Biomedical subjects

J E Turner

Publications and source records attributed to J E Turner.

At least 37 records · Page 2Linked to original sources

PubMed2000

The development of the use of long-term knowledge to assist short-term recall.

The influence of item familiarity upon memory span was examined in adults and children aged 5, 7, and 10 years by comparing the recall of words and nonwords. Using a probed recall task, both item recall and position recall were tested. The effect of familiarity upon item recall was found to develop with age, from no effects in the 5-year-olds to significant effects in the older children and adults. By contrast, no effect of familiarity was found at any age when recall of position was required. Dissociations between word length effects and familiarity effects supported the conclusion that the familiarity effect does not result from rehearsal. Several explanations for the source of the familiarity effect were examined, and the familiarity effect was attributed to a strategic redintegration or reconstruction process, which is necessary for item recall but not for position recall.

Adult↗

PubMed2000

Effect of maturity on degradation kinetics of sod-seeded cereal grain forage grown in northern Arkansas.

Wheat (Triticum aestivum L.), oat (Avena sativa L.), and rye (Secale cereale L.) were overseeded into a dormant bermudagrass (Cynodon dactylon (L.) Pers.) sod and harvested at 3-wk intervals throughout March, April, May, and early June. Plant growth stage was documented for each forage on each harvest date, and harvested forages were evaluated for forage quality characteristics. Degradation kinetics of DM and NDF for these forages were evaluated by the in situ method. Fractional degradation rates for DM and NDF in all three species were relatively rapid for vegetative forage (> or =0.086 h(-1)) but declined rapidly by the heading stage of development and stabilized thereafter. Forage quality declined and forages were more resistant to ruminal degradation as plants entered the reproductive stages of growth. Based on these findings, growth stage is an effective predictor of most characteristics of in situ DM and NDF disappearance. The relationships between these degradation parameters and growth stage were typically explained with quadratic or cubic models. Clearly, forage quality characteristics of overseeded rye deteriorated more rapidly with phenological development and growth stage than quality characteristics of overseeded wheat and oat grown in the same environment. For rye, this problem is further complicated by its accelerated phenological development. These factors combine to permit a very narrow harvest window in early spring, relative to the other cereal grains evaluated. Acceptable forage quality may persist for an extended period in wheat and oat; this suggests that producers wishing to utilize these forages may lengthen the harvest window by planting more than one species, either as a mixture or preferably in independent stands.

Animal Feed↗

PubMed1999

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.

Adaptation, Physiological↗

PubMed1999

Retina-derived fetuin (RDF): analysis by immunocytochemistry, reverse transcriptase-polymerase chain reaction and Southern blot.

PURPOSE: This study was undertaken to determine the presence of retina-derived fetuin (RDF) protein and its message in retinal tissues and retinal pigment epithelial (RPE) cells. The techniques utilized in this study included light micros-copy, immunochemistry, Western blot, reverse transcriptase-polymerase chain reaction (RT-PCR) and Southern blot. METHODS: Retinal tissues and sections from embryonic, early postnatal and adult normal rats and retinal pigment epithe-lial (RPE) cells from postnatal rats were immunostained for fetuin with a polyclonal fetuin antibody and a peroxidase conjugated-secondary antibody using immunocytochemistry and Western blot analyses. The cDNA generated from RNA isolated from early postnatal rat retinas and RPE was probed with primers for rat fetuin, amplified by PCR and the PCR products were analyzed by Southern blot. RESULTS: Fetuin (RDF) was immunolocalized to cells of the neuroepithelium in retinas of early postnatal rats and most prominently in the nuclei and perinuclear region of cultured neonatal rat RPE cells. In adult retinas, ganglion cells, inner segments of photoreceptor cells, some components of the outer plexiform layer, ganglion cells and optic nerve processes were immunoreactive for the fetuin protein. As shown by Western blot, fetuin (RDF) was higher in embryonic and early postnatal retinas than in late postnatal retinas, indicating that this protein may be developmentally regulated. Using RT-PCR, the message for rat fetuin was demonstrated in the retina and RPE of normal postnatal rats. Southern blot confirmed that the PCR product from the retina and RPE was generated from rat fetuin mRNA as well as from rat liver, the primary source of fetuin. CONCLUSIONS: Fetuin, termed retina-derived fetuin (RDF), is reported for the first time in retinal tissues. Fetuin is a cysteine protease inhibitor that may play a role in support of neuronal cell survival during early retinal development and the maintenance of neuronal activity. RDF may interact with other growth factors and cytokines in providing trophic support for neurons and possibly other cells of the developing retina.

