Comparing the RETeval® portable ERG device with more traditional tabletop ERG systems in normal subjects and selected retinopathies.

PURPOSE: Our study aimed to determine if ISCEV standard-like ERGs recorded with the LKC RETeval® portable ERG unit compared to those obtained using the more traditional tabletop unit. METHODS: ERGs recorded from normal subjects and patients affected with retinal ON and OFF pathway anomalies were compared. Analysis included peak time and amplitude measurements as well as time-frequency domain analysis with the discrete wavelet transform of waveforms obtained with the two systems. RESULTS: Although both systems were similarly able to record reliable and highly reproducible ERG responses, there were major discrepancies in ERG responses between the portable and tabletop units, pointing toward a weaker stimulation of the retinal OFF pathway with the portable RETeval® unit. CONCLUSION: The portable RETeval® unit appears to be able to record highly reproducible and diagnostically useful clinical ERGs, albeit with some significant differences in waveform composition compared to those obtained with more standard tabletop systems. Given the unknown origin of these waveform discrepancies, if left uncorrected, these differences could potentially lead to erroneous interpretation when used in the clinical context and/or compared to ERGs recorded using more traditional table top units. Clearly, more research is warranted before handheld devices, such as the RETeval®, can be homologated as a diagnostically sound ERG devices.

Organic visual loss measured by kinetic perimetry and retinal electrophysiology in children with functional amblyopia.

PURPOSE: To demonstrate an organic (retinal) amblyogenic defect in functional amblyopes not responding to treatment. METHODS: Twenty-four children (Mean age: 5.9 ± 1.8 years; range: 4-10 years) with functional amblyopia were recruited for this study. All these children underwent complete ophthalmic and orthoptic evaluation. In addition, Kinetic Goldman Visual Fields (KGVF), Spectral Domain Optical Coherence Tomography (SD-OCT), full field flash electroretinograms (ffERG) and multifocal electroretinograms (mfERG) were also performed. Ratios were subsequently derived by comparing the amplitudes obtained from the amblyopic eye (AE) to the good eye (GE) for the a- and b-waves of the ffERG, as well as for the ring analysis of the mfERG. RESULTS: KGVF showed a central scotoma of varying size (3°-7°) and density (absolute to relative), with increasing target size in 14/24 patients whose best post-treatment vision in the AE ranged from 20/100 to 20/40. The scotoma decreased in size and density with improving vision until a plateau of recovery was reached. The remaining 10/24 patients with a vision ≥ 20/30 showed no scotoma. SD-OCT showed no significant difference between the AE and GE. ffERG and mfERG were obtained in 18/24 patients. The ffERG AE/GE ratio was abnormal in 7 patients, 5 of which had large scotomas on KGVF. The mfERG ring 1 AE/GE ratio was significantly (p < .05) attenuated in 9/18 patients out of which 3 were no longer amblyopic. However, there was no significant difference (p > .05) in ring 1 AE/GE amplitude ratio between those who achieved 20/50-20/40 (.81 ± .26) and those with ≥ 20/25(.86 ± .25). CONCLUSIONS: The combined findings of central scotoma on KGVF and mfERG anomalies in patients who did not achieve optimal vision with treatment suggest an underlying organic defect impairing macular function.

Ring analysis of multifocal oscillatory potentials (mfOPs) in cCSNB suggests near-normal ON-OFF pathways at the fovea only.

PURPOSE: To investigate the center-periphery distribution of ON and OFF retinal responses in complete congenital stationary night blindness (cCSNB). METHODS: Photopic full-field flash ERGs (photopic ffERGs) and OPs (photopic ffOPs) and slow m-sequence (to enhance OP prominence) mfERGs (and filtered mfOPs) evoked by a 37 hexagon stimulus array were recorded from normal subjects and cCSNB patients. Discrete wavelet transform (DWT) analysis of photopic ffERGs and mfERGs was also performed in order to assess the contribution of the ON and OFF retinal pathways (i.e., OFF-to-ON ratio) in both cohorts. RESULTS: As expected, the photopic ffERG (and ffOPs) responses in cCSNB were devoid of the first two of the three OPs (i.e., OP2 and OP3 and OP4) normally seen on the ascending limb of the b-wave. A similar finding was also noted in the mfERGs (and mfOPs) of ring 4. In contrast, the mfERGs (and mfOPs) of ring 1 included all three OPs. DWT analysis revealed that while in normal subjects, the OFF-to-ON ratio of mfERGs slightly increased from rings 1 to 4 (from 0.61 ± 0.03 to 0.78 ± 0.04; p < 0.05; median: from 0.62 to 0.79; p < 0.05), in cCSNB this ratio increased significantly more [from 0.73 ± 0.13 (ring 1) to 1.18 ± 0.17 (ring 4); p < 0.05; median: 0.78 to 1.22; p < 0.05], hence from a normal ON-dominated ratio (central ring) to an OFF-dominated ratio (peripheral ring). CONCLUSIONS: Our results show a clear discrepancy of ON and OFF mfOP components in cCSNB. Responses originating from the most central ring (i.e., ring 1) disclosed a near-normal electrophysiological contribution (as revealed with the presence of OP2, OP3 and OP4 as well as with the DWT OFF-to-ON ratio) of the retinal ON and OFF pathways in mfERG (and mfOPs) responses compared to responses from the more peripheral ring (and ffOP) which are devoid of the ON OPs (i.e., OP2 and OP3).

A longitudinal study of retinopathy in the PEX1-Gly844Asp mouse model for mild Zellweger Spectrum Disorder.

Zellweger Spectrum Disorder (ZSD) is an autosomal recessive disease caused by mutations in any one of 13 PEX genes whose protein products are required for peroxisome assembly. Retinopathy leading to blindness is one of the major untreatable handicaps faced by patients with ZSD but is not well characterized, and the requirement for peroxisomes in retinal health is unknown. To address this, we examined the progression of retinopathy from 2 to 32 weeks of age in our murine model for the common human PEX1-p.Gly843Asp allele (PEX1-p.Gly844Asp) using electrophysiology, histology, immunohistochemistry, electron microscopy, biochemistry, and visual function tests. We found that retinopathy in male and female PEX1-G844D mice was marked by an attenuated cone function and abnormal cone morphology early in life, with gradually decreasing rod function. Structural defects at the inner retina occurred later in the form of bipolar cell degradation (between 13 and 32 weeks). Inner segment disorganization and enlarged mitochondria were seen at 32 weeks, while other inner retinal cells appeared preserved. Visual acuity was diminished by 11 weeks of age, while signal transmission from the retina to the brain was relatively intact from 7 to 32 weeks of age. Molecular analyses showed that PEX1-G844D is a subfunctional but stable protein, contrary to human PEX1-G843D. Finally, C26:0 lysophosphatidylcholine was elevated in the PEX1-G844D retina, while phopshoethanolamine plasmalogen lipids were present at normal levels. These characterization studies identify therapeutic endpoints for future preclinical trials, including improving or preserving the electroretinogram response, improving visual acuity, and/or preventing loss of bipolar cells.

The effects of bandpass filtering on the oscillatory potentials of the electroretinogram.

