Retinoid Supplementation Research Articles

Actions of Retinoic Acid in the Pathophysiology of HIV Infection.

The vitamin A metabolite all-trans retinoic acid (RA) plays a key role in tissue homeostasis and mucosal immunity. RA is produced by gut-associated dendritic cells, which are among the first cells encountered by HIV. Acute HIV infection results in rapid reduction of RA levels and dysregulation of immune cell populations whose identities and function are largely controlled by RA. Here, we discuss the potential link between the roles played by RA in shaping intestinal immune responses and the manifestations and pathogenesis of HIV-associated enteropathy and similar conditions observed in SIV-infected non-human primate models. We also present data demonstrating the ability of RA to enhance the activation of replication-competent viral reservoirs from subjects on suppressive anti-retroviral therapy. The data suggest that retinoid supplementation may be a useful adjuvant for countering the pathologic condition of the gastro-intestinal tract associated with HIV infection and as part of a strategy for reactivating viral reservoirs as a means of depleting latent viral infection.

713b A Multi-Systems Approach with Big Data from Adverse Event Reporting and Health Records, Clinical Biospecimens, and Experimental Animal Data Reveal That Retinoids Impact Asparaginase-Associated Pancreatitis Outcomes in Leukemic Patients

Asparaginase is a crucial cornerstone chemotherapy for the most common cancer in children, acute lymphoblastic leukemia (ALL). However, a major concern is that 5-10% of all asparaginase users develop the painful, life-threatening complication of pancreatitis. Asparaginase-associated pancreatitis (AAP) is a more severe form of pancreatitis, and discontinuation of asparaginase is associated with inferior disease-free survival in high-risk cancer patients. Thus, there is a major unmet need to mitigate AAP. In this study, we took a multi-systems approach that combined adverse event reporting data and electronic health records (EHR), clinical biospecimens, and experimental data to identify factors that impact AAP. An analysis of 14 million records from the FDA Adverse Events Reporting System (FAERS) demonstrated the novel finding that vitamin A supplementation to asparaginase was correlated with reduction in the risk of AAP. To home in the role of systemic vitamin A exposure, versus topical agents, an analysis of over 64 million records from an EHR database (TriNetX Research) identified a protective effect of non-topical vitamin A supplementation on AAP. To pursue this further, we performed a case control study of 26 ALL patients who developed pancreatitis at a median of 3 months after the start of chemotherapy and compared them with 24 well-matched control ALL patients who did not develop pancreatitis over the course of leukemia therapy. We performed non-targeted metabolomics of plasma before and after induction chemotherapy. The striking difference between the two groups was that the precursors of vitamin A were reduced in the pancreatitis cases post-induction compared to the controls. In mice, we found that single agent asparaginase administration by itself led to reduce retinol levels in serum. Analysis of transcriptomic data from leukemic cells demonstrated that the transcriptional signature from asparaginase exposure was potentially reversed by retinoid interventions. The finding was similar in transcriptomic data from a cerulein model of pancreatitis. Taken together, these data suggest that reductions in retinol levels may be correlated with AAP, that asparaginase causes those reductions, and that retinoid supplementation may prevent AAP. The major novelty of the current work is that it provides crucial insight into mitigating the adverse event of AAP and elucidates a broader role for retinoids in preventing the sterile inflammation of the pancreas. Fig 2Vitamin A metabolite levels between pancreatitis patients and controls. The y-axis represents log2 post/pre. * q<0.1. View Large Image Figure Viewer Download Hi-res image

Advanced late-onset retinitis pigmentosa with dominant-acting D477G RPE65 mutation is responsive to oral synthetic retinoid therapy

ObjectivesNo therapeutic interventions are currently available for autosomal dominant retinitis pigmentosa (adRP). An RPE65 Asp477Gly transition associates with late-onset adRP, reduced RPE65 enzymatic activity being one feature associated with this...

Vitamin A and beta (β)-carotene supplementation for cystic fibrosis.

