Search

Disruption of adipocyte de novo lipogenesis (DNL) by deletion of fatty acid synthase (FASN) in mice induces browning in inguinal white adipose tissue (iWAT). However, adipocyte FASN knockout (KO) increases acetyl-coenzyme A (CoA) and malonyl-CoA in addition to depletