Characterization of Polarimetric Properties in Various Brain Tumor Types Using Wide-Field Imaging Mueller Polarimetry.

Neuro-oncological surgery is the primary brain cancer treatment, yet it faces challenges with gliomas due to their invasiveness and the need to preserve neurological function. Hence, radical resection is often unfeasible, highlighting the importance of precise tumor margin delineation to prevent neurological deficits and improve prognosis. Imaging Mueller polarimetry, an effective modality in various organ tissues, seems a promising approach for tumor delineation in neurosurgery. To further assess its use, we characterized the polarimetric properties by analysing 45 polarimetric measurements of 27 fresh brain tumor samples, including different tumor types with a strong focus on gliomas. Our study integrates a wide-field imaging Mueller polarimetric system and a novel neuropathology protocol, correlating polarimetric and histological data for accurate tissue identification. An image processing pipeline facilitated the alignment and overlay of polarimetric images and histological masks. Variations in depolarization values were observed for grey and white matter of brain tumor tissue, while differences in linear retardance were seen only within white matter of brain tumor tissue. Notably, we identified pronounced optical axis azimuth randomization within tumor regions. This study lays the foundation for machine learning-based brain tumor segmentation algorithms using polarimetric data, facilitating intraoperative diagnosis and decision making.

Near-real-time Mueller polarimetric image processing for neurosurgical intervention.

PURPOSE: Wide-field imaging Mueller polarimetry is a revolutionary, label-free, and non-invasive modality for computer-aided intervention; in neurosurgery, it aims to provide visual feedback of white matter fibre bundle orientation from derived parameters. Conventionally, robust polarimetric parameters are estimated after averaging multiple measurements of intensity for each pair of probing and detected polarised light. Long multi-shot averaging, however, is not compatible with real-time in vivo imaging, and the current performance of polarimetric data processing hinders the translation to clinical practice. METHODS: A learning-based denoising framework is tailored for fast, single-shot, noisy acquisitions of polarimetric intensities. Also, performance-optimised image processing tools are devised for the derivation of clinically relevant parameters. The combination recovers accurate polarimetric parameters from fast acquisitions with near-real-time performance, under the assumption of pseudo-Gaussian polarimetric acquisition noise. RESULTS: The denoising framework is trained, validated, and tested on experimental data comprising tumour-free and diseased human brain samples in different conditions. Accuracy and image quality indices showed significant ( p < 0.05 ) improvements on testing data for a fast single-pass denoising versus the state-of-the-art and high polarimetric image quality standards. The computational time is reported for the end-to-end processing. CONCLUSION: The end-to-end image processing achieved real-time performance for a localised field of view ( ≈ 6.5 mm 2 ). The denoised polarimetric intensities produced visibly clear directional patterns of neuronal fibre tracts in line with reference polarimetric image quality standards; directional disruption was kept in case of neoplastic lesions. The presented advances pave the way towards feasible oncological neurosurgical translations of novel, label-free, interventional feedback.

Robustness of the wide-field imaging Mueller polarimetry for brain tissue differentiation and white matter fiber tract identification in a surgery-like environment: an ex vivo study.

During neurooncological surgery, the visual differentiation of healthy and diseased tissue is often challenging. Wide-field imaging Muller polarimetry (IMP) is a promising technique for tissue discrimination and in-plane brain fiber tracking in an interventional setup. However, the intraoperative implementation of IMP requires realizing imaging in the presence of remanent blood, and complex surface topography resulting from the use of an ultrasonic cavitation device. We report on the impact of both factors on the quality of polarimetric images of the surgical resection cavities reproduced in fresh animal cadaveric brains. The robustness of IMP is observed under adverse experimental conditions, suggesting a feasible translation of IMP for in vivo neurosurgical applications.

Effects of formalin fixation on polarimetric properties of brain tissue: fresh or fixed?

