RESUMEN
In recent years, enantioselective electrocatalysis has surfaced as an increasingly-effective platform for sustainable molecular synthesis. Despite indisputable progress, strategies that allow the control of two distinct stereogenic elements with high selectivity remain elusive. In contrast, we, herein, describe electrochemical cobalt-catalyzed C-H activations that enable the installation of chiral stereogenic centers along with a chiral axis with high levels of enantio- and diastereoselectivities. The developed electrocatalysis strategy allowed for C-H/N-H activations/annulations with cyclic and non-cyclic alkenes providing expedient access to various central as well as atropo-chiral dihydroisoquinolinones paired to the valuable hydrogen evolution reaction. Studies on the atropo-stability of the obtained compounds demonstrated that the exceedingly mild conditions ensured by the electrocatalytic process were key for the achieved high stereoselectivities.
RESUMEN
First examples of anionic cyclic (alkyl)(amino)carbenes (Ani-cAACs) that contain borane substituents have been synthesized. The nature of the borane substituents allows a modulation of the σ-donor or π-acceptor abilities compared to their neutral analogues. A B(CN)3 -substituted Ani-cAAC has been generated and used in situ. The corresponding C2 F5 BF2 -Ani-cAAC 6 was obtained in high yield on a multigram scale. First reactions of these novel ligands with elemental selenium and chloro(triphenylphosphine)gold(I) led to the anionic selenium adducts 7 and 8 and the Ani-cAAC gold complex 9. The properties of these compounds and data derived from theoretical calculations provide an insight into the electronic and steric properties of these novel anionic cAACs. Especially the ease of synthesis and the combination of properties such as negative charge, large buried volume, and good σ-donor and π-acceptor ability renders Ani-cAACs unique and promising new building blocks.
RESUMEN
The 1-methyl-3-(tricyanoborane)imidazolin-2-ylidenate anion (2) was obtained in high yield by deprotonation of the B(CN)3 -methylimidazole adduct 1. Regarding charge and stereo-electronic properties, anion 2 closes the gap between well-known neutral NHCs and the ditopic dianionic NHC, the 1,3-bis(tricyanoborane)imidazolin-2-ylidenate dianion (IIb). The influence of the number of N-bonded tricyanoborane moieties on the σ-donating and π-accepting properties of NHCs was assessed by quantum chemical calculations and verified by experimental data on 2, IIb, and 1,3-dimethylimidazolin-2-ylidene (IMe, IIa). Therefore NHC 2, which acts as a ditopic ligand via the carbene center and the cyano groups, was reacted with alkyl iodides, selenium, and [Ni(CO)4 ] yielding alkylated imidazoles 3 and 4, the anionic selenium adduct 5, and the anionic nickel tricarbonyl complex 8, respectively. The results of this study prove that charge, number of coordination sites, buried volume (%Vbur ) and σ-donor and π-acceptor abilities of NHCs can be effectively fine-tuned via the number of tricyanoborane substituents.
RESUMEN
Cervical cancer (CC) arises from premalignant cervical intraepithelial neoplasia (CIN) induced by a persistent infection with human papillomaviruses. The multi-stepwise disease progression is driven by genetic and epigenetic alterations. Our previous studies demonstrated a clear downregulation of inter-α-trypsin-inhibitor-heavy chain 5 (ITIH5) at mRNA and protein levels in CC compared to CIN2/3 and normal cervical tissue. Initial in vitro functional analyses revealed a suppressive effect of ITIH5 on relevant mechanisms for cancer progression in conventional two dimensional (2D) cell culture model systems. Based on these studies, we aimed to investigate the functional relevance of ITIH5 in multicellular tumor spheroid (MCTS) models, which resemble in vivo tumors more closely. We successfully established CC cell line-derived MCTS using the hanging-drop technique. ITIH5 was ectopically overexpressed in HeLa and SiHa cells and its functional relevance was investigated under three dimensional (3D) culture conditions. We found that ITIH5 re-expression significantly suppressed tumor spheroid growth and spheroid invasiveness of both HeLa and SiHa spheroids. Immunohistochemical (IHC) analyses revealed a significant reduction in Ki-67 cell proliferation index and CAIX-positive areas indicative for hypoxia and acidification. Furthermore, we observed an increase in cPARP-positive cells suggesting a higher rate of apoptosis upon ITIH5 overexpression. An effect of ITIH5 expression on the susceptibility of cervical MCTS towards cytostatic drug treatment was not observed. Collectively, these data uncover pronounced anti-proliferative effects of ITIH5 under 3D cell culture conditions and provide further functional evidence that the downregulation of ITIH5 expression during cervical carcinogenesis could support cancer development.