Animals↗

PubMed1999

In situ disappearance of neutral detergent insoluble nitrogen from alfalfa and eastern gamagrass at three maturities.

In situ digestion kinetics of neutral detergent insoluble nitrogen (NDIN) from alfalfa (Medicago sativa L.) harvested at one-tenth bloom and eastern gamagrass (Tripsacum dactyloides L.) harvested at the boot (GGB), anthesis (GGA), and physiological maturity (GGM) stages of growth were determined with nonlinear regression techniques. Whole-plant tissue and associated leaf and stem fractions were incubated in the ventral rumen simultaneously. On a wholeplant basis, potential extents of degradation were particularly high (> or =904 g/kg NDIN) for GGB and GGA, relative to those of GGM and alfalfa (772 and 658 g/kg NDIN, respectively). For all plant parts, degradation rates of NDIN were faster (P<.05) for alfalfa than for all gamagrass forages. Degradation rate of NDIN did not differ (P>.05) across maturities for any gamagrass tissue type. These results indicate 1) that phenological development and lignification do not limit the rate of NDIN degradation in gamagrass forages but do markedly limit the potential extent of NDIN availability and 2) that most of the NDIN in these forages is potentially available in the rumen and can contribute to the ruminal N supply. Our secondary objective was to compare estimates of N escaping ruminal degradation that were determined on the basis of NDIN degradation kinetics (NDIN method) with those determined traditionally, on the basis of total residual N. The NDIN method mathematically eliminates all neutral detergent soluble N from consideration as part of the pool of dietary N potentially escaping the rumen intact. Estimates of rumen escape nitrogen determined on the basis of degradation rates of NDIN were consistently less than corresponding estimates that were determined on the basis of total residual N. When ruminal escape N that was determined with the NDIN method was regressed on corresponding estimates with the total residual N method, the slopes of the regression lines were .53 and .66 for assumed passage rates of .02 and .06 h(-1), respectively. For the forages evaluated in this study, these results indicate that neutral detergent soluble N may make important contributions to the pool of N escaping ruminal degradation.

Animals↗

PubMed1999

Degradability of forage proteins by in situ and in vitro enzymatic methods.

The overall objective of these two studies was to evaluate the efficacy of using the proteolytic enzyme from Streptomyces griseus to estimate concentrations of ruminally degradable protein (RDP) in a wide array of forages. In the first study, alfalfa and prairie hays that previously had been evaluated in vivo for RDP were incubated in a replicated 3 x 3 factorial combination of enzyme concentrations (6.6, 0.66, and 0.066 activity units/ml of incubation medium) and incubation times (2, 4, and 48 h). Two treatment combinations (6.6 activity units for 4 h and 0.066 activity units for 48 h) yielded respective RDP estimates for alfalfa and prairie hay that were close to the known in vivo values. In the second study, 20 diverse forages were evaluated for RDP by using the in situ technique. These forages also were evaluated for RDP with the two enzyme concentrations identified in the first study, but incubation times were expanded to include 1, 2, 3, 4, and 5 h at the high concentration and 24, 30, 36, 42, 48, and 54 h at the low concentration. At the high enzyme concentration, r2 statistics from linear regressions of enzymatic estimates of RDP on corresponding estimates obtained by the in situ procedure were high (r2 > or = 0.898) at all incubation times; in addition, slopes (range = 0.88 to 1.00) and intercepts (range = -9.4 to 3.5%) approached unity and 0, respectively. At the lower enzyme concentration, r2 statistics were still good (> 0.81), but slopes (0.59 to 0.67) and intercepts (18.5 to 21.9%) for all incubation times did not meet the respective goals of unity and 0.

Animal Feed↗

PubMed1999

The clinical significance of beta hemolytic streptococci of the milleri group in oral abscesses.

Aspirated exudate from 50 patients with oral abscesses were cultured and bacterial growth was assessed qualitatively and semi-quantitatively. Bacteria were recovered from all specimens. The mean number of species isolated per specimen was 3.6 Both facultative and obligate anaerobes were isolated from 39 (78%) specimens. The most commonly isolated facultative anaerobic species was Streptococcus virdans group, while pigmented gram negative bacilli were the most commonly isolated obligate anaerobes. Beta-hemolytic Streptococci of the "milleri" groups were recovered from 11 (22%) of the abscesses. Eight of those isolates were determined to carry Lancefield group F antigen, 2 group C, and 1 isolate was not groupable for any Lancefield antigen. These abscesses may be sequelae of oral infectious disease or trauma. Patients with oral abscesses infected with beta hemolytic Streptococci are apt to be male, and less than 35 years old. They are also likely to have a more rapid onset of infection and to require hospitalization for intensive treatment of a life threatening condition.

Adolescent↗

PubMed1998

RPE secreted proteins and antibody influence photoreceptor cell survival and maturation.

Proteins in media conditioned by retinal pigment epithelial cells (RPE-CM) and an antibody against these proteins (RPE-SP) were tested for their respective effects on rat retinal development in vitro and in vivo. Proteins of RPE-CM were separated in denaturing gels and evaluated by Western blot analysis. Retinal explants from postnatal day 2 (P2) rats were cultured in RPE-CM only or CM diluted with the RPE-SP antibody and, after 7 days, the explants were dissociated into single cells that were immunostained for opsin. RPE-CM or antibody was also injected into the vitreous of postnatal day 7 (P7) Long-Evans rats and analyzed 7 and 21 days later. Electrophoretic analysis of RPE-CM predominantly showed 60-70 kDa proteins; when these proteins were probed with RPE-SP antibody by Western blot, immunoreactive proteins were restricted to this narrow molecular weight range. In P2 retinal explant cultures supplemented with RPE-CM, long ganglion cell-like neurites were detected in 3 days. This activity was nullified in explant cultures grown in RPE-CM titrated with antibody, and these explants appeared to degenerate within 5 days. Over 80% of dissociated retinal cells from explants 7 days after treatment with RPE-CM expressed opsin, compared to only 20% of cells from explants grown in defined medium or serum. Retinas of P14 rats injected intravitreally with RPE-CM at P7 had increased numbers of ectopic photoreceptor cells within the inner nuclear layer when compared to retinas of sham-injected eyes. In contrast, retinas of eyes injected intravitreally with RPE-SP antibody exhibited shorter outer (OS) and inner (IS) segments and thinner outer nuclear (ONL) and outer plexiform (OPL) layers than retinas of sham-injected eyes. In conclusion, proteins in RPE-CM appeared to accelerate and maximize the development of rat photoreceptor cells in vitro, while intravitreal injections of its antibody caused an apparent retardation of outer segment maturation. These results suggest that a protein(s) secreted by RPE plays a key role in normal retinal development, particularly in photoreceptor cell survival and outer segment maturation.

Animals↗

PubMed1998

Monte Carlo simulation of diffusion and reaction in water radiolysis--a study of reactant 'jump through' and jump distances.

In Monte Carlo simulations of water radiolysis, the diffusion of reactants can be approximated by "jumping" all species randomly, to represent the passage of a short period of time, and then checking their separations. If, at the end of a jump, two reactant species are within a distance equal to the reaction radius for the pair, they are allowed to react in the model. In principle, the possibility exists that two reactants could "jump through" one another and end up with a separation larger than the reaction radius with no reaction being scored. Ignoring this possibility would thus reduce the rate of reaction below that intended by such a model. By making the jump times and jump distances shorter, any error introduced by 'jump through' is made smaller. This paper reports numerical results of a systematic study of 'jump through' in Monte Carlo simulations of water radiolysis. With a nominal jump time of 3 ps, it is found that more than 40% of the reactions of the hydrated electron with itself and of the H atom with itself occur when reactions during 'jump through' are allowed. For all other reactions, for which the effect is smaller, the contributions of 'jump through' lie in the range 1%-16% of the total. Corrections to computed rate constants for two reactions are evaluated for jump times between 0.1 and 30 ps. It is concluded that jump-through corrections are desirable in such models for jump times that exceed about 1 ps or even less. In a separate study, we find that giving all species of a given type the same size jump in a random direction yields results that are indistinguishable from those when the jump sizes are selected from a Gaussian distribution. In this comparison, the constant jump size is taken to be the root-mean-square jump size from the Gaussian distribution.

Models, Chemical↗

PubMed1998

Investigation of a Monte Carlo model for chemical reactions.

