance in Elovl4 mice adds to the value of the hESC-RPE as the cell choice for cell-based therapy to treat macular disease (in this case, a subset of patients with Stargardt's disease caused by mutation in the Elovl4 gene) [16, 25]. Stargardt's macular dystrophy is one of the most frequent forms of juvenile macular degeneration. Although some rescue can be achieved by growth factor delivery such as direct injection [26] or factor-releasing cells (encapsulated cells) such as ARPE19 cells transduced to produce ciliary neurotrophic factor [27, 28] or Schwann cells [29], these approaches cannot replace the other functions of RPE cells. Because hESC-RPE cells have a molecular profile more closely resembling native RPE than do ARPE-19 [18], they may be able to take on a broader range of RPE functions than ARPE-19 beyond simple factor delivery. The mechanism whereby hESC-RPE cells rescue vision is not totally clear. One possibility might be that the hESC-RPE cells replace crucial functions of the host RPE, because a previous study has shown, for example, that hESC-RPE cells are able to phagocytose latex beads in vitro [18]. However, because the location of photoreceptor rescue extends beyond the area of donor cell distribution, part of the rescue effect is likely also to be mediated by a diffusible trophic factor effect.
In this study, the deterioration of function with time may be related to the inadequacy of the immunosuppressive protocol used in long-term discordant (human-to-rodent) xenografts. Previous studies have shown that that donor cells decline even under triple immunosuppression [30]. Another confounding factor is that, in RCS rats, leaky blood vessels appear during the course of degeneration [23, 24], which may abrogate the immune-privileged status of the eye and compromise cyclosporine efficacy. It is not known whether some form of immunosuppression (possibly local and/or topical administration using eyedrops) might be needed when the present cells are used in a clinical setting (under those circumstances, they would be close allogeneic rather than the xenogeneic grafts of this study). An alternative is that transplants might need to be repeated to obtain a sustained therapeutic benefit. It is very encouraging, however, that even at P240 (i.e., 220 days after transplantation), some donor cells survive, photoreceptors are rescued, and a level of visual function is preserved.
- Top of page
- Abstract
- Acknowledgements
- Supporting Information
In summary, we showed the long-term safety of hESC-derived RPE cells in immune-deficient animals, as well as their long-term function in two different animal models of disease using GMP conditions suitable for clinical trials. This represents an important step toward the possibility of using hESC-derived cells to slow the progress of specific macular degenerative diseases.
- Top of page
- Abstract
- Acknowledgements
- Supporting Information
We thank Benjamin Cottam and Jie Duan for technical support.
- Top of page
- Abstract
- Acknowledgements
- Supporting Information
Raymond Lund and Peter Francis acted as consultants for and were employed by Advanced Cell Technology.
- Top of page
- Abstract
- Acknowledgements
- Supporting Information
- 1Embryonic stem cell lines derived from human blastocysts. Science 1998; 282: 1145–1147., , et al.
- 2Retinal pigment epithelium and photoreceptor transplantation frontiers. In: RyanSJ, ed. Retina. 4th ed. Philadelphia: Elsevier Mosby, 2006: 2597–2613., , , et al.
- 3Prevalence of age-related macular degeneration in the United States. Arch Ophthalmol 2004; 122: 564–572., , et al.
- 4An international classification and grading system for age-related maculopathy and age-related macular degeneration. The International Arm Epidemiological Study Group. Surv Ophthalmol 1995; 39: 367–374., , et al.
- 5The epidemiology of age-related macular degeneration. Am J Ophthalmol 2004; 137: 486–495., , et al.
- 6The genetics of age-related macular degeneration: a review of progress to date. Surv Ophthalmol 2006; 51: 316–363., , et al.
- 7Mutations in MERTK, the human orthologue of the RCS rat retinal dystrophy gene, cause retinitis pigmentosa. Nat Genet 2000; 26: 270–271., , et al.
- 8Subretinal transplantation of genetically modified human cell lines attenuates loss of visual function in dystrophic rats. Proc Natl Acad Sci U S A 2001; 98: 9942–9947., , et al.
- 9Human embryonic stem cell-derived cells rescue visual function in dystrophic RCS rats. Cloning Stem Cells 2006; 8: 189–199., , et al.
- 10Morphological changes in the Royal College of Surgeons rat retina during photoreceptor degeneration and after cell-based therapy. J Comp Neurol 2005; 491: 400–417., , .
- 11Autologous iris pigment epithelial cell transplantation in monkey subretinal region. Curr Eye Res 2000; 20: 268–275., , et al.
- 12Long-term preservation of cortically dependent visual function in RCS rats by transplantation. Nat Neurosci 2002; 5: 53–56., , et al.
- 13Retinal degeneration and RPE transplantation in Rpe65(-/-) mice. Invest Ophthalmol Vis Sci 2002; 43: 3307–3311., , .
- 14Mutation of the receptor tyrosine kinase gene Mertk in the retinal dystrophic RCS rat. Hum Mol Genet 2000; 9: 645–651., , et al.
- 15Inherited retinal dystrophy in the rat. J Cell Biol 1962; 14: 73–107., .
- 16A 5-bp deletion in ELOVL4 is associated with two related forms of autosomal dominant macular dystrophy. Nat Genet 2001; 27: 89–93., , et al.
- 17Impaired RPE survival on aged submacular human Bruch's membrane. Exp Eye Res 2005; 80: 235–248., , et al.
- 18Derivation and comparative assessment of retinal pigment epithelium from human embryonic stem cells using transcriptomics. Cloning Stem Cells 2004; 6: 217–245., , et al.
- 19Protection of visual functions by human neural progenitors in a rat model of retinal disease. PLoS ONE 2007; 2: e338., , et al.
- 20Time course of deterioration of rod and cone function in RCS rat and the effects of subretinal cell grafting: a light- and dark-adaptation study. Vision Res 2005; 45: 343–354., ,
- 21Grafting of ARPE-19 and Schwann Cells to the Subretinal Space in RCS Rats. Invest Ophthalmol Vis Sci 2005; 46: 2552–2560., , et al.
- 22Retinal pigmented epithelium determination requires the redundant activities of Pax2 and Pax6. Development 2003; 130: 2903–2915., , et al.
- 23Ganglion cell loss in RCS rat retina: a result of compression of axons by contracting intraretinal vessels linked to the pigment epithelium. J Comp Neurol 1998; 392: 58–77., , et al.
- 24Evolving neurovascular relationships in the RCS rat with age. Curr Eye Res 2003; 27: 183–196., , et al.