Significance: Imaging Mueller polarimetry (IMP) appears as a promising technique for real-time delineation of healthy and neoplastic tissue during neurosurgery. The training of machine learning algorithms used for the image post-processing requires large data sets typically derived from the measurements of formalin-fixed brain sections. However, the success of the transfer of such algorithms from fixed to fresh brain tissue depends on the degree of alterations of polarimetric properties induced by formalin fixation (FF). Aim: Comprehensive studies were performed on the FF induced changes in fresh pig brain tissue polarimetric properties. Approach: Polarimetric properties of pig brain were assessed in 30 coronal thick sections before and after FF using a wide-field IMP system. The width of the uncertainty region between gray and white matter was also estimated. Results: The depolarization increased by 5% in gray matter and remained constant in white matter following FF, whereas the linear retardance decreased by 27% in gray matter and by 28% in white matter after FF. The visual contrast between gray and white matter and fiber tracking remained preserved after FF. Tissue shrinkage induced by FF did not have a significant effect on the uncertainty region width. Conclusions: Similar polarimetric properties were observed in both fresh and fixed brain tissues, indicating a high potential for transfer learning.

A motor neuron disease mouse model reveals a non-canonical profile of senescence biomarkers.

To evaluate senescence mechanisms, including senescence-associated secretory phenotype (SASP), in the motor neuron disease model hSOD1-G93A, we quantified the expression of p16 and p21 and senescence-associated ß-galactosidase (SA-ß-gal) in nervous tissue. As SASP markers, we measured the mRNA levels of Il1a, Il6, Ifna and Ifnb. Furthermore, we explored whether an alteration of alternative splicing is associated with senescence by measuring the Adipor2 cryptic exon inclusion levels, a specific splicing variant repressed by TAR DNA-binding protein (TDP-43; encoded by Tardbp). Transgenic mice showed an atypical senescence profile with high p16 and p21 mRNA and protein in glia, without the canonical increase in SA-ß-gal activity. Consistent with SASP, there was an increase in Il1a and Il6 expression, associated with increased TNF-R and M-CSF protein levels, with females being partially protected. TDP-43 splicing activity was compromised in this model, and the senolytic drug Navitoclax did not alter the disease progression. This lack of effect was reproduced in vitro, in contrast to dasatinib and quercetin, which diminished p16 and p21. Our findings show a non-canonical profile of senescence biomarkers in the model hSOD1-G93A.

Polarimetric visualization of healthy brain fiber tracts under adverse conditions: ex vivo studies.

We suggest using the wide-field imaging Mueller polarimetry to contrast optically anisotropic fiber tracts of healthy brain white matter for the detection of brain tumor borders during neurosurgery. Our prior studies demonstrate that this polarimetric imaging modality detects correctly the in-plane orientation of brain white matter fiber tracts of a flat formalin-fixed thick brain specimen in reflection geometry [IEEE Trans. Med. Imaging39, 4376 (2020)10.1109/TMI.2020.3018439]. Here we present the results of ex vivo polarimetric studies of large cross-sections of fresh calf brain in reflection geometry with a special focus on the impact of the adverse measurement conditions (e.g. complex surface topography, presence of blood, etc.) on the quality of polarimetric images and the detection performance of white matter fiber tracts and their in-plane orientation.

Graphene on glassy carbon microelectrodes demonstrate long-term structural and functional stability in neurophysiological recording and stimulation.

Objective.There is a growing interest in the use of carbon and its allotropes for microelectrodes in neural probes because of their inertness, long-term electrical and electrochemical stability, and versatility. Building on this interest, we introduce a new electrode material system consisting of an ultra-thin monoatomic layer of graphene (Gr) mechanically supported by a relatively thicker layer of glassy carbon (GC).Approach.Due to its high electrical conductivity and high double-layer capacitance, Gr has impressive electrical and electrochemical properties, two key properties that are useful for neural recording and stimulation applications. However, because of its two-dimensional nature, Gr exhibits a lack of stiffness in the transverse direction and hence almost non-existent flexural and out-of-plane rigidity that will severely limit its wider use. On the other hand, GC is one of carbon's important allotropes and consists of three-dimensional microstructures of Gr fragments with a natural molecular similarity to Gr. Further, GC has exceptional chemical inertness, good electrical properties, high electrochemical stability, purely capacitive charge injection, and fast surface electrokinetics coupled with lithography patternability. This makes GC an ideal candidate for addressing Gr's lack of out-of-plane rigidity through providing a matching sturdier and robust mechanical backing. Combining the strengths of these two allotropes of carbon, we introduce a new neural probe that consists of ∼1 nm thick layer of patterned Gr microelectrodes supported by another layer of 3-5µm thick patterned GC.Main results. We present the fabrication technology for the newGr on GC(graphene on glassy carbon) microelectrodes and the accompanying pattern transfer technology on flexible substrate and report on the bond between these two allotropes of carbon through FTIR, surface morphology through SEM, topography through atomic force microscopy, and microstructure imaging through scanning transmission electron microscopy. A long-term (18 weeks)in vivostudy of the use of theseGr on GCmicroelectrodes assessed the quality of the electrocorticography-based neural signal recording and stimulation through electrophysiological measurements. The probes were demonstrated to be functionally and structurally stable over the 18 week period with minimal glial response-the longest reported so far for Gr-based microelectrodes.Significance.TheGr on GCmicroelectrodes presented here offers a compelling case for expanding the potentials of Gr-based technology in the broad areas of neural probes.

