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INTRODUCTION: Caused by mutation or deletion of the CHM gene, choroideremia is a rare X-linked recessive chorioretinal dystrophy characterized by progressive degeneration of the retinal pigment epithelium, photoreceptors, and the choriocapillaris. There are few published reports of choroideremia associated with complex syndromic phenotypes due to large or contiguous gene deletions. METHODS: Case report and review of literature. RESULTS: We present a case of a 46-year-old male with a prior clinical diagnosis of syndromic retinitis pigmentosa, who was found to have syndromic choroideremia associated with a novel multi-gene deletion of 13.5 megabase pairs. This deletion encompassing 18 genes is one of the largest deletions reported in the literature. A total of 18 male cases of choroideremia associated with confirmed large or contiguous gene deletions have been published to date. Previously reported deletions range in size from 4 to 15 megabase pairs, and observed phenotypes include cleft lip and palate, ptosis, obesity, metabolic diseases, developmental delay, and hearing loss. DISCUSSION: The contribution of our case aims to expand our understanding of Xq21 deletions and prompts further investigation of genes found in this locus. Furthermore, it highlights the importance of including syndromic choroideremia on the differential diagnosis in the workup of other syndromic retinopathies, particularly those that feature obesity, hearing loss, or intellectual disability.
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The first steps in vision take place in photoreceptor cells, which are highly compartmentalized neurons exhibiting significant structural variation across species. The light-sensitive ciliary compartment, called the outer segment, is located atop of the cell soma, called the inner segment. In this study, we present an ultrastructural analysis of human photoreceptors, which reveals that, in contrast to this classic arrangement, the inner segment of human rods extends alongside the outer segment to form a structure hereby termed the "accessory inner segment". While reminiscent of the actin-based microvilli known as "calyceal processes" observed in other species, the accessory inner segment is a unique structure: (1) it contains an extensive microtubule-based cytoskeleton, (2) it extends far alongside the outer segment, (3) its diameter is comparable to that of the outer segment, (4) it contains numerous mitochondria, and (5) it forms electron-dense structures that likely mediate adhesion to the outer segment. Given that the spacing of extrafoveal human photoreceptors is more sparse than in non-primate species, with vast amounts of interphotoreceptor matrix present between cells, the closely apposed accessory inner segment likely provides structural support to the outer segment. This discovery expands our understanding of the human retina and directs future studies of human photoreceptor function in health and disease.
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BACKGROUND: Pemigatinib is an inhibitor of the fibroblast growth factor receptor (FGFR), recently approved for the treatment of cholangiocarcinoma. FGFR retinopathy is a newly recognized entity, with only two other FGFR inhibitors reported to cause serous retinopathy. Herein, we describe the first published report of a multifocal serous retinopathy secondary to pemigatinib. CASE PRESENTATION: A 67-year-old male with stage 4A metastatic colon adenocarcinoma undergoing systemic therapy with pemigatinib was found to have developed bilateral multifocal serous retinopathy. Fundus autofluorescence showed corresponding multifocal hypoautofluorescent foci, whereas fluorescein angiography and indocyanine green angiography were unremarkable. Subretinal fluid resolved rapidly after discontinuation of pemigatinib. CONCLUSIONS: Multifocal serous retinopathy appears to be a class effect of FGFR inhibitors. FGFR retinopathy clinically resembles MEK retinopathy-both feature multifocal subretinal fluid, low visual significance, and quick resolution. However, given that FGFR inhibitors have a broader molecular range than MEK inhibitors, further characterization of FGFR retinopathy is necessary to generate management guidelines.