RESUMEN
There is a demand for ultra low-loss metal films with high-quality single crystals and perfect surface for nanophotonics and quantum information processing. Many researches are devoted to alternative materials, but silver is by far theoretically the most preferred low-loss material at optical and near-IR frequencies. Usually, epitaxial growth is used to deposit single-crystalline silver films, but they still suffer from unpredictable losses and well-known dewetting effect that strongly limits films quality. Here we report the two-step approach for e-beam evaporation of atomically smooth single-crystalline metal films. The proposed method is based on the thermodynamic control of film growth kinetics at atomic level, which allows depositing state-of-art metal films and overcoming the film-surface dewetting. Here we use it to deposit 35-100 nm thick single-crystalline silver films with the sub-100pm surface roughness and theoretically limited optical losses, considering an ideal material for ultrahigh-Q nanophotonic devices. Utilizing these films we experimentally estimate the contribution of grain boundaries, material purity, surface roughness and crystallinity to optical properties of metal films. We demonstrate our «SCULL¼ two-step approach for single-crystalline growth of silver, gold and aluminum films which open fundamentally new possibilities in nanophotonics, biotechnology and superconductive quantum technologies. We believe it could be readily adopted for the synthesis of other extremely low-loss single-crystalline metal films.
RESUMEN
This study investigates the influence of photopigment bleaching on autofluorescence lifetimes in the fundus in 21 young healthy volunteers. Three measurements of 30° retinal fields in two spectral channels (SSC: 498-560 nm, LSC: 560-720 nm) were obtained for each volunteer using fluorescence lifetime imaging ophthalmoscopy (FLIO). After dark-adaptation by wearing a custom-made lightproof mask for 30 minutes, the first FLIO-measurement was recorded (dark-adapted state). Subsequently, the eye was bleached for 1 minute (luminance: 3200 cd/m2), followed by a second FLIO-measurement (bleached state). Following an additional 10 minute dark adaptation using the mask, a final FLIO-measurement was recorded (recovered state). Average values of the fluorescence lifetimes were calculated from within different areas of a standardized early treatment diabetic retinopathy study (ETDRS) grid (central area, inner and outer rings). The acquisition time in the bleached state was significantly shortened by approximately 20%. The SSC did not show any significant changes in fluorescence lifetimes with photopigment bleaching, only the LSC showed small but significant bleaching-related changes in the fluorescence lifetimes τ1 and τ2 from all regions, as well as the mean fluorescence lifetime in the central area. The fluorescence lifetime differences caused by bleaching were by far less significant than pathological changes caused by eye diseases. The magnitudes of fluorescence lifetime changes are <10% and do not interfere with healthy or disease related FLIO patterns. Thus, we conclude that bleaching is not a relevant confounder in current clinical applications of FLIO.
RESUMEN
We demonstrate a broadband multilayer anti-reflection (AR) coating applied to a quantum cascade laser (QCL) facet. The bandwidth of the AR coating was optimized for the range of 8.0 to 12.0 µm to cover the entire emission spectrum of a broad-gain QCL. The laser facets are high-reflectance and AR-coated for use in an external cavity QCL. The AR coating is composed of layers of yttrium fluoride, zinc sulfide, and germanium, all deposited using electron-beam evaporation, and the modal reflectance of the laser facet across the wavelength range was reduced to 0.75%. The external cavity laser performance in a Littrow-type configuration is also described.
RESUMEN
We describe an external-cavity quantum cascade laser with an intra-cavity out-coupling optical system, tunable from 8.4 µm to 10.8 µm. The new optical configuration allows higher output power compared to a conventional Littrow-type external cavity approach, while keeping the broad tunability of wavelength. We achieved 1 W output power at the maximum in pulse-mode operation, which is more than twice the optical power using the Littrow design with the same gain chip.