PURPOSE: In order to study the OPs, the ERG signal must be filtered to eliminate the low-frequency waves known as the a- and b-waves. Unfortunately, the ISCEV ERG standard does not give clear guidelines on how to proceed apart from indicating that frequencies below 75 Hz should be filtered out when recording scotopic OPs, while no suggestions are offered for the photopic OPs. The purpose of this study was thus to characterize more extensively the effects of various digital filters on the photopic OP waveforms in order to suggest the most appropriate filtering method to record them. METHODS: Filtered OPs (N = 9600 tracings) were extracted from a photopic ERG databank of 40 normal subjects [intensity: 4.4 cd s m-2; background: 30 cd m-2] using 240 different combinations of five digital filters types (Bessel; Butterworth; Elliptic; Chebyshev type 1 and 2), eight bandwidth ranges (50-300; 75-300; 100-300; 125-300; 50-1000; 75-1000; 100-1000; 125-1000 Hz), three filter orders (1, 2 and 5) and with/without phase lag corrections that were generated using MATLAB 2015b. The peak time and the percentage of OPs (sum of OP amplitudes on the b-wave amplitude) were calculated in the time domain (TD%OP). RESULTS: The timing of the OPs was less affected than the amplitude by the different filters used. Depending on the filter used, the resulting OPs were either severely depressed (16.16% of broadband OP content) or slightly reduced (93.63%). The filters that most successfully eliminated the slow components of the ERG (i.e., < 12% of broadband value) were the Bessel, the Butterworth and the Chebyshev type 1 filters and out of the latter, the Butterworth filter was that which most faithfully reproduced the high-frequency OPs (i.e., > 96%). CONCLUSION: Our results vividly demonstrate the need to better define the characteristics of the filter that is used to record the OPs as it does have a significant impact on the resulting waveform.

ISCEV extended protocol for the stimulus-response series for light-adapted full-field ERG.

The International Society for Clinical Electrophysiology of Vision (ISCEV) standard for full-field electroretinography (ERG) describes a minimum protocol for clinical testing but encourages additional ERG testing when appropriate. This ISCEV extended protocol describes methods to record and evaluate a light-adapted (LA) ERG stimulus-response series with increasing flash strengths. The LA ERG stimulus-response series (also referred to as the luminance-response or intensity-response series in the published literature) can characterise generalised cone system function more comprehensively than the ISCEV standard LA ERGs alone. The amplitude of LA ERG a-waves, arising from cones and cone off-bipolar cells, typically shows a saturating function. The LA ERG b-wave amplitudes, which arise primarily from activity of retinal bipolar cells, show an amplitude peak followed by a nonzero plateau (the "photopic hill" phenomenon). This ISCEV extended protocol specifies a stimulus-response series suitable to evaluate generalised dysfunction affecting the LA retina, to aid in distinguishing between the on- and off-responses of the cone system and to monitor ERG changes in these characteristics. The LA ERG stimulus-response series for a- and b-waves is recorded to a sequence of nine flash stimuli ranging from 0.03 to 300 cd s m-2, superimposed on a standard background of 30 cd m-2. A shorter protocol is also presented to measure the mid-range of the function (the "photopic hill") using 5 flash stimuli.

Revealing a retinal facilitatory effect with the multifocal ERG.

PURPOSE: We have previously shown that the amplitude of the mfERG response obtained to a single (large) hexagon is significantly smaller than that obtained when summating all the mfERG responses evoked to an array of 7-61 hexagons covering the same retinal area. The purpose of this study was to confirm our initial findings in normal subjects of different ages and in selected patients. METHODS: Binocular mfERGs (1, 7, 19, 37 and 61 hexagon arrays; Espion V6.0.54 Diagnosys LLC) were recorded from 40 normal subjects (25 aged 18-25, and 15 aged 3-12). Individual mfERG waveforms evoked in response to the multi-hexagon arrays (7, 19, 37 and 61) were summated, and the amplitude of the resulting composite mfERG waveform was compared to that measured in the response evoked to the single (large) hexagon stimulus to yield the amplitude ratio (i.e., 7:1 X100, 19:1X100, etc.). RESULTS: In normal subjects, the 7:1 ratio was 119.5 ± 9.2%, a value that gradually decreased to reach 109.4 ± 20.6% with the 61:1 ratio and a finding that was similar across all ages. CONCLUSION: The present study indicates a significant enhancement in amplitude of the summed mfERG composite waveform evoked to the 7 hexagon stimulus array (and to a lesser extent to the 19, 37 and 61 stimuli) compared to the 1 hexagon array, possibly mediated through the retinal lateral pathway (horizontal or amacrine cells), a claim that awaits confirmation. Preliminary results obtained from patients treated with Plaquenil suggest that this new method of mfERG analysis might probe a feature of macular function different from that investigated with the more usual method of mfERG ring ratio.

