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Received 8 April 2010 | Accepted 12 July 2010 | Published 15 July 2010
The fate of Müller's glia following experimental retinal detachment: nuclear migration, cell division, and subretinal glial scar formation
Geoffrey P. Lewis, Ethan A. Chapin, Gabriel Luna, Kenneth A. Linberg, Steven K. Fisher
Neuroscience Research Institute, University of California, Santa Barbara, CA
Correspondence to: Geoffrey P. Lewis, Neuroscience Research Institute, University of California Santa Barbara, Santa Barbara, CA, 93106. Phone: (805) 893-3611; FAX: (805) 893-2005; email: G_lewis@lifesci.ucsb.edu.
Purpose: To study the fate of Müller's glia following experimental retinal detachment, using a "pulse/chase" paradigm of bromodeoxyuridine (BrdU) labeling for the purpose of understanding the role of Müller cell division in subretinal scar formation.
Methods: Experimental retinal detachments were created in pigmented rabbit eyes, and 3 days later 10 µg of BrdU was injected intravitreally. The retinas were harvested 4 h after the BrdU was administered (i.e., day 3) or on days 4, 7, and 21 post detachment. The tissue was fixed, embedded in agarose, and sectioned at 100 µm. The sections were labeled with various combinations of probes, including anti-vimentin and anti-S100 (as markers for Müller cells), anti-BrdU, anti-phosphohistone H3 (to identify mitotic cells), and the isolectin B4 (to identify macrophages and microglia). Images were captured using an Olympus Fluoview 500 confocal microscope. To aid in our understanding of how Müller cell nuclei undergo cell division, two additional procedures were used: 1) electron microscopy of normal cat and rabbit retinas and 2) a new method using 5-fluorouracil and subsequent anti-BrdU labeling to detect all Müller cell nuclei, using confocal imaging.
Results: Three days after detachment, anti-vimentin labeled all Müller cells, some of which were also labeled with anti-BrdU. On day 4, many of the anti-BrdU-labeled Müller cell nuclei appeared in columns with one labeled nucleus in the inner nuclear layer and another directly sclerad to it in the outer nuclear layer. By day 7, most anti-BrdU-labeled nuclei were observed in subretinal scars. At 3 weeks, some anti-BrdU-labeled nuclei that remained within the retina did not express vimentin or S100. Anti-phosphohistone H3-labeled (i.e., mitotic) cells, some of which were also labeled with anti-BrdU, were only observed in the outer nuclear layer on day 4, and these nuclei were surrounded by an accumulation of vimentin filaments. Isolectin B4-labeled microglia and macrophages also incorporated BrdU and were observed throughout the retina and in subretinal scars during all times of detachment. Electron microscopy and immunofluorescence labeling of the 5-fluorouracil-injected eyes revealed the presence of a unique structural relationship between Müller cell nuclei and intermediate filament proteins.
Conclusions: Following retinal detachment, many Müller cell nuclei initially migrate to the outer retina, undergo mitosis, and eventually reside in subretinal glial scars, suggesting a possible link between the early division of Müller cells and the process of subretinal gliosis. In addition, a subpopulation of anti-BrdU-labeled cells, presumably once Müller cells, appears to stop expressing well accepted Müller cell marker proteins, suggesting a potential dedifferentiation of some of these cells over time. Additionally, Müller cell nuclei may use intermediate filaments as a "track" for migration into the outer retina and later as an important component of cell division by the accumulation of vimentin filaments around the mitotic nuclei.
Injury to the retina, as elsewhere in the central nervous system (CNS), results in the activation of glial cells and the formation of glial scars [1,2]. In retinal detachment (RD), a form of traumatic injury where the retina becomes separated from the underlying retinal pigment epithelial (RPE) layer, the Müller cell, generally considered a specialized radial astrocyte, is the predominant glial cell type involved. Following RD, Müller cells actively proliferate and hypertrophy within the retina and onto either retinal surface where they form structures similar to those formed by reactive astrocytes in the brain and spinal cord [3]. Such cellular scars or "membranes" present on the subretinal or epiretinal (vitreal) surface are considered part of the spectrum of "fibrocontractive retinal disorder" termed proliferative vitreoretinopathy (PVR) [4]. Despite numerous attempts to reduce the incidence of PVR, no effective pharmacological treatment has been found to date [5,6]. This may be because PVR is a complex disease involving cell proliferation, growth, spreading, and contractility. Dividing cells have been observed in membranes removed from patients with PVR ([7–9] and unpublished data); however, data from animal models as well as from membranes removed from human patients clearly show a role for Müller cell expansion as a critical part of the response.
Müller cell reactivity can ultimately result in loss of vision because of glial scars on the retinal surfaces. When scars are present in the subretinal space, the regeneration of photoreceptor outer segments is prevented [10]. Subretinal glial scars can also cause visual distortion by preventing proper flattening of the retina or causing macular wrinkling, both of which may require surgical removal of the scar tissue [11]. This is not an insignificant problem since subretinal glial scars in humans have been estimated to occur in 15.7% of all human retinal detachments [12]. When a glial scar grows on the vitreal surface of the retina, the cells can contract, causing retinal folds and re-detachment of the retina. Indeed, epiretinal membrane formation and subsequent contraction detachment remains the most common reason for failure of retinal reattachment surgery and occurs in 5%–12% of cases [13–17]. Although advances in surgical management have improved the ability to ultimately reattach the retina after the occurrence of a contraction detachment, the visual prognosis remains poor, with only 11%–25% of patients achieving a visual acuity of 20/100 [18]. Glial scar formation, therefore, is a serious complication of retinal injury. However, the early events in scar formation are poorly understood.
PVR is generally thought to be a condition involving uncontrolled cell proliferation, although the exact role of proliferation and how it relates to the hypertrophy of Müller cells and ultimate glial scar formation in the injured retina is unclear. It has been demonstrated that within a few hours of detachment, Müller cells upregulate transcription factors related to both proliferation and hypertrophy [19], and at one day after detachment, both events are underway, as evidenced by anti-Ki67 and anti-glial fibrillary acidic protein (GFAP) labeling, respectively [20,21]. Proliferation peaks 3–4 days after detachment, is reduced at day 7, but continues at low levels for as long as the retina remains detached [22,23]. Hypertrophy and growth of Müller cells into the subretinal space also appear to occur as an ongoing event since longer detachment durations result in larger subretinal glial scars.
The question remains, however, as to the role, if any, cellular proliferation has in the formation of glial scars. That is, are glial scars formed only by the population of cells that proliferate? Use of the anti-proliferative agent 5-fluorouracil (5-FU) did not significantly reduce the occurrence of PVR in human patients [6], although issues of timing and dosage make these results difficult to interpret. Moreover, the process of cell division of Müller cells, complex cells that span the entire width of the retina, has not been elucidated in the mammalian retina. To address these issues, we used an intraocular pulse of bromodeoxyuridine (BrdU), given at the peak of the proliferative response after retinal detachment in the rabbit, and followed the distribution of the labeled nuclei over time. We show here that the Müller cell nuclei that incorporate BrdU on day 3 appear to migrate into the outer retina where they undergo mitosis. Some Müller cells with anti-BrdU-labeled nuclei then grow beyond the outer limiting membrane (OLM) to begin the process of glial scar formation. The glial scars expand with increasing detachment time, and many anti-BrdU-labeled nuclei come to reside in the scars. These data suggest a significant role for Müller cell proliferation, beginning early after detachment, in contributing to the formation of subretinal glial scars. In addition, immunocytochemical observations of anti-GFAP and anti-vimentin labeling along with electron microscopic data suggest a link between the migration of Müller cell nuclei and the intermediate filament cytoskeleton.
Retinal detachments were created in adult New Zealand Red pigmented rabbits (Myrtle's Rabbitry, Thompsons Station, TN) by infusing a solution of sodium hyaluronate (Healon, 0.25% in balanced salt solution; Pharmacia, Piscataway, NJ) between the neural retina and RPE via a fine glass pipette. The Healon is necessary to prevent spontaneous reattachment of the retina. The pipette was inserted into the eye via an incision that was made several millimeters below the pars plana to prevent the pipette from touching the lens. Approximately one-half of the inferior retina was detached in the right eye, leaving the superior attached regions as internal controls. The left eyes served as non-detached controls. On day 3 after detachment, the eyes were injected with 10 µg BrdU (Sigma, St Louis, MO) in 50 µl balanced salt solution (BSS; Alcon, Ft. Worth, TX). Animals were euthanized either 4 h after the BrdU injection on day 3, or on days 4, 7, or 21 post detachment, using sodium pentobarbital (120 mg/ml, IV). Three animals were used at each time point. Following enucleation, the eye was fixed in 4% paraformaldehyde for at least 24 h (in 0.1 M sodium cacodylate buffer, pH 7.4; Electron Microscopy Sciences, Fort Washington, PA) and stored until used. All experimental procedures conformed to the ARVO statement for the Use of Animals in Ophthalmic and Vision Research, the Guide for the Care and Use of Laboratory Animals, and the guidelines of the Animal Resource Center of the University of California, Santa Barbara.