- 25Evaluation of the ELOVL4 gene in patients with age-related macular degeneration. Ophthalmic Genet 2001; 22: 233–239., , et al.
- 26Multiple growth factors, cytokines, and neurotrophins rescue photoreceptors from the damaging effects of constant light. Proc Natl Acad Sci U S A 1992; 89: 11249–11253., , et al.
- 27Transplantation of Schwann cell line clones secreting GDNF or BDNF into the retinas of dystrophic Royal College of Surgeons rats. Invest Ophthalmol Vis Sci 2004; 45: 267–274., , et al.
- 28Intraocular gene transfer of ciliary neurotrophic factor prevents death and increases responsiveness of rod photoreceptors in the retinal degeneration slow mouse. J Neurosci 1998; 18: 9282–9293., , et al.
- 29Schwann cell grafting into the retina of the dystrophic RCS rat limits functional deterioration. Royal College of Surgeons. Invest Ophthalmol Vis Sci 2000; 41: 518–528., , et al.
- 30Triple immune suppression increases short-term survival of porcine fetal retinal pigment epithelium xenografts. Invest Ophthalmol Vis Sci 2003; 44: 4044–4053., , et al.
Supporting Information
- Top of page
- Abstract
- Acknowledgements
- Supporting Information
Additional supporting information available online.
|Supporting Information Table 1
Please note: Wiley Blackwell is not responsible for the content or functionality of any supporting information supplied by the authors. Any queries (other than missing content) should be directed to the corresponding author for the article.


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Wisconsin River Pro Rodeo
Bareback riding, steer wrestling, tie down roping, saddle bronc riding, team roping, women's barrel racing and America's favorite...bull riding. Dances following the Fri & Sat performances.


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Goat, sheep producers gather to share ideas
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An educational "roundup" for sheep and goat producers will be held Aug. 23 and 24 in Raleigh at the Wake Commons Conference Center and Oakwood Farm. Goat and sheep producers in the Southeast will gather to share and learn more about the latest issues related to the dairy and meat goat and sheep industry.
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Hart-Fox Battery Charges I-Cubs
Two things of note from the Iowa Cubs' road win in Salt Lake City yesterday:
1. Jake Fox played catcher after a game at 3B earlier on the trip. Is his versatility on display for potential trade suitors?
2. Kevin Hart threw three innings of perfect relief highlighted by five strikeouts in easily his most encouraging outing of the season.
In more routine news, Fox also slammed his 17th homer of the year, drove in four more runs and was named the PCL Player of the Week for last week. Ho-hum...MW
Re: Hart-Fox Battery Charges I-Cubs
If he can channel Lloyd McClendon in his "ability" to catch for 3-4 batters when a team is losing and a double-switch is needed, and to stand once every three weeks at 3B when David Wright needs a day off or there's a blowout, he would be a good fit for the Delgado-less Mets. But do the Mets have anybody we'd want back in return?
Re: Hart-Fox Battery Charges I-Cubs
Even the Mets would have to have someone better than Neal Cotts...
Re: Hart-Fox Battery Charges I-Cubs
heck ... we might have better than Neal Cotts (I wouldn't be against Casey Lambert getting a look ... I still think his future is in the pen, and I think he has pen ready stuff right now).
As for Fox, I think an AL team is probably the best fit, for obvious reasons. He can DH, play some first, occasionally pinch in the corner OF, act as the emergency catcher, and every blue moon, play 3rd.
The Mets plan seems to be, based on an article that came out last week (forget where) to go with what they have and see how far along FMart is come June. If he's ready, Dan Murphy could pinch at first. If not, maybe they'd investigate the trade market, but this being New York, and Omar, I'm not sure they'd go after a minor leaguer unless Fox got some big league AB's.
Re: Hart-Fox Battery Charges I-Cubs
Why the big hurry to trade Jake Fox? This team has lost its best hitter for an extended period of time and are trying to patch it up with three guys who weigh a total of 450 pounds. Why not bring Fox up, cut Patton loose, and put Fox out there for a couple 7 inning games a week at 1B and 3B? Is Patton that good that the Cubs can afford to carry a spare part just to have a reliever next season? I find it almost embarrassing that Jake Fox still sits at Iowa with those numbers while we tread water with an underperforming infield (minus the shortstop). At worst we'd have a pretty good lefty/righty pinch hit tandem with Hoffpauir and Fox that might negate a bullpen move or two.
Re: Hart-Fox Battery Charges I-Cubs
The Cubs aren't treading water, and a good month of hitting in AAA doesn't make you Aramis Ramirez.
Re: Hart-Fox Battery Charges I-Cubs
No, Fox is not the hitter that Ramirez is, but it's a possible asset to use in place of a known non-asset (Patton). Fox > Miles? Fox > Fontenot? Fox > Scales? Why not find out? Strike while the iron is hot.
Maybe it's early to call this stretch "treading water", we'll see what they do against teams a notch above Houston and San Diego before that terms settles in. Piniella seems to think the term fits.
Re: Hart-Fox Battery Charges I-Cubs
just to be clear, I'm in no hurry to trade Fox, was just responding to the post.
Re: Hart-Fox Battery Charges I-Cubs
While it is much, much too early to evaluate the DeRosa trade, I was curious to see how the three pitchers the Cubs got back were doing so far.
John Gaub is a 6'2" 200lb lefty, pitching in relief for AA Tennessee. In 11.2 IP he has 20 Ks, a 1.54 ERA, and a .154 average against.
Christopher Archer is a 6'3" 180lb righty, starting for A Peoria. In 25 IP he has a 1.80 ERA with 30 Ks and a .181 average against.
Jeff Stevens is 6'2" 205lb righy, pitching in relief for AAA Iowa. In 16 IP he has a 2.16 ERA with 13 Ks and a .143 average against.
Re: Hart-Fox Battery Charges I-Cubs
It was never as bad a deal as people made it out to be, although it was certainly disappointing when it happened as DeRosa was a good team guy. Haven't heard velocity reports on Gaub - up until his junior year, he was in the low-mid 90's, but then got hurt, iirc, and since then, was only low 90's. Off the top, a plus slider. Very solid LOOGY prospect.
Stevens is sort of in that Kevin Hart mold for me - capable of filling some role in the bigs, but not sure how key of a role it would be.
Archer was the prize of that trade, with the plus fast/plus curve. Love how he's done so far, and love his starting potential. Definitely has work to do with, um, the change, along with command, but Archer and Carpenter have been excellent down in Peoria (wouldn't mind seeing Carpenter get a look in Daytona soon - Archer has a ways to go and is young enough to keep him in Peoria for the whole year, imo).