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Purpose: Herpes simplex virus type I (HSV-1) infection of corneal epithelial cells activates ataxia telangiectasia mutated (ATM), an apical kinase in the host DNA damage response pathway, whose activity is necessary for the progression of lytic HSV-1 infection. The purpose of this study is to investigate the mechanism of ATM activation by HSV-1 in the corneal epithelium, as well as its functional significance. Methods: Mechanistic studies were performed in cultured human corneal epithelial cell lines (hTCEpi, HCE), as well as in esophageal (EPC2) and oral (OKF6) cell lines. Transfection-based experiments were performed in HEK293 cells. HSV-1 infection was carried out using the wild-type KOS strain, various mutant strains (tsB7, d120, 7134, i13, n208), and bacterial artificial chromosomes (fHSVΔpac, pM24). Inhibitors of ATM (KU-55933), protein synthesis (cycloheximide), and viral DNA replication (phosphonoacetic acid) were used. Outcomes of infection were assayed using Western blotting, qRT-PCR, immunofluorescence, and comet assay. Results: This study demonstrates that HSV-1-mediated ATM activation in corneal epithelial cells relies on the viral immediate early gene product ICP4 and requires the presence of the viral genome in the host nucleus. We show that ATM activation is independent of viral genome replication, the ICP0 protein, and the presence of DNA lesions. Interestingly, ATM activity appears to be necessary at the onset of infection, but dispensable at the later stages. Conclusions: This study expands our understanding of HSV-1 virus-host interactions in the corneal epithelium and identifies potential areas of future investigation and therapeutic intervention in herpes keratitis.
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ADN Viral/genética , Epitelio Corneal/metabolismo , Infecciones Virales del Ojo/virología , Herpesvirus Humano 1/genética , Queratitis Herpética/virología , Replicación Viral/fisiología , Células Cultivadas , Daño del ADN , Replicación del ADN , Epitelio Corneal/patología , Epitelio Corneal/virología , Infecciones Virales del Ojo/metabolismo , Infecciones Virales del Ojo/patología , Humanos , Queratitis Herpética/metabolismo , Queratitis Herpética/patologíaRESUMEN
A systematic analytic treatment of local fluctuations in the regularized Laplacian growth problem is given. The interface dynamics is stabilized by a short-distance cutoff â preventing the cusps production in a finite time. The regularization mechanism results in the violation of the incompressibility condition of the viscous fluid on a microscale in the vicinity of the moving interface, thus producing local fluctuations of pressure. Dissipation of fluctuations with time is described by universal Dyson Brownian motion, which reduces to the complex viscous Burgers equation in the hydrodynamic approximation. Because of the intrinsic instability of the interface dynamics, tiny fluctuations of pressure generate universal complex patterns with well developed fjords and fingers in a long time asymptotic.
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A one-parametric stochastic regularized dynamics of the interface in the Hele-Shaw cell is introduced. The short-distance regularization suggested by the aggregation model stabilizes the growth by preventing the formation of cusps at the interface and makes the interface dynamics chaotic. The introduced stochastic growth process generates universal complex patterns with the well-developed fjords of oil separating the fingers of water. In a long time asymptotic, by coupling a conformal field theory to the stochastic growth process, we introduce a set of observables (the martingales), whose expectation values are constant in time. The martingales are closely connected to degenerate representations of the Virasoro algebra and can be written in terms of conformal correlation functions. A direct link between Laplacian growth and conformal Liouville field theory with the central charge c≥25 is proposed.
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We develop statistical mechanics for stochastic growth processes and apply it to Laplacian growth by using its remarkable connection with a random matrix theory. The Laplacian growth equation is obtained from the variation principle and describes adiabatic (quasistatic) thermodynamic processes in the two-dimensional Dyson gas. By using Einstein's theory of thermodynamic fluctuations we consider transitional probabilities between thermodynamic states, which are in a one-to-one correspondence with simply connected domains occupied by gas. Transitions between these domains are described by the stochastic Laplacian growth equation, while the transitional probabilities coincide with a free-particle propagator on an infinite-dimensional complex manifold with a Kähler metric.