Monte Carlo computer simulations are in use at a number of laboratories for calculating time-dependent yields, which can be compared with experiments in the radiolysis of water. We report here on calculations to investigate the validity and consistency of the procedures used for simulating chemical reactions in our code, RADLYS. Model calculations were performed of the rate constants themselves. The rates thus determined showed an expected rapid decline over the first few hundred ps and a very gradual decline thereafter out to the termination of the calculations at 4.5 ns. Results are reported for different initial concentrations and numbers of reactive species. Generally, the calculated rate constants are smallest when the initial concentrations of the reactants are largest. It is found that inhomogeneities that quickly develop in the initial random spatial distribution of reactants persist in time as a result of subsequent chemical reactions, and thus conditions may poorly approximate those assumed from diffusion theory. We also investigated the reaction of a single species of one type placed among a large number of randomly distributed species of another type with which it could react. The distribution of survival times of the single species was calculated by using three different combinations of the diffusion constants for the two species, as is sometimes discussed in diffusion theory. The three methods gave virtually identical results.

Computer Simulation↗

PubMed1998

Product yields from irradiated glycylglycine in oxygen-free solutions: Monte Carlo simulations and comparison with experiments.

The radiation chemistry of photon-irradiated aqueous solutions of biological molecules may be considered under four distinct time regimes: physical transport (< or = 10(-15) s); prechemical conversion of H2O+, H2O*, and subexcitation electrons into free radicals and molecular products (10(-15) s to 10(-12) s); chemical reactions within individual electron tracks (10(-12) s to 10(-6) s); and chemical reactions within overlapping tracks (>10(-6) s). We have previously reported of the use of the Monte Carlo radiation transport/chemistry codes OREC and RADLYS to model the radiolysis of glycylglycine in oxygen-free solution to a time of 1 micros. These simulations successfully predicted the yields of free ammonia, an end product created solely in the reaction of the hydrated electron with the solute within individual tracks. Other measurable products are only partially created during intratrack reactions, and thus one must additionally consider the late, intertrack chemistry of this system. In this paper, we extend our simulations of glycylglycine radiolysis to model for the first time the events which occur during this late chemistry stage. The model considers the product rates of the reactants in bulk solution by using previously available microsecond intratrack yields given by single-track OREC/RADLYS simulations and an x-ray dose rate of 2.80 Gy min(-1) as used in a companion experimental program. These rates are then applied in a series of coupled, differential rate equations that describe the solution chemistry of glycylglycine radiolysis. Product yields are reported as a function of time over a total irradiation period of 10(4) s. Excellent overall agreement is seen between the theoretical predictions and measurements of five radiolysis end products: free ammonia, acetylglycine, diaminosuccinic acid, aspartic acid, and succinic acid. The model also gives the explicit contributions of intratrack and intertrack reactions to the various end products. For example, the model predicts that approximately 56% and 93% of succinic acid and aspartic acid, respectively, are produced during intertrack reactions at a solute concentration of 0.05 M; these contributions drop to 0.07% and 11%, respectively, at 1.2 M.

Computer Simulation↗

PubMed1998

Immunocytochemical characterisation of proteins secreted by retinal pigment epithelium in retinas of normal and Royal College of Surgeons dystrophic rats.

In a previous study, an antigen consisting of proteins secreted by retinal pigment epithelial (RPE) cells was injected into a sheep and the specificity of the resulting antiserum was shown by Western blotting and its effects on retinal development were determined in vitro and in vivo. In the present study, the distribution of these secreted proteins was determined by light microscopy immunocytochemistry in cultured neonatal rat RPE cells and retinas of normal and Royal College of Surgeons (RCS) dystrophic rats and cerebrum of normal adult rats. Immunolabelling for these RPE-secreted proteins was detected in cytoplasmic vesicles surrounding nuclei and within processes of cultured normal and transformed rat RPE. In retinas of late postnatal and adult rats, dense immunostaining was found in the cytoplasm of RPE cells and ganglion cell bodies. In addition to RPE and ganglion cells, scattered photoreceptors within the thin outer nuclear layer and small structures within the debris zone were also densely immunoreactive in retinas of 2-mo-old RCS dystrophic rats. The numbers of immunostained ganglion cells appeared to decrease in retinas of older RCS rats, although the immunoreactivity within the RPE appeared to increase in density. No other neuron within the retina, i.e. bipolar, amacrine or horizontal, was immunoreactive for RPE-secreted proteins. In the cerebral cortex of adult rats, immunoreactivity for RPE-secreted proteins was primarily detected within large perikarya of pyramidal neurons and smaller granule neurons. In conclusion, we report an immunocytochemical analysis of an antiserum raised against secreted proteins of rat RPE. This antiserum recognised proteins within secretory-like vesicles of cultured neonatal normal and transformed rat RPE and showed a specificity for RPE and ganglion cells in normal rat retinas, that appeared to be developmentally regulated, and neuron perikarya in adult rat cerebrum.

