The effects of short-term, progressive exercise training on disease activity in smouldering multiple myeloma and monoclonal gammopathy of undetermined significance: a single-arm pilot study.

BACKGROUND: High levels of physical activity are associated with reduced risk of the blood cancer multiple myeloma (MM). MM is preceded by the asymptomatic stages of monoclonal gammopathy of undetermined significance (MGUS) and smouldering multiple myeloma (SMM) which are clinically managed by watchful waiting. A case study (N = 1) of a former elite athlete aged 44 years previously indicated that a multi-modal exercise programme reversed SMM disease activity. To build from this prior case study, the present pilot study firstly examined if short-term exercise training was feasible and safe for a group of MGUS and SMM patients, and secondly investigated the effects on MGUS/SMM disease activity. METHODS: In this single-arm pilot study, N = 20 participants diagnosed with MGUS or SMM were allocated to receive a 16-week progressive exercise programme. Primary outcome measures were feasibility and safety. Secondary outcomes were pre- to post-exercise training changes to blood biomarkers of MGUS and SMM disease activity- monoclonal (M)-protein and free light chains (FLC)- plus cardiorespiratory and functional fitness, body composition, quality of life, blood immunophenotype, and blood biomarkers of inflammation. RESULTS: Fifteen (3 MGUS and 12 SMM) participants completed the exercise programme. Adherence was 91 ± 11%. Compliance was 75 ± 25% overall, with a notable decline in compliance at intensities > 70% V̇O2PEAK. There were no serious adverse events. There were no changes to M-protein (0.0 ± 1.0 g/L, P =.903), involved FLC (+ 1.8 ± 16.8 mg/L, P =.839), or FLC difference (+ 0.2 ± 15.6 mg/L, P =.946) from pre- to post-exercise training. There were pre- to post-exercise training improvements to diastolic blood pressure (- 3 ± 5 mmHg, P =.033), sit-to-stand test performance (+ 5 ± 5 repetitions, P =.002), and energy/fatigue scores (+ 10 ± 15%, P =.026). Other secondary outcomes were unchanged. CONCLUSIONS: A 16-week progressive exercise programme was feasible and safe, but did not reverse MGUS/SMM disease activity, contrasting a prior case study showing that five years of exercise training reversed SMM in a 44-year-old former athlete. Longer exercise interventions should be explored in a group of MGUS/SMM patients, with measurements of disease biomarkers, along with rates of disease progression (i.e., MGUS/SMM to MM). REGISTRATION: https://www.isrctn.com/ISRCTN65527208 (14/05/2018).

Exercise-induced amplification of mitogen-stimulated oxidative burst in whole blood is strongly influenced by neutrophil counts during and following exercise.

This study characterized the effect of moderate- or vigorous-intensity exercise on leukocyte counts, using fingertip sampling, and mitogen-stimulated oxidative burst, measured in whole blood with a point-of-care test. In a randomized crossover design, 13 healthy adults (mean ± SD age: 22 ± 2 years; seven male, six female) cycled for 30-min, once at 52 ± 5% V˙ O2peak and on another occasion at 74 ± 9% V˙ O2peak . Blood was sampled at baseline, immediately post-exercise, and 15- and 60-min post-exercise. The leukocyte differential and mitogen-stimulated Reactive Oxygen Species (ROS) production were assessed. Lymphocytes increased immediately post-exercise and decreased below pre-exercise levels 15- and 60-min later. Lymphocyte mobilization immediately post-exercise was 59 ± 36% greater with vigorous- compared to moderate-intensity exercise (p < 0.01). Neutrophils increased immediately after exercise (38 ± 19%, p < 0.01) remaining elevated 60-min later (50 ± 34%, p < 0.01; averaged across intensities) and did not differ between intensities (p = 0.259). Mitogen-stimulated ROS production was amplified immediately (+32 ± 37%, p < 0.01) and 60-min post-exercise (+56 ± 57%, p < 0.01; averaged across intensities) compared to rest and did not differ with intensity (p = 0.739). Exercise-induced amplification of ROS production was abolished when correcting for neutrophil, monocyte and platelet counts and correlated most strongly with neutrophil mobilization immediately (r = 0.709, p < 0.01) and 60-min after vigorous exercise (r = 0.687, p < 0.01). Leukocyte kinetics can be assessed using fingertip blood sampling in exercise settings. Exercise-induced amplification of oxidative burst is detectable with a point-of-care test, but results are strongly influenced by neutrophil counts, which may not be routinely quantified.