RESUMEN
Fundus autofluorescence (FAF) imaging is a well-established method in ophthalmology; however, the fluorophores involved need more clarification. The FAF lifetimes of 20 post mortem porcine eyes were measured in two spectral channels using fluorescence lifetime imaging ophthalmoscopy (FLIO) and compared with clinical data from 44 healthy young subjects. The FAF intensity ratio of the short and the long wavelength emission (spectral ratio) was determined. Ex vivo porcine fundus fluorescence emission is generally less intense than that seen in human eyes. The porcine retina showed significantly (p<0.05) longer lifetimes than the retinal pigment epithelium (RPE): 584 ± 128 ps vs. 121 ± 55 ps 498-560 nm, 240 ± 42 ps vs. 125 ± 20 ps at 560-720 nm. Furthermore, the lifetimes of the porcine RPE were significantly shorter (121 ± 55 ps and 125 ± 20 ps) than those measured from human fundus in vivo (162 ± 14 ps and 179 ± 13 ps, respectively). The fluorescence emission of porcine retina was shifted towards a shorter wavelength compared to that of RPE and human FAF. This data shows the considerable contribution of fluorophores in the neural retina to total FAF intensity in porcine eyes.
RESUMEN
We describe a design concept for broadband anti-reflection coatings for the mid-infrared that allows thinner film thickness and characterize its properties in the mid-infrared range from 7 µm to 12 µm. Two demonstrator coatings comprising zinc sulfide, yttrium fluoride, and germanium, all of which are deposited by electron beam evaporation on indium phosphide substrates, are optimized for the 7-12 µm and the 10-12 µm ranges. They exhibit less than 1.0% reflection over the target wavelength ranges with notably thin coating thickness.
RESUMEN
New complexes of Zn(ii), Pd(ii) and Pt(ii) with Schiff bases are prepared in a one-step condensation of 4-(dibutylamino)-2-hydroxybenzaldehyde and 4,5-diaminophthalonitrile in the presence of a metal salt. The complexes possess efficient absorption in the blue-green part of the spectrum with molar absorption coefficients up to 98 000 M-1 cm-1. The Pt(ii) complex shows very strong red phosphorescence in anoxic solutions at room temperature with a quantum yield of 65% in toluene which places it among the brightest emitters available for this spectral range. The phosphorescence of the Pd(ii) complex under the same conditions is very weak (Φ < 1%) but is enhanced to Φ > 10% upon immobilization into polymers. Optical thermometers based on self-referenced lifetime read-out are prepared upon immobilization of the dyes into gas-blocking poly(vinylidene chloride-co-acrylonitrile). At 25 °C, the materials based on Pd(ii) and Pt(ii) complexes show sensitivities of -2.1 and -0.52%τ/K, respectively. Application of the sensors for imaging of temperature on surfaces (planar optode) and for monitoring of fast temperature fluctuations (fiber-optic microsensor) is demonstrated. Immobilized into a gas-permeable matrix, the Pt(ii) complex also performs as a promising oxygen-sensing material. The new systems are also attractive for imaging of oxygen or temperature with the help of multi-photon microscopy, due to a good match with the biological optical window and much better brightness under two photon excitation compared to that of the conventional Pt(ii) meso-tetra-(pentafluorophenyl)porphyrin.
RESUMEN
PURPOSE: To investigate fundus autofluorescence (FAF) lifetimes in geographic atrophy (GA) with a focus on macular pigment (MP) and foveal sparing. METHODS: The study included 35 eyes from 28 patients (mean age 79.2 ± 8.0 years) with GA. A 30° retinal field, centred at the macula, was investigated using fluorescence lifetime imaging ophthalmoscopy (FLIO). The FLIO technology is based on a Heidelberg Engineering Spectralis system. Decays of FAF were detected in a short (498-560 nm, SSC) and long (560-720 nm, LSC) spectral channel. The mean fluorescence lifetime, τm , was calculated from a three-exponential approximation of the FAF decays. Macular optical coherence tomography (OCT) scans as well as fundus photography were recorded. RESULTS: Review of FLIO data reveals specific patterns of significantly prolonged τm in regions of GA (SSC 616 ± 343 ps, LSC 615 ± 154 ps) as compared to non-atrophic regions. Large τm differences between the fovea and atrophic areas correlate with better visual acuity (VA). Shorter τm at the fovea than within other non-atrophic regions indicates sparing, which was identified in 16 eyes. Seventy per cent of patients treated with lutein supplementation showed foveal sparing, whereas the rate among non-supplemented patients was 22%. CONCLUSION: Using FLIO, we present a novel way to detect foveal sparing, investigate MP, and analyse variability of τm in different foveal regions (including the prognostic valuable border region) in GA. These findings support the potential utility of FLIO in monitoring disease progression. The findings also highlight the possibly protective effect of lutein supplementation, with implication in recording the presence and distributional pattern of MP.