Animals↗

PubMed1998

Gonadotrophin profiles and dioestrous pulsatile release patterns in mares as determined by collection of jugular blood at 4 h intervals throughout an oestrous cycle.

In mares, dioestrous FSH profiles based on once-a-day sampling are variable; however, the pulsatility of plasma FSH, which has been suggested by limited windows of intensive sampling, may contribute to this variability. Jugular blood from six mares was sampled at 4 h intervals throughout an ovulatory cycle to determine cyclic FSH and LH patterns more accurately and to measure gonadotrophin pulse frequency during dioestrus. Synchronous pulses of FSH and LH occurred regularly in all mares between day 4 and day 12 (ovulation = day 0) with a mean (+/- SEM) frequency of 1.9 +/- 0.1 (FSH) or 1.6 +/- 0.1 (LH) pulses day-1. LH pulse amplitude declined (P < 0.0001) between day 4 and day 10, but FSH pulse amplitude remained large and stable, dipping slightly but not significantly on day 6. Daily mean FSH concentrations exceeded (P < 0.0001) early oestrous values between day 4 and day 5, and between day 7 and day 10. However, significantly different patterns were obtained when once-a-day sampling was simulated by selecting samples collected at 08:00 h or noon. LH was higher during the periovulatory surge than during dioestrus (P < 0.0001) and profiles were similar whether daily means or selected samples were used. It is concluded that: (1) the marked pulsatility of plasma FSH during dioestrus makes once-a-day sampling misleading for determining FSH profiles; (2) the dioestrous pattern of large, slow FSH pulses was consistent among mares, unlike that of the daily mean FSH profiles; and (3) no discrete FSH 'surges' were observed during dioestrus, although FSH pulse amplitude tended to undergo alternate increases and decreases. A period of higher amplitude FSH pulses preceded ovulation by 10.2 +/- 0.7 days, which corresponds to the approximate time the ovulatory follicle emerges. Therefore, it is possible that the signal for follicular recruitment in mares is intermittent excursions of plasma FSH above a threshold value.

Animals↗

PubMed1998

Contrasting effects of age of acquisition and word frequency on auditory and visual lexical decision.

In four experiments, we examined the effects of frequency and age of acquisition on auditory and visual lexical decision. Word frequency affected visual, but not auditory, lexical decision speed (Experiments 1 and 3). Age of acquisition affected lexical decision speed in both modalities (Experiments 2 and 4). We suggest that previous reports of effects of frequency on auditory lexical decision may be due to a confounding of frequency with age of acquisition, and we discuss the implications of these findings for theories of auditory and visual word recognition.

Adult↗

PubMed1998

The dos and don'ts of an oral mucosal biopsy performed by the general dentist.

The recognition of a pathologic entity is only the first step in the chain of events which must be performed for accurate diagnoses and appropriate patient management. Other steps include selection of the type of biopsy to be performed; proper biopsy technique, instrumentation and specimen handling; appropriate fixation; detailed documentation; and postoperative patient instructions. If the clinician performs each step carefully and correctly, the patient is ensured of receiving an accurate diagnosis in the shortest possible period of time.

Biopsy↗

PubMed1997

Basic fibroblast growth factor treatment delays age-related photoreceptor degeneration in Fischer 344 rats.