Immunocytochemistry
Following at least 24 h of fixation, retinal pieces approximately 4 mm square, were excised from three detached regions within each eye. The tissue was rinsed in PBS (10X PBS stock: 250 mls distilled water, 21.92 g NaCl, 0.67 g NaH2PO4, 2.88 g NaH2PO4; stir well, dilute 1:10 in distilled water, adjust pH to 7.4. Yields 0.086M PBS), embedded in low-melt agarose (5%; Sigma), and sectioned at 100 µm using a vibratome (Technical Products International, Polysciences, Warrington, PA). Sections were incubated in normal donkey serum (1:20) in PBS, 0.5% BSA, 0.1% Triton X-100, and 0.1% azide (PBS, BSA [PBTA]) overnight at 4 °C on a rotator. The following day the sections were pretreated with 2 N HCL for 1 h as an antigen retrieval step for the BrdU. After rinsing in PBTA, the primary antibodies and lectin were added and incubated overnight at 4 °C on a rotator in PBTA. Anti-BrdU (1:200; Accurate Chemical and Scientific Corp., Westbury, NY) was used to detect cells in S-phase; anti-phosphohistone H3 (1:100; Lake Placid, NY) was used to detect cells undergoing mitosis; anti-vimentin (1:500; Dako, Carpinteria, CA) and anti-S100 (1:1000; Dako) were used to identify Müller cells; and isolectin B4, Griffonia simplicifolia (1:50; Vector Labs, Burlingame, CA), was used to label microglia and macrophages. Following rinsing of the primary antibodies in PBTA, the secondary probes (streptavidin CY5, donkey antirat CY3, and donkey antimouse CY2; Jackson ImmunoResearch, West Grove, PA) were added together, each at 1:200 in PBTA, overnight at 4 °C on a rotator. On the final day, the sections were rinsed in PBTA, mounted on glass slides, using 5% n-propyl gallate in glycerol, and viewed on an Olympus Fluoview 500 laser scanning confocal microscope (Center Valley, PA). Each image represents a projection from a z-stack of 6–10 images collected at 0.5-µm increments.
Müller cell nuclei and the cytoskeleton
To systematically study the relationship between intermediate filaments and Müller cell nuclei, we took advantage of the observation that Müller cells appear to specifically take-up 5-FU, which can then be detected with anti-BrdU. Since uracil is normally converted to uridine by cells, presumably the same process occurs with 5-FU after injection into the vitreous, and the product is then detected by anti-BrdU. Why this appears to preferentially label Müller cells remains a mystery but may be related to Müller cells' close apposition to the vitreous cavity, the site of 5-FU deposition. In this experiment, 5-FU (0.5 mg in 50-µl balanced salt solution) was injected intravitreally into "normal" rabbit eyes (n=3). Three days later animals were euthanized (120 mg/kg Euthasol, intravenous, Henry Schein, Melville, NY), and the eyes fixed as described above. The retinas were then sectioned and labeled with anti-BrdU as well as anti- GFAP (1:400; Dako) and/or anti-vimentin (1:500; Dako), as described below.
Electron microscopy of archived cat and rabbit tissue was also examined to study the ultrastructure of Müller cell nuclei. Both nondetached cat and rabbit retinas and cat retinas detached for 50 days were fixed in 1% glutaraldehyde plus 1% paraformaldehyde for 24 h after which the tissue was rinsed in isotonic phosphate buffer, postfixed for 2 h in 2% osmium tetroxide in phosphate buffer, dehydrated in a graded ethanol series, processed through propylene oxide, and embedded in Spurr's resin. The retinas were then sectioned at 90 nm in thickness, stained with uranyl acetate and lead citrate, and viewed using a JEOL 1230 electron microscope.
Fate of injected bromodeoxyuridine
In control nondetached rabbit retinas, anti-vimentin labeling of the Müller cells extends from the inner limiting membrane (ILM) into the outer nuclear layer (ONL; Figure 1A, green). Based on previous data showing that intraretinal proliferation peaks at 3 days in the rabbit [20], BrdU was injected into the vitreous at this time and the tissue examined 4 h later. At this time point, anti-vimentin labeling increased, filling the entire Müller cell from the ILM to the OLM (Figure 1B). Numerous anti-BrdU-labeled Müller cell nuclei could also be seen in the inner nuclear layer (INL; Figure 1B, red). At day 4, 24 h after BrdU injection, labeled nuclei frequently appeared in radial "columns" across the retina, such that one labeled nucleus in the ONL directly overlies another in the INL (Figure 1C, arrows). In some cases two nuclei could be observed directly adjacent to one another, always in a "vertical" arrangement, giving the impression that they occur in the same Müller cell (Figure 1D, arrows). We never observed two labeled nuclei lying "side-by-side" in the INL. Also at the 4-day detachment time, the first sign of Müller cell growth beyond the OLM into the subretinal space was observed (Figure 1E, bracket). Invariably, there was an anti-BrdU-labeled nucleus associated with this early stage of growth, itself often appearing to extend partially beyond the OLM. At 7 days following detachment, many anti-BrdU-labeled nuclei were present within, or adjacent to, large subretinal glial scars (Figure 1F, brackets), although in regions away from the scars, the retina appeared mostly devoid of labeled nuclei (Figure 1F, left half of the image). Subretinal scars, however, were never observed without containing anti-BrdU-labeled nuclei.
Microglia and macrophages also undergo cell division in the retina after detachment [22]. To ascertain that the majority of BrdU labeling was occurring in Müller cells, we used the isolectin B4 to identify the former cell types. As shown previously, the cells within the retina are primarily resident microglia, whereas those in the subretinal space are mostly macrophages that have entered the eye from the blood [24]. Isolectin B4-labeled microglia in attached regions from an eye with a retinal detachment appear as an array of fine stellate processes confined primarily to the inner plexiform layer (IPL; Figure 1A, blue). Following detachment, the microglia round-up and migrate into the outer retina, whereas macrophages appear to remain in the subretinal space (Figure 1B-F; also see ref. #26, for quantitative data). Some macrophages (in the subretinal space) and microglia (in the retina) can be observed labeled with anti-BrdU (Figure 1D-F, arrowheads). In comparison to Müller cells, the number of anti-BrdU-labeled immune cells is very low (Figure 1D). That, combined with the prominent anti-vimentin labeling of Müller cells makes their confusion unlikely. Some retinal regions were relatively devoid of immune cells, while others contained many, but there was no apparent correlation between the presence of these cells and the extent of glial scar formation.
The columnar pattern of BrdU-labeled nuclei across the retina suggests that both nuclei at this stage occur within the same Müller cell, although it is difficult to be certain from the confocal two-dimensional projection images in Figure 1 (C-E). However, by processing the z-stack of images using bioView software (version 1.1.18, free download from the Center for bio-image informatics, UCSB, Santa Barbara, CA) to provide varying contrasts, it appears likely that the vertical pairs of BrdU-labeled nuclei are indeed associated with a single Müller cell "stalk" of intermediate filaments (Figure 2A,B). Figure 2A shows a pair of nuclei lined up directly along one distinct column of intermediate filaments, presumably within a single Müller cell, whereas the image in Figure 2B is rotated slightly so that the lower nucleus in the INL appears off to the side of the major intermediate filament bundle.
Twenty-one days after detachment, many of the anti-BrdU-labeled nuclei in regions away from a subretinal scar appeared unassociated with anti-vimentin-labeled cells (Figure 3A-D, arrows; day 21), especially when viewed through the z-series of images. Some of the nuclei could also be observed in the ganglion cell layer (GCL) at this time (Figure 3C). Vimentin, however, is a cytoskeletal protein and hence does not necessarily fill the entire cell. Therefore, anti-S100, a calcium-binding protein, was used to label the cytoplasm of Müller cells. Since anti-S100 labels the entire cytoplasm with heavy labeling of the cell body, any anti-BrdU-labeled nuclei in S100 positive cells should reflect the overlay of the two fluorochromes (in this case, red+blue=pink). Interestingly, many BrdU-labeled nuclei do not appear to lie within anti-S100-labeled cells (Figure 3A-D, arrows), i.e., the nuclei appear red with no overlying blue or green labeling. By turning off the BrdU (red) channel, it became apparent that there was little to no S100 or vimentin in the cells associated with this subpopulation of BrdU-positive nuclei, although they always lie adjacent to areas of heavy anti-S100 or anti-vimentin labeling (Figure 3B,D, arrows). In an area where the band of intermediate filaments overlays the nucleus of a BrdU- and S100-positive cell, the resulting triple labeling produces a white signal (Figure 3A; arrowhead).