Cell Stress Induces Mislocalization of Transcription Factors with Mitochondrial Enrichment.

Previous evidence links the formation of extranuclear inclusions of transcription factors, such as ERK, Jun, TDP-43, and REST, with oxidative, endoplasmic-reticulum, proteasomal, and osmotic stress. To further characterize its extranuclear location, we performed a high-content screening based on confocal microscopy and automatized image analyses of an epithelial cell culture treated with hydrogen peroxide, thapsigargin, epoxomicin, or sorbitol at different concentrations and times to recreate the stresses mentioned above. We also performed a subcellular fractionation of the brain from transgenic mice overexpressing the Q331K-mutated TARDBP, and we analyzed the REST-regulated mRNAs. The results show that these nuclear proteins exhibit a mitochondrial location, together with significant nuclear/extranuclear ratio changes, in a protein and stress-specific manner. The presence of these proteins in enriched mitochondrial fractions in vivo confirmed the results of the image analyses. TDP-43 aggregation was associated with alterations in the mRNA levels of the REST target genes involved in calcium homeostasis, apoptosis, and metabolism. In conclusion, cell stress increased the mitochondrial translocation of nuclear proteins, increasing the chance of proteostasis alterations. Furthermore, TDP-43 aggregation impacts REST target genes, disclosing an exciting interaction between these two transcription factors in neurodegenerative processes.

Nuclear lipidome is altered in amyotrophic lateral sclerosis: A pilot study.

Nucleocytosolic transport, a membrane process, is impaired in motor neurons in amyotrophic lateral sclerosis (ALS). This study analyzes the nuclear lipidome in motor neurons in ALS and examines molecular pathways linked to the major lipid alterations. Nuclei were obtained from the frozen anterior horn of the lumbar spinal cord of ALS patients and age-matched controls. Lipidomic profiles of this subcellular fraction were obtained using liquid chromatography and mass spectrometry. We validated the mechanisms behind presumable lipidomic changes by exploring ALS surrogate models including human motor neurons (derived from ALS lines and controls) subjected to oxidative stress, the hSOD-G93A transgenic mice, and samples from an independent cohort of ALS patients. Among the differential lipid species, we noted 41 potential identities, mostly belonging to phospholipids (particularly ether phospholipids, as plasmalogens), as well as diacylglycerols and triacylglycerides. Decreased expression of alkyldihydroxyacetonephosphate synthase (AGPS)-a critical peroxisomal enzyme in plasmalogen synthesis-is found in motor neuron disease models; this occurs in parallel with an increase in the expression of sterol carrier protein 2 (SCP2) mRNA in ALS and Scp2 levels in G93A transgenic mice. Further, we identified diminished expression of diacylglycerol-related enzymes, such as phospholipase C ßI (PLCßI) and protein kinase CßII (PKCßII), linked to diacylglycerol metabolism. Finally, lipid droplets were recognized in the nuclei, supporting the identification of triacylglycerides as differential lipids. Our results point to the potentially pathogenic role of altered composition of nuclear membrane lipids and lipids in the nucleoplasm in the anterior horn of the spinal cord in ALS. Overall, these data support the usefulness of subcellular lipidomics applied to neurodegenerative diseases.