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A point source on a plane constantly emits particles which rapidly diffuse and then stick to a growing cluster. The growth probability of a cluster is presented as a sum over all possible scenarios leading to the same final shape. The classical point for the action, defined as a minus logarithm of the growth probability, describes the most probable scenario and reproduces the Laplacian growth equation, which embraces numerous fundamental free boundary dynamics in nonequilibrium physics. For nonclassical scenarios we introduce virtual point sources, in which presence the action becomes the Kullback-Leibler entropy. Strikingly, this entropy is shown to be the sum of electrostatic energies of layers grown per elementary time unit. Hence the growth probability of the presented nonequilibrium process obeys the Gibbs-Boltzmann statistics, which, as a rule, is not applied out from equilibrium. Each layer's probability is expressed as a product of simple factors in an auxiliary complex plane after a properly chosen conformal map. The action at this plane is a sum of Robin functions, which solve the Liouville equation. At the end we establish connections of our theory with the τ function of the integrable Toda hierarchy and with the Liouville theory for noncritical quantum strings.
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BACKGROUND/AIMS: Herpes simplex virus (HSV) type I keratitis remains a leading cause of corneal morbidity, despite the availability of effective antiviral drugs. Improved understanding of virus-host interactions at the level of the host DNA damage response (DDR), a known factor in the development of HSV-1 keratitis, may shed light on potential new therapeutic targets. This report examines the role of checkpoint kinase 2 (Chk2), a DDR mediator protein, in corneal epithelial HSV-1 infection. METHODS: A small-molecule inhibitor of Chk2 (Chk2 inhibitor II) was applied to HSV-1-infected cultured human corneal epithelial cells (hTCEpi and HCE) as well as to explanted and organotypically cultured human and rabbit corneas. Infection levels were assessed by plaque assay and real-time PCR. RNAi-mediated depletion of Chk2 was performed to confirm the effect of the inhibitor. RESULTS: Inhibition of the Chk2 kinase activity greatly suppresses the cytopathic effect, genome replication and infectious progeny production in vitro and ex vivo. CONCLUSION: This report demonstrates the critical role of Chk2 kinase in the establishment of HSV-1 corneal epithelial infection. These data contribute to our understanding of herpesvirus-host interactions and underscore the significance of DDR activation in HSV-1 keratitis.
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Quinasa de Punto de Control 2/metabolismo , Epitelio Corneal/virología , Herpesvirus Humano 1/fisiología , Queratitis Herpética/virología , Replicación Viral , Animales , Western Blotting , Células Cultivadas , Quinasa de Punto de Control 2/antagonistas & inhibidores , Efecto Citopatogénico Viral , Electroforesis en Gel de Poliacrilamida , Activación Enzimática , Inhibidores Enzimáticos/farmacología , Epitelio Corneal/efectos de los fármacos , Técnica del Anticuerpo Fluorescente Indirecta , Humanos , Queratitis Herpética/enzimología , Técnicas de Cultivo de Órganos , Fosforilación , Conejos , Reacción en Cadena en Tiempo Real de la PolimerasaRESUMEN
Age-related macular degeneration (AMD) is the most common cause of blindness among older adults in developed countries, and retinal iron accumulation may exacerbate the disease. Iron can upregulate the production of amyloid precursor protein (APP). Since amyloid-ß (Aß), a byproduct of APP proteolysis, is found in drusen, the histopathological hallmark of AMD, we tested the role of iron in regulating APP and Aß levels in the retinal pigment epithelial cell line ARPE-19. We found that treatment with ferric ammonium citrate (FAC) increases APP at the translational level. FAC treatment also results in increased generation of APP C-terminal fragments C83 and C99, the products of APP proteolysis by α- and ß-secretase, respectively, as well as levels of Aß42, a highly aggregative amyloid species. Additionally, retinal tissue sections from a patient with aceruloplasminemia, a disease causing iron overload in the retinal pigment epithelium (RPE), showed increased Aß deposition in the RPE and drusen. Overall, our results suggest that RPE iron overload could contribute to Aß accumulation in the retina.