"May Top 10"
Someone asked AzPhil's thoughts in a post yesterday, and I just wanted to ask it again. I was messing around with a list, just my own fan's take, for fun. As with all lists, more for discussion value. But I was messing around and in a quick run through, a possible top 10 I came up with is below.
1. Josh Vitters, A-. Power has been showing up.
2. Jeff Samardzija, B+. Easy to forget his 3rd full pro season. Control in AAA has been sharpened but more work. I hated the call up..
3. Jay Jackson, B. Slow start, but has gotten it going. I was one of those that had him highly ranked entering this year (had him 3rd, same order as above) and don't see a reason to change it.
4. Dan McDaniel, B. I was huge on him last year, but this has been better than I thought. Yes, I'll take his value ahead of Cashner. Cashner's got better stuff, but I don't think it's by enough to offset McDaniel's starting potential.
5. Andrew Cashner, B. I'm still not huge on Cashner due to the control.
6. Ryan Flaherty, B. Hasn't had the huge season, but I'm not worried as the discipline is there.
7. Tony Thomas, B. Very disappointed with the SB%. That's just eh. Other than that (and defensive concerns), excellent season that's sort of gone under the radar nationally, IMO.
8. Starlin Castro, B-. If he simply stayed afloat after the huge jump, I would've been happy. The fact that he has a 7 game hit streak and has gotten his offense going is just pudding.
9. Marcos Mateo, B-. Control has been better last couple of times out.
10. Dae-Eun Rhee, B-. Wary of ranking him higher until he gets back out there, but that's me being conservative.
Anyhow was wondering what folks thought on that quick run through. I think the system is improving, and I think there's hope, particularly lower down. Had a bunch of names that I pondered in the last few top 10 slots (Jose Ascanio, Hak-ju Lee, Chris Carpenter, Tyler Colvin, Jake Fox, Casey Coleman, Steve Clevenger, Welington Castillo).
Re: "May Top 10"
Unfortunately I fear that Vitters is really over-rated at this point.
His walk rate has been Ryan Harvey-esque up to this point.
Even Brian Dopirak put up big Midwest league numbers in his 2nd full pro season.
As Corey Patterson,Felix Pie,Harvey,Dopirak have all shown us over the years. If you can't control the strike zone, you can't be a consistent offensive force at the major league level.
Re: "May Top 10"
the low walk rate concerns me, but almost all the scouting reports suggest he controls the zone real well and that it's a case of the pitching at the lower levels basically not being challenging enough. At some point, one wonders if his plate coverage can really compensate, or if he can learn to take more walks as he mvoes up, but I'm not too worried yet. That's why I think, within a month's time, I'd really like to see vitters at Daytona. Keedy doesn't interest me all that much, tbh, and Rosa's future may be at first anyways (big fan of Rosa, but he had an injury and those swing problems this year so he has struggled). I was more concerned about Vitters' power, to be honest.
Only time will tell.
Re: "May Top 10"
I really think for guys at that level it's more about what they are capable of rather than what they actually do and Vitters is very nicely built and has nice potential. Now it's up to Cubs coaching to teach him how to be a real ball player.
Re: "May Top 10"
Vitters's low BB rate bothers me a little, but less so since his K rate is pretty low. The dude seems to be hitting everything he's swinging at.
Ryan Harvey, for example, swung and missed a lot. The numbers seem to say that Vitters is swinging a lot, but not missing anything.
Re: "May Top 10"
Ryan Searle should at least be in the conversation.
Re: "May Top 10"
yeah, he's been a nice surprise. after the ba comments about him, i wasn't sure what to expect.
Re: Hart-Fox Battery Charges I-Cubs
Completely unrelated question. I used to watch the Daily Rewind on MLB.com all the time. It would be a 25 minute video with 2-3 minutes of highlights per game, that would recap every game from the day before. Basically, Sportscenter without all of the other crap I don't really care that much about. It's been a couple of summers since I watched it regularly, but I can't find it now. Does anyone know if MLB.com did away with it?
Re: Hart-Fox Battery Charges I-Cubs
Dopirak this year in AA:
Did they really fix his swing?
Re: Hart-Fox Battery Charges I-Cubs
May 9,
MANCHESTER – Brian Dopirak clobbered the Defenders in a career night, leading the Fisher Cats to an 18-6 slugfest win at Merchantsauto.com Stadium last night.
The hulking slugger homered twice, doubled twice, tripled and finished with eight RBIs in the 5-for-6 outing. All eyes were on Dopirak in the eighth inning to see if he'd single for the first-ever cycle in franchise history.
He doubled, leaving him with a mere 15 total bases, easily the best single-game hitting performance in Fisher Cats history.
Re: Hart-Fox Battery Charges I-Cubs
Re: Hart-Fox Battery Charges I-Cubs
If you get the new MLB Network on your cable/satellite line up, they have a show like that, all extended highlights of each game, late at night and rebroadcast the following day over and over until 11 am with no analysis. It's called Quick Pitch.
Re: Hart-Fox Battery Charges I-Cubs
Good to know, but it won't help my particular situation. I no longer have cable, basically because they took away WGN. I kept it mainly just to watch some of the Cubs games on real TV rather than MLB.TV. Now I am just watching them all online and canceled my cable. But I did enjoy watching sportscenter during breakfast to see what else was going on in baseball. Now it seems my only option is to watch each highlight individually on MLB or watch each games very short highlights (with commercials to start) on ESPN, which is simply not doable. I want something I can play and leave on for a while I am getting ready in the morning.
Re: Hart-Fox Battery Charges I-Cubs
What's the problem? The MLB highlights play automatically once you start with one.
Re: MLB.com Daily Rewind
looks like it's gone...closest option would be to bookmark this page:
and then click on each 2 minute highlight package for each game in the morning...hopefully they continue to order it by the latest posted.
reasons why the Sun Times cannot die soon enuf
content like this from BY TONI GINNETTI [email protected] today:
(Corey) "Patterson might not be a star, but he is a major-leaguer, now playing for the Washington Nationals"
Re: reasons why the Sun Times cannot die soon enuf
He is not a major leaguer and he is not playing for the Nats. He hit 7th and played LF for the Syracuse Chiefs last night.
195/263/264 in 87 AB's.
Re: reasons why the Sun Times cannot die soon enuf
He's a five tools player: hammer, wrench, screwdriver, blowtorch and sickle.
Re: Hart-Fox Battery Charges I-Cubs
Seriously-- is Jake Fox going for the best AAA season of all-time?
.429/.513/.968 (1.462)
I realize that there's not really any place to put him-- the only positions he can play (poorly) are already blocked by people making a lot of money (LF Soriano, RF Bradley, 1B Lee).
But those offensive numbers are SICK.
I suppose it might make sense to try him out at 3B, which is what Iowa did a couple nights ago (Fox caught last night and had 2 PBs), but I guess the Cubs aren't feeling they need him there right now (which I understand because, well, the Cubs are playing fairly well at the moment).
I wonder would be see Fox only if Lee had/has to go on the DL, or if Bradley or Soriano get injured (not that I'm advocating more injuries, just wondering).
Mike Wellman-- are you able to watch Fox in the field? What's he like out there?
My guess is that Fox is an insurance policy right now and if he continues to rake, he might be worth a missing part or two around June/July.
Re: Hart-Fox Battery Charges I-Cubs
ROMERO, welcome to the site - lol!
Thanks Mike for the "Heart-full" report. Unfortunately, to this point, as soon as Kevin gets to a major-league ballpark, and makes it to the mound, he forgets his control - and that hitters in the MLB sit on fastballs with no movement.
He does not suffer from lack of talent. With Hart, its all "upstairs". If the team had a sports psychologist, like the Red Sox and a couple other teams, it would do him some good, imo.
I want to nominate "toonsterwu" for the most weird "handle" of the month!
Re: Hart-Fox Battery Charges I-Cubs
Yeah, it's weird.
It's Say-"It's Weird"-In-A-Nic-Cage-Voice Day in my office.
Re: Hart-Fox Battery Charges I-Cubs
Fox at third for a month? Count me in.
...just to see if he can outhit Fontenot. It would put Fontenot in a role he's more useful at (backup IF) and gives Miles even less playing time which I'm all for. Unfortunately it would probably end the warm and fuzzy feelings of Bobby Scales being up here.
Might be an ugly glove but I see it as an improvement for the offense until ARam returns. He can't be too much worse than Ryan Braun was (or this years Matt Gamel experience at 3B) and it just might upgrade his trade value (to an AL team) as well as helping as righty DH in the interleague games coming soon.
Re: Hart-Fox Battery Charges I-Cubs
I endorse the Mat Gamel comparisons to Jake Fox. I'm a big believer in pitching & defense over offense, but at this point you almost have to give him a chance and see what he can do. He's absolutely destroying the PCL.
And PS --
Bobby Scales is here to stay. ;)
Re: Hart-Fox Battery Charges I-Cubs
I hate to burst the Bobby Scales bubble, but he can't field 3b, and if teams consistently start throwing him breaking balls he's already proven he can't lay off them and can't hit their either. So he's a mediocre 2b, which Hendry seems to collect by the bunch.
Re: Hart-Fox Battery Charges I-Cubs
Sounds to me like you relish it.
Re: Hart-Fox Battery Charges I-Cubs
No, I was happy for the guy getting his shot, and his hitting has been a pleasant surprise, but opposing pitchers seem to be having brain cramps when they pitch to him. His first week he showed repeatedly he can't lay off a breaking ball and can't hit them, looking terrible taking weak half swings. But they keep throwing him fastballs and he's been doing a good job hitting those.
If I were pitching to him and could get my breaking balls over the plate for strikes, I would never throw him a fastball. And those scouting reports will get around.
Re: Hart-Fox Battery Charges I-Cubs
I just don't think there's a bubble to burst. He's performed well for a 54 year old backup who's been called upon to play regularly due to an injury. Does anyone really think he's the answer to anything?
Re: Hart-Fox Battery Charges I-Cubs
He's definitely the answer to the question: what does regression to the mean look like?
Re: Hart-Fox Battery Charges I-Cubs
What do Bobby Scales and Felix Pie have in common?
They both strike out in a quarter of their at bats.
Re: Hart-Fox Battery Charges I-Cubs
They're both 5-tool former top prospects who were destined to be superstars?
Oh, wait, that's just Pie. Scales is a backup.
Re: Hart-Fox Battery Charges I-Cubs
Don't worry, TRN is just looking for a way to reconcile Scale's success so far b/c he looked at Scale's minor league stats and didn't see it coming.
I did.
Re: Hart-Fox Battery Charges I-Cubs
Yes, Ryno, the amazing super scout who knows more than all the major league GM's.
He has a nice stroke, you're right about that. He also has very poor bat speed. In the majors he's just a guess hitter, and as we will see over the next few weeks guess hitters with JuCo level bat speed won't be very successful in the big leagues.
A new Inbox will be posted later Monday -- I know the hirejimessian people will be happy to read that. I get a lot of repeat mailbag questions so will try to address some here: The Cubs abandoned the high tech batting helmets they used last year because they were always breaking. They also have dumped the red-billed caps because it's easier to deal with one. There's nothing I can do to get more games on WGN or to realign the divisions. And Matt P. of Tampa will be happy to know the equipment guys are working on straightening out the crooked "C" on some of the batting helmets.
glad Muskat knows her fans...
Re: Heh
Let's see, who can we hate today? There's always Muskat.
She did a nice job the other day when she interviewed the Japanese coach who helped Fukudome in the offseason.
The walking Pulitzers in the dailies missed that story. I guess they've been distracted by other things, like the suspicions surrounding Theriot.
Re: Hart-Fox Battery Charges I-Cubs
Regarding the Bobby Scales talk and breaking balls and such...
He's outhitting Fontenot big time.
End of story.
Re: Hart-Fox Battery Charges I-Cubs
The Real Neal:
What do Bobby Scales and Felix Pie have in common?
They both strike out in a quarter of their at bats.
I'll go out on a limb and wager $5 virtual dollars that Scales will outproduce Pie at the big league level this year in terms of OPS.
And it's no mistake or mystery. Scales knows how to hit and Pie does not. Sure Pie might have more ability or raw talent, but when you're swiping the bat around like a retard that doesn't really matter much.
Re: Hart-Fox Battery Charges I-Cubs
there's not a GM out there who'd take scales over fontenot, though.
scales don't project for much. he has todd-walker-like defense (the older version) and is pretty much a slap hitter who's gonna swing a ton.
he's got 4xbase hits out of his 8 hits and that's a hell of a lucky start. it's a bit misleading, though.
Re: Hart-Fox Battery Charges I-Cubs
I hope Scales becomes our Pujols, but I suspect he's only a darker Dan Rohn.
Speaking of Pujols, Garry Meier on WGN Radio today was having people come up with their best Pujols nicknames. One of the few he said he could read on the air was Albert "Cleans My" Pujols.
Carrie Muscat: Can Harden work deeper?
Carrie's mailbox article this week has an interesting name. Hmmm, Harden deeper, better leave that one alone.