Degeneración valvular en pacientes con bioprótesis aórtica porcina evaluada mediante PET con 18Ffluoruro de sodio / Valve degeneration in patients with aortic porcine bioprosthesis evaluated through 18Fsodium fluoride PET / Degeneração valvular em pacientes com bioprótese aórtica suína avaliada mediante PET com fluoreto18F

Introducción: la degeneración valvular protésica es un problema clínico; los métodos de imagen convencionales permiten diagnosticarla en las últimas etapas. La tomografía por emisión de positrones (PET) con 18Ffluoruro puede detectar de manera precoz la degeneración subclínica. Objetivo: correlacionar parámetros de deterioro estructural protésico por PET con parámetros hemodinámicos ecocardiográficos al año de la sustitución valvular aórtica (SVA) por bioprótesis porcina. Métodos: estudio prospectivo ad hoc de un ensayo clínico. Se reclutaron pacientes sometidos a SVA por bioprótesis porcina en dos centros nacionales entre el 01/01/2019 y el 13/02/2020. Se realizaron controles clínicos y ecocardiográficos. Se seleccionaron aleatoriamente 19 sujetos a los que se les realizó PET 18Ffluoruro de sodio con angiotomografía al año de la SVA. Se midió la captación del trazador en la válvula (SUVavV) y aurícula derecha (SUVavA), calculando el índice SUVavV/SUVavA, que se comparó con los gradientes ecocardiográficos medio y máximo al año, mediante análisis de correlación de Spearman. Resultados: de 140 sujetos sometidos a SVA se realizó PET a 19, a los 16,3 meses (15,9-16,9) luego de la SVA. La mediana del índice SUVavV/SUVavA fue de 1,17 (1,11-1,27). Se encontró una correlación negativa moderada entre la captación de 18Ffluoruro y el gradiente medio (coeficiente de correlación -0,516, p = 0,028) y máximo (coeficiente -0,589, p = 0,010) al año. Conclusiones: en el seguimiento de los pacientes en los que se le realizó una sustitución valvular aórtica con bioprótesis, encontramos valores bajos de captación en el PET y gradientes ecocardiográficos normales con una correlación negativa moderada entre estos hallazgos

Introduction: prosthetic valve degeneration is a relevant clinical disorder; conventional imaging methods allow diagnosis in the later stages. 18Ffluoride positron emission tomography (PET) can detect subclinical degeneration earlier. Objective: correlate parameters of prosthetic structural deterioration by PET with echocardiographic hemodynamic parameters one year after aortic valve replacement (AVR) by porcine bioprosthesis. Methods: prospective ad hoc study of a clinical trial. Patients undergoing AVR by porcine bioprosthesis were recruited in two national centers between 01/01/2019 and 02/13/2020. Clinical and echocardiographic controls were carried out. 19 subjects were randomly selected and underwent 18Fsodium fluoride PET with CT angiography one year after AVR. Tracer uptake in the valve (SUVavV) and right atrium (SUVavA) was measured, creating the SUVavV/SUVavA index, which was compared with the mean and maximum gradients at one year, using Spearman's correlation analysis. Results: of a total of 140 subjects, PET was performed on 19 at 16.3 months (15.9-16.9) after the AVR. The median SUVavV/SUVavA ratio was 1.17 (1.11-1.27). A moderate negative correlation was found between. 18Ffluoride uptake and the mean gradient (correlation coefficient -0.516, p = 0.028) and maximum (coefficient of -0.589, p = 0.010) at one year. Conclusions: we found low uptake values in PET, echocardiographic gradients in normal range and no positive correlation between both parameters. It is the first national report with these imaging techniques