Evidences Suggesting that Distinct Immunological and Cellular Responses to Light Damage Distinguishes Juvenile and Adult Rat Retinas.

To unravel the mechanisms behind the higher resistance to light damage of juvenile (JR) versus adult (AR) rats, Sprague Dawley rats were exposed to a bright luminous environment of 10, 000 lux. The light-induced retinopathy (LIR) was assessed with histology, electroretinography and immunohistochemistry (IHC). In JR, 2 days of exposure induced the typical LIR, while >3 days added little LIR. IHC revealed a subtle migration of microglia (Iba1 marker) from the inner to the outer retina after 3 days of exposure in JR contrasting with the stronger reaction seen after 1 day in AR. Similarly, in JR, the Müller cells expressed less intense GFAP, CNTF and FGF2 staining compared to AR. Our results suggest that in JR the degree of retinal damage is not proportional to the duration of light exposure (i.e., dose-independent retinopathy), contrasting with the dose-dependent LIR reported in AR. The immature immune system in JR may explain the delayed and/or weaker inflammatory response compared to AR, a finding that would also point to the devastating contribution of the immune system in generating the LIR phenotype, a claim also advanced to explain the pathophysiology of other retinal degenerative disorders such as Age-related Macular Degeneration, Diabetic Retinopathy and Retinitis Pigmentosa.

Evaluating the neuroprotective effect of 17ß-estradiol in rodent models of oxidative retinopathies.

PURPOSE: To determine the neuroprotective effect of estrogen on the structure and function of the retina exposed to an oxidative stress. METHODS: Male Sprague-Dawley rat pups were exposed to either hyperoxia (O2E: from P8 to P14) or bright light (LE: from P14 to P28) with or without 17 ß-estradiol (ßE2) treatment. Retinal structure (histology) and function (ERG) were assessed at selected time points. RESULTS: In the O2E model, ßE2 injections caused a significant reduction of the ERG and a significantly thinner OPL compared to untreated oxygen-exposed group (O2-exposed) rats. In contrast, in the LE model ßE2, treatment was beneficial to the retinal structure (thicker ONL) and function (better preserved ERG amplitudes) compared to untreated light-exposed group (light-exposed rats). CONCLUSION: Our results show that in conditions where the primary target of the oxidative stress is the outer retina (i.e., the photoreceptors) estrogen can protect the retina, while in situations where the inner retina (or retinal vasculature) is the main site of oxidative damage, estrogen may potentiate the detrimental effect of oxidative stress on the retina.

Choroidal Involution Is Associated with a Progressive Degeneration of the Outer Retinal Function in a Model of Retinopathy of Prematurity: Early Role for IL-1ß.

Retinopathy of prematurity (ROP), the most common cause of blindness in premature infants, has long been associated with inner retinal alterations. However, recent studies reveal outer retinal dysfunctions in patients formerly afflicted with ROP. We have recently demonstrated that choroidal involution occurs early in retinopathy. Herein, we investigated the mechanisms underlying the choroidal involution and its long-term impact on retinal function. An oxygen-induced retinopathy (OIR) model was used. In vitro and ex vivo assays were applied to evaluate cytotoxic effects of IL-1ß on choroidal endothelium. Electroretinogram was used to evaluate visual function. We found that proinflammatory IL-1ß was markedly increased in retinal pigment epithelium (RPE)/choroid and positively correlated with choroidal degeneration in the early stages of retinopathy. IL-1ß was found to be cytotoxic to choroid in vitro, ex vivo, and in vivo. Long-term effects on choroidal involution included a hypoxic outer neuroretina, associated with a progressive loss of RPE and photoreceptors, and visual deterioration. Early inhibition of IL-1ß receptor preserved choroid, decreased subretinal hypoxia, and prevented RPE/photoreceptor death, resulting in life-long improved visual function in IL-1 receptor antagonist-treated OIR animals. Together, these findings suggest a critical role for IL-1ß-induced choroidal degeneration in outer retinal dysfunction. Neonatal therapy using IL-1 receptor antagonist preserves choroid and prevents protracted outer neuroretinal anomalies in OIR, suggesting IL-1ß as a potential therapeutic target in ROP.