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Precursor de Proteína beta-Amiloide/genética , Regulación de la Expresión Génica , Hierro/metabolismo , Degeneración Macular/genética , Biosíntesis de Proteínas , ARN Mensajero/genética , Epitelio Pigmentado de la Retina/metabolismo , Precursor de Proteína beta-Amiloide/biosíntesis , Western Blotting , Células Cultivadas , Ensayo de Inmunoadsorción Enzimática , Humanos , Inmunohistoquímica , Degeneración Macular/metabolismo , Degeneración Macular/patología , Masculino , Persona de Mediana Edad , Reacción en Cadena en Tiempo Real de la Polimerasa , Epitelio Pigmentado de la Retina/patologíaRESUMEN
PURPOSE: Proangiogenic protein VEGF-A contributes significantly to retinal lesions and neovascularization in diabetic retinopathy (DR). In preclinical DR, hyperglycemia can upregulate VEGF-A in retinal cells. The VEGF-A promoter is responsive to the transcription factor specificity protein 1 (Sp1). The O-GlcNAc modification is driven by glucose concentration and has a profound effect on Sp1 activity. This study investigated the effects of hyperglycemia on Sp1-mediated expression of VEGF-A in the retinal endothelium and pigment epithelium. METHODS: Hyperglycemia-exposed ARPE-19 (human retinal pigment epithelial cells) and TR-iBRB (rat retinal microendothelial cells) were assayed for levels of VEGF-A by qRT-PCR, Western blot, and ELISA. Small molecule inhibitors of O-GlcNAc transferase (OGT) or O-GlcNAcase (OGA) were used to manipulate O-GlcNAc levels. Vascular endothelial growth factor-A protein and transcript were measured in cells depleted of OGT or Sp1 by shRNA. The proximal VEGF-A promoter was analyzed for glucose sensitivity by luciferase assay. Chromatin immunoprecipitation (ChIP) was used to assess Sp1 occupancy on the VEGF-A promoter. RESULTS: Hyperglycemia increased VEGF-A promoter activity and upregulated VEGF-A transcript and protein. Elevation of O-GlcNAc by OGA inhibitors was sufficient to increase VEGF-A. O-GlcNAc transferase inhibition abrogated glucose-driven VEGF-A. Cellular depletion of OGT or Sp1 by shRNA significantly abrogated glucose-induced changes in VEGF-A. ChIP analysis showed that hyperglycemia significantly increased binding of Sp1 to the VEGF-A promoter. CONCLUSIONS: Hyperglycemia-driven VEGF-A production is mediated by elevated O-GlcNAc modification of the Sp1 transcription factor. This mechanism may be significant in the pathogenesis of preclinical DR through VEGF-A upregulation.
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Hiperglucemia/metabolismo , N-Acetilglucosaminiltransferasas/fisiología , Retina/metabolismo , Factor de Transcripción Sp1/fisiología , Factor A de Crecimiento Endotelial Vascular/metabolismo , Animales , Línea Celular , Modelos Animales de Enfermedad , Células Endoteliales/metabolismo , Células Epiteliales/metabolismo , Humanos , N-Acetilglucosaminiltransferasas/antagonistas & inhibidores , Ratas , Epitelio Pigmentado de la Retina/metabolismo , Activación Transcripcional , Regulación hacia Arriba/efectos de los fármacosRESUMEN
The purpose of our studies was to examine the relationship between iron and melanogenesis in retinal pigment epithelial cells, as prior observations had suggested that iron may promote melanogenesis. This relationship has potential clinical importance, as both iron overload and hyperpigmentation are associated with age-related macular degeneration (AMD). Human fetal retinal pigment epithelial cells and ARPE-19 cells were treated with iron in the form of ferric ammonium citrate, after which quantitative RT-PCR and electron microscopy were performed. Melanogenesis genes tyrosinase, tyrosinase-related protein 1, Hermansky-Pudlak Syndrome 3, premelanosome protein and dopachrome tautomerase were upregulated, as was the melanogenesis-controlling transcription factor, microphthalmia-associated transcription factor (MITF). Iron-treated cells had increased pigmentation and melanosome number. Multiple transcription factors upstream of MITF were upregulated by iron.