To determine if Müller cells were undergoing mitosis in addition to undergoing DNA synthesis in S phase, we labeled sections with both anti-phosphohistone H3 (green) and anti-BrdU (red) at all detachment time points (Figure 4). Many anti-phosphohistone H3-labeled cells were observed, but only at the 4-day time point. Some of these cells were also double labeled with anti-BrdU (Figure 4A,B; insets have the red BrdU channel turned off). Significantly, while anti-BrdU-labeled cells were observed throughout the retina, anti-phosphohistone H3-labeled mitotic figures were observed only in the ONL, suggesting that Müller cell nuclei incorporate BrdU in the INL before migrating to the ONL to divide. The fact that anti-phosphohistone H3-labeled mitotic figures were observed only at 4 days suggests that most of the cell division occurs as a single "wave" within a few days following detachment. Additionally, the anti-phosphohistone H3-labeled cells often appear surrounded by dense anti-vimentin labeling, presumably representing a localized accumulation of intermediate filaments (Figure 4A,C,D, blue, arrows). To appreciate that this "cage" of intermediate filaments completely envelops the mitotic figure formed as the cell divides, Figure 4C,D shows one image tilted slightly in two different directions (anti-phosphohistone H3, green, arrow; anti-vimentin, blue). The top and bottom images of the z-stack were omitted to avoid obscuring the anti-phosphohistone H3 labeling within the cage of vimentin filaments.
Structural mechanism for nuclear migration
Since mitotic Müller cells were only observed in the ONL we sought a possible mechanism to explain nuclear migration into the outer retina. In many of the images labeled with anti-BrdU and anti-vimentin, we observed what appeared to be a narrowing of the band of intermediate filaments and their apparent association with a "groove" or "notch" in the Müller cell nucleus (Figure 5A,B). Electron microscopy of tissue from both rabbit and cat retinas clearly demonstrate the deep indentation of the Müller cell nuclei and the presence of intermediate filaments within the notch formed by the indentation (Figure 5C,D, arrows). The prevalence of this nuclear notch and its relationship to the intermediate filament cytoskeleton is demonstrated by confocal imagining of tissue from normal (nondetached) rabbit retinas that had been treated with 5-FU and labeled with anti-BrdU to visualize all Müller cell nuclei. In both radial sections (Figure 6A,B) and wholemount preparations (Figure 6C-F), the notch can be clearly observed in every Müller cell nucleus (red labeling) within a field. In addition, vimentin filaments (green) invariably localize on the side of the cell facing the notch and in many instances can be seen to fill the notch itself. While anti-GFAP labeling also localized to this region (Figure 6C,D, blue), unlike vimentin filaments, GFAP filaments are not present in all the nuclei in the nondetached retina. When present, however, they too bundle with vimentin at the site of the notch.
The original goal of this study was to determine if Müller cell proliferation is an important mechanism in the formation of subretinal glial scars. The data, however, provide new information about the reactivity of these highly differentiated glial cells as well as suggest a specific role for their proliferation in glial scar formation. BrdU, injected into the vitreous at the height of the proliferative response following detachment, provided a means to follow the fate of the labeled nuclei over time. Our interpretation of the results is summarized in Figure 7 showing a Müller cell nucleus with its unique structural relationship to the cells' intermediate filament cytoskeleton (based on the image in Figure 5A). Within 24 h after BrdU injection, anti-BrdU-labeled Müller cell nuclei migrate along the intermediate filament cytoskeleton to the outer retina (Figure 7, step 1). Once there, vimentin filaments assemble around the nucleus, which then undergoes mitosis, as indicated by the anti-phosphohistone H3-labeling data (Figure 7, step 2). Assuming that every BrdU-labeled Müller cell nucleus undergoes division in the outer retina and based on the columnar pattern observed frequently at day 4, the parent Müller cell may occur in a binucleate phase for a short time, about the same time Müller cells begin to grow processes into the subretinal space. Some of these cells may remain binucleate, with one nucleus returning to its normal location in the INL (Figure 7, step 3) and the other nucleus migrating into the processes forming the subretinal scar (Figure 7, steps 4 and 5). Other Müller cells seem to bud off, forming a daughter cell containing one of the nuclei but not labeling with such typical Müller cell markers as anti-vimentin or anti-S100, as if they occur in some stage of dedifferentiation (Figure 7, step 6). Taken together, these data suggest that early Müller cell proliferation is in some way mechanistically coupled to the eventual formation of large subretinal glial scars.
Overall, our observations are remarkably similar to those observed in the injured zebrafish retina [25], although the final outcome is probably very different. Following light damage to the fish retina, which destroys the photoreceptors, Müller cell nuclei migrate to the outer retina where they re-enter the cell cycle. The daughter cell in this case, however, buds off from the parent Müller cell and becomes a retinal progenitor cell, as evidenced by Pax6 expression, which can dedifferentiate and eventually replace lost photoreceptors. A similar phenomenon has also been shown in the chicken retina in response to damage or exogenous growth factors [26]. In this case Müller cells dedifferentiate, proliferate, begin expressing transcription factors normally found in embryonic retinal progenitors, and produce both neurons and glial cells. In the detached rabbit retina, however, we have no evidence that cells in the subretinal space are anything other than Müller cells since they label with typical Müller cell markers. While we have yet to successfully label the BrdU-labeled nuclei that appear in the outer retina with typical markers to progenitor cells as shown in the chicken retina [26], the availability of new progenitor cell markers suitable for use in the mammalian retina in the future will allow addressing this issue.
The observation of a special structural relationship between Müller cell nuclei and the intermediate filament cytoskeleton is also of interest. The observations are certainly suggestive that intermediate filaments may anchor the nuclei in place in the normal retina, provide a "track" along which they migrate in the injured retina, and provide a means of isolating the nuclei in a cage-like structure in the outer retina during mitosis. A similar relationship has been observed between migrating pigment granules and the vimentin cytoskeleton in Xenopus melanocytes [27]. It is possible that this cage of filaments isolates the nucleus from the cell and allows for nuclear division without the need for the entire cell to round-up since we found no evidence that these cells retract their inner processes during division.
Some BrdU-labeled nuclei observed here also occur in cells known to participate in other reactions to detachment. For example, a few labeled Müller cell nuclei were observed near the vitreal border in longer term detachments (e.g., 21 days; Figures 3C), and it may be the Müller cells associated with these nuclei that ultimately contribute to epiretinal membranes on the vitreal surface of the retina. In addition, since there are no astrocytes in this region of the rabbit retina (i.e., distant from the medullary rays), the only other cell types that incorporate BrdU are the microglia and macrophages. The robust labeling of these cells with anti-BrdU is more evidence for a significant inflammatory response to retinal detachment [28–30], and while they have been shown to migrate to the photoreceptor layer after detachment [24], their role if any in glial scar formation is unclear.
The data presented here suggest that Müller cell proliferation is a critical step for the formation of glial scars in the retina and that an essential trigger for this scar formation takes place during the period of early proliferation that occurs within a few days of the injury. Clinical trials using anti-proliferative drugs to prevent proliferative diseases in the eye have not proven successful; however, this may be due to the timing of treatment since these drugs are typically given only for a short time during the surgical removal of scar tissue that has already formed, that is, late in the disease state. Anti-proliferative drugs used in prior clinical studies also tend to have a very short half-life in the eye. Two separate studies in this animal model, however, have recently demonstrated that the intravitreal administration, at the time of detachment surgery, of two compounds with a relatively long half-life in the eye effectively reduce both intraretinal proliferation and the formation of subretinal glial scars [31,32]. Those studies support the data presented in this paper that Müller cell proliferation, initiated within a few days after detachment, is critical to the formation of subretinal glial scars.
This work is supported NIH grant R01EY000888; NSF grant 0941717.