Oocyte activation and passage through the metaphase/anaphase transition of the meiotic cell cycle is blocked in clams by inhibitors of HMG-CoA reductase activity.

Cell cycle progression for postembryonic cells requires the activity of 3-hydroxy-3-methylglutaryl coenzyme A reductase (HMG-R), the enzyme which catalyzes the production of the isoprenoid precursor, mevalonate. In this study, we examine the requirements of HMG-R activity for cell cycle progression during the meiotic and early mitotic divisions using oocytes and dividing embryos from the surf clam, Spisula solidissima. Using two different inhibitors of HMG-R, we find that the activity of this enzyme appears to be required at three distinct points of the cell cycle during meiosis. Depending on the stage at which these inhibitors are added to synchronous clam cultures, a reversible cell cycle block is triggered at the time of activation or at metaphase of either meiosis I or II, whereas there is not block to the mitotic cell cycle. Inhibition of HMG-R activity in activated oocytes does not affect the transient activation of p42MAPK but results in a block at metaphase of meiosis I that is accompanied by the stabilization of cyclins A and B and p34cdc2 kinase activity. Our results suggest that metabolites from the mevalonate biosynthetic pathway can act to influence the process of activation, as well as the events later in the cell cycle that lead to cyclin proteolysis and the exit from M phase during clam meiosis.

Comparisons of calculations with PARTRAC and NOREC: transport of electrons in liquid water.

Monte Carlo computer models that simulate the detailed, event-by-event transport of electrons in liquid water are valuable for the interpretation and understanding of findings in radiation chemistry and radiation biology. Because of the paucity of experimental data, such efforts must rely on theoretical principles and considerable judgment in their development. Experimental verification of numerical input is possible to only a limited extent. Indirect support for model validity can be gained from a comparison of details between two independently developed computer codes as well as the observable results calculated with them. In this study, we compare the transport properties of electrons in liquid water using two such models, PARTRAC and NOREC. Both use interaction cross sections based on plane-wave Born approximations and a numerical parameterization of the complex dielectric response function for the liquid. The models are described and compared, and their similarities and differences are highlighted. Recent developments in the field are discussed and taken into account. The calculated stopping powers, W values, and slab penetration characteristics are in good agreement with one another and with other independent sources.

The response to prism deviations in human infants.

Previous research has suggested that infants are unable to make a corrective eye movement in response to a small base-out prism placed in front of one eye before 14-16 weeks [1]. Three hypotheses have been proposed to explain this early inability, and each of these makes different predictions for the time of onset of a response to a larger prism. The first proposes that infants have a 'degraded sensory capacity' and so require a larger retinal disparity (difference in the position of the image on the retina of each eye) to stimulate disparity detectors [2]. This predicts that infants might respond at an earlier age than previously reported [1] when tested using a larger prism. The second hypothesis proposes that infants learn to respond to larger retinal disparities through practice with small disparities [3]. According to this theory, using a larger prism will not result in developmentally earlier responses, and may even delay the response. The third hypothesis proposes that the ability to respond to prismatic deviation depends on maturational factors indicated by the onset of stereopsis (the ability to detect depth in an image on the basis of retinal disparity cues only) [4] [5], predicting that the size of the prism is irrelevant. To differentiate between these hypotheses, we tested 192 infants ranging from 2 to 52 weeks of age using a larger prism. Results showed that 63% of infants of 5-8 weeks of age produced a corrective eye movement in response to placement of a prism in front of the eye when in the dark. Both the percentage of infants who produced a response, and the speed of the response, increased with age. These results suggest that infants can make corrective eye movements in response to large prismatic deviations before 14-16 weeks of age. This, in combination with other recent results [6], discounts previous hypotheses.

Targeting of Th1-associated chemokine receptors CXCR3 and CCR5 as therapeutic strategy for inflammatory diseases.

CXCR3 and CCR5 are chemokine receptor that are predominantly expressed on the surface of Th1 polarized T cells. In a variety of human and experimental autoimmune diseases the enhanced expression of CXCR3 and CCR5 binding chemokine ligands is followed by the recruitment of CXCR3- and CCR5-positive T cells, indicating an important role for these chemokine receptors in T cell-mediated tissue damage. In this review, we summarize a number of in vivo studies available on the neutralization of CXCR3 and CCR5 in inflammatory disease, and specifically focus on the potential therapeutic effects of CXCR3 and CCR5 blockade in human autoimmune disease and organ transplantation.

Interventional laser surgery for oral potentially malignant disorders: a longitudinal patient cohort study.

Oral squamous cell carcinoma (OSCC) is a lethal disease, with rising incidence. There were 6767 new OSCC cases and 2056 deaths in the UK in 2011. Cancers are preceded by oral potentially malignant disorders (PMDs), recognizable mucosal diseases harbouring increased SCC risk, offering clinicians a 'therapeutic window' to intervene. Contemporary practice remains unable to predict lesion behaviour or quantify malignant transformation risk. No clear management guidelines exist and it is unclear from the literature whether early diagnosis and intervention prevents cancer. Between 1996 and 2014, 773 laser treatments were performed on 590 PMD patients in Newcastle maxillofacial surgery departments. The efficacy of the intervention was examined by review of the clinicopathological details and clinical outcomes of the patients (mean follow-up 7.3 years). Histopathology required up-grading in 36.1% on examining excision specimens. Seventy-five percent of patients were disease-free, mostly younger patients with low-grade dysplasia; 9% exhibited persistent disease and were generally older with proliferative verrucous leukoplakia. Disease-free status was less likely for erythroleukoplakia (P=0.022), 'high-grade' dysplasia (P<0.0001), and with lichenoid inflammation (P=0.028). Unexpected OSCC was identified in 12.0%, whilst 4.8% transformed to malignancy. Interventional laser surgery facilitates definitive diagnosis and treatment, allows early diagnosis of OSCC, identifies progressive disease, and defines outcome categories. Evidence is lacking that intervention halts carcinogenesis. Multicentre, prospective, randomized controlled trials are needed to confirm the efficacy of surgery.

Transplantation to the diseased and damaged retina.

Retinas of Royal College of Surgeons (RCS) dystrophic rats undergo a dramatic loss of photoreceptor cells as a result of defective retinal pigment epithelial (RPE) cells. These retinas are therefore a valuable model in the investigation of the role of the RPE on photoreceptor-cell survival and development. Also, rat retinas damaged by excessive light serve as a suitable environment to study survival of transplanted photoreceptor cells. Even though photoreceptor cells are lost in these retinas, a normal inner retinal structure is retained. Both models have recently been used in successful RPE-cell and/or photoreceptor-cell transplantation studies designed to replace defective or lost cells due to retinal disease or damage. These new approaches in the field of retinal transplantation offer unique and novel opportunities for the development of possible therapeutic strategies in human eye disease, and for improving our understanding of the normal relationships between retinal cells.

Carcinogenic activity of a chlorinated polyether polyurethan.

On the basis of the results of an earlier study, a particular polyurethan sample (Y-238) was selected for further evaluation of its carcinogenic potential. This sample was subjected to physical and chemical tests for elucidation of its chemical structure, molecular weight, and molecular weight distribution. Additional biological tests were conducted on male NBR rats by implanting various quantities of the sample i.p., while others received an intrabronchus implant. Tumors, assessed histologically as malignant, were observed following both routes of implantation. The most common neoplasms of the pulmonary site was epidermoid carcinoma, while fibrosarcoma was the most common neoplasm in the peritoneal cavity. Data from the i.p. implantation suggested a dose-related incidence of cancers.

Carcinogenesis from polyurethans.

Seventeen polyurethans, containing various substituent groups, and a polyethylene were implanted i.p. in groups of male black Bethesda rats and were evaluated for carcinogenesis over a 2-year period. Thirteen of the polyurethans and the polyethylene were similarly studied in females. Tumor development in these animals was expressed in terms of the incidence in the at-risk population, and the tumorigenic latent period was approximated for each sample. Twenty months after implantation, the relative tumorigenicity (area under the corrected cumulative tumor mortality versus time curve) in the males ranged from 0 (for the unimplanted controls) to 6.18 (for Y-238); for female rats this range was 0.29 (for unimplanted controls) to 5.72 (for Y-238). Estimated latent periods in the males ranged from 5 months (for Y-304) to 16 months (for Y-303), and 22.5 months for the unimplanted controls; for the females, the range was from 9 months (for Y-290) to 13.5 months (for Y-217), and 14 months for the unimplanted controls. The relative tumorigenicity of each sample was also compared to its in vitro activation energy for thermal decomposition. These data are discussed in terms of solid-state versus chemical carcinogenesis.

Exercise-induced B cell mobilisation: Preliminary evidence for an influx of immature cells into the bloodstream.

The number of peripheral blood B lymphocytes doubles during acute exercise, but the phenotypic composition of this response remains unknown. In two independent exercise studies, using complimentary phenotyping strategies, we investigated the mobilisation patterns of distinct B cell subsets. In study one, nine healthy males (mean±SD age: 22.1±3.4years) completed a continuous cycling bout at 80% V̇O2MAX for 20min. In study two, seven healthy experienced cyclists (mean±SD age: 29.9±4.7years) completed a 30min cycling trial at a workload corresponding to +5% of the individual blood lactate threshold. In study one, CD3-CD19+ B cell subsets were classified into immature (CD27-CD10+), naïve (CD27-CD10-), memory (CD27+CD38-), plasma cells/plasmablasts (CD27+CD38+) and finally, recently purported 'B1' cells (CD27+ CD43+ CD69-). In study two, CD20+ B cells were classified into immature (CD27-IgD-), naïve (CD27-IgD+), and IgM+/IgG+/IgA+ memory cells (CD27+IgD-). Total B cells exhibited a mean increase of 88% (study one) and 60% (study two) during exercise. In both studies, immature cells displayed the greatest increase, followed by memory cells, then naïve cells (study one: immature 130%>mature 105%>naïve 84%; study two: immature 110%>mature 56%>naïve 38%). Our findings show that, unlike T cells and NK cells, B cell mobilisation is not driven by effector status, and, for the first time, that B cell mobilisation during exercise is comprised of immature CD27- IgD-/CD10+ cells.

Mesophase formation by ceramides and cholesterol: a model for stratum corneum lipid packing?

Previous X-ray diffraction and electron microscopy experiments have suggested that there is an unusual double bilayer structure formed by stratum corneum lipids, with a lamellar spacing of about 131 A (White, S.H., Mirejovsky, D. and King, G.I. (1988) Biochemistry 27, 3725-3732; Hou, S.Y.E., Mitra, A.K., White, S.H., Menon, G.K., Ghadially, R. and Elias, P.M. (1991) J. Invest. Dermatol. 96, 215-223; Bouwstra, J.A., De Vries, M.A., Gouris, G.S., Bras, W., Brussee, J. and Ponec, M. (1991) J. Controlled Release 15, 209-220). Two contradictory models have been proposed for this structure. In the Downing model, used to explain electron microscopy observations, acylceramides are vital, acting as a 'lynch-pin' and holding the lipid layers together (Swartzendruber, W.C., Kitko, D.J., Madison, K.C. and Downing, D.T. (1989) J. Invest. Dermatol. 92, 251-257). Alternatively, to explain X-ray diffraction results from intact corneum, protein intercallation into the lipid bilayers is suggested, since an electron dense region wider than can be accounted for by lipid headgroups alone, is required (Bouwstra, J.A., De Vries, M.A., Gouris, G.S., Bras, W., Brussee, J. and Ponec, M. (1991) J. Controlled Release 15, 209-220). Thus, existing models require the presence of either acylceramides or protein. We describe how a similar structure can be prepared in vitro using mixtures of cholesterol and ceramides. Cholesterol induces a novel double-bilayer structure in ceramides II, and IV. This result is in conflict with the existing literature which cites acylceramides, or protein as instrumental in maintaining the in vivo structure of the phase. Characterisation has been carried out using optical microscopy and synchrotron X-ray diffraction.

A comparison of two commercially available ELISA methods for the quantification of human plasma heat shock protein 70 during rest and exercise stress.

This study compared resting and exercise heat/hypoxic stress-induced levels of plasma extracellular heat shock protein 70 (eHSP70) in humans using two commercially available enzyme-linked immunosorbent assay (ELIS)A kits. EDTA plasma samples were collected from 21 males during two separate investigations. Participants in part A completed a 60-min treadmill run in the heat (HOT70; 33.0 ± 0.1 °C, 28.7 ± 0.8 %, n = 6) at 70 % V̇O2max. Participants in part B completed 60 min of cycling exercise at 50 % V̇O2max in either hot (HOT50; 40.5 °C, 25.4 relative humidity (RH)%, n = 7) or hypoxic (HYP50; fraction of inspired oxygen (FIO2) = 0.14, 21 °C, 35 % RH, n = 8) conditions. Samples were collected prior to and immediately upon termination of exercise and analysed for eHSP70 using EKS-715 high-sensitivity HSP70 ELISA and new ENZ-KIT-101 Amp'd(™) HSP70 high-sensitivity ELISA. ENZ-KIT was superior in detecting resting eHSP70 (1.54 ± 3.27 ng · mL(-1); range 0.08 to 14.01 ng · mL(-1)), with concentrations obtained from 100 % of samples compared to 19 % with EKS-715 assay. The ENZ-KIT requires optimisation prior to running samples in order to ensure participants fall within the standard curve, a step not required with EKS-715. Using ENZ-KIT, a 1:4 dilution allowed for quantification of resting HSP70 in 26/32 samples, with a 1:8 (n = 3) and 1:16 (n = 3) dilution required to determine the remaining samples. After exercise, eHSP70 was detected in 6/21 and 21/21 samples using EKS-715 and ENZ-KIT, respectively. eHSP70 was increased from rest after HOT70 (p < 0.05), but not HOT50 (p > 0.05) or HYP50 (p > 0.05) when analysed using ENZ-KIT. It is recommended that future studies requiring the precise determination of resting plasma eHSP70 use the ENZ-KIT (i.e. HSP70 Amp'd(®) ELISA) instead of the EKS-715 assay, despite additional assay development time and cost required.

Monitoring changes in thioredoxin and over-oxidised peroxiredoxin in response to exercise in humans.

INTRODUCTION: Peroxiredoxin (PRDX) and thioredoxin (TRX) are antioxidant proteins that control cellular signalling and redox balance, although their response to exercise is unknown. This study aimed to assess key aspects of the PRDX-TRX redox cycle in response to three different modes of exercise. METHODS: Healthy males (n = 10, mean ± SD: 22 ± 3 yrs) undertook three exercise trials on separate days: two steady-state cycling trials at moderate (60% [Formula: see text]O2MAX; 27 min, MOD) and high (80% [Formula: see text]O2MAX; 20 min, HIGH) intensities, and a low-volume high-intensity interval training trial (10 × 1 min 90% [Formula: see text]O2MAX, LV-HIIT). Peripheral blood mononuclear cells were assessed for TRX-1 and over-oxidised PRDX (isoforms I-IV) protein expression before, during, and 30 min following exercise (post + 30). The activities of TRX reductase (TRX-R) and the nuclear factor kappa B (NF-κB) p65 subunit were also assessed. RESULTS: TRX-1 increased during exercise in all trials (MOD, + 84.5%; HIGH, + 64.1%; LV-HIIT, + 205.7%; p < 05), whereas over-oxidised PRDX increased during HIGH only (MOD, - 28.7%; HIGH, + 202.9%; LV-HIIT, - 22.7%; p < .05). TRX-R and NF-κB p65 activity increased during exercise in all trials, with the greatest response in TRX-R activity seen in HIGH (p < 0.05). DISCUSSION: All trials stimulated a transient increase in TRX-1 protein expression during exercise. Only HIGH induced a transient over-oxidation of PRDX, alongside the greatest change in TRX-R activity. Future studies are needed to clarify the significance of heightened peroxide exposure during continuous high-intensity exercise and the mechanisms of PRDX-regulatory control.