Introdução: a degeneração da válvula protética é um problema clínico; os métodos convencionais de imagem permitem o diagnóstico nas fases posteriores. A tomografia por emissão de pósitrons (PET) com fluoreto18F pode detectar a degeneração subclínica precocemente. Objetivo: correlacionar parâmetros de deterioração estrutural protética por PET com parâmetros hemodinâmicos ecocardiográficos após um ano da troca valvar aórtica (SVA) por bioprótese suína. Métodos: estudo ad hoc prospectivo de um ensaio clínico. Pacientes submetidos a SVA por bioprótese suína foram recrutados em dois centros nacionais entre 01/01/2019 e 13/02/2020. Foram realizados controles clínicos e ecocardiográficos. 19 indivíduos foram selecionados aleatoriamente que foram submetidos a PET com fluoreto de sódio 18F com angiotomografia um ano após AVS. A captação do traçador na válvula (SUVavV) e átrio direito (SUVavA) foi medida, criando o índice SUVavV/SUVavA, que foi comparado com os gradientes médio e máximo em um ano, usando a análise de correlação de Spearman. Resultados: de um total de 140 indivíduos submetidos a SVA, PET foi realizado em 19, em 16,3 meses (15,9-16,9) após a SVA. A proporção média de SUVavV/SUVavA foi de 1,17 (1,11-1,27). Uma correlação negativa moderada foi encontrada entre a captação de fluoreto18F e o gradiente médio (coeficiente de correlação -0,516, p = 0,028) e máximo (coeficiente de -0,589, p = 0,010) em um ano. Conclusões: encontramos valores baixos de captação na PET, gradientes ecocardiográficos dentro da normalidade, sem correlação positiva entre os dois parâmetros. É o primeiro trabalho nacional com essas técnicas de imagem

Method to calibrate a full-Stokes polarimeter based on variable retarders.

We present a calibration method for a full-Stokes polarimeter. The polarimeter uses two liquid-crystal variable retarders (LCVR) and a linear polarizer to measure the four Stokes parameters. The calibration method proposed in this paper calculates the errors in the experimental setup by fitting the experimental intensity measurements for a set of calibration samples to a theoretical polarimeter with errors. The errors calculated in the method include the axes alignment errors and the errors in the retardance values of both LCVRs. The resulting calibration parameters are verified by measuring the polarization state of a light beam passing through a rotating linear polarizer, a half-wave plate, and a quarter-wave plate and comparing with the predictions for an ideal, error-free polarimeter. It is found that an average reduction in rms error of 55.8% can be obtained with the proposed method.

Synthesis and DNA binding profile of monomeric, dimeric, and trimeric derivatives of crystal violet.

Monomeric, dimeric, and trimeric derivatives of the triphenylmethane dye crystal violet (1a-1f) have been synthesized for the purpose of evaluating their affinity and sequence selectivity for duplex DNA. Competitive ethidum displacement assays indicate that 1a-1f have apparent association constants for CT DNA in the range of 1.80-16.2 × 107 M-1 and binding site sizes of 10-14 bp. Viscosity experiments performed on ligand 1f confirmed that these dyes associate with duplex DNA by a non-intercalative mode of binding. Circular dichroism and competition binding studies of the tightest binding ligand 1e with known major and minor groove binding molecules suggest that these dye derivatives likely occupy the major groove of DNA. Data from the binding of 1e to polynucleotides indicate close to an order of magnitude preference for associating with AT rich homopolymers over GC rich homopolymers, suggesting a shape-selective match of the sterically bulky ligand with DNA containing a wider major groove.

Cryptic exon splicing function of TARDBP interacts with autophagy in nervous tissue.

TARDBP (TAR DNA binding protein) is one of the components of neuronal aggregates in sporadic amyotrophic lateral sclerosis (ALS) and frontotemporal lobar degeneration. We have developed a simple quantitative method to evaluate TARDBP splicing function that was applied to spinal cord, brainstem, motor cortex, and occipital cortex in ALS (n = 8) cases compared to age- and gender-matched control (n = 17). Then, we quantified the abundance of a TARDBP-spliced cryptic exon present in ATG4B (autophagy related 4B cysteine peptidase) mRNA. Results of these analyses demonstrated that the loss of this TARDBP function in spinal cord, brainstem, motor cortex, and occipital cortex differentiated ALS from controls (area under the curve of receiver operating characteristic: 0.85). Significant correlations were also observed between cryptic exon levels, age, disease duration, and aberrant mRNA levels. To test if TARDBP function in splicing is relevant in ATG4B major function (autophagy) we downregulated TARDBP expression in human neural tissue and in HeLa cells, demonstrating that TARDBP is required for maintaining the expression of ATG4B. Further, ATG4B overexpression alone is sufficient to completely prevent the increase of SQSTM1 induced by TARDBP downregulation in human neural tissue cells and in cell lines. In conclusion, the present findings demonstrate abnormal alternative splicing of ATG4B transcripts in ALS neural tissue in agreement with TARDBP loss of function, leading to impaired autophagy. ABBREVIATIONS: ALS: amyotrophic lateral sclerosis; ATG4B: autophagy related 4B cysteine peptidase; AUC: area under the curve; FTLD: frontotemporal lobar degeneration; iPSC: induced pluripotent stem cells; ROC: receiver operating characteristic; TARDBP: TAR DNA binding protein; RT-qPCR: quantitative RT-PCR.