Fisher SK, Lewis GP, Linberg KA, Verardo MR. Cellular remodeling in mammalian retina: results from studies of experimental retinal detachment. Prog Retin Eye Res. 2005; 24:395-431. [PMID: 15708835]
Jones BW, Marc RE. Retinal remodeling during retinal degeneration. Exp Eye Res. 2005; 81:123-37. [PMID: 15916760]
Fisher SK, Lewis GP. Müller cell and neuronal remodeling in retinal detachment and reattachment and their potential consequences for visual recovery: a review and reconsideration of recent data. Vision Res. 2003; 43:887-97. [PMID: 12668058]
Machemer R, Aaberg TM, Freeman HM, Irvine AR, Lean JS, Michels RM. An updated classification of retinal detachment with proliferative vitreoretinopathy. Am J Ophthalmol. 1991; 112:159-65. [PMID: 1867299]
Charteris DG, Aylward GW, Wong D, Groenewald C, Asaria RH, Bunce C. PVR Study Group. A randomized controlled trial of combined 5-fluorouracil and low-molecular-weight heparin in management of established proliferative vitreoretinopathy. Ophthalmology. 2004; 111:2240-5. [PMID: 15582080]
Wickham L, Bunce C, Wong D, McGurn D, Charteris DG. Randomized controlled trial of combined 5-Fluorouracil and low-molecular-weight heparin in the management of unselected rhegmatogenous retinal detachments undergoing primary vitrectomy. Ophthalmology. 2007; 114:698-704. [PMID: 17398320]
Heidenkummer HP, Kampik A, Petrovski B. Proliferative activity in epiretinal membranes. The use of the monoclonal antibody Ki-67 in proliferative vitreoretinal diseases. Retina. 1992; 12:52-8. [PMID: 1565872]
Tsanou E, Loachim E, Stefaniotou M, Gorezis S, Charalabopoulos K, Bagli H, Peschos D, Psilas K, Agnantis NJ. Immunohistochemical study of angiogenesis and proliferative activity in epiretinal membranes. Int J Clin Pract. 2005; 59:1157-61. [PMID: 16178982]
Zhang X, Barile G, Chang S, Hays A, Pachydaki S, Schiff W, Sparrow J. Apoptosis and cell proliferation in proliferative retinal disorders: PCNA, Ki-67, caspase-3, and PARP expression. Curr Eye Res. 2005; 30:395-403. [PMID: 16020270]
Anderson DH, Guerin CJ, Erickson PA, Stern WH, Fisher SK. Morphological recovery in the reattached retina. Invest Ophthalmol Vis Sci. 1986; 27:168-83.3943943. [PMID: 3943943]
Machemer R. Surgical approaches to subretinal strands. Am J Ophthalmol. 1980; 90:81-5. [PMID: 7395961]
Miura M, Ideta H. Factors related to subretinal proliferation in patients with primary rhegmatogenous retinal detachment. Retina. 2000; 20:465-8. [PMID: 11039420]
Greven CM, Sanders RJ, Brown GC, Annesley WH, Sarin LK, Tasman W, Morgan TM. Pseudophakic retinal detachments. Anatomic and visual results. Ophthalmology. 1992; 99:257-62. [PMID: 1553218]
Girard P, Mimoun G, Karpouzas I, Montefiore G. Clinical risk factors for proliferative vitreoretinopathy after retinal detachment surgery. Retina. 1994; 14:417-24. [PMID: 7899716]
Bonnet M, Fleury J, Guenon S, Yaniali A, Dumas C, Hajjar C. Cryopexy in primary rhegmatogenous retinal detachment: a risk factor for postoperative proliferative vitreoretinopathy? Graefes Arch Clin Exp Ophthalmol. 1996; 234:739-43. [PMID: 8986445]
Duquesne N, Bonnet M, Adeleine P. Preoperative vitreous hemorrhage associated with rhegmatogenous retinal detachment: a risk factor for postoperative proliferative vitreoretinopathy? Graefes Arch Clin Exp Ophthalmol. 1996; 234:677-82. [PMID: 8950587]
Speicher MA, Fu AD, Martin JP, von Fricken MA. Primary vitrectomy alone for repair of retinal detachments following cataract surgery. Retina. 2000; 20:459-64. [PMID: 11039419]
Lewis H, Aaberg TM, Abrams GW. Causes of failure after initial vitreoretinal surgery for severe proliferative vitreoretinopathy. Am J Ophthalmol. 1991; 111:8-14. [PMID: 1985496]
Geller SF, Lewis GP, Fisher SK. FGFR1, signalling, and AP-1 expression following retinal detachment: reactive Müller and RPE cells. Invest Ophthalmol Vis Sci. 2001; 42:1363-9. [PMID: 11328752]
Geller SF, Lewis GP, Anderson DH, Fisher SK. Use of the MIB-1 antibody for detecting proliferating cells in the retina. Invest Ophthalmol Vis Sci. 1995; 36:737-44. [PMID: 7890504]
Lewis GP, Matsumoto B, Fisher SK. Changes in the organization of cytoskeletal proteins during retinal degeneration induced by retinal detachment. Invest Ophthalmol Vis Sci. 1995; 36:2404-16. [PMID: 7591630]
Fisher SK, Erickson PA, Lewis GP, Anderson DH. Intraretinal proliferation induced by retinal detachment. Invest Ophthalmol Vis Sci. 1991; 32:1739-48. [PMID: 2032796]
Lewis GP, Sethi CS, Linberg KA, Charteris DG, Fisher SK. Experimental retinal reattachment: A new perspective. Mol Neurobiol. 2003; 28:159-75. [PMID: 14576454]
Lewis GP, Sethi CS, Carter KM, Charteris DG, Fisher SK. Microglial cell activation following retinal detachment: a comparison between species. Mol Vis. 2005; 11:491-500. [PMID: 16052164]
Bernardos RL, Barthel LK, Meyers JR, Raymond PA. Late-stage neuronal progenitors in the retina are radial Müller glia that function as retinal stem cells. J Neurosci. 2007; 27:7028-40. [PMID: 17596452]
Fischer AJ, Reh TA. Potential of Müller glia to become neurogenic retinal progenitor cells. Glia. 2003; 43:70-6. [PMID: 12761869]
Chang L, Barlan K, Chou Y-H, Grin B, Lakonishok M, Serpinskaya AS, Shumaker DK, Herrmann H, Gelfand VI, Goldman RD. The dynamic properties of intermediate filaments during organelle transport. J Cell Sci. 2009; 122:2914-23. [PMID: 19638410]
Farjo R, Peterson WM, Naash MI. Expression profiling after retinal detachment and reattachment: a possible role for aquaporin-0. Invest Ophthalmol Vis Sci. 2008; 49:511-21. [PMID: 18234993]
Hollborn M, Francke M, Iandiev I, Buhner E, Foja C, Kohen L, Reichenbach A, Wiedemann P, Bringmann A, Uhlmann S. Early activation of inflammation- and immune response-related genes after experimental detachment of the porcine retina. Invest Ophthalmol Vis Sci. 2008; 49:1262-73. [PMID: 18326757]
Banna CD, Lewis GP, Fisher SK. Time-course analysis using microarrays reveals multiple signaling pathways activated in response to experimental retinal detachment. ARVO Annual Meeting; 2009; May 3–7; Fort Lauderdale (FL)
Lewis GP, Chapin EA, Byun J, Luna G, Sherris D, Fisher SK. Müller cell reactivity and photoreceptor cell death is reduced following experimental retinal detachment using an inhibitor of the Akt/mTOR pathway. Invest Ophthalmol Vis Sci. 2009; 50:4429-4435. [PMID: 19369237]
Zahn G, Klatt K, Lewis GP, Vossmeyer D, Stragies R, Heier JS, Daniel PE, , Jr Adamis AP, Chapin EA, Fisher SK, Holz FG, Loeffler KU, Knolle J. Assessment of the Integrin alpha5B1 antagonist JSM6427 in proliferative retinopathy using in vitro assays and a rabbit model of retinal detachment. Invest Ophthalmol Vis Sci. 2010; 51:1028-35. [PMID: 19815730]


Chiefly New England and Inland North. hero sandwich.
In barrels around the grinder's edge, artillery simulators blasted away.
By now we were in a state of maximum disorientation, just trying to stay on the grinder with the others.
He is host of "The David Webb Show" on Sirius XM Patriot and "The Grinder" business talk show on AM 970 New York City.
Put the paste, back in the grinder or in the mortar, grind or pound again, then filter again with another third of the water.
My duties at the office of Stillwell, Grinder & Co. were varied.
Walter Carew's wife would scarcely have sought out the Grinder's daughter for her friend and bosom companion.
My uncle came along and began to sharpen a scythe on the grinder and I was turnin' it for him.
A grinder, on the contrary, is one who 'grinds' or reads with an unusual degree of application.
Old English grindere "one who grinds (grain);" agent noun from grind (v.). Meaning "molar tooth" is late 14c. (Old English had grindetoð). Meaning "machine for milling" is from 1660s; of persons, from late 15c. Large sandwich sense is from 1954, though the exact signification is uncertain (perhaps from the amount of chewing required to eat one).


The MSI GTX 980Ti 6GD5T OC is the new flagship graphics card engineered and optimized for 4K gaming. It's based on the state-of-the-art new generation NVIDIA Maxwell architecture, and features 2816 CUDA Cores and gigantic 6GB, 384-bit GDDR5 memory. It delivers unrivalled graphics performance to take on virtual reality (VR) and 4K gaming at high settings, while also being acoustic-, thermal- and power-efficient for an evolved gaming experience like never before. Highest-grade components and meticulous craftsmanship ensure unbeatable stability and reliability even in the heat of gaming.