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Compuestos Férricos/farmacología , Melaninas/biosíntesis , Melanosomas/metabolismo , Compuestos de Amonio Cuaternario/farmacología , Epitelio Pigmentado de la Retina/efectos de los fármacos , Regulación hacia Arriba/fisiología , Western Blotting , Proteínas Portadoras/genética , Células Cultivadas , Edad Gestacional , Humanos , Péptidos y Proteínas de Señalización Intracelular , Oxidorreductasas Intramoleculares/genética , Glicoproteínas de Membrana/genética , Monofenol Monooxigenasa/genética , Oxidorreductasas/genética , ARN Mensajero/genética , Reacción en Cadena en Tiempo Real de la Polimerasa , Epitelio Pigmentado de la Retina/embriología , Epitelio Pigmentado de la Retina/metabolismo , Donantes de Tejidos , Antígeno gp100 del Melanoma/genéticaRESUMEN
PURPOSE: Herpes keratitis (HK) is the leading cause of cornea-derived and infection-associated blindness in the developed world. Despite the availability of effective antivirals, some patients develop refractory disease, drug-resistant infection, and topical toxicity. A nonpharmaceutical treatment modality may offer a unique advantage in the management of such cases. This study investigated the antiviral effect of nonthermal dielectric barrier discharge (DBD) plasma, a partially ionized gas that can be applied to organic substances to produce various biological effects. METHODS: Human corneal epithelial cells and explanted corneas were infected with herpes simplex virus type 1 (HSV-1) and exposed to culture medium treated with nonthermal DBD plasma. The extent of infection was measured by plaque assay, quantitative PCR, and Western blot. Corneal toxicity assessment was performed with fluorescein staining, histologic examination, and 8-OHdG detection. RESULTS: Application of DBD plasma-treated medium to human corneal epithelial cells and explanted corneas produced a dose-dependent reduction of the cytopathic effect, viral genome replication, and the overall production of infectious viral progeny. Toxicity studies showed lack of detrimental effects in explanted human corneas. CONCLUSIONS: Nonthermal DBD plasma substantially suppresses corneal HSV-1 infection in vitro and ex vivo without causing pronounced toxicity. TRANSLATIONAL RELEVANCE: Nonthermal plasma is a versatile tool that holds great biomedical potential for ophthalmology, where it is being investigated for wound healing and sterilization and is already in use for ocular microsurgery. The anti-HSV-1 activity of DBD plasma demonstrated here could be directly translated to the clinic for use against drug-resistant herpes keratitis.
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PURPOSE: Herpes keratitis (HK) remains the leading cause of cornea-derived blindness in the developed world, despite the availability of effective antiviral drugs. Treatment toxicity and the emergence of drug resistance highlight the need for additional therapeutic approaches. This study examined ataxia telangiectasia mutated (ATM), an apical kinase in the host DNA damage response, as a potential new target for the treatment of HK. METHODS: Small molecule inhibitor of ATM (KU-55933) was used to treat herpes simplex virus type 1 (HSV-1) infection in three experimental models: (1) in vitro--cultured human corneal epithelial cells, hTCEpi, (2) ex vivo--organotypically explanted human and rabbit corneas, and (3) in vivo--corneal infection in young C57BL/6J mice. Infection productivity was assayed by plaque assay, real-time PCR, Western blot, and disease scoring. RESULTS: Robust ATM activation was detected in HSV-1-infected human corneal epithelial cells. Inhibition of ATM greatly suppressed viral replication in cultured cells and in explanted human and rabbit corneas, and reduced the severity of stromal keratitis in mice. The antiviral effect of KU-55933 in combination with acyclovir was additive, and KU-55933 suppressed replication of a drug-resistant HSV-1 strain. KU-55933 caused minimal toxicity, as monitored by clonogenic survival assay and fluorescein staining. CONCLUSIONS: This study identifies ATM as a potential target for the treatment of HK. ATM inhibition by KU-55933 reduces epithelial infection and stromal disease severity without producing appreciable toxicity. These findings warrant further investigations into the DNA damage response as an area for therapeutic intervention in herpetic ocular diseases.
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Inhibidores Enzimáticos/farmacología , Epitelio Corneal/virología , Herpesvirus Humano 1/fisiología , Queratitis Herpética/prevención & control , Morfolinas/farmacología , Pironas/farmacología , Aciclovir/farmacología , Animales , Antivirales/farmacología , Proteínas de la Ataxia Telangiectasia Mutada/antagonistas & inhibidores , Proteínas de la Ataxia Telangiectasia Mutada/metabolismo , Western Blotting , Células Cultivadas , Modelos Animales de Enfermedad , Combinación de Medicamentos , Epitelio Corneal/enzimología , Femenino , Humanos , Inmunohistoquímica , Queratitis Herpética/enzimología , Queratitis Herpética/virología , Ratones , Ratones Endogámicos C57BL , Técnicas de Cultivo de Órganos , Conejos , Reacción en Cadena en Tiempo Real de la Polimerasa , Ensayo de Placa Viral , Replicación Viral/fisiologíaRESUMEN
Herpes keratitis is one of the most severe pathologies associated with the herpes simplex virus-type 1 (HSV-1). Herpes keratitis is currently the leading cause of both cornea-derived and infection-associated blindness in the developed world. Typical presentation of herpes keratitis includes infection of the corneal epithelium and sometimes the deeper corneal stroma and endothelium, leading to such permanent corneal pathologies as scarring, thinning, and opacity. Corneal HSV-1 infection is traditionally studied in two types of experimental models. The in vitro model, in which cultured monolayers of corneal epithelial cells are infected in a Petri dish, offers simplicity, high level of replicability, fast experiments, and relatively low costs. On the other hand, the in vivo model, in which animals such as rabbits or mice are inoculated directly in the cornea, offers a highly sophisticated physiological system, but has higher costs, longer experiments, necessary animal care, and a greater degree of variability. In this video article, we provide a detailed demonstration of a new ex vivo model of corneal epithelial HSV-1 infection, which combines the strengths of both the in vitro and the in vivo models. The ex vivo model utilizes intact corneas organotypically maintained in culture and infected with HSV-1. The use of the ex vivo model allows for highly physiologically-based conclusions, yet it is rather inexpensive and requires time commitment comparable to that of the in vitro model.
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Enfermedades de la Córnea/virología , Endotelio Corneal/virología , Herpes Simple/virología , Herpesvirus Humano 1/fisiología , Enfermedad Aguda , Animales , Enfermedades de la Córnea/patología , Modelos Animales de Enfermedad , Endotelio Corneal/patología , Herpes Simple/patología , Herpesvirus Humano 1/genética , Humanos , ConejosRESUMEN
BACKGROUND: NASP (Nuclear Autoantigenic Sperm Protein) is a histone chaperone that is present in all dividing cells. NASP has two splice variants: tNASP and sNASP. Only cancer, germ, transformed, and embryonic cells have a high level of expression of the tNASP splice variant. We examined the consequences of tNASP depletion for prostate cancer PC-3 cells. METHODS: tNASP was depleted from prostate cancer PC-3 cells, cervical cancer HeLa cells, and prostate epithelial PWR-1E cells using lentivirus expression of tNASP shRNA. Cell cycle changes were studied by proliferation assay with CFSE labeling and double thymidine synchronization. Gene expression profiles were detected using RT(2)Profiler PCR Array, Western and Northern blotting. RESULTS: PC-3 and HeLa cells showed inhibited proliferation, increased levels of cyclin-dependant kinase inhibitor p21 protein and apoptosis, whereas non-tumorigenic PWR-1E cells did not. All three cell types showed decreased levels of HSPA2. Supporting in vitro experiments demonstrated that tNASP, but not sNASP is required for activation of HSPA2. CONCLUSIONS: Our results demonstrate that PC-3 and HeLa cancer cells require tNASP to maintain high levels of HSPA2 activity and therefore viability, while PWR-1E cells are unaffected by tNASP depletion. These different cellular responses most likely arise from changes in the interaction between tNASP and HSPA2 and disturbed tNASP chaperoning of linker histones. This study has demonstrated that tNASP is critical for the survival of prostate cancer cells and suggests that targeting tNASP expression can lead to a new approach for prostate cancer treatment.
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Adenocarcinoma/patología , Apoptosis/efectos de los fármacos , Autoantígenos/genética , Proliferación Celular/efectos de los fármacos , Proteínas Nucleares/genética , Neoplasias de la Próstata/patología , ARN Interferente Pequeño/farmacología , Adenocarcinoma/genética , Apoptosis/genética , Autoantígenos/fisiología , Ciclo Celular/efectos de los fármacos , Línea Celular Tumoral , Regulación hacia Abajo/efectos de los fármacos , Regulación hacia Abajo/genética , Técnicas de Silenciamiento del Gen , Células HeLa , Chaperonas de Histonas/antagonistas & inhibidores , Chaperonas de Histonas/genética , Chaperonas de Histonas/fisiología , Humanos , Masculino , Proteínas Nucleares/antagonistas & inhibidores , Proteínas Nucleares/fisiología , Neoplasias de la Próstata/genéticaRESUMEN
Thermal plasmas and lasers have been widely used in medicine to cut, ablate and cauterize tissues through heating; in contrast, non-thermal plasma produces no heat, so its effects can be selective. In order to exploit the potential for clinical applications, including wound healing, sterilization, blood coagulation, and cancer treatment, a mechanistic understanding of the interaction of non-thermal plasma with living tissues is required. Using mammalian cells in culture, it is shown here that non-thermal plasma created by dielectric barrier discharge (DBD) has dose-dependent effects that range from increasing cell proliferation to inducing apoptosis. It is also shown that these effects are primarily due to formation of intracellular reactive oxygen species (ROS). We have utilized γ-H2AX to detect DNA damage induced by non-thermal plasma and found that it is initiated by production of active neutral species that most likely induce formation of organic peroxides in cell medium. Phosphorylation of H2AX following non-thermal plasma treatment is ATR dependent and ATM independent, suggesting that plasma treatment may lead to replication arrest or formation of single-stranded DNA breaks; however, plasma does not lead to formation of bulky adducts/thymine dimers.
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Apoptosis/efectos de la radiación , Proliferación Celular/efectos de la radiación , Rayos Láser , Animales , Células Cultivadas , Daño del ADN/efectos de la radiación , Relación Dosis-Respuesta en la Radiación , Mamíferos , Especies Reactivas de Oxígeno/metabolismoRESUMEN
Biomarkers for early detection of cancer have great clinical diagnostic potential. Numerous reports have documented the generation of humoral immune responses that are triggered in response to changes in protein expression patterns in tumor tissues and these biomarkers are referred to as tumor associated antigens (TAAs). Using a high-throughput technology, we previously identified 65 proteins as diagnostically useful TAAs by profiling the humoral immune responses in ovarian cancer (OVCA) patients. Here we determined the expression status of some of those TAAs in tissues from OVCA patients. The protein expression patterns of 4 of those 65 antigens, namely NASP, RCAS1, Nijmegen breakage syndrome1 (NBS1) and eIF5A, along with p53 and Her2 (known molecular prognosticators) and two proteins that interact with NBS1, MRE11 and RAD50, were assessed by immunohistochemistry (IHC). NASP and RCAS1 proteins were more frequently expressed in ovarian cancer tissues than with normal ovarian tissue and serous cystadenomas and MRE11 was less frequently expressed. When evaluated simultaneously, only NASP and MRE11 remained statistically significant with sensitivity of 66% and specificity of 89%. None of these proteins' expression levels were prognostic for survival. Together, our results indicate that occurrence of humoral immune responses against some of these TAAs in OVCA patients is triggered by antigen protein overexpression.