People with cystic fibrosis (CF) and pancreatic insufficiency are at risk of a deficiency in fat-soluble vitamins, including vitamin A. Vitamin A deficiency predominantly causes eye and skin problems, while excessive levels of vitamin A can harm the respiratory and skeletal systems in children and interfere with the metabolism of other fat-soluble vitamins. Most CF centres administer vitamin A as supplements to reduce the frequency of vitamin A deficiency in people with CF and to improve clinical outcomes such as growth, although the recommended dose varies between different guidelines. Thus, a systematic review on vitamin A and vitamin A-like supplementation (carotenes or other retinoids) in people with CF would help guide clinical practice. This is an update of an earlier Cochrane Review. To determine if supplementation with vitamin A, carotenes or other retinoid supplements in children and adults with CF reduces the frequency of vitamin A deficiency disorders, improves general and respiratory health and affects the frequency of vitamin A toxicity. We searched the Cochrane Cystic Fibrosis and Genetic Disorders Group Cystic Fibrosis Trials Register compiled from electronic database searches and handsearching of journals and conference abstract books. Additionally we searched several ongoing trials registries, including ClinicalTrials.gov, the WHO International Clinical Trials Registry Platform and the International Standard Randomised Controlled Trial Number Registry.Most recent database searches: 01 June 2018. All randomised or quasi-randomised controlled studies comparing all preparations of oral vitamin A, carotenes or retinoids (or in combination), used as a supplement compared to placebo at any dose, for at least three months, in people with CF (diagnosed by sweat tests or genetic testing) with and without pancreatic insufficiency. Two authors individually assessed study quality and extracted data on outcome measures. The authors assessed the quality of the evidence using the GRADE system. Investigators were contacted to retrieve missing quantitative data. No studies of vitamin A or other retinoid supplementation were eligible for inclusion. However, one randomised study of beta (β)-carotene supplementation involving 24 people with CF who were receiving pancreatic enzyme substitution was included. The study compared successive β-carotene supplementation periods (high dose followed by low dose) compared to placebo. The results for the low-dose supplementation period should be interpreted with caution, due to the lack of a wash-out period after the high-dose supplementation.The included study did not report on two of the review's primary outcomes (vitamin A deficiency disorders and mortality); results for our third primary outcome of growth and nutritional status (reported as z score for height) showed no difference between supplementation and placebo, mean difference (MD) -0.23 (95% confidence interval (CI) -0.89 to 0.43) (low-quality evidence). With regards to secondary outcomes, supplementation with high-dose β-carotene for three months led to significantly fewer days of systemic antibiotics required to treat pulmonary exacerbations, compared to controls, MD -15 days (95% CI -27.60 to -2.40); however, this was not maintained in the second three-month section of the study when the level of β-carotene supplementation was reduced, MD -8 days (95% CI -18.80 to 2.80) (low-quality evidence). There were no statistically significant effects between groups in lung function (low-quality evidence) and no adverse events were observed (low-quality evidence). Supplementation affected levels of β-carotene in plasma, but not vitamin A levels. The study did not report on quality of life or toxicity. Since no randomised or quasi-randomised controlled studies on retinoid supplementation were identified, no conclusion on the supplementation of vitamin A in people with CF can be drawn. Additionally, due to methodological limitations in the included study, also reflected in the low-quality evidence judged following the specific evidence grading system (GRADE), no clear conclusions on β-carotene supplementation can be drawn. Until further data are available, country- or region-specific guidelines regarding these practices should be followed.

Impact of LCA-Associated E14L LRAT Mutation on Protein Stability and Retinoid Homeostasis.

Vitamin A (all-trans-retinol) is metabolized to the visual chromophore (11-cis-retinal) in the eyes and to all-trans-retinoic acid, a hormone like compound, in most tissues. A key enzyme in retinoid metabolism is lecithin:retinol acyltransferase (LRAT), which catalyzes the esterification of vitamin A. The importance of LRAT is indicated by pathogenic missense and nonsense mutations, which cause devastating blinding diseases. Retinoid-based chromophore replacement therapy has been proposed as treatment for these types of blindness based on studies in LRAT null mice. Here, we analyzed the structural and biochemical basis for retinal pathology caused by mutations in the human LRAT gene. Most LRAT missense mutations associated with retinal degeneration are localized within the catalytic domain, whereas E14L substitution is localized in an N-terminal α-helix, which has been implicated in interaction with the phospholipid bilayer. To elucidate the biochemical consequences of this mutation, we determined LRAT(E14L)'s enzymatic properties, protein stability, and impact on ocular retinoid metabolism. Bicistronic expression of LRAT(E14L) and enhanced green fluorescence protein revealed instability and accelerated proteosomal degradation of this mutant isoform. Surprisingly, instability of LRAT(E14L) did not abrogate the production of the visual chromophore in a cell-based assay. Instead, expression of LRAT(E14L) led to a rapid increase in cellular levels of retinoic acid upon retinoid supplementation. Thus, our study unveils the potential role of retinoic acid in the pathology of a degenerative retinal disease with important implications for the use of retinoid-based therapeutics in affected patients.