Performance is just part of the story. The new Maxwell architecture is engineered with the new VXGI (Voxel Global Illumination) technology to solve some of the most complex lighting and graphics challenges in visual computing. Scenes are significantly more lifelike as light interacts more realistically in the game environment. Super Resolution technology lets you enjoy the detail and brilliance of 4K gaming even on 1080p displays. The G-Sync technology ensures smooth, tear-free images by synchronizing display frame rate to GPU (require displays that support G-Sync). An array of NVIDIA GTX technologies are also built in, like GPU Boost, NVIDIA 3D Vision, NVIDIA Surround, NVIDIA PhysX, and GeForce Experience. These technologies together provide the most immersive and competitive gaming experience possible.
This MSI card is overclocked to 1102/1190MHz Base/Boost Clock (1000/1075MHz for the reference card), and boasts Military Class 4 Components for unbeatable durability and reliability. An evolved cooling solution featuring two large fans enables efficient and quiet cooling even at high loading level.
Superb Cooling The Armor 2X thermal design incorporates two large fans and huge heat sink for better heat dissipation, while the Propeller Blade technology produces 20% more airflow and less noise than traditional fan design. Airflow Control technology deflects more airflow to heatpipes and improves airflow efficiency, which achieves the effect of increasing the heatsink area to enhance dissipation efficiency.
Military Class 4 Components Certified to MIL-STD-810G standards, this card is built with Hi-c CAP, Solid CAP, and New SFC components for top stability, durability and quality you can count on in the harshest gaming environments.
Multi-Frame sampled Anti-Aliasing (MFAA) With the launch of Maxwell the Multi-Frame Anti-Aliasing (MFAA) is introduced to the world. This new, Maxwell-exclusive anti-aliasing technique improves edge quality with a minimal performance cost, and helps you enjoy anti-aliased games at ultra high resolutions like 4K.
Dynamic Super Resolution (DSR) Dynamic Super Resolution renders a game at a higher, more detailed resolution and intelligently shrinks the result back down to the resolution of your monitor, giving you 4K-quality graphics on an HD screen.
Voxel Global Illumination (VXGI) Voxel Global Illumination (VXGI) is a stunning advancement, delivering incredibly realistic lighting, shading and reflections to next-generation games and game engines.
NVIDIA G-SYNC NVIDIA G-SYNC is a groundbreaking new display technology that delivers the smoothest and fastest gaming experience ever. G-SYNC's revolutionary performance is achieved by synchronizing display refresh rates to the GPU in your GeForce GTX-powered PC, eliminating screen tearing and minimizing display stutter and input lag. The result: scenes appear instantly, objects look sharper, and gameplay is super smooth, giving you a stunning visual experience and a serious competitive edge.
Other Thoughts: I would always recommend MSI GPUs and MBs.
Yet to let me down.
Thank you for sharing your experience with the MSI product! Glad to hear you are still enjoying your card! We certainly appreciate your patronage over the years! If you ever require technical assistance, please feel free to contact our technical support department directly at 626.271.1004 from M to F, 9am to 6pm Pacific Standard Time.
Pros: Fans aren't too loud.
Made out of durable material.
Cons: Runs REALLY hot, under full load with fans at 100% and not overclocked the temperatures reach 91C, doesn't look like the card will last long.
Doesn't have a back plate.
Other Thoughts: I wouldn't recommend this particular model from MSI, if you are a buyer look for other models like the gaming 6g one.
Thank your for sharing you experience with us. We always value your feedback, and I'd like to apologize for your unfortunate experience with the GPU. This model by default does not come with a backplate. A backplate does not necessarily improve GPU cooling; temperatures are mostly dependent on the GPU's heatpipes in conjunction with its fans. Does your setup have the proper amount of ventilation required to help move and exhaust air out of the case? I would recommend checking your case to see if the fans are balanced and/or are inducing unwanted anomalies that do not properly cool the card.
Additionally, I'd highly advise you to call our tech support department, as they are much more qualified to provide you with answers that will help you maximize the cooling on the 980 Ti. You can reach them through phone or email.
I want a second one!
Cons: This card loses performance in games that do not support DX11 or DX12. DX9 games are most notorious for the apparent lack of performance.
Bare that in mind when you use this card.
Note: This card doesn't overclock particularly well, but it will overclock.
Other Thoughts: Now if you want to see what makes this card so Beast; Go play Star Citizen.
Very High Detail: FPS does change and is noticeable.
Other Advantages: I can now stream SC on twitch at 60FPS @1280x720 with near perfect fluidity; What you see on my twitch stream is how smooth the game feels to me.
I am working on tweaking my broadcaster to stream 1920x1080 @ 60fps on twitch. It is possible with this card.
Do not try to run Haeven Benchmarks; The score you will get is unrealistic because the software doesn't support Quad Core CPU's It only supports Dual Cores.
Some software is designed to use a certain number of cores, the rest are just wasted on benchmark tests.
Star Citizen for example will support Hexa Core: So SC is a good benchmark to make for the newer Haswell Cores.
Update: 08/11/2016 - I recently upgraded the cooling solution for this card. This card doesn't break a sweat even when overclocked. And when I say it doesn't break a sweat I mean that literally.
Liquid cooled with NZXT Kraken G10 + NZXT Kraken X31 Liquid cooler (Upgraded radiator fan to 117CFM fans - THIS MADE A HUGE temp difference) When I was running on air cooled 30-40 degrees C idle, and up to 80 degrees C overclocked. Only downside to the Kraken upgrade. It's a HUGE pain to install. If you plan to liquid cool 2 cards. Do them at the same time, not at different times. You'll understand why when you install the Kraken G10.
I do have one question to Newegg: Why are you selling this card for $820? When I bought mine for $650, and people can get 1070's and 1080's for $400-$700 with 5-20% more performance? You are crazy if you buy this card from newegg for $800. It's a good card, but not worth $800 especially now that the 1070 and 1080's are out.
Pros: This card is a beast! 2k 1440Hz ultra everything 60+ fps! Runs hot like a performance card should!
Other Thoughts: If you have a well cooled rig this will run around 72° even though Nvidia approves it up to 105°.
Pros: It's a 980 Ti... It's good.
OC out of the box is nice.
Cons: Runs a bit hot, like everyone else has mentioned. I found the best way to work around this is to use MSI Afterburner and set up custom fan speed curves.
It's a bit loud when the fan is running at higher speeds, but if you have a headset/earbuds/surround speakers, it's not noticable at all.
Other Thoughts: 9.5/10 would recommend.
Oh, and I was upgrading my old 760, and I broke the PCIe 3.0 slot somehow, but I threw it into a PCIe 2.0 slot and it works just fine. If this happens to anyone else, don't worry, I did research and apparently it will only cut performance by a few percent, but that's even only for high end games and benchmark tests.
Cons: Temps get to around 77C in benchmarks but, never go above 72C while gaming.
Other Thoughts: Very large cooler, make sure your case has the clearance to fit the card.
Pros: Excellent product from M.S.I ,The fans are super quite cant hear them even on full load gaming,runs everything buttery smooth,coming from 2 670's in sli this single card beats those hands down,noticed stuttering in tom clancy's the division then i popped this bad boy in and no hiccups at all,absolutely amazing card.
Cons: none,runs great and is very quite cant hear it over my case fans!
Other Thoughts: I was going to opt for an evga card but i seen the price of this and had to jump on it,paid less and have about the same core clock as the ftw,i would definitely recommend this for gamers. also the white goes great with my corsair air 540 snow edition!


Mats Wilander believes conditions will need to be perfect for Roger Federer is to add to his record-breaking haul of 20 Grand Slam titles, as he suggests the 37-year-old legend is a fading force at the top of the game.
Federer's defence of his Australian Open crown ended with a shock defeat against rising Greek star Stefanos Tsitsipas, with Wilander telling Tennis365.com that the Swiss maestro is now starting to show signs of weakness at the back end of his career.
"We thought this was going to be the best chance for him to win a major until we saw how slow the balls are out here and they've made a huge difference," Wilander told us. "He needs the perfect conditions to be a threat to win a Slam, and that could be at Wimbledon, or could be at the US Open, but to me it's all starting to depend on conditions.
Wilander believes Federer will try to continue playing through to next year's Olympics, as he backed his decision to play the clay-court season in 2019 after skipping that section of the calendar for the last few years.
"Not playing on clay last year was an obvious mistake because he lost at Wimbledon in a tight match and would've benefitted from more match practice," adds Wilander.
"2020 is a season that's definitely on his calendar because of the Olympics and I think he's building now to be at his very best in the only event he hasn't won. I think playing the clay season is more about being better and improving.
Meanwhile, Federer is set for a big rankings drop after his early exit at the Australian Open, with the former world No 1 set to be ranked down in sixth place when the ATP rankings are released next Monday.