Asunto(s)
Angiografía con Fluoresceína/métodos , Fóvea Central/patología , Atrofia Geográfica/diagnóstico , Monitoreo Fisiológico/métodos , Oftalmoscopía/métodos , Tomografía de Coherencia Óptica/métodos , Agudeza Visual , Anciano , Estudios Transversales , Progresión de la Enfermedad , Femenino , Estudios de Seguimiento , Fondo de Ojo , Atrofia Geográfica/fisiopatología , Humanos , Masculino , Estudios Prospectivos , Reproducibilidad de los Resultados , Factores de TiempoRESUMEN
Aim was to identify methylated genes with functional involvement in cisplatin-resistance development of epithelial ovarian cancer (EOC). Genome-wide analyses of hypermethylated CpG-islands in resistant cell lines in combination with qRT-PCR analyses were used to identify epigenetically silenced genes. EOC-Type-II tumors were analyzed for gene methylation and expression and TCGA data were interrogated in-silico. Experiments revealed 37 commonly hypermethylated genes in resistant cells of which Tribbles 2 (TRIB2) showed the most pronounced downregulation on mRNA level and was characterized further. TRIB2 showed a reactivation after 5'-Aza-Cytidine treatment in resistant cells but a cisplatin-dependent, prominent upregulation on mRNA level in sensitive cells, only. Re-expression in resistant A2780 cells increased the sensitivity to cisplatin and other DNA-damaging agents, but not taxanes. Contrary, knockdown of TRIB2 increased resistance to cisplatin in sensitive cells. TRIB2 was involved in the induction of a cisplatin-dependent cell cycle arrest and apoptosis by influencing p21 and survivin expression. An increased Pt-DNA-adduct formation in TRIB2 re-expressing cells did not translate in higher levels of dsDNA damage (yH2AX-foci). Thus, TRIB2 is potentially involved in the signal transduction from nucleotide excision repair of intrastrand cross links. Importantly, patient stratification of two homogenous cohorts of EOC-Type-II patients from Jena (n = 38) and the TCGA (n = 149) by TRIB2 mRNA expression consistently revealed a significantly decreased PFS for patients with low TRIB2 levels (log-rank p < 0.05). Tumors from resistant patients expressed the lowest levels of TRIB2. Downregulation of TRIB2 contributes to platin-resistance and TRIB2 expression should be validated as prognostic and predictive marker for EOC.
Asunto(s)
Proteínas Quinasas Dependientes de Calcio-Calmodulina/metabolismo , Cisplatino/farmacología , Daño del ADN , Péptidos y Proteínas de Señalización Intracelular/metabolismo , Neoplasias Glandulares y Epiteliales/tratamiento farmacológico , Neoplasias Glandulares y Epiteliales/genética , Neoplasias Ováricas/tratamiento farmacológico , Neoplasias Ováricas/genética , Antineoplásicos/farmacología , Apoptosis/efectos de los fármacos , Proteínas Quinasas Dependientes de Calcio-Calmodulina/biosíntesis , Proteínas Quinasas Dependientes de Calcio-Calmodulina/genética , Carcinoma Epitelial de Ovario , Línea Celular Tumoral , Inhibidor p21 de las Quinasas Dependientes de la Ciclina/biosíntesis , Aductos de ADN/biosíntesis , Metilación de ADN , Resistencia a Antineoplásicos/genética , Femenino , Fase G2 , Humanos , Péptidos y Proteínas de Señalización Intracelular/biosíntesis , Péptidos y Proteínas de Señalización Intracelular/genética , Puntos de Control de la Fase M del Ciclo Celular , Neoplasias Glandulares y Epiteliales/metabolismo , Neoplasias Ováricas/metabolismo , Proteoma/metabolismo , Células Tumorales CultivadasRESUMEN
PURPOSE: To discriminate non-proliferative diabetic retinopathy (NPDR) patients from healthy controls by fluorescence lifetime imaging ophthalmoscopy (FLIO). METHODS: A prototype FLIO (Heidelberg-Engineering, Heidelberg, Germany) was used to examine the retina of 33 patients and 28 controls. As increased fluorescence of the diabetic lens is known, the lenses of 34 patients and 24 controls were investigated as well. Time-resolved decay was detected in two spectral channels (ch1: 498-560 nm, ch2: 560-720 nm) and approximated by a series of three exponential functions yielding in lifetimes (τ1 , τ2 , τ3 ), amplitudes (α1 , α2 , α3 ) and their amplitude-weighted means (τm ). RESULTS: Significant differences between patients and controls were found for all fundus lifetime components (τm , τ1 -τ3 ) as for the amplitude α3 in both spectral channels. Channel 1 showed the largest differences: the average of mean fluorescence lifetime τm in the macula was 259 ± 137 ps in the patients versus 147 ± 69 ps in the controls. A logistic regression model allowed discrimination between study and control group with a sensitivity of 90.09% and a specificity of 71.4% (area under the curve: 0.865). Significantly shorter τm in the patients group than in the control group was detected in channel 2 in the crystalline lens (1587 ± 326 ps versus 1854 ± 384 ps, p = 0.006). CONCLUSIONS: Fundus Fluorescence lifetimes are significantly increased in NPDR while lens lifetimes are shorter in the patient group. Lifetime changes might be indicative for the accumulation of advanced glycation end products (AGEs) which enables detection of the disease with high sensitivity and specificity possibly bearing diagnostic merit.