Hepatotoxicity of bisphenol A under conditions of differential supplementation with retinoids.

Classical xenoestrogenic in vivo effects of bisphenol A (2,2-bis(4-hydroxyphenyl)propane, BPA) are well-described in the literature, however the molecular mechanisms of BPA-induced hepatotoxicity are not fully characterized. The work is aimed to assess biochemical markers of BPA induced hepatotoxicity under conditions of differential supplementation with retinoids. We demonstrate that the absence of hepatic retinyl esters as the main form of vitamin A storage provides for a resistance to BPA induced liver damage. Retinoid supplementation increases the hepatotoxic effects of bisphenol A, evidenced in higher indexes of oxidative damage of lipids, proteins and non-protein thiol groups as well as increase of serum alanine aminotransferase activity and myeloperoxidase activity in liver parenchyma. The absence of hepatotoxicity signs when hepatic retinoid stores are depleted and their presence during normal or excessive retinoid supplementation suggest that hepatic retinoid availability is one of the factors determining the hepatotoxicity of bisphenol A.

MP87-19 THE IMPACT OF STEROID USE ON ARTIFICIAL URINARY SPHINCTER REOPERATION

You have accessJournal of UrologyUrodynamics/Lower Urinary Tract Dysfunction/Female Pelvic Medicine: Male Incontinence: Therapy I1 Apr 2016MP87-19 THE IMPACT OF STEROID USE ON ARTIFICIAL URINARY SPHINCTER REOPERATION Roderick Clark, Jennifer Winick-Ng, Andrew McClure, and Blayne Welk Roderick ClarkRoderick Clark More articles by this author , Jennifer Winick-NgJennifer Winick-Ng More articles by this author , Andrew McClureAndrew McClure More articles by this author , and Blayne WelkBlayne Welk More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2016.02.2369AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES Artificial urinary sphincters (AUS) are used treat male stress incontinence, often following radical prostatectomy. Previous research has suggested that 1 in 4 men require a second AUS-related operation after 5 years, due to AUS complications or recurrent incontinence. Corticosteroids are associated with impaired wound healing and infections in the surgical setting, and are commonly used in the treatment of metastatic prostate cancer, and for various inflammatory diseases. The objective of this study was to determine if oral steriod use was associated with an increased risk of AUS related reoperation. METHODS We used administrative data from the province of Ontario, Canada to conduct a retrospective cohort study. Our population was men >65 years of age, who underwent implantation of an incident AUS between April 1st 2002 and December 31st 2013. Our primary outcome was the first reoperation on the AUS for any reason. Our primary exposure was defined as a filled prescription for an oral corticosteroid at the time of AUS implantation, or during followup (but prior to any AUS reoperation); we considered men to be exposed from the first day of the prescription to 180 days after the last day of medication. Our primary analysis was a cox proportional hazards model with steroid usage as a time varying covariate. RESULTS We identified 747 men, (median age 71, IQR 68-75), of which 592 (79.3%) had a prior radical prostatectomy, and 29 (3.9%) had prior primary radiation therapy for prostate cancer. The median cohort follow-up was 3.2 (IQR 1.3-5.9) years. 175/747 (23.4%) were exposed to corticosteroids during the study period (median duration of prescription was 22.0 days, IQR 5.0-133.0). We identified an initial AUS reoperation in 176/747 men (23.5%) a median of 1.4 years (IQR 0.4-3.0) after implantation. In our primary analysis, with adjustment for age, radiation exposure, and year of implantation, the hazard ratio for subsequent reoperation on the AUS among steroid users was 1.75 (95% CI 1.08-2.84, p=0.02). CONCLUSIONS To our knowledge this is the first observational study to demonstrate the significant increased risk in reoperation among men taking oral steriods. This risk factor should be considered in patient counselling, and further studies evaluating the specific utilization patterns of steroids and their association with AUS complications may be warranted. Given the potential cost and morbidity associated with AUS reoperation, potential strategies for reducing the harmful effects of steroids on wound healing (such as retinoid supplementation) could be further studied among AUS patients on steriod therapy. © 2016FiguresReferencesRelatedDetails Volume 195Issue 4SApril 2016Page: e1122 Advertisement Copyright & Permissions© 2016MetricsAuthor Information Roderick Clark More articles by this author Jennifer Winick-Ng More articles by this author Andrew McClure More articles by this author Blayne Welk More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...