The DTL ERG electrode comes in different shapes and sizes: Are they all good?

PURPOSE: Although the DTL fiber electrode has been in use in the ERG field for more than four decades, its composition was never clearly defined. We compared five different types of conductive (DTL type) yarn (differing in terms of mass, number of filaments, and crimping degree) in order to determine whether we could identify one that would be better suited for the recording of ERGs. METHODS: Photopic flash ERGs were recorded from five subjects using the following DTL electrodes: 27/7, 22/1, 11/1, 11/1*2, and 22/1*2. Data analysis included amplitude and peak time measurements of the a- and b-waves in the time domain (TD) as well as measurements of specific frequency descriptors of the ERG waveform in the time-frequency domain using the discrete wavelet transform (DWT) approach. The degree of comfortableness was also assessed in 12 subjects with two surveys (Likert 5-point and the ranking scale). RESULTS: Comparisons of TD and DWT parameters did not permit to identify the best DTL electrode, all yielding comparable measures. There was a slight trend for the largest electrode (22/1*2) to yield the largest response, but this was at the expense of comfort, the 22/1*2 electrode being rated as the least comfortable. CONCLUSIONS: Given the minimal impact the different electrodes had on the amplitude of the signal, we believe that comfort should dictate our choice. It would appear from our results that use of a multifilament electrode is the best choice since one can get an electrode whose size is optimized for the recording of large responses while minimizing the foreign-body sensation due to the small size of each of the filaments that compose this multifilament electrode.

Witnessing the first sign of retinitis pigmentosa onset in the allegedly normal eye of a case of unilateral RP: a 30-year follow-up.

PURPOSE: A patient initially presented with constricted visual field, attenuated retinal vasculature, pigmentary clumping and reduced ERG in OS only, suggestive of unilateral retinitis pigmentosa (RP). This patient was subsequently seen on eight occasions (over three decades), and, with time, the initially normal eye (OD) gradually showed signs of RP-like degeneration. The purpose of this study was to evaluate which clinical modality (visual field, funduscopy or electroretinography) could have first predicted this fate. METHODS: At each time points, data obtained from our patient were compared to normative data using Z tests. RESULTS: At initial visit, all tests were significantly (p < 0.05) altered in OS and normal in OD. Visual field and retinal vessel diameter in OD reduced gradually to reach statistical significance at the 5th visit and 6th visit (21 and 22 years after the first examination, respectively). In OD, the amplitude of the scotopic and photopic ERGs reduced gradually and was significantly smaller than normal at the 2nd visit (after 11 years) and 3rd visit (after 18 years), respectively. When the photopic ERG was analyzed using the discrete wavelet transform (DWT), we were able to detect a significant change at the 2nd visit (after 11 years) instead of the 3rd visit (18 years). CONCLUSIONS: Our study allowed us to witness the earliest manifestation of an RP disease process. The ERG was the first test to detect significant RP changes. A significantly earlier detection of ERG anomalies was obtained when the DWT was used, demonstrating its advantage for early detection of ERG changes.

GPR179 is required for depolarizing bipolar cell function and is mutated in autosomal-recessive complete congenital stationary night blindness.