Prospective pretransplant ultrasound screening in 206 patients for acquired renal cysts and renal cell carcinoma.

BACKGROUND: Numerous studies have reported an increased prevalence of renal cell carcinoma in association with acquired cystic kidney disease (ACKD). In 1995, the clinical practice guidelines of the American Society of Transplant Physicians for evaluation of renal transplant candidates recommend not screening for ACKD and renal cell carcinoma, on the basis of the low frequency of cancer and reported regression of ACKD after transplantation. The objective of this study was to prospectively evaluate the prevalence of ACKD and renal cancer during renal transplant evaluation. METHODS: A total of 206 consecutive adult patients evaluated for renal transplantation underwent a routine renal ultrasound. Patients with a suspicious ultrasound underwent a contrasted computed tomographic scan of the kidneys followed by excision of kidneys with solid, enhancing (>10 Hounsfield units) lesions. RESULTS: Sixty-three (30.6%) of 206 patients had ACKD, with a greater proportion being male, African-American, and dialysis-dependent for a longer duration. Eight patients (3.8%) had histologically proven localized cancer (six unilateral, two bilateral), seven in association with ACKD and one in association with autosomal dominant polycystic kidney disease. With a mean follow-up of 14 months (range, 3-33 mo), there has been no recurrence. The positive predictive value of a solid lesion on ultrasound was 100% (8 of 8 patients). CONCLUSION: With the high prevalence (3.4%) of renal cell carcinoma in association with ACKD and the concern that immunosuppression accelerates the growth of preexisting cancers, we continue to recommend ultrasound screening of the native kidneys before renal transplantation.

RPE transplants stabilize retinal vasculature and prevent neovascularization in the RCS rat.

Previous reports indicate that in the Royal College of Surgeons (RCS) rat a decline in the retinal vessel density accompanies the loss of the normal architecture of the deep bed. This begins at about three months with neovascularization that originates in the deep vessel bed and develops in the direction of the retinal pigment epithelial (RPE) cells by four months. A surgical technique has been developed recently for the transplantation of healthy RPE cells into the subretinal space of the RCS rat, resulting in the rescue of photoreceptor cells. This permits evaluation of the possibility that such transplants also protect the retinal vessels. We report for the first time: (1) the stabilization of the normal retinal vasculature by maintenance of the density and architecture of the deep vessel bed; and (2) prevention of neovascularization of the RPE by the surgical transplantation of healthy RPE cells into the subretinal space of the RCS rat. More specifically, we show a maintenance of the deep vessel bed density under the transplant in contrast to a significant reduction in the vessel density that had taken place in corresponding areas in nongrafted and sham injected controls at four months of age. The vessel density in the transplanted group is statistically different from the nongrafted and the sham injected groups. We also report a significant decline in the number of neovascularization profiles around the transplant site of the RPE-grafted RCS retina. We also note that the pathological changes in the vasculature of the RCS rat occur in a predictable central to peripheral gradient.

Donor age influences on the success of retinal grafts to adult rat retina.

The rat retina can be successfully grafted within a long time period which extends into the first 2 weeks of postnatal life. Postnatal grafts taken 1-2 days (PN 1-2) after birth demonstrate no significant differences in their ability to form successful grafts. However, grafting success begins to diminish gradually starting between PN 2-4 and reaches a low point in organization and survival by PN 14. PN 21 grafts rapidly degenerate by 1 to 2 days after transplantation. Although early postnatal retinal tissue can be successfully grafted, E 15 embryonic retinas make better grafts for their ability to form consistent laminae and to integrate with host tissue in a fresh lesion paradigm.

Retinal pigment epithelial cell transplants in retinal degeneration slow mice do not rescue photoreceptor cells.