Tonight's lineups
Sori, Riot, Fukudome, Lee, Bradley, Fontenot, Soto, Miles, Lilly vs. Ryan, Rasmus, Pujols, Molina, Duncan, Stavinoha, Barden, Pineiro, Thurston
Re: Tonight's lineups
I'm feelin it. I think Aaron Miles goes DEEP tonight. Deeper than Harden.
Re: Hart-Fox Battery Charges I-Cubs
4th straight game with a HR for Vitters.
Re: Hart-Fox Battery Charges I-Cubs
Many are saying that McGuire's tuteledge of Chris Duncan is in part responsible for the player's resurgence this year. The dude is raking big time.
So far, he's not come up with a positive test result, either.
Re: Hart-Fox Battery Charges I-Cubs
mcguire on a ballfield outside his private residence?
i'll believe it when i see it.
Re: Hart-Fox Battery Charges I-Cubs
Beliefe it. Mc Guire is a private batting instructor for two Cards, and two A's. I think Giambi is one of them as well.
Re: Hart-Fox Battery Charges I-Cubs
oh, i'm just playing around.
still, the guy hates going anywhere in public. i know he's still doing offseason drills/camps/whatever with players on his property, though.
Re: Hart-Fox Battery Charges I-Cubs
Who the fuck is McGuire?
Re: Hart-Fox Battery Charges I-Cubs
the only thing that could have made tonight's game worse would be a crapload of errors.
lilly didn't pitch that bad. his curve was pretty damn nice tonight.
Re: Hart-Fox Battery Charges I-Cubs
Yes that was embarassingly feeble, maybe Piniero had the night of his life but the guy doesn't usually have dominate stuff and usually bad control on top of that.
I hope this isn't the kind of game that sends the offense into a funk. Just have to move on tomorrow and hope Carpenter isn't 'old Carpenter' tomorrow.
Re: Hart-Fox Battery Charges I-Cubs
Given the crappy Cub performance vs Joel Fuggin Piniero, The Blackhawks losing in OT, 3-2...
Kerry Wood caps a bad night...
enters bottom 9th vs KC with a 5-2 lead.
Guillen GO 4-3
Jacobs HR
Teahen HR
Olivo BB
DeJesus Triple, RBI...Blown save 5-5
Bloomquist SF. Loss 6-5.
Re: Hart-Fox Battery Charges I-Cubs
Maybe the Indians will get so sick of him that they'll trade him back to us for Aaron Miles and Neal Cotts, and eat about 75% of his salary.
Ahh, wishful thinking.
Re: Hart-Fox Battery Charges I-Cubs
Hey, at least the Farns got the W.
Re: Hart-Fox Battery Charges I-Cubs
i think larussa took pinearo off the scrap heat there should be no way he shuts us out oh wait a minute he did.
scales,fontenot,miles two of them have to start each night
cubs definately need a better option for third,i would guess fontenots struggles are worse, because he is playing third.
i would have iowa start fox at third base a few games in a row,his numbers are much better then other three mentioned.
which pitcher goes on friday?
Re: Hart-Fox Battery Charges I-Cubs
If we swing at the first pitch all the way through the lineup all game long, it almost doesn't matter who's pitching. No way to really know if Piniero had a great night, the way the Cubs hitters handed it to him. Jeez
Re: Hart-Fox Battery Charges I-Cubs
Cubs hitters musta had a bachelor party or some shit to go to. Jeez. It was like Dusty was managing.
Re: Hart-Fox Battery Charges I-Cubs
yeah i just saw where he threw 92 pitches in nine innings.
maybe we need to dust off freel and let him start a few in a row at third.
Re: Hart-Fox Battery Charges I-Cubs
where am i going to find fontenots batting splits for playing third vs second ?
Re: Hart-Fox Battery Charges I-Cubs
Only a marginal difference:
213/340/685 at 2nd
203/424/707 at 3rd
4 out of 5 of his homers at 3rd.
Re: Hart-Fox Battery Charges I-Cubs
Re: Hart-Fox Battery Charges I-Cubs
New RSS feed headline:
Piniella: 'We need Fontenot to hit' (AP)
Um, just take him out of the lineup and replace him with Scales. Who plays baseball like it runs his groin?
Why the fuck is this so hard for managers?
And I hate it when they say, "He hasn't been there".
I was there dude. Played wide out and got my face creamed into the soil. So don't ever say that again.
I mean, I love Lou but I don't like him talking like he's a fucking God cuz he was there and I wasn't.
Re: Hart-Fox Battery Charges I-Cubs
Okay then
Re: Hart-Fox Battery Charges I-Cubs
Need Fox Now
this is ridiculous that we have Miles, Scales, Freel, Johnson, Hill on the bench - that has to be the lightest hitting bench of all time. Hoff is the only respectable bat on the bench.
Fox was a catcher all through college and his first seasons in the minors - I can't believe he can't be serviceable there given the bench is calling every pitch. Give Soto/Lee some days off and get Hoff and Fox some at bats!
Recent comments
I don't hate Coghlan as a PHer but he's leading off today which is as bad as Austin Jackson doing so last year. He doesn't even have the speed.
TLS has options. He will be up again in Sept, and maybe earlier.
Very odd -- TLS .308/.419 in July (in 26 AB). Strange move for a team struggling to score runs. Hard to imagine Coghlan doing better than that. Maybe they like Coghlan better as a PH?
Sorry to nitpick -- but the first game wasn't a save.
Wow La Stella sent to Iowa and Coghlan brought up. I'm not a huge Stella fan but he has been hitting lately and is to me better than Coghlan. Neither are good defensively but I guess Coghlan can play the outfield allowing Bryant to stay at 3rd more where he's better.
Not 2nd to last but Cubs are definitely bottom of the pack in the 2nd half in oWAR and wRC+. #1 in dWAR though which is certainly a good sign for the playoffs.
I was going to say that you wouldn't want Heyward to achieve Soriano's OBP--.317 with the Cubs--but now I see that he's already done that. Pitchers have lost their fear of him.
Oh, well, at least Chapman seems to be as good as advertised.
Thanks to E-man and Quiet Man for the link on Chapman in a previous post. It's funny because I never really noticed Chapman's delivery until I saw him doing it in a blue uniform. Two saves in a Cubs uniform already. I hope they don't need him today, despite that smooth delivery.
Cubs are middle of the pack in average, 5th in OPS, 2nd in walks, 4th in runs scored in MLB. Their runs scored ranks 17th in the MLB in July.
Boston, leading everything, continues to rake in July.