Permitted experimental errors for optimized variable-retarder Mueller-matrix polarimeters.

An optimized Mueller-matrix polarimeter is simulated. The polarimeter is optimized by finding the configurations of the polarization state generator and polarization state analyzer that give the minimum condition number. Noise is included in the measurement of the polarimeter intensities, and the eigenvalue calibration procedure is used to reduce the errors in the final Mueller matrix. Controlled errors are introduced to the polarimeter configuration, and the error in the final measured Mueller matrix is calculated as a function of these configuration errors. It is found that the alignment of the retarder axes in the polarimeter is much more important than the use of the ideal, optimized retardance values. In particular, the misalignment of the retarders farthest from the sample is the error source with the highest impact in the precision of the polarimeter.

Experimental limits for eigenvalue calibration in liquid-crystal Mueller-matrix polarimeters.

A numerical study is carried out to find the experimental conditions necessary for the eigenvalue calibration procedure to work correctly in a liquid-crystal variable-retarder-based Mueller-matrix polarimeter. Using the error between the simulated experimental Mueller matrix in a polarimeter with errors and the expected ideal Mueller matrices for four calibration samples, the maximum experimental errors are estimated for a successful eigenvalue calibration. It is found that the retarder axes' orientations have smaller permitted errors than the retardation values.

Calibration and data extraction in nonoptimized Mueller matrix polarimeters.

We present a method for calibration and data extraction for a nonoptimized Mueller matrix polarimeter. The advantage of this type of method is a reduction in measurement time for multiwavelength systems or in systems with slow response times. The calibration process requires the measurement of four known polarization devices. Here we use free-space transmission, a horizontal and a vertical linear polarizer, and a quarter-wave retarder with its fast axis at 30° to the horizontal. Experimental measurements of rotating quarter-wave and half-wave retarders show that accurate results can be obtained with the proposed method.

Contrast based circular approximation for accurate and robust optic disc segmentation in retinal images.

A new method for automatic optic disc localization and segmentation is presented. The localization procedure combines vascular and brightness information to provide the best estimate of the optic disc center which is the starting point for the segmentation algorithm. A detection rate of 99.58% and 100% was achieved for the Messidor and ONHSD databases, respectively. A simple circular approximation to the optic disc boundary is proposed based on the maximum average contrast between the inner and outer ring of a circle centered on the estimated location. An average overlap coefficient of 0.890 and 0.865 was achieved for the same datasets, outperforming other state of the art methods. The results obtained confirm the advantages of using a simple circular model under non-ideal conditions as opposed to more complex deformable models.

Early and gender-specific differences in spinal cord mitochondrial function and oxidative stress markers in a mouse model of ALS.

INTRODUCTION: Amyotrophic lateral sclerosis (ALS) is a motor neuron disease with a gender bias towards major prevalence in male individuals. Several data suggest the involvement of oxidative stress and mitochondrial dysfunction in its pathogenesis, though differences between genders have not been evaluated. For this reason, we analysed features of mitochondrial oxidative metabolism, as well as mitochondrial chain complex enzyme activities and protein expression, lipid profile, and protein oxidative stress markers, in the Cu,Zn superoxide dismutase with the G93A mutation (hSOD1-G93A)- transgenic mice and Neuro2A(N2A) cells overexpressing hSOD1-G93A. RESULTS AND CONCLUSIONS: Our results show that overexpression of hSOD1-G93A in transgenic mice decreased efficiency of mitochondrial oxidative phosphorylation, located at complex I, revealing a temporal delay in females with respect to males associated with a parallel increase in selected markers of protein oxidative damage. Further, females exhibit a fatty acid profile with higher levels of docosahexaenoic acid at 30 days. Mechanistic studies showed that hSOD1-G93A overexpression in N2A cells reduced complex I function, a defect prevented by 17ß-estradiol pretreatment. In conclusion, ALS-associated SOD1 mutation leads to delayed mitochondrial dysfunction in female mice in comparison with males, in part attributable to the higher oestrogen levels of the former. This study is important in the effort to further understanding of whether different degrees of spinal cord mitochondrial dysfunction could be disease modifiers in ALS.