Comprehensive genotyping reveals RPE65 as the most frequently mutated gene in Leber congenital amaurosis in Denmark.

Leber congenital amaurosis (LCA) represents the most severe form of inherited retinal dystrophies with an onset during the first year of life. Currently, 21 genes are known to be associated with LCA and recurrent mutations have been observed in AIPL1, CEP290, CRB1 and GUCY2D. In addition, sequence analysis of LRAT and RPE65 may be important in view of treatments that are emerging for patients carrying variants in these genes. Screening of the aforementioned variants and genes was performed in 64 Danish LCA probands. Upon the identification of heterozygous variants, Sanger sequencing was performed of the relevant genes to identify the second allele. In combination with prior arrayed primer extension analysis, this led to the identification of two variants in 42 of 86 cases (49%). Remarkably, biallelic RPE65 variants were identified in 16% of the cases, and one novel variant, p.(D110G), was found in seven RPE65 alleles. We also collected all previously published RPE65 variants, identified in 914 alleles of 539 patients with LCA or early-onset retinitis pigmentosa, and deposited them in the RPE65 Leiden Open Variation Database (LOVD). The in silico pathogenicity assessment of the missense and noncanonical splice site variants, as well as an analysis of their frequency in ~60 000 control individuals, rendered 864 of the alleles to affect function or probably affect function. This comprehensive database can now be used to select patients eligible for gene augmentation or retinoid supplementation therapies.

Pathogenesis of Endometriosis: Roles of Retinoids and Inflammatory Pathways.

Endometriosis is a nonmalignant, but potentially metastatic, gynecological condition manifested by the extrauterine growth of inflammatory endometrial implants. Ten percent of reproductive-age women are affected and commonly suffer pelvic pain and/or infertility. The theories of endometriosis histogenesis remain controversial, but retrograde menstruation and metaplasia each infer mechanisms that explain the immune cell responses observed around the ectopic lesions. Recent findings from our laboratories and others suggest that retinoic acid metabolism and action are fundamentally flawed in endometriotic tissues and even generically in women with endometriosis. The focus of our ongoing research is to develop medical therapies as adjuvants or alternatives to the surgical excision of these lesions. On the basis of concepts put forward in this review, we predict that the pharmacological actions and anticipated low side-effect profiles of retinoid supplementation might provide a new treatment option for the long-term management of this chronic and debilitating gynecological disease.

Vitamin A Kinetics in Neonatal Rats vs. Adult Rats: Comparisons from Model-Based Compartmental Analysis ,

A critical role for vitamin A (VA) in development is well established, but still relatively little is known about whole-body VA metabolism in early postnatal life. Recently, methods of mathematical modeling have begun to shed light on retinol kinetics in the postnatal growth period and on the effect of retinoid supplementation on retinol kinetics. Comparison of kinetic parameters from tracer studies in neonatal rats with those previously determined in models of VA metabolism in the adult suggests both similarities and differences in the relative transfer rates of plasma retinol to extrahepatic tissues, resulting in similarities and differences in kinetic parameters and inferences about physiologic processes. Similarities between neonatal and adult models include the capacity for efficient digestion and absorption of VA; characteristics of a high-response system; extensive retinol recycling among liver, plasma, and extrahepatic tissues; and comparable VA disposal rates. Differences between neonatal and adult models include that, in neonates, retinol turnover is faster and retinol recycling is much more extensive; there is a greater role for extrahepatic tissues in the uptake of chylomicron VA; and the intestine plays an important role in chylomicron VA uptake, especially in neonatal rats treated with a supplement containing VA. In summary, retinol kinetic modeling in the neonatal rat has provided a first view of whole-body VA metabolism in this age group and suggests that VA kinetics in neonatal rats differs in many ways from that in adults, perhaps reflecting an adaption to the lower VA concentration found in neonates compared with adults.