Lester has given up one run or fewer in 11 starts this year. So I'd answer yes. Three of his last four starts were duds, yes. Not worried about him at all. Arrieta is concerning, to be sure. I look it at as, what he was doing was historic, so the regression was bound to happen. He showed in his start against the Mets that he's still capable, and I'd bet on him regaining his form.
marlins getting a.cashner and c.rea from SD
This is not an attack...But In what stat are the Cubs second from last in MLB, in hitting? I don't see this.
How odd the stats:
Cubs 1st in pitching in MLB
2nd to last in hitting.
In May they were 1st in both.
trevor clifton (high-A) could be interesting in a couple seasons...could even become a high/middle rotation guy. he's got a lot of pluses in his pitching except control.
eric leal's (high-A) progression through the minors should be worth watching even if only projects to be an mid/end-rotation starter.
we also gotta keep a long-distance eye on guys like jose paulino (ss-A) and preston morrison (A).
As Johnny Bach used to say when it was time for the MJ Bulls to crank up the defensive pressure: "Release the Dobermans!", and Jordan, Pippen, Rodman and Harper would just suffocate the other team.
I'd love to add Carl Jr. to that group - he's got the stuff.
Historically, the Moneyball guys have been great at identifying productive hitters. Not so much with pitchers.


text: Kids' toys run the gamut from good-looking to garish, educational to, well, questionably useful in a young one's life. But you can never go wrong with a classic set of blocks, and millergoodman's graphic geometric prints make these basics even more fun. Your tot can turn these bright rubberwood pieces over—and over and over—in the box to create a new design (the Shapemaker Flickr pool has some finished masterpieces), then rearrange them again for a brand new look.


text:
Talk:Soccer in New South Wales
|WikiProject Australia / Association football
|(Rated Start-class)
|WikiProject Football
|(Rated Start-class, Low-importance)
Northern NSW Football[edit]
This article should not conflate NNSW with NSW for football purposes, as for all intents and purposes they are separate states of equal standing. The article gives the impression that Football NSW is the governing body for the entire state and the NSWPL is the premier competition.
Article should be split into two separate articles for the two different state federations. 22.214.171.124 (talk) 10:10, 29 November 2009 (UTC)
Image copyright problem with Image:Football NSW logo.jpg[edit]
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Stay Connected Anywhere
TOPEKA, Kan. (WIBW)-- Topekans settled indoors for a musical salute to our nation.
With that great rejoicing of freedom, Grace Cathedral kicked off their Independence Day Celebration and Service with music and prayer.
Boy Scout Troop 18 from Tecumseh Posted The Colors for todays event. A special community choir got attendees in a patriotic spirit.


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Lair Clean Up
|Collect 150 B.I.N.K.E.Y. in the Lair as Megamind
The Lair is your home base or the hub that you visit between levels. At the very beginning of the game, just run around and collect all the B.I.N.K.E.Y. (blue glowing things) you see and this will unlock.
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Ross Harries presents Scrum V with highlights of all the weekend's Pro 12 action and a look ahead to Wales's next Six Nations game against Italy.
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The diversity is good in a bad way, and by that I mean that the LGBT groups are rather hostile to any masculine appearing male or feminine appearing female. Differences of sexuality are very well accepted, but there is some racial hostility among some of the white students from upstate New York and nonwhites from everywhere else. Racial jokes are common. Prepare to be verbally attacked if you're heterosexual and you and your partner breath in the wrong direction.
Everyone is accepting of different views/backgrounds.
Everyone is super nice and non-judgmental.
There are people from all over the world here. It is exciting to meet people from other places and learn about their background and different customs. Everyone is accepting of each other here; it does not matter whether one is black, white, Jewish, Muslim, or Catholic. The diversity is what makes Wells so special.
Considering that Wells is such a small school, there aren't many different diverse groups, but we work with what we have, and I consider this school to be the a great school
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Rumours of a replacement for the Toyota Supra have been circulating for some time now but the Japanese manufacturer has announced there'll be 'some form of performance concept at Detroit.'
The idea of a successor to the Supra was first floated with Toyota FT-HS at the Detroit Motor Show back in 2007, although the car never made it into production.
We are yet to see any images of Toyota's latest concept but the brands California-based CLATY design studio has "had a hand in its design", an insider from the manufacturer revealed. Toyota has also been tight-lipped about the powertrain but Internet rumours suggest a high-performance hybrid system is likely.
Earlier in the year Toyota signed an agreement with BMW, which would see the two manufacturers team up and develop new sports car architecture and hybrid technology. This rear-wheel drive coupe is expected to be the first sign of things to come from the tie up.


text:
Chilled Cantaloupe Soup
- 1 large chilled cantaloupe, seeded and cut into 1-inch chunks
- 1/3 cup orange juice
- 4 lime wedges
- Place the cantaloupe in a blender, add the orange juice, and puree until well blended.
- Divide the soup evenly among 4 bowls. Squeeze the limes over the soup. Serve chilled.


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Underwater 'Star Trek' and 11 other great sci-fi shows on Netflix
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Abrams' Fringe and Joss Whedon's Firefly are both on the streaming service in full, so you can universe jump and shiny shindig ...
J.J. Abrams got his favorite Easter Egg into 'Star Wars'
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Walter and William Bell worked on programs at a firm called Kelvin Genetics in "Fringe ...
20 years ago today: 'Jumanji' opened in theaters
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Remembering the Jewishness of Leonard Nimoy and the Vulcan salute
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And don't forget 'Fringe' in your Leonard Nimoy tributes
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And also don't forget "Fringe," which offered Nimoy one of his last major roles, playing the brilliant William Bell, colleague to John Noble ...
Interview: 'Sleepy Hollow' star John Noble on Season 2 and what drives War, Henry and Jeremy
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what a great concept for a maternity portrait!
Fantastic picture! Love how real it is.
This photo has a special kind of humour and ligthness, and I really love pics of pregnant women. The model are very cute. Congrats.
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On Second Thought is a one-hour, daily news talk show that airs at 9 a.m. ET weekdays.
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The Georgia Department of Transportation said Thursday it will work with federal authorities to reconsider a proposed route for Highway 411. The freeway would connect Rome to Interstate 75. Governor Nathan Deal said he received a letter from the National Highway Administration asking for the state to come up with alternatives.
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Thu., October 4, 2012 2:18am
Mine Prompts 411 Connector Route Shift
State transportation planners want to adjust the route for a proposed highway connecting the Rome and Cartersville areas more directly to I-75. The changes come in response to a federal ruling this summer that an 1867 manganese mine on Dobbins Mountain has historic value, halting federal money to the roadway because it would damage the area.