Asunto(s)
Retinopatía Diabética/diagnóstico por imagen , Productos Finales de Glicación Avanzada/metabolismo , Imagen Óptica/métodos , Anciano , Glucemia/metabolismo , Retinopatía Diabética/metabolismo , Femenino , Hemoglobina Glucada/metabolismo , Voluntarios Sanos , Humanos , Masculino , Persona de Mediana Edad , Oftalmoscopía/métodos , Curva ROC , Reproducibilidad de los Resultados , Sensibilidad y EspecificidadRESUMEN
In this study the influence of hydrogen peroxide (H2 O2 ) on the redox state, NADH protein binding, and mitochondrial membrane potential in Müller cells is investigated. Cultures of permanent human Müller cells MIO-M1 were exposed to H2 O2 in 75 µM and 150 µM concentration for two hours. Fluorescence emission spectra and lifetimes were measured by two-photon microscopy (excitation wavelength: 740 nm) at the mitochondria which were identified in the microscopic images by their fluorescence properties (spectra and intensity). Two hours of H2 O2 exposure did not impair viability of MIO-M1 cells in culture. Whereas the ratio of flavine- to NADH fluorescence intensity did not change under either H2 O2 concentration, the mean lifetime was significantly different between controls, not exposed to H2 O2 , and the 150 µM H2 O2 exposure (972 ± 63 ps vs. 1152 ± 64 ps, p = 0.014). One hour after cessation of the H2 O2 exposure, the value retuned to that of the control (983 ± 36 ps). A hyperpolarization of the mitochondrial membrane under 150 µM H2 O2 was found. These findings suggest a shift form free to protein-bound NADH in mitochondria as well as a hyperpolarization of their inner membrane which could be related to an impairment of Müller cell function despite their preserved viability. Exposure of human Müller cells to hydrogen peroxide for two hours results in a reversible change of protein binding of mitochondrial NADH upon unchanged redox ratio. The mitochondrial membrane potential is increased during exposure.
Asunto(s)
Células Ependimogliales/metabolismo , Potencial de la Membrana Mitocondrial , Estrés Oxidativo , Células Cultivadas , Células Ependimogliales/citología , Humanos , Peróxido de Hidrógeno , Microscopía Fluorescente , MitocondriasRESUMEN
PURPOSE: To investigate the impact of macular pigment (MP) on fundus autofluorescence (FAF) lifetimes in vivo by characterizing full-thickness idiopathic macular holes (MH) and macular pseudo-holes (MPH). METHODS: A total of 37 patients with MH and 52 with MPH were included. Using the fluorescence lifetime imaging ophthalmoscope (FLIO), based on a Heidelberg Engineering Spectralis system, a 30° retinal field was investigated. FAF decays were detected in a short (498-560 nm; ch1) and long (560-720 nm; ch2) wavelength channel. τm , the mean fluorescence lifetime, was calculated from a three-exponential approximation of the FAF decays. Macular coherence tomography scans were recorded, and macular pigment's optical density (MPOD) was measured (one-wavelength reflectometry). Two MH subgroups were analysed according to the presence or absence of an operculum above the MH. A total of 17 healthy fellow eyes were included. A longitudinal FAF decay examination was conducted in nine patients, which were followed up after surgery and showed a closed MH. RESULTS: In MH without opercula, significant τm differences (p < 0.001) were found between the hole area (MHa) and surrounding areas (MHb) (ch1: MHa 238 ± 64 ps, MHb 181 ± 78 ps; ch2: MHa 275 ± 49 ps, MHb 223 ± 48 ps), as well as between MHa and healthy eyes or closed MH. Shorter τm , adjacent to the hole, can be assigned to areas with equivalently higher MPOD. Opercula containing MP also show short τm . In MPH, the intactness of the Hele fibre layer is associated with shortest τm . CONCLUSIONS: Shortest τm originates from MP-containing retinal layers, especially from the Henle fibre layer. Fluorescence lifetime imaging ophthalmoscope (FLIO) provides information on the MP distribution, the pathogenesis and topology of MH. Macular pigment (MP) fluorescence may provide a biomarker for monitoring pathological changes in retinal diseases.