Noncanonical Autophagy Promotes the Visual Cycle

Phagocytosis and degradation of photoreceptor outer segments (POS) by retinal pigment epithelium (RPE) is fundamental to vision. Autophagy is also responsible for bulk degradation of cellular components, but its role in POS degradation is not well understood. We report that the morning burst of RPE phagocytosis coincided with the enzymatic conversion of autophagy protein LC3 to its lipidated form. LC3 associated with single-membrane phagosomes containing engulfed POS in an Atg5-dependent manner that required Beclin1, but not the autophagy preinitiation complex. The importance of this process was verified in mice with Atg5-deficient RPE cells that showed evidence of disrupted lysosomal processing. These mice also exhibited decreased photoreceptor responses to light stimuli and decreased chromophore levels that were restored with exogenous retinoid supplementation. These results establish that the interplay of phagocytosis and autophagy within the RPE is required for both POS degradation and the maintenance of retinoid levels to support vision.

Retinoid supplementation of differentiating human neural progenitors and embryonic stem cells leads to enhanced neurogenesis in vitro

Retinoids are important molecules involved in the development and homeostasis of the nervous system. As such, various retinoid derivatives are often found in culture media and supplement formulations to support the growth and maintenance of neural cells. However, all-trans-retinoic acid (ATRA) and its associated derivatives are light sensitive and are highly susceptible to isomerisation. This can lead to variability in retinoid concentrations and the nature of the retinoid species present in culture solutions which in turn can influence biological activity and introduce inconsistency. We have previously described the development of the synthetic retinoid derivative, EC23, as a chemically and light stable alternative that does not degrade and has biological activity similar to ATRA. In this study we demonstrate that the addition of exogenous retinoid can significantly enhance neuronal differentiation of both human neuroprogenitor and human embryonic stem cells. In the former, both ATRA and EC23 induced increased maturation and stabilisation of the axonal cytoskeleton. However, EC23 was particularly potent at lower nanomolar concentrations resulting in significantly greater neurogenesis than ATRA. In ES cells enhanced motor neuron marker expression was also detected in response to both retinoids when incorporated into an established protocol for neuronal differentiation. We propose that synthetic retinoid EC23 represents a valuable addition to the formulation of new and existing culture supplements to enhance neuronal differentiation whilst enabling improved consistency.

Nrl-Knockout Mice Deficient in Rpe65 Fail to Synthesize 11-cisRetinal and Cone Outer Segments

To define rod and cone function further in terms of visual cycle mechanism, the retinal phenotype resulting from Rpe65 (retinoid isomerase I) deficiency in Nrl(-)(/)(-) mice having a single class of photoreceptors resembling wild-type cones was characterized and outcomes of retinoid supplementation evaluated. Rpe65(-)(/)(-)/Nrl(-)(/)(-) mice were generated by breeding Rpe65(-)(/)(-) and Nrl(-)(/)(-) strains. Retinal histology, protein expression, retinoid content, and electroretinographic (ERG) responses were evaluated before and after treatment with 11-cis retinal by intraperitoneal injection. Results Retinas of young Rpe65(-)(/-)/Nrl(-)(/-) mice exhibited normal lamination, but lacked intact photoreceptor outer segments at all ages examined. Rpe65, Nrl, and rhodopsin were not detected, and S-opsin and M/L-opsin levels were reduced. Retinyl esters were the only retinoids present. In contrast, Nrl(-)(/)(-) mice exhibited decreased levels of retinaldehydes and retinyl esters, and elevated levels of retinols. ERG responses were elicited from Rpe65(-)(/-)/Nrl(-)(/-) mice only at the two highest intensities over a 4-log-unit range. Significant retinal thinning and outer nuclear layer loss occurred in Rpe65(-)(/-)/Nrl(-)(/-) mice with aging. Administration of exogenous 11-cis retinal did not rescue retinal morphology or markedly improve ERG responses. The findings provide clarification of reported cone loss of function in Rpe65(-)(/-)/Nrl(-)(/-) mice, now showing that chromophore absence results in destabilized cone outer segments and rapid retinal degeneration. The data support the view that rod-dominant retinas do not have a cone-specific mechanism for 11-cis retinal synthesis and have potential significance for therapeutic strategies for rescue of cone-rich retinal regions affected by disease in the aging human population.