text:
Capital Swim and Culture
View of Wellington from the balcony where I stayed
It's the annual swim meet between TAMS and Different Strokes (DSW). I've got my friend Ros staying so we take a break from walking around Waiheke and fly to Wellington. I'm staying with Rangimoana and Bill who live within walking distance of Freyberg Pool which juts out into the harbour from the trendy Oriental Parade. It is here that we will race. My hosts have a newly acquired pet called Caf – a long haired Chihuahua which Bill refers to as 'Pussy Dog'. Also staying is a guy called Hans – an interesting Kiwi Bear who now lives in Berlin doing a drag act called Princez Hans. As it's Friday night it's fish and chips for tea. The shop has got the order wrong and there's too much to eat. Never mind, I'm supposed to 'carb up' for tomorrow.
Pussy Dog
Ros has friends to visit nearby and also wants to watch the swimming so I collect her in the morning and we walk to the pool, pausing to have coffee on the sea-front. Once again, I've forgotten which events I've entered in this 33.3 metre pool and as we've got team relays which count towards the challenge cup, I think that 6 events might be too much. What to scratch? The warm up decides it as I finish off with a long length of Butterfly, I think 'Nah, not going to do the 133m Individual Medley.'
Freyberg Pool Oriental Bay
The next shock is that I'm in heat 4 of 4 for the 67m freestyle and heat one of the next event, 100m Backstroke. All I have to do is change from lane 3 to land 2. I hold back on the freestyle but it's still a struggle doing the backstroke. Kevin from my club and a few years younger just beats me by a second. It's his first race of the day. My family have also come to watch and cheer. Cousin Marie and partner Anne then later my niece with her partner and 8 month son. It's quite an occasion and with only 42 competitors, it all goes very quickly. There's last minute reorganisation of the relays and I move from the Mixed Medley to the 4x67m freestyle relay plus the 15×33.3m relay which counts towards the Challenge Cup. There are some very young, fast and beautiful guys from Australia who've flown in for this meet – they are mostly from a Melbourne gay club called the Glamourhead Sharks. They actually win the relay but as it's between Wellington and Auckland clubs, that doesn't count. The Wellington swimmers are younger and faster than us and we have to graciously admit defeat and in the process get to hug and kiss everyone. There are no medals or age group categories, it's just a fun swim, but I do take note of my times.
Wellington is experiencing a heat wave and the harbour is looking great. When it's like this, Wellington harbour is one of the best and when it's cold windy and wet, one of the worst. I go off with friend and family to a harbour-side bar for a beer and a meal. There's a play on at the National Museum, Te Papa, and Anne has nipped out during the swimming to get tickets for us. Helen Pearse-Otane's play 'The Ragged' is the first of a quartet following the life of a family from the 1840 new British Colony at Port Nicholson, Wellington through the years and into the future. This is the early days of the New Zealand Company, acting independently of the British Government and headed by the unscrupulous Edward Gibbon Wakefield. An illiterate immigrant (Samuel) from Manchester, England, has paid for land. When he rows out of the harbour to Te Miti on the south coast he finds that the local tribe has not sold it. He decides to stay and is adopted by the chief. There is a Pakeha (white man) slave who is trouble and following the murders of the chief's sons and grandchildren, the slave is served up to the self-important dignitaries from the new settlement. Samuel marries the chief's daughter-in-law and thus a new family is begun and he has his land after all. It's a powerful piece of work, beautifully spoken in Maori and English. Veteran director and actor Jim Moriarty has directed strongly and also plays the ailing Maori chief. At the end, we are invited to comment on the experience. After several enthusiastic replies, my friend Rangimoana, who works at the museum, gets up and does a long speech in Maori, congratulating and acknowledging each performer and their origins. I get the gist and everyone is moved by his powerful oration. We take Ros back with us for wine and talk, into the night.
Oriental Bay
Sunday is the day of the harbour swim, which also happens in Oriental Bay. Some of my team-mates are doing this 3K event and I go down to see if I can spot any of them. I arrive as the last few swimmers are emerging from the sea but I've missed most of it. I do, however, manage to meet up with Coach Cynthia and others who have swum. After the prize – giving we go to the same harbour-side bar for beer and food. I'm off for the rest of the weekend to spend some time with my great nephew, George. Ros and I meet up on Monday in the departure lounge of Wellington Airport for our return flight to Auckland.
Author Christopher PrestonPosted on February 11, 2015 Categories All, Swimming
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redpajama_set_name: RedPajamaCommonCrawl


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Elizabeth b. 1614
October 26, 2019 October 26, 2019 / Kirstin Miller
Elizabeth 10th great grandma on RootsMagic tree.
Elizabeth was born in about 1614, her last name isn't known. In 1630 she married Thomas Judd. The birth and marriage places of Elizabeth and Thomas aren't known. The Judds were part of the 1620-1640 'Great Migration' documented by genealogist Robert Charles Anderson, he's written volumes. Elizabeth and family left England for America in 1634, this is shown in a land grant in Cambridge on August 4, 1634, "lots granted in Westend, To Tho: Judd 4 Ackrs". Elizabeth and family stayed in the Massachusetts colony for awhile, then with Reverend Thomas Hooker, left to start a new settlement in the Connecticut Colony, fort called New Towne then called Hartford. Elizabeth and Thomas had 9 children, 3 daughters and 6 sons. Two daughters married Loomis brothers. two sons and one daughter married a Steele sibling, one son married Mary Howkins. One son, Benjamin married Mary Lewis, their 7th great grandson was Faber Miller.
At Kenyon Street AngelFire website, an annotated map.
The single available record with Elizabeth's name is Dr John Winthrop's medical notebook, "8 July 1669, John Winthrop Jr. treated -Jud Elis[abeth] above 60 years wife of [blank] Jud Senior of Farmington-". Elizabeth lived until about 1678. Her burial place is unknown.
The Records of the town of Cambridge (formerly Newtowne) Massachusetts. 1630-1703. Cambridge, 1901, Page 9.
Great migration, immigrants to New England 1634-1635, Page 121-126 Thomas Judd at American Ancestors.
Original distribution of the lands in Hartford among the settlers, 1639, Page 549, 550, page 320.
Kenyon Street an AngelFire website last updated in 2013. The annotated map shadows Thomas and Elizabeth's land top left next to the Charter Oak. This map was made by a neighborhood association, people living in the area. Mark Twain's house is part of the neighborhood.
Hannah Judd b. 1681
April 19, 2019 / Kirstin Miller
Hannah Judd 8th great grandma on RootsMagic tree
Hannah was a middle child of Benjamin and Mary Lewis Judd. She was born, then baptized on March 13, 1681 in Farmington, Connecticut.
Marriage of Gershom and Hannah, children's births
On May 4 1710 Hannah married Gershom Smith he was from nearby Glastonbury, Connecticut. Gershom and Hannah's records show only 2 kids: Hannah the oldest and Gershom a son who died at 16. The Smith family stayed in Glastonbury. Gershom died in 1747 at age 67 and is buried there. Widow Smith (Hannah) moved to her daughter's home. Hannah's daughter, also Hannah, was married to Richard Risley and living in Tolland, Connecticut about 30 miles northeast.
Hannah is on a land record dated 1756: "On 3 Nov. 1756, Richard and Hannah Risiey, with Widow Hannah Smith, all of Glastonbury, sold land where said Risley now dwells". Hannah was 75. There's no record of her death or burial. Husband Gershom has a headstone at Glastonbury, Hannah may be buried there or may be buried in Tolland with her daughter's Risley family.
At American Ancestors. The American genealogist volume 25 page 130. New England marriages to 1700 database Volume 2 page 1391.
At FamilySearch.org. Connecticut Marriages, 1640-1939 FHL 001316154 Digital Folder 007730404 Image 00087 (87 of 784) marriage of Gershom and Hannah, children's births
Thomas Judd b. 1608
January 18, 2019 January 18, 2019 / Kirstin Miller
Thomas Judd 10th great grandfather on RootsMagic tree.
Thomas Judd was born in England, about 1608. He landed in Cambridge, Massachusetts Bay in 1634. In Cambridge he was admitted to the church and made a freeman in May of 1636. Thomas left Cambridge for Connecticut, first Hartford, probably with Puritan minister Thomas Hooker. Both Hooker and Thomas Judd are landowners on a map of Hartford in the 1640s, these would be original settlers. The map was "prepared from the original records by vote of the town" and created in the 1800s. Thomas Judd is in the bottom left corner No. 154. Other Miller family landowners on this map in this same area in 1640 are Jeremy Adams, Thomas Bliss and Richard Risley.
UCONN libraries provides a digital copy of this map and details.
A list of the landowners 'freeholders' here
More details on the map here.
Thomas left Hartford for Farmington where he held lots of town service positions including, in August of 1658 "to communicate the mind of the court to the Indians". Church records of Farmington, Connecticut name Thomas as the second Deacon of the church. "The number of such as are in full communion in the church in Farmington March 1 1679/80. Deacon Judd. Benjamin Judd and his wife. John Judd and his wife. William Judd and his wife". Finally Thomas moved to Northampton, Massachusetts where he is buried and has a headstone credited to a descendant: Sylvester Judd of 1858.