Thu., August 2, 2012 12:29pm
Feds Rule Dobbins Mine Is Historic
A former mine in northwest Georgia could force state transportation officials to pick another route for the proposed U.S. Highway 411 connector in Bartow County. A federal official notified the state this week that the mine is historically significant and eligible for listing on the National Register of Historic Places.


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Casino Royale [Import]
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John Huston was only one of five directors on this expensive, all-star 1967 spoof of Ian Fleming's 007 lore. David Niven is the aging Sir James Bond, called out of retirement to take on the organized threat of SMERSH and pass on the secret-agent mantle to his idiot son (Woody Allen). An amazing cast (Orson Welles, Peter Sellers, Deborah Kerr, etc.) is wonderful to look at, but the film is not as funny as it should be, and the romping starts to look mannered after awhile. The musical score by Burt Bacharach, however, is a keeper. --Tom Keogh
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Casino Royale (1967) arrives at blu ray with MPEG-4 AVC 1080p 2.34:1 encode. The bit rate was actually quite high at AVC 36 MBPS on a BD-50 disc. Consider the age of this movie, the picture actually looks very smooth and film-like. Black levels are excellent, especially in all the scenes in the casino with all the gentlemen in their tuxedos, or the Berlin sequences, with its many dark interiors. Detail is generally quite good, but the image is somewhat soft, which maybe due to the original design of the photography. (4/5)
The DTS-HD Master Audio 5.1 lossless track sounds terrific. The beautiful music of Burt Bacharach floats in the air and never intrudes into the action of the movie. I still own the original motion picture soundtrack vinyl album, which was considered as an audiophile collector's item in the past. Of course, listening to Dusty Springfield's rendition of The Look Of Love is also worth the price of admission. This song, by Burt Bacharach (music) and Hal David (lyrics) was nominated for an Oscar as Best Original Song, but lost to the song Talk To The Animals from the movie Doctor Dolittle. (4/5)
With gadgets, gaming and girls galore, this movie is a brilliant parody of - itself. The outstanding part of this movie is the casts, including David Niven, Peter Sellers, Ursula Andress, Joanna Pettet, Orson Welles, Woody Allen, Deborah Kerr, William Holden, Charles Boyer and John Huston.
It has a budget of $12 million, but grossed $41 million worldwide, which is considered outstanding at that time.Read more ›
With gadgets, gaming and girls galore, this camp classic is not only the coolest of the spy films, but it's also a brilliant parody of itself!
Will the real "James Bond" please stand up? When secret agency chief "M" [John Huston] is killed, James Bond [David Niven] is thrust out of spy retirement to help smash SMERSH, the band of hit men who are likely responsible. And to protect his real identity, Bond's name is given to numerous other agents, including Evelyn Tremble [Peter Sellers] and Bond's neurotic nephew, Jimmy [Woody Allen]. With five directors, a cast of Hollywood icons that also includes Ursula Andress, Charles Boyer, Peter O'Toole, Jacqueline Bisset and Orson Welles, a soundtrack by Burt Bacharach and a frisky, farcical script, Casino Royale is Bond. Psychedelic Bond.
FILM FACT: Burt Bacharach's musical score was praised, earning him an Academy Award® nomination for the song "The Look of Love" and was performed by Dusty Springfield.
Cast: Peter Sellers, Ursula Andress, David Niven, Orson Welles, Joanna Pettet, Daliah Lavi, Woody Allen, Deborah Kerr, William Holden, Charles Boyer, John Huston, Kurt Kasznar, George Raft, Jean-Paul Belmondo, Terence Cooper, Barbara Bouchet, Angela Scoular, Gabriella Licudi, Tracey Crisp, Elaine Taylor, Jacqueline Bisset, Alexandra Bastedo, Anna Quayle, Derek Nimmo, Ronnie Corbett, Colin Gordon, Bernard Cribbins, Tracy Reed, John Bluthal, Geoffrey Bayldon, John Wells, Duncan Macrae, Graham Stark, Chic Murray, Jonathan Routh, Richard Wattis, Vladek Sheybal, Percy Herbert, Penny Riley, Jeanne Roland, Jennifer Baker (uncredited), Susan Baker (uncredited), R.S.M.Read more ›
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DVD does not work in North America. Is set for Europe. I contacted the seller over a week ago and they never got back to me.Published 3 months ago by Amazon Customer
very disappointed because I could not play it. It was UK format not North American.Published 5 months ago by Jayne Rutledge,Jayne Rutledge
the movie is very bad but for any fan of james bond u need it in your colectionPublished 7 months ago by simon
mauvais achat, impossible de lire le dvd avec mon lecteur.Published 8 months ago by Martin Blondeau
The original Casino Royale is must have for the ultimate Bond collection.Published 10 months ago by Amazon Customer
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Published: May 7, 2010
Thirty-one members of Cardinal Mooney National Honor Society and their adviser, Peggy Moore, worked with representatives of Lien Forward Ohio/V&M Star to collect more than 100 bags of trash and recyclables during a recent Youth Day of Caring. Prepared to bag some of the debris, above, Cardinal Mooney High School senior Joe Stoops joins others working during the clean-up day along the West Federal Street corridor near Girard.
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on July 13, 2012 - 12:01 AM
If you Google "The downfall of Yes," you'll come across a YouTube video that some enterprising fan of the seminal progressive rock band concocted, using the now well-known footage from a film about Hitler in which der Fuhrer is having a meltdown, in German, at the expense of a few of his underlings. In this version, a YouTube auteur created subtitles in which the raving Hitler is now freaking out about lineup changes in Yes -- specifically, the departure of singer Jon Anderson and the addition of Benoit David to the band's ranks.
This is all in bad taste -- nothing about Hitler is funny. But it does raise an issue that, as "classic" bands continue touring well into their 60s, affects fans and potential fans of that band.
When is a band no longer a band, but simply a brand?
This summer, we'll see several major groups with half, or even less than half, of their original members on stage. The Guess Who, a perennial favorite on the Buffalo summer concert schedule, might well be called "the Guess Who It Isn't," since singer Burton Cummings and guitarist Randy Bachman are no longer in the house. Kiss will arrive at Darien Lake in September with only founders Gene Simmons and Paul Stanley on board, stand-ins Tommy Thayer and Eric Singer handling the musical figures pioneered by original members Ace Frehley and Peter Criss.
And then there's Yes.
When the band plays Artpark's Tuesday in the Park series this week, Jon Anderson will not be in attendance. Anderson's replacement, the aforementioned Benoit David, won't be singing for Yes, either. Third generation prog-rock outfit Glass Hammer vocalist Jon Davison will be manning the microphone, following David's as yet unexplained sudden departure. Which makes Davison a replacement for the replacement.