Retinoic Acid Modulates Hepatic Iron Homeostasis in Rats by Attenuating the RNA-Binding Activity of Iron Regulatory Proteins ,

Vitamin A deficiency has been widely associated with perturbations of iron homeostasis, a consequence that can be reversed by retinoid supplementation. Despite the numerous studies that demonstrate an interaction between these 2 nutrients, the mechanistic basis for this relation has not been well characterized. Because iron regulatory proteins (IRP) have been established as central regulators of iron homeostasis, we investigated the potential role of IRP in the regulation of iron homeostasis under conditions of vitamin A deficiency and supplementation with all-trans-retinoic acid (atRA). Rats were fed a control diet or a diet deficient in either vitamin A or iron or both micronutrients. Four parallel groups of rats were supplemented with atRA daily (30 μmol/kg body weight) during the final week of this study. As expected, iron-deficient (−Fe) rats exhibited a decrease in hepatic nonheme iron levels and a subsequent increase in IRP RNA-binding activity, resulting in diminished ferritin abundance. Interestingly, atRA supplementation inhibited the increase in IRP RNA-binding activity in −Fe rats to a level that was not significantly (P = 0.139) different from control values, and it partially restored ferritin abundance. This inhibition of IRP RNA-binding activity by atRA supplementation was also associated with a 40% reduction in transferrin receptor abundance. Taken together, these results indicate that IRP represent a mechanistic link between vitamin A and the regulation of iron homeostasis, a key finding toward further understanding this important nutrient-nutrient interaction.

Retinoids Increase Lung Elastin Expression But Fail to Alter Morphology or Angiogenesis Genes in Premature Ventilated Baboons

Retinoids regulate elastin synthesis by alveolar myofibroblasts and affect angiogenesis pathways, both of which are processes critical for alveolar development. Retinoids accelerate alveolarization in rodents and are now used therapeutically in premature infants at risk of bronchopulmonary dysplasia (BPD). This study examined the effects of retinoid supplementation on alveolar elastin expression and deposition and angiogenesis-related signaling in a primate model of BPD. Premature baboons delivered at 125 d of gestation after maternal steroid treatment were given surfactant and ventilated with minimal supplemental oxygen for 14 d with (n = 5) and without (n = 5) supplemental vitamin A (5000 U/kg/d) and compared with 140-d unventilated controls. Ventilatory efficiency index (VEI) and oxygenation index (OI) were not statistically different between ventilated treatment groups. Expression of vascular endothelial growth factor A (VEGF-A), fms-related tyrosine kinase 1 (Flt-1), and tyrosine kinase with immunoglobulin-like and EGF-like domains 1 (TIE-1) was repressed by premature delivery and mechanical ventilation and was not altered by retinoid supplementation. Retinoid supplementation did not enhance alveolar angiogenesis. Elastin expression was repressed by premature delivery and extended ventilation, and retinoid supplementation increased elastin expression specifically in alveolar myofibroblasts within alveolar walls. These results suggest that the small decrease in mortality among premature infants receiving retinoid supplementation may not be mediated through enhanced alveolar development.

Rat hepatic stellate cells become retinoid unresponsive during activation

Hepatic stellate cells (HSC) play an essential role in fibrogenesis. Many stimuli cause HSC to activate, lose their Vitamin A and produce collagen. It is unclear whether Vitamin A loss causes activation, potentiates it or is simply an event in the cascade of activation changes. We determine if exogenous retinoids prevent the activation of freshly isolated rat HSC activated by plating on plastic. We also determine if retinoids: (1) reverse HSC activation; (2) maintain/restore HSC intracellular retinoid levels; (3) maintain expression of HSC nuclear receptors for retinoic acid (RAR) in HSC that are becoming activated or are chronically activated. Markers of activation in freshly isolated HSC were decreased by either retinol or retinoic acid without increases in HSC retinoid concentration. mRNA levels for RAR-alpha, RAR-beta and RAR-gamma, the nuclear receptors for retinoic acid, decreased during activation of freshly isolated HSC even with retinoid supplementation. RAR-alpha, RAR-beta and RAR-gamma mRNA and RAR-beta protein was undetectable in chronically activated HSC and remained absent after retinoic acid supplementation. Activation markers in chronically activated HSC were only slightly decreased after retinoid exposure. We conclude that exposure of HSC to extracellular retinoids diminishes some markers of activation but does not prevent HSC activation.