Oral intake of genetically engineered high-carotenoid corn ameliorates hepatomegaly and hepatic steatosis in PTEN haploinsufficient mice.

Non-alcoholic fatty liver disease (NAFLD) is the most common form of chronic liver disease. Here we show that a mouse model of haploinsufficiency in the lipid and protein phosphatase and tensin homolog protein (PTEN(+/-)) exhibits hepatomegaly, increased liver lipogenic gene expression (SREBP-1C and PPARγ) and hepatic lesions analogous to human NAFLD. The livers of PTEN(+/-) mice also contained lower levels of retinoic acid (RA) than normal, similarly to human NAFLD patients. The RA signaling pathway thus offers a novel therapeutic target for the treatment of NAFLD although the impact of nutrition in this context is unclear. We therefore fed PTEN(+/-) mice for 36weeks a diet containing genetically engineered high-carotenoid corn (HCAR) to investigate its potential beneficial effects on the hepatic symptoms of NAFLD. The HCAR diet reduced hepatomegaly and promoted the repartitioning of fatty acids in the liver, away from triacylglycerol storage. At the molecular level, the HCAR diet clearly reduced lipogenic gene expression, boosted catabolism, and increased hepatic RA levels. These results set the stage for human trials to evaluate the use of high-carotenoid foods for the reduction or prevention of steatosis in NAFLD.

Interplay between TDP-43 and docosahexaenoic acid-related processes in amyotrophic lateral sclerosis.

BACKGROUND: Docosahexaenoic acid (DHA), a key lipid in nervous system homeostasis, is depleted in the spinal cord of sporadic amyotrophic lateral sclerosis (sALS) patients. However, the basis for such loss was unknown. METHODS: DHA synthetic machinery was evaluated in spinal cord samples from ALS patients and controls by immunohistochemistry and western blot. Further, lipid composition was measured in organotypic spinal cord cultures by gas chromatography and liquid chromatography coupled to mass spectrometry. In these samples, mitochondrial respiratory functions were measured by high resolution respirometry. Finally, Neuro2-A and stem cell-derived human neurons were used for evaluating mechanistic relationships between TDP-43 aggregation, oxidative stress and cellular changes in DHA-related proteins. RESULTS: ALS is associated to changes in the spinal cord distribution of DHA synthesis enzymatic machinery comparing ten ALS cases and eight controls. We found increased levels of desaturases (ca 95% increase, p<0.001), but decreased amounts of DHA-related ß-oxidation enzymes in ALS samples (40% decrease, p<0.05). Further, drebrin, a DHA-dependent synaptic protein, is depleted in spinal cord samples from ALS patients (around 40% loss, p<0.05). In contrast, chronic excitotoxicity in spinal cord increases DHA acid amount, with both enhanced concentrations of neuroprotective docosahexaenoic acid-derived resolvin D, and higher lipid peroxidation-derived molecules such as 8-iso-prostaglandin-F2-α (8-iso-PGF2α) levels. Since α-tocopherol improved mitochondrial respiratory function and motor neuron survival in these conditions, it is suggested that oxidative stress could boost motor neuron loss. Cell culture and metabolic flux experiments, showing enhanced expression of desaturases (FADS2) and ß-oxidation enzymes after H2O2 challenge suggest that DHA production can be an initial response to oxidative stress, driven by TDP-43 aggregation and drebrin loss. Interestingly, these changes were dependent on cell type used, since human neurons exhibited losses of FADS2 and drebrin after oxidative stress. These features (drebrin loss and FADS2 alterations) were also produced by transfection by aggregation prone C-terminal fragments of TDP-43. CONCLUSIONS: sALS is associated with tissue-specific DHA-dependent synthetic machinery alteration. Furthermore, excitotoxicity sinergizes with oxidative stress to increase DHA levels, which could act as a response over stress, involving the expression of DHA synthetic enzymes. Later on, this allostatic overload could exacerbate cell stress by contributing to TDP-43 aggregation. This, at its turn, could blunt this protective response, overall leading to DHA depletion and neuronal dysfunction.