This study was undertaken to investigate the potential of basic fibroblast growth factor (bFGF) to delay photoreceptor cell loss in retinas of Fischer 344 rats which exhibit an age-related peripheral retinopathy. Eight male 16-month-old Fischer 344 rats were injected intravitreally with 2.0 micrograms bFGF in the right eye, while the vehicle was injected into the left eye and all rats were killed at two months post-injection. Eight other Fischer rats were injected with either bFGF or vehicle at 16 and again at 18 months, then killed two months later. After enucleation, eyes were processed for light and electron microscopic and morphometric analyses. The distance from the ora serrata to the point where the outer nuclear layer (ONL) in the superior retina was two cells in thickness, called the die-back zone, that represents the extent of the peripheral retinopathy, was compared in bFGF- and vehicle-injected Fischer rats. These measurements revealed that the die-back zone in retinas of eyes injected twice with bFGF was significantly reduced (P < 0.01) when compared to this zone in retinas of age-matched rats that received vehicle injections. However, there was no significant difference (P > 0.05) in this degeneration zone in retinas of 18-month-old rats that received a single-bFGF injection when compared to retinas of respective vehicle-injected rats. In addition, the ONL at 2 and 3 mm from the ora serrata in the superior retina of bFGF-injected 20-month-old Fischer rats was significantly thicker than corresponding regions in retinas of vehicle-control rats. Furthermore, the photoreceptor cells in the superior retina of 20-month-old rats which received two injections of bFGF had normal-appearing inner and outer segments, while the few photoreceptors had short inner and outer segments in vehicle-injected retinas of Fisher rats. These findings reveal that injections of bFGF into eyes of Fischer 344 rats significantly delays the progress of photoreceptor cell degeneration suggesting that exogenous bFGF may act as a survival-promoting factor in these aged retinas.

Aging↗

PubMed1997

Cranial polyneuropathy--Ramsay Hunt's syndrome: case report and discussion.

Ramsay Hunt's syndrome is an infectious cranial polyneuropathy caused by varicella zoster, the herpetic virus that also causes chickenpox and shingles. Its symptoms include facial paralysis, ear pain, and an auricular rash. Oral lesions are also present in most cases. This syndrome can affect any cranial nerve and usually affects multiple nerves, causing central, cervical, and peripheral effects. This article reports the case of a 35-year-old white female who was treated by the oral surgery service of a large urban hospital, after first reporting to the emergency clinic. Her reported symptoms of unilateral left-side facial paralysis, auricular pain, and trigeminal hyperesthesia were confirmed by clinical examination. An initial short low-dose steroid regimen was unsuccessful. A second daily dosage of 50 mg of prednisone was successful in 21 days. No permanent sequelae were evident or reported after treatment.

Adult↗

PubMed1997

A transformed neonatal rat retinal pigment epithelial cell line: secreted protein analysis and fibroblast growth factor and receptor expression.

PURPOSE: A newly-derived transformed neonatal rat retinal pigment epithelial (tnrRPE) cell line was investigated: for secreted proteins by electrophoresis, and for basic and acidic fibroblast growth factor (FGF) by immunocytochemistry, Northern blot and reverse transcriptase-polymerase chain reaction (RT-PCR). The FGFR-1 (flg) receptor, which is recognized by aFGF and bFGF, was studied by RT-PCR. METHODS: Retinal pigment epithelial (RPE) cells were isolated from 6-day-old pigmented normal Long Evans rats, and became spontaneously transformed after the second passage. RESULTS: RPE cells at the 5th through 28th passages expressed the epithelial cell marker cytokeratin and cellular retinaldehyde binding protein (CRALBP), an RPE cell marker, but were negative for glial fibrillary acidic protein (GFAP), as shown by immunofluorescence. Secreted proteins of late passage tnrRPE cells were in a narrow molecular weight range of 60-80kDa, while early passage cells exhibited multiple proteins from 20-200kDa. These tnrRPE cells increased by 17-30 fold over a 4-day culture period. At 5th and 28th passage, immunostaining for bFGF and aFGF was dense within nuclei, but light and diffuse within the cytoplasm of transformed RPE cells. As shown by Northern blot, similar levels of message for bFGF were detected in 5th and 30th passage RPE cells. As shown by Northern blot, similar levels of message for bFGF were detected in 5th and 30th passage RPE cells. Furthermore, as shown by RT-PCR, bFGF mRNA was found in freshly isolated and transformed neonatal rat RPE cells. However, the message for FGFR-1(flg) receptor was detected only in the transformed RPE cells. CONCLUSIONS: This study demonstrated a neonatal rat RPE cell line that proliferated rapidly in vitro, expressed high levels of message for hFGF and FGFR-1(flg) receptor, and continued to express RPE-cell characteristics. Importantly, mRNA levels of confluent cultures of these cells were sufficient for bFGF mRNA blot analysis, which eliminates the necessity for PCR and for using excessive numbers of animals for such studies.

Animals↗