I've been conducting an informal poll all summer long, trying to take the temperature of the seemingly limitless flow of area prog-rock fans I encounter at shows, in clubs, or even while walking my dogs. I ask them if they'll go to the Yes show, considering all the seismic shifts in the lead singer department.
Three quarters of these folks immediately scoff, say "No way!" and proclaim their eternal allegiance to Anderson as "the only true voice of Yes music." (Anderson was rather unceremoniously ousted from Yes following a severe respiratory illness that prevented him from filling tour obligations a few years back. He has made it more than plain that he's none too pleased with this state of affairs.)
Most of the rest say they'll go, simply because the music remains vital and, frankly, awesome, no matter who is singing. Throw in the inviting Artpark setting, and the low-dough ticket price of $5 advance, $10 after Sunday, and the general feeling among this portion of the crowd is, "Why not?"
Yes has a history at Artpark, we should note. It was on this very site, on the stage of the indoor amphitheater, that the band played a historic concert -- with Anderson singing, and a symphony orchestra joining the band -- back in 2001. So Yes an awful lot to live up to on Tuesday.
I've been asked many times this summer if I plan to be at the Tuesday show and what my take on the whole Anderson/David/Davison mess is. Well... hell yeah, I'm going! Most definitely, I'd prefer Anderson to be in Yes, where he belongs. But founder/bassist Chris Squire, guitarist Steve Howe, drummer Alan White, and keyboardist Geoff Downes are all going to be on the Artpark stage. Squire and Howe in particular are, in my view, two of the finest rock musicians extant. White is no slouch, either, and Downes has a history with Yes, having played on the evergreen "Drama" album -- interestingly, during the brief time that Anderson split with the band for the first time, in the early 1980s.
How much of a substance can be whittled away without completely changing the character of that substance? It's tough to say. There are certainly bands that tour under their original banners with dubious lineups playing the classics, presumably before a percentage of audience members who are either oblivious to the changes, or simply don't care, as long as they hear "the hits." Observe the career of Journey: The band continues to pull in more than respectable numbers as a live act, even though singer Arnel Pineda is clearly attempting to clone departed singer Steve Perry's parts. Journey fans don't appear to be too broken up about any of this.
Progressive rock fans are a different breed, however. They don't appreciate being messed with, and they are not easy to fool. I'm hoping Yes will convince them of its collective sincerity on Tuesday. And then I'm hoping the band wraps up this tour and gives Jon Anderson a call.
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I'm using:
Mozilla/5.0 (Macintosh; U; Intel Mac OS X 10.6; en-US; rv:22.214.171.124) Gecko/20091221 Firefox/3.5.7
When I click the title element for adding a new question the text What's your programming question? Be specific, doesn't disappear until I click on another element on the page and then back to that element again.
This just started happening in the last day or two.


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Sep 16
James Ethan Clark & The Renegades make their first appearance in Sanford, NC at the renowned Temple Theatre for Tuesdays At The Temple on August 5, 2014. Doors open at 6pm for this 7pm show. General admission is $10. Call the Box Office at (919) 774-4155 or click attached link for tickets.
Jul 5
Saturday, July 5th @ The Chop Shop Adam Pitts / Kryss Dula / American Mannequins / Carson Hill / Follow Apollo / Revelus / Kick the Robot / The Business People / James Ethan Clark and The Renegades / Gnarly Charlies / Grown Up Avenger Stuff / The Quixote Project / Skytown Riot / Chasing Pedestrians / (i am) isis / Beitthemeans / Robert Johnson, Jr. / Perfect Mark / Something Clever
May 6
Representing the Southeast at North America's best music and arts fest, 2014 Canadian Music Week in Toronto, ON!
Mar 15
Join us March 15th for our Annual Irish Street party. The shenanigans will start at 11am and run through till 2am. There will be music inside The Pour House all day and on Blount Street! We will be adding additional bands to the lineup, so keep checking back often. The Blount Street Party will be FREE admission all day, so start making your plans now.
Feb 22
As part of the Millennium Music Conference, we will be on stage around 11PM, bring all your folks with you!!
Sounds Like: Ryan Adams, Cary Brothers, My Morning Jacket, Neil Young, Kings Of Leon
Label: Fallen Tree Publishing
Manager: Jeff Lynn
Bio: As a writer and performer, James Ethan Clark draws inspiration from an eclectic melting pot of genres ranging from progressive rock, folk and soul. Hailing from North Carolina, Clark released his debut record in April 2013. Since the release of "Southern Hotel", Clark has gained a notable interna...See Full Bio
"Artist-on-the-rise James Ethan Clark internationally released his freshman LP entitled "Southern Hotel" on Apr. 2. The young alternative folk-rock solo artist uses his soulful voice, guitar, piano and harmonica on the ten song debut album to prove that his new sound is play-worthy. The self-narrative of Clark's life as a young man, many of his songs tell a tale of a brokenhearted boy coming into his own as strong-willed man. Hailing from North Carolina, Clark recorded the album, his first full-length recording, in Nashville with the help of a few guest artists." (Full Review Attached)


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Alrighty. I've been pressing forward at a steady pace with getting the CO modified to a set schedule. I told my ex we had to modify. I have asked him to come to an agreement with me. I have given him the names of mediators and have stayed on top of him to get this done.
Well, after days and days of heated text discussions, him telling me things I would get banned for repeating, him telling me he wants primary custody and I am a selfish b*tch, and me sticking to my guns, my meeting with a lawyer today and telling my ex the gig is up.... The Heavens have opened up and prayers were answered and I believe we have an agreement and we are going to put it into a court order.
I will put Bean into a M/W preschool program and ex will pick Bean up on Wednesdays at 4:30. He will have Bean until Saturday at 4:00 each week. My ex will put in a hardship request for a set schedule and will be off Th/F/Sat each week. He has already cleared this part.
I will be responsible for all medical appointments and will only schedule medical appointments on my time. He does not want to be involved in medical appointments.
The attorney I talked to today has agreed to review and draft the order for us to file with the courts. I LOVED this attorney...if I have to modify with a lawyer in the future I will use his services.
So...please say a prayer it all stays in place. I thanked my ex for cooperating (loosely speaking) and am walking on egg shells hoping to get this finalized before something falls through or he has another episode.
I am happy with this. Bean gets preschool, I get a set schedule and I get 1.25 days of each week-end.
Thank you all for giving me your perspective and welcoming me onto your board. Your advice is great and you all seem like nice ladies! :)