He didn't leave a will at his death but there is a probate record, 15 pages, handwritten mostly land deeds and an inventory.
This page from the will lists the children and what they receive, Benjamin Judd 4th on the list is the Miller ancestor through Mary Ella Gaines, grandma of Faber Miller who married Gladys Cable.
Connecticut, Wills and Probate Records, 1609-1999 at Ancestry . com
Find a grave https://www.findagrave.com/memorial/28130545
Church Records of Farmington in Connecticut The New England historical and genealogical register, 1874 Volume 28 at Archive.org


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Country profile: Guinea-Bissau
One of world's poorest and least politically stable nations, Guinea-Bissau also a staging post for Europe-bound cocaine.
13 Apr 2012 08:24 GMT
After taking power in a 1980 coup, Joao Bernardo Vieira was Guinea-Bissau's off-and-on president for 29 years [EPA]
Reports of an attempted coup in Guinea-Bissau mark the latest chapter in a turbulent political history punctuated by military power grabs and the assassination of the country's longest ruling president.
In addition to frequent upheaval, the small West African nation is considered one of the world's poorest nations with a GDP per capita of $1,100, according to the CIA World Factbook, and suffers low literacy rates and a severe lack of access to healthcare.
Owing to its political and economic weakness and favourable geography, which includes an archipelago of Atlantic islands, Guinea-Bissau has also become an important hub for Europe-bound drugs from Latin America, with cocaine believed to be the country's most lucrative trade.
After centuries as a Portuguese colony, Guinea-Bissau gained independence in 1974 following a more than 15-year revolution against Lisbon's rule led by the African Party for the Independence of Guinea and Cape Verde (PAIGC).
Luis Cabral, whose half-brother had founded and led PAIGC until his assassination in 1973, became the country's first president, but he was toppled in a 1980 military coup led by Joao Bernardo Vieira.
Plots and purges
Vieira's rule was characterised by suppression of opposition and purges of rivals, according to the CIA World Factbook, including the thwarting of alleged coup plots in 1983, 1984 and 1993.
Moves towards democracy saw the country stage elections for the first time in 1994, with Vieira elected to the presidency.
But an army uprising triggered a civil war in 1998, with Vieira removed from power by the military a year later.
Another military coup, this one bloodless, took place in 2003, and the military allowed legislative elections to be held seven months later in 2004, although a series of military mutinies delayed presidential elections scheduled for later in the year.
Presidential elections were finally held in June 2005, with Vieira voted back into power. But he was shot dead by soldiers in a 2009 attack, shortly after a bomb blast had killed the head of the country's military.
Elections in 2009 saw Malam Bacai Sanha, a former interim president during the country's civil war, elected to the presidency, but Sanha died in a Paris hospital due to what are believed to be diabetes-related problems in January this year.
Army chiefs said in December 2011, while Sanha was in Paris, that they had thwarted an attempted coup , arresting the country's navy chief and dozens of soldiers accused of stockpiling weapons.
Sanha's death prompted early elections in March, with a runoff vote between outgoing prime minister Carlos Gomes Junior and former president Kumba Yala set for the end of April.
Drugs hub
Leaked US diplomatic cables released by WikiLeaks in 2011 highlighted Guinea-Bissau's burgeoning reputation as a hub for Latin American drug producers seeking access to European markets.
Cartels identified the country's uninhabited islands as a prime place to land small planes loaded with drugs, which are then transported to Europe in smaller quantities.
Traffickers, according to analysts, had bought off key members of the government and the military, creating a "narcostate".
According to a 2007 cable written by the US embassy in Guinea-Bissau entitled "No confidence in government, high confidence in drugs", politicians were routinely "purchased with drug money".
Helder Proenca, Guinea- Bissau's defence minister at the time, was "widely believed to be a drug kingpin", the cable said.
Proenca was subsequently alleged to have been part of the 2009 coup plot that killed Vieira and was shot dead by soldiers later in the same year.


Many plants produce fruits in order to help in the dispersal of their seeds. It would be pointless for a plant to drop all its seeds directly beneath, by its roots, since all the seedlings would then compete with each other and their parent. Plants have therefore evolved various mechanisms by which their seeds can be dispersed. Some plants, such as sycamore, have dry spiky or leathery fruits which may be dispersed by the wind, others such as burdock have barbs that attach themselves to animals' fur or our clothing, but the fruits that we are interested in here are those fleshy fruits which the plant has made attractive for animals to eat.
As the seeds mature, the ovary and sometimes also the receptacle swell up around the seeds to become a juicy, fleshy and often sweet fruit. The seeds inside usually have a tough outer coat so that when the animal eats the fruit it will either, if the seeds are small, deposit the seeds unharmed in its faeces some distance from the parent plant e.g. the pips in soft fruit, or if the seed is large, as in plums, the fruit may be carried away and the seed dropped somewhere. Normally the fruit tastes awful before the seeds are ripe, so it will not get eaten before the seeds are mature enough to germinate. Seeds and fruit therefore ripen at the same time to ensure that the seeds are at the right stage when taken by an animal. Either way the animal has "paid" for its nice meal by doing the plant a favour in dispersing it seeds.
One of the wonderful things about fruits is that we are actually eating something offered to us by the plant. Assuming that we are living a fairly natural lifestyle -i.e. not eating imported fruit that could never grow in this climate, eating fresh fruit that is in season, not eating seedless varieties, not throwing fruit seeds or cores into the rubbish bin where they have no chance of re-entering the natural cycle - then you could say that the plant would want us to eat the fruit. In turn the plant has done us a favour by producing this nice juicy, tasty morsel full of naturally filtered water, natural sugars, flavours, vitamins and minerals and other nutrients. Fruits are amongst the most delicious foods on Earth. Most are light and easy to digest and they are a natural convenience food because there is normally no preparation needed.
Most fruits are very easy to digest because food is stored in the fruit as sugars. (Unripe fruits are often high in starch and organic acids, which makes them taste very sour, and these are converted to sugars only as the fruit ripens). Most fruits contain no fats at all though some, such as olives and avocados have quite a high fat content. Whilst it is not really feasible to grow these fruits in temperate areas, we can grow Elaeagnus and Hippophae berries which contain Essential Fatty Acids (EFA's).
Most fruits also contain no protein. Protein and fats are present in the seed but this is usually not eaten (e.g. in plums) or passes straight through (pips in berries) so fruits are NOT a complete food and could not really be recommended as the sole part of the diet. Also, unless you eat Special Berries (see below) all year round, you will go quite short of minerals. Most fruits are particularly low in calcium and magnesium which are necessary for healthy teeth and bones.
Fruits are also very low in zinc which is essential for the function of the immune system, the formation of skin, the healing of wounds, brain function and it is essential for the function of the reproductive organs. But do not despair, this deficiency can be made up by eating plenty of green leaves (see the section 'Green Gold' ).
Fruits are best eaten fresh, ripe and raw. There is normally no need to cook them - unless you have a glut and want to preserve some by bottling it (which can be done without sugar if your jar has a good seal), or if there are windfall apples that are not ripe enough to eat raw. But it is preferable if cooked fruit only forms a small part of the diet, as it is so much healthier and more satisfying to eat them raw.
A word of warning about fruits for those who are not use to eating much of it: Like leaves, fruits are a very strongly cleansing food, capable of stirring up and removing toxins that were long buried in the body. (These toxins come from certain foods, especially "junk food" with chemical additives, an excess of meat, dairy products etc. and environmental toxins). However unlike leaves, which actually help to carry toxins out of the body as well, fruits often tend to stir them up without carrying them away. So if you know that your diet has not been all it could have been over the last few years and that your system is liable to be rather toxic, then please approach fruit with caution.
Do not suddenly eat a lot of fruit in one go (especially of the more acid berries) until you know that your system can cope with it. What is liable to happen is that a lot of toxins will be stirred up into what is known as a Healing Crisis and you will feel ill. Perhaps you will blame the healthy diet for making you ill and perhaps even go to the doctor for some pills. These will only suppress the symptoms and drive the toxins deeper into the body, as well as adding more toxins, -so you will be worse off than before. So unless you understand about healing crises and know how to manage them, please go easy on the fruit if you are not used to it. Many Naturopaths have used short-term fruit diets for healing with a great deal of success. This is because of the wonderful cleansing and healing properties of fruits, especially if accompanied by green leaves or leaf juices. However this tends to be too drastic for most people nowadays because of the general decrease in health and nutrition of the overall population. This is still possible to do but we would only recommend it under the guidance of a qualified Naturopath or Nutritional Therapist.
Fruits contain quite good levels of vitamins but are quite low in many important minerals. On their own, they will not provide you with enough minerals. Indeed, if you just ate fruit and nothing else for a while, you might lose all your teeth. So perhaps it would be all right to eat a lot of fruit as long as one eats a lot of leaves as well? If you study the tables of mineral composition of various different fruits, three main groups emerge.