Asunto(s)
Mácula Lútea/patología , Monitoreo Fisiológico/métodos , Oftalmoscopía/métodos , Perforaciones de la Retina/diagnóstico , Epitelio Pigmentado de la Retina/patología , Anciano , Estudios Transversales , Femenino , Estudios de Seguimiento , Humanos , Masculino , Estudios Prospectivos , Reproducibilidad de los Resultados , Tomografía de Coherencia Óptica/métodos , Agudeza VisualRESUMEN
INTRODUCTION: A substantial part of the human genome originates from transposable elements, remnants of ancient retroviral infections. Roughly 8% of the human genome consists of about 400,000 LTR elements including human endogenous retrovirus (HERV) sequences. Mainly, the interplay between epigenetic and post-transcriptional mechanisms is thought to silence HERV expression in most physiological contexts. Interestingly, aberrant reactivation of several HERV-H loci appears specific to colorectal carcinoma (CRC). RESULTS: The expression of HERV-H Gag proteins (Gag-H) was assessed using novel monoclonal mouse anti Gag-H antibodies. In a flow cytometry screen four antibody clones were tested on a panel of primary CRC cell lines and the most well performing ones were subsequently validated in western blot analysis. Finally, Gag-H protein expression was analyzed by immune histology on cell line cytospins and on clinical samples. There, we found a heterogeneous staining pattern with no background staining of endothelial, stromal and infiltrating immune cells but diffuse staining of the cytoplasm for positive tumor and normal crypt cells of the colonic epithelium. CONCLUSION: Taken together, the Gag-H antibody clone(s) present a valuable tool for staining of cells with colonic origin and thus form the basis for future more detailed investigations. The observed Gag-H protein staining in colonic epithelium crypt cells demands profound analyses of a potential role for Gag-H in the normal physiology of the human gut.
Asunto(s)
Anticuerpos Monoclonales/inmunología , Colon/inmunología , Colon/virología , Retrovirus Endógenos/inmunología , Productos del Gen gag/inmunología , Secuencia de Aminoácidos , Animales , Línea Celular , Neoplasias Colorrectales/inmunología , Neoplasias Colorrectales/virología , Citoplasma/inmunología , Citoplasma/virología , Femenino , Genoma Humano/genética , Células HEK293 , Humanos , Mucosa Intestinal/inmunología , Mucosa Intestinal/virología , Ratones , Ratones Endogámicos BALB C , Proteínas Recombinantes/inmunologíaRESUMEN
PURPOSE: To investigate the interrelationship between the oxygen supply of the retina and its regulation with the severity of primary open-angle glaucoma (POAG). METHODS: Central retinal artery (CRAE) and vein (CRVE) diameters and oxygen saturation of peripapillary retinal vessels in 41 patients suffering from POAG (64.1 ± 12.9 years) and 40 healthy volunteers (63.6 ± 14.1 years) were measured using the retinal vessel analyzer. All measures were taken before and during flicker light stimulation. The mean retinal nerve fiber layer thickness (RNFLT) was determined by OCT and the visual field mean defect (MD) was identified using perimetry. RESULTS: In glaucoma patients, CRAE (r = -0.48 p = 0.002) and CRVE (r = -0.394 p = 0.014) at baseline were inversely related to MD, while arterial and venous oxygen saturation showed no significant dependence on the severity of the damage. However, the flicker light-induced change in arterio-venous difference in oxygen saturation was correlated with the MD (r = 0.358 p = 0.027). The diameters of arteries and veins at baseline decreased with reduction of the mean RNFLT (arteries: r = 0.718 p < 0.001; veins: r = 0.685 p < 0.001). CONCLUSION: Vessel diameters showed a strong correlation with RNFLT and MD. This, as well as the reduction of stimulation-induced change in arterio-venous oxygen saturation difference with visual field loss, may be explained by a reduction of the retinal metabolic demand with progressive loss of neuronal tissue in glaucoma.