Complete congenital stationary night blindness (cCSNB) is a clinically and genetically heterogeneous group of retinal disorders characterized by nonprogressive impairment of night vision, absence of the electroretinogram (ERG) b-wave, and variable degrees of involvement of other visual functions. We report here that mutations in GPR179, encoding an orphan G protein receptor, underlie a form of autosomal-recessive cCSNB. The Gpr179(nob5/nob5) mouse model was initially discovered by the absence of the ERG b-wave, a component that reflects depolarizing bipolar cell (DBC) function. We performed genetic mapping, followed by next-generation sequencing of the critical region and detected a large transposon-like DNA insertion in Gpr179. The involvement of GPR179 in DBC function was confirmed in zebrafish and humans. Functional knockdown of gpr179 in zebrafish led to a marked reduction in the amplitude of the ERG b-wave. Candidate gene analysis of GPR179 in DNA extracted from patients with cCSNB identified GPR179-inactivating mutations in two patients. We developed an antibody against mouse GPR179, which robustly labeled DBC dendritic terminals in wild-type mice. This labeling colocalized with the expression of GRM6 and was absent in Gpr179(nob5/nob5) mutant mice. Our results demonstrate that GPR179 plays a critical role in DBC signal transduction and expands our understanding of the mechanisms that mediate normal rod vision.

Functional decomposition of the human ERG based on the discrete wavelet transform.

The morphology of the electroretinogram (ERG) can be altered as a result of normal and pathological processes of the retina. However, given that the ERG is almost solely assessed in terms of its amplitude and timing, defining the shape of the ERG waveform so that subtle, physiologically driven, morphological changes can be systematically and reproducibly detected remains a challenging problem. We examined if the discrete wavelet transform (DWT) could meet this challenge. Normal human photopic ERGs evoked to a broad range of luminance intensities (to yield waveforms of various shapes, amplitudes, and timings) were analyzed using DWT descriptors of the ERG. Luminance-response curves that were generated using the various DWT descriptors revealed distinct (p < 0.05) luminance-dependence patterns, indicating that the stimulus luminance differently modulates the various time-frequency components of the ERG and thus its morphology. The latter represents the first attempt to study the luminance-dependence of ERG descriptors obtained with the DWT. Analyses of ERGs obtained from patients affected with ON or OFF retinal pathway anomalies were also presented. We show here for the first time that distinct time-frequency descriptors can be specifically associated to the function of the ON and OFF cone pathway. Therefore, in this study, the DWT revealed reproducible, physiologically meaningful and diagnostically relevant descriptors of the ERG over a wide range of signal amplitudes and morphologies. The DWT analysis thus represents a valuable addition to the electrophysiologist's armamentarium that will improve the quantification and interpretation of normal and pathological ERG responses.

Ischemic neurons prevent vascular regeneration of neural tissue by secreting semaphorin 3A.

The failure of blood vessels to revascularize ischemic neural tissue represents a significant challenge for vascular biology. Examples include proliferative retinopathies (PRs) such as retinopathy of prematurity and proliferative diabetic retinopathy, which are the leading causes of blindness in children and working-age adults. PRs are characterized by initial microvascular degeneration, followed by a compensatory albeit pathologic hypervascularization mounted by the hypoxic retina attempting to reinstate metabolic equilibrium. Paradoxically, this secondary revascularization fails to grow into the most ischemic regions of the retina. Instead, the new vessels are misdirected toward the vitreous, suggesting that vasorepulsive forces operate in the avascular hypoxic retina. In the present study, we demonstrate that the neuronal guidance cue semaphorin 3A (Sema3A) is secreted by hypoxic neurons in the avascular retina in response to the proinflammatory cytokine IL-1ß. Sema3A contributes to vascular decay and later forms a chemical barrier that repels neo-vessels toward the vitreous. Conversely, silencing Sema3A expression enhances normal vascular regeneration within the ischemic retina, thereby diminishing aberrant neovascularization and preserving neuroretinal function. Overcoming the chemical barrier (Sema3A) released by ischemic neurons accelerates the vascular regeneration of neural tissues, which restores metabolic supply and improves retinal function. Our findings may be applicable to other neurovascular ischemic conditions such as stroke.

Role of receptor-mediated endocytosis in the antiangiogenic effects of human T lymphoblastic cell-derived microparticles.