1). The large cultivated fruits such as apples, pears and tomatoes.
Although these form the bulk of most people's fruit intake, they are pretty low in minerals and vitamins. They have been selectively bred, sometimes over thousands of years, in order to produce a bigger size, milder flavour, more sugar and higher water content. Unfortunately the nutrient content has sadly lost out. Hence the wild crab apples and pears - though often inedible due to their acidity and astringency - are actually more nutritious than the cultivated ones. A clear example of this can be seen if we look at the tomato. The wild tomato is no larger than a cherry, yet a single fruit contains more vitamins and minerals than a single much larger cultivated tomato. When comparing the size of each fruit you begin to realise just how much nutrient we have lost from many of our cultivated foods!
2). Small berries such as strawberries, raspberries, blackberries, blackcurrants and gooseberries.
keep you regular!) and lower in calories.
These are normally wild plants that have not been bred up and hence not been devitalised and demineralised in that way. They are much richer in nutrients than the other fruits and also often have great medicinal value. A few examples are mentioned below: -.
Rosehips. Whilst wild rosehips are extremely nutritious, most are small and fiddly to harvest and use because the layer of flesh covering the hairy seeds is very thin. However, Rosa rugosa has particularly large hips which can be eaten much more easily (and certain varieties have larger fruits than others). Another good source of vitamin C, it also contains some EFA's and is also very high in carotene or provitamin A, the precursor of vitamin A. While true vitamin A or retinol, which is only found in animal products, is toxic in large amounts, carotene is found in plant foods and is totally non-toxic. The body only makes as much vitamin A from it as it needs. Ripe Rosa rugosa fruits are also extremely delicious - true ambrosia!
Hawthorns. Whilst the fruits of the British two native hawthorns are edible few people are going to want to eat many of them. However, there are several species with cherry-size fruits that are absolutely delicious to eat. Again these fruits are high in bioflavonoids and fairly high in vitamin C. They also have several useful medicinal properties including making a good heart tonic whilst reducing high blood pressure and cholesterol levels in the blood. See our leaflet on the hawthorns for more information.
Barberries (Berberis). These fruits usually have a rather acid flavour, but are pleasant nonetheless and go very well in muesli. They have a particularly beneficial effect on the urinary system, that they cleanse and stimulate.
It is particularly important that we take in vitamin C because so much of our food is cooked nowadays and thus largely devoid of vitamin C. Also, during evolution we lost the ability to manufacture Vitamin C in our own bodies so we are at a disadvantage to most other animals that can. This is one of the reasons why we need to eat lots of fresh raw food. Vitamin C is also useful in the treatment of many diseases ranging from colds to cancer.
Elaeagnus and Hippophae are very rich in bioflavonoids, which are beneficial plant pigments that assist in the utilisation of vitamin C and are said to improve the function of the brain. Bioflavonoids are also very beneficial to our health in many other ways. For example, the bioflavonoids found in the fruits of all Elaeagnus and Hippophae species have been shown to be an effective preventative for and treatment of cancer.
EFA's are essential for the formation of healthy cell membranes and necessary for many body functions especially that of the immune system. Hence these fruits are extremely beneficial, but are of particular benefit for those with diseases of the immune system such as MS, ME, rheumatoid arthritis, juvenile onset diabetes, cancer and AIDS. This is especially true of the Hippophae. Considering also that high levels of vitamin C, B vitamins and minerals are also beneficial to the immune system, these berries are a particularly good food, tonic and medicine for anyone with an under functioning immune system.
So the general conclusion is that all fruit is good for you but some are more nutritious than others. They are not as nutritious as leaves, but they do taste better because the plant has deliberately laced the fruit with sugars and pleasant flavours to tempt us.
develop their full flavour, sweetness or nutritional value because the plant hasn't had enough time to put all it wanted to in the fruit. Fruit that has been transported halfway across the globe is normally picked under ripe and sprayed to stop it rotting. It is also not in harmony with the climatic zone in which we live so is much less good for us, as well as contributing to unnecessary pollution in the transportation process. The ideal way to eat fruit is to eat it straight from the plant. Once it starts to age it loses its nutritional value in the same way as other parts of the plant do.
With careful planning it is possible to pick fresh fruit from your garden for 10 or even eleven months of the year. The season starts with the evergreen Elaeagnus in early April. These will continue to crop until the middle or end of May when Mahonias and the first strawberries ripen. The variety of fruits then increases considerably with the greatest abundance coming in late summer and early autumn. There are, however, many different fruits that can be harvested in late autumn and early winter. These include the Hippophae, which can be harvested until the middle of January, and various Gaultherias, which can be picked in early spring. However, these late harvests are usually quite small in quantity - more a nibble than a real meal.
The apple is the best example for this climate. There are many late-ripening varieties that will keep for varying amounts of time in storage, the latest storing for a whole year or more in good condition. These late keeping apples are normally not actually ready to be eaten when they are harvested. When in storage, the starches and acids in the fruit are gradually converted to sugars and the fruit becomes ripe over a period of weeks or months. If the late apples were allowed to ripen fully on the tree they would get damaged by the weather and eaten by the birds before they ripened properly, so they would not store at all. Whilst not the most nutritious fruit, they are very good for ensuring a year round supply of fresh fruit in this climate.
Certain pear varieties will keep for several months in a similar way to apples though each variety ripens over a much shorter period so you can end up with a sudden glut of fruit which will quickly go over-ripe and then rot if they are not eaten in time.
Contrary to popular opinion, kiwi fruits can be grown very successfully in much of Britain. If the fruits are picked in November, they will keep in a cool store for several months. Kiwis are actually very nutritious, being very high in vitamin C and also good sources of carotene and folic acid (because of their green colour).
Golden berries are closely related to tomatoes, but have a much richer flavour. Any green fruits left on the plant when the first frosts come along can be stored in a cool place and will ripen slowly over the next few months.
The bletted fruits. Certain fruits, such as Mespilus germanica (the medlar) and Sorbus domestica, (the service tree) do not ripen fully in this climate. They are picked in late autumn and will then ripen slowly in store. They do not become edible raw until they are almost at the point of going rotten (a process known as bletting), but when at this stage taste like a delicious tropical fruit. However, you do need to be careful because if you overdo the bletting the fruits will ferment in your stomach and cause digestive problems.
Therefore with the summer and autumn berries, and stored apples (and possibly pears and kiwis) over the winter and spring, it is very possible to be totally self sufficient in fruit in this climate.
Fruits are amongst the most delectable and delicious foods on Earth, and unlike other foods, are actually deliberately produced by the plant to be eaten. This is not pure altruism on their behalf, but symbiosis or co-operation, because in return they want their seeds to be dispersed. They are also the ideal healthy convenience or snack food because you can just pick them up and eat them - no preparation is usually needed - just make sure they are ripe.
Most conventional shop bought fruit is a reasonable source of vitamins, depending on how fresh it is, but is not very rich in minerals. The ideal suppliers of vitamins and minerals are the green leaves (See Green Gold leaflet).
The small wild and semi wild berries are far more nutrient-rich because they are closer to their natural state and have not had the nutrients bred out of them. Some berries such as Elaeagnus, Hippophae, Rosa Rugosa and Crataegus are particularly nutrient-rich and have numerous beneficial health giving properties. It is a good idea to include more berries in your diet, especially the Special Berries. The easiest and best way to do this is to grow them yourself because this ensures their availability, freshness and enables you to pick them at their best (assuming the birds don't get there first!) (See leaflet on alternative fruits for more details).
While being a very enjoyable and healthy food, please bear in mind that fruits are far from being a complete food (unless you are a Fruit Bat!) so do not rely on them too heavily - or your teeth might fall out! (this is particularly true of the imbalanced commercial fruit).
In most fruits food is stored in the form of sugars so they are a very light and easy to digest food. Digestive problems will respond quite well to a temporary diet of fruit to give it a rest from heavy foods.
A few fruits do contain fats.
Most fruits have no or virtually no protein. This is stored in their seeds.
Fruits are very cleansing and detoxifying and in this way are very helpful in the treatment of diseases. If you have a cold, flu or other infection, a fruit (and leaf) diet will clear it out of you faster. They are also very helpful in the treatment of chronic diseases. But unless you are absolutely sure of what you are doing, and can cope with the possibility of a flare up (Healing Crisis) without going to the doctor for suppressive drugs, then using fruit for healing purposes must be done under the supervision of a qualified practitioner.
Well, I hope this leaflet has given you some new insights into fruit and that you will consider growing and eating more of these wonderful foods.
The database has more details on these plants: Elaeagnus x ebbingei, Fragaria x ananassa, Hippophae salicifolia, Malus domestica, Mespilus germanica, Pyrus communis sativa, Ribes nigrum, Ribes uva crispa, Rosa rugosa, Rubus idaeus, Sorbus domestica.


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