Enhanced UV-mediated free radical generation; DNA and mitochondrial damage caused by retinol supplementation.

Retinoid supplementation has been therapeutically used against various human disorders. We and others have demonstrated that retinol treatment causes free radical generation and increased iron uptake, iron storage and oxidative damage, both in vitro and in vivo. Here, we investigate the possible synergistic effect of retinol on UV-mediated free radical generation, oxidative damage to biomolecules and decreased cellular viability in primary cultured mammalian cells. Retinol treatment (7 microM) resulted in a threefold increase in UV-mediated free radical generation and a 40%, increase in lipoperoxidation. DNA fragmentation and mitochondrial oxidative damage also increased significantly in retinol-supplemented UV-irradiated cultured cells as compared to UV-irradiated control cells, which were only treated with the solvent used to deliver the retinol (0.1% ethanol). All measurements were restored to control values when an iron chelator, 1,10-phenanthroline (100 microM), or an OH* scavenger, mannitol (1 mM), was co-administrated. Rather than protecting against free radical generation, retinol seems to enhance UV-mediated oxidative damage and decreases cellular viability in cultured cells. We suggest that retinol-enhanced iron uptake and storage and increased reactive oxygen species generated by the Fenton reaction may act synergistically with UV-irradiation in causing oxidative damage to cells.

Retinoid-Induced Modulation of Immunoglobulin M Secretion by Bovine Mononuclear Leukocytes In Vitro

Effects of trans and cis isomers of retinol and retinoic acid on IgM secretion by bovine peripheral blood mononuclear leukocytes were evaluated in vitro. Mononuclear leukocyte cultures that were unstimulated or stimulated by pokeweed mitogen were supplemented with isomers of retinol and retinoic acid at 10–10 to 10–6M. Concentrations of polyclonal IgM in supernatants from 14-d cultures were measured by an ELISA. Cultures stimulated by pokeweed mitogen consistently secreted more IgM than parallel, unstimulated cultures. Retinoid supplementation did not affect basal IgM secretion by unstimulated cultures. However, each retinoid affected IgM secretion by cultures stimulated by mitogen. The nature of the effect was dependent on the concentration of the specific retinoid. All-trans-retinoic acid enhanced secretion at 10–10M and inhibited secretion at 10–6M. The other retinoids, however, did not inhibit IgM secretion at any concentration. Each retinoid enhanced IgM secretion at one or more concentrations, although enhancement was produced by much lower concentrations of retinoic acid isomers than retinol isomers. These results indicate that retinol and retinoic acid modulate polyclonal IgM secretion by cultures of bovine mononuclear leukocytes stimulated by mitogen. Future research will determine which subsets of the mononuclear leukocyte population are affected and whether trans-retinoic acid is the metabolite that produces these effects.

Effect of vitamin A on the growth and differentiation of rat external auditory canal epithelium in organ culture

Intact rat external ear canal explants were maintained either in retinoid-deficient or retinoid-supplemented medium for 7, 10, 14, and 21 days. The morphology of the external ear canal epidermis was well maintained, including the presence of sebaceous glands even after 21 days in culture with retinoid-deficient medium. However, after culturing in retinoid-supplemented medium, the external ear canal epithelium showed both a loss of keratohyalin granules and the formation of keratin. Extensive microvilli formation occurred, even though the desquamation process continued from the superficial layer after 7, 10, and 14 days in culture with retinoid-enriched medium. After 21 days in culture with retinoid-supplemented medium, the ear canal epithelium contained well-developed Golgi apparatus and secretory granules as well. It was concluded that the ear canal epithelium was transformed into a secretory-like mucosal epithelium by retinoid supplementation.