Microparticles possess therapeutic potential regarding angiogenesis. We have demonstrated the contribution of apoptotic human CEM T lymphocyte-derived microparticles (LMPs) as inhibitors of angiogenic responses in animal models of inflammation and tumor growth. In the present study, we characterized the antivascular endothelial growth factor (VEGF) effects of LMPs on pathological angiogenesis in an animal model of oxygen-induced retinopathy and explored the role of receptor-mediated endocytosis in the effects of LMPs on human retinal endothelial cells (HRECs). LMPs dramatically inhibited cell growth of HRECs, suppressed VEGF-induced cell migration in vitro experiments, and attenuated VEGF-induced retinal vascular leakage in vivo. Intravitreal injections of fluorescently labeled LMPs revealed accumulation of LMPs in retinal tissue, with more than 60% reductions of the vascular density in retinas of rats with oxygen-induced neovascularization. LMP uptake experiments demonstrated that the interaction between LMPs and HRECs is dependent on temperature. In addition, endocytosis is partially dependent on extracellular calcium. RNAi-mediated knockdown of low-density lipoprotein receptor (LDLR) reduced the uptake of LMPs and attenuated the inhibitory effects of LMPs on VEGF-A protein expression and HRECs cell growth. Intravitreal injection of lentivirus-mediated RNA interference reduced LDLR protein expression in retina by 53% and significantly blocked the antiangiogenic effects of LMPs on pathological vascularization. In summary, the potent antiangiogenic LMPs lead to a significant reduction of pathological retinal angiogenesis through modulation of VEGF signaling, whereas LDLR-mediated endocytosis plays a partial, but pivotal, role in the uptake of LMPs in HRECs.

Interspecific prediction of photosynthetic light response curves using specific leaf mass and leaf nitrogen content: effects of differences in soil fertility and growth irradiance.

BACKGROUND AND AIMS: Previous work has shown that the entire photosynthetic light response curve, based on both Mitscherlich and Michaelis-Menten functions, could be predicted in an interspecific context through allometric relations linking the parameters of these functions to two static leaf traits: leaf nitrogen (N) content and leaf mass per area (LMA). This paper describes to what extent these allometric relations are robust to changes in soil fertility and the growth irradiance of the plants. METHODS: Plants of 25 herbaceous species were grown under controlled conditions in factorial combinations of low/high soil fertility and low/high growth irradiance. Net photosynthetic rates per unit dry mass were measured at light intensities ranging from 0 to 700 µmol m(-2) s(-1) photosynthetically active radiation (PAR). KEY RESULTS: The differing growth environments induced large changes in N, LMA and in each of the parameter estimates of the Mitscherlich and Michaelis-Menten functions. However, the differing growth environments induced only small (although significant) changes in the allometric relationships linking N and LMA to the parameters of the two functions. As a result, 88 % (Mitcherlich) and 89 % (Michaelis-Menten) of the observed net photosynthetic rates over the full range of light intensities (0-700 µmol m(-2) s(-1) PAR) and across all four growth environments could be predicted using only N and LMA using the same allometric relations. CONCLUSIONS: These results suggest the possibility of predicting net photosynthetic rates in nature across species over the full range of light intensities using readily available data.

Resting state electroretinography: An innovative approach to intrinsic retinal function monitoring.

The electroretinogram (ERG) represents the biopotential evoked by the retina in response to a light stimulus. The flash evoked ERG (fERG) is the ERG modality most frequently used clinically to diagnose and monitor retinal disorders. We hereby present a new method to record spontaneous retinal activity, without the use of a flash stimulus, that we named the resting-state ERG (rsERG). The recordings were done in normal subjects under light- and dark-adaptation and with different background light conditions (i.e., variations of wavelength and intensity). Additionally, rsERG recordings were obtained in five patients with retinopathies. The signals were subsequently analyzed in the frequency domain, extracting both periodic (i.e., frequency peaks) and aperiodic (i.e., background trend) components of the signal. The later was further assessed through a multifractal analysis using Wavelet Leaders. Results show that, irrespective of the recording conditions used, the rsERG always includes the same 90 Hz component; a frequency component also present in the fERG response, suggesting a retinally-intrinsic origin. However, in addition, the fERGs also includes a low-frequency component which is absent in the rsERGs, a finding supporting a retinally-induced origin. Comparing rsERGs with fERGs in selected patients with various retinal disorders indicates that the two retinal signals are not always similarly affected (either as a result of underlying retinal pathology or otherwise), suggesting an added value in the assessment of retinal function. Thus, the rsERG could have a similar role in clinical visual electrophysiology as that of the resting-state EEG in neurology namely, to quantify changes in spontaneous activity that result from a given disease processes.