Asunto(s)
Glaucoma de Ángulo Abierto/fisiopatología , Oxígeno/sangre , Vasos Retinianos/patología , Trastornos de la Visión/fisiopatología , Campos Visuales/fisiología , Anciano , Antihipertensivos/uso terapéutico , Femenino , Glaucoma de Ángulo Abierto/tratamiento farmacológico , Humanos , Masculino , Persona de Mediana Edad , Fibras Nerviosas/patología , Estimulación Luminosa , Células Ganglionares de la Retina/patología , Tomografía de Coherencia Óptica , Pruebas del Campo VisualRESUMEN
PURPOSE: To characterize the macular region and to investigate the influence of the macular pigment (MP) on fundus autofluorescence (FAF) lifetimes in vivo. METHODS: Forty-eight healthy subjects with a mean age of 24.1 ± 3.6 years (range, 20-37 years) were included. A 30° retinal field was investigated using the fluorescence lifetime imaging ophthalmoscope (FLIO), based on a Heidelberg Engineering Spectralis system, detecting FAF decays in a short (498-560 nm; ch1)- and a long (560-720 nm; ch2)-wavelength channel. The mean fluorescence lifetime τm was calculated from a 3-exponential approximation of the FAF decays. Macular pigment optical density (MPOD) was measured by one-wavelength reflectometry, and macular optical coherence tomogram (OCT) scans were recorded. Correlations between τm and MPOD were analyzed. RESULTS: The τm showed shortest values at the macular region with a mean of 82 ps (ch1) and 126 ps (ch2). We found a strong correlation of τm to the MPOD (ch1: r = -0.760; ch2: r = -0.663; P < 0.001), as well as a topologic agreement of shortest τm with highest MPOD. CONCLUSIONS: Macular pigment, which is known to have very short fluorescence decays, considerably contributes to the macular autofluorescence (AF). This study gives indirect evidence for a strong impact of MP on macular τm, although no direct measurement of MP autofluorescence lifetimes in vivo is possible at this point. Potentially, imaging the FAF lifetimes could lead to a novel methodology for the detection of macular pigment properties and pathology-induced changes in the living human retina.
Asunto(s)
Pigmento Macular/química , Epitelio Pigmentado de la Retina/metabolismo , Tomografía de Coherencia Óptica/métodos , Adulto , Femenino , Angiografía con Fluoresceína , Fondo de Ojo , Humanos , Masculino , Oftalmoscopía , Valores de Referencia , Epitelio Pigmentado de la Retina/citología , Estudios Retrospectivos , Adulto JovenRESUMEN
Fluorescence lifetime imaging ophthalmoscopy (FLIO) is a new technique for measuring the in vivo autofluorescence intensity decays generated by endogenous fluorophores in the ocular fundus. Here, we present a software package called FLIM eXplorer (FLIMX) for analyzing FLIO data. Specifically, we introduce a new adaptive binning approach as an optimal tradeoff between the spatial resolution and the number of photons required per pixel. We also expand existing decay models (multi-exponential, stretched exponential, spectral global analysis, incomplete decay) to account for the layered structure of the eye and present a method to correct for the influence of the crystalline lens fluorescence on the retina fluorescence. Subsequently, the Holm-Bonferroni method is applied to FLIO measurements to allow for group comparisons between patients and controls on the basis of fluorescence lifetime parameters. The performance of the new approaches was evaluated in five experiments. Specifically, we evaluated static and adaptive binning in a diabetes mellitus patient, we compared the different decay models in a healthy volunteer and performed a group comparison between diabetes patients and controls. An overview of the visualization capabilities and a comparison of static and adaptive binning is shown for a patient with macular hole. FLIMX's applicability to fluorescence lifetime imaging microscopy is shown in the ganglion cell layer of a porcine retina sample, obtained by a laser scanning microscope using two-photon excitation.