PURPOSE: To determine if retinal pigment epithelial cells are in any way involved in the degeneration of photoreceptor cells in the retinal dystrophy mouse model, retinal degeneration slow (rds); to determine if normal retinal pigment epithelial cell transplants can affect outer segment development in the retina. METHODS: Retinal pigment epithelial cells of neonatal normal pigmented C3H mice were isolated and transplanted into retinas of postnatal day 33 albino rds mice. Then eyes of 4-month-old rds mice, retinal pigment epithelial cell-transplanted and sham and non-treated control mice, were processed for light and electron microscopy and the thickness of the outer nuclear layer were measured and compared. RESULTS: Measurements of outer nuclear layer thickness in the transplant and control groups revealed that normal retinal pigment epithelial cell transplants did not cause photoreceptor cell rescue in rds mice. In addition, outer segments were not seen in retinal pigment epithelial cell-transplanted rds retinas. CONCLUSIONS: This study supports the conclusions of other investigators that the photoreceptor cell is the primary site of the genetic defect that results in retinal dystrophy in the rds mouse model.

A primary role for RPE transplants in the inhibition and regression of neovascularization in the RCS rat.

PURPOSE: Healthy retinal pigment epithelial (RPE) cell transplants in retinas of postnatal day 27 Royal College of Surgeons (RCS) rat are capable of preventing photoreceptor cell degeneration, inhibiting the degeneration of the retinal vascular bed and the subsequent neovascularization of the RPE that occurs in retinas injected with saline. In the present study, the authors have tested the hypothesis that RPE transplants might also prevent vascularization of the RPE layer in RCS retinas when photoreceptor cells either had just degenerated or, at 3 months, disappeared. METHOD: Retinas of 3- and 6 month-old RCS rats were injected with either healthy neonatal RPE cells or vehicle and were examined at 6 and 8 months, respectively. These retinas were studied using horseradish peroxidase visualization of the vasculature in retinal wholemount preparations and by light microscopy. RESULTS: The number of neovascular profiles in wholemount preparations of RCS retinas that had received healthy RPE transplants at 3 months and were analyzed at 6 months were significantly decreased when compared to sham-injected retinas of age-matched RCS rats, 2.09 +/- 0.94 and 15.28 +/- 1.34 profiles per mm2, respectively (P < 0.001). In addition, when comparing retinas transplanted or sham injected at 6 months and examined at 8 months, significantly fewer neovascular profiles were found in the transplant group, 9.32 +/- 1.02 and 15.42 +/- 0.84 profiles per mm2, respectively (P < 0.002). CONCLUSION: These data provide still further evidence for the role of healthy RPE in maintaining the homeostasis of the normal retinal vasculature in the retina of the RCS rat. The relationships between the RPE and the retinal vasculature are important when considering that alterations in the vascularization of the retina play a major role in some of the most sight-debilitating diseases, such as the wet form of age-related macular degeneration and proliferative diabetic retinopathy.

Müller and retinal pigment epithelial (RPE) cell expression of NGFI-A and c-fos mRNA in response to medium conditioned by the RPE.

Retinal pigment epithelial (RPE) cells secrete a factor(s) which promotes Müller and RPE cell survival and proliferation in vitro. These influences may play developmental and functional roles as well as contribute to ocular pathologies such as proliferative vitreoretinopathy (PVR). In the past few years, a number of immediate early genes (IEGs) have been identified. Many IBGs encode transcription factors, the expression of which is altered by stimuli such as growth factors. Since an RPE-derived factor(s) elicits proliferation of Müller and RPE cells, we investigated the expression of two IEGs, NGFI-A and c-fos, in both cell types after treatment with medium conditioned by the RPE (RPE-CM). We found that Müller and RPE cells had increased levels of NGFI-A mRNA following treatment with RPE-CM; in contrast, only a slight increase in c-fos mRNA was induced in RPE, but not Müller cells. Immunolabeling for NGFI-A protein revealed nuclear staining in both cell types which corresponded with the increased mRNA levels in RPE-CM-treated cultures. This in vitro study demonstrates a potential mechanism by which RPE-secreted factors may exert autocrine or paracrine effects on retinal cells in vivo. Specifically, NGFI-A may be the primary target of a second messenger system that is regulated by an RPE-derived factor(s).