Complete deficiency of methylenetetrahydrofolate reductase in mice is associated with impaired retinal function and variable mortality, hematological profiles, and reproductive outcomes.

Severe deficiency of methylenetetrahydrofolate reductase (MTHFR) with homocystinuria can result in early demise or later-onset neurological impairment, including developmental delay, motor dysfunction, and seizures. We previously characterized BALB/c Mthfr (-/-)mice as a model for this disorder and have recently backcrossed the disrupted allele onto the C57Bl/6 background to examine the variable phenotypes in MTHFR deficiency. Compared with BALB/c Mthfr (-/-)mice, C57Bl/6 Mthfr (-/-)mice have enhanced survival rates (81% vs 26.5%). Four-day-old BALB/c mutant pups had lower body, brain, and spleen weights relative to their wild-type counterparts compared with C57Bl/6 mutants. Pregnant BALB/c Mthfr (+/-)mice had increased resorptions and embryonic delays compared with wild-type littermates, whereas these outcomes in C57Bl/6 c Mthfr (+/-)mice were similar to those of wild-type C57Bl/6 mice. BALB/c-mutant pups had altered hematological profiles (higher hematocrit, hemoglobin, and white blood cell counts, with lower platelet counts) compared with C57Bl/6 mutants. Mutants of both strains had similar degrees of hepatic steatosis, hepatic activity of betaine:homocysteine methyltransferase, and altered cerebellar histology. Electroretinograms (ERG) in C57Bl/6 Mthfr (-/-)mice revealed decreased amplitude of scotopic and photopic waves in 6-week-old mice, with normalized ERGs at 13 weeks. Plasma homocysteine was modestly higher in C57Bl/6 compared with BALB/c mice. Our results emphasize the variable presentation of MTHFR deficiency in different genetic backgrounds and suggest that plasma homocysteine is not a predictor of severity. In addition, our novel findings of decreased spleen weights, thrombocytopenia, and impaired retinal function warrant investigation in patients with severe MTHFR deficiency or other forms of homocystinuria.

Longitudinal assessment of retinal structure and function reveals a rod-cone degeneration in a guinea pig model initially presented as night blind.

We have previously reported a naturally occurring retinopathy in a population of guinea pigs, where the affected animals presented a defect of the rod-mediated vision. The purpose of this study was to investigate if the mutants were affected with a stationary or degenerative retinopathy and to identify the cellular origin of this unique disorder. Electroretinogram (ERG) [postnatal day 1 (P1) to P450], light (LM) and electron microscopy (EM) [P5, P150, P450], and immunohistochemistry [P30, P150, P450] were evaluated from normal and mutant animals. Irrespective of age, the scotopic ERGs of mutants could only be evoked by bright flashes, and the resulting ERGs were of photopic waveform. Interestingly, the amplitude of the cone and the rod/cone a-waves was always of smaller amplitude in mutants, but this difference tended to decrease with age. In contrast, the b-waves were of larger amplitude than normal in photopic ERGs obtained prior to age 25 (days) and prior to age 10 for rod/cone ERGs. LM revealed, in mutants, an absence of the outer segment layer (OSL) with a reduction in the outer nuclear layer (ONL) thickness. EM disclosed the presence of cone outer segment (OS) while no rod OS could be evidenced. Immunohistochemistry revealed the presence of rhodopsin, both cone opsins as well as normal synaptophysin immunoreactivity. Finally, neither the retinal structure nor the function in the mutants achieved normal development. Results suggest that mutant animals are suffering from a degenerative retinal disorder that affects the structure and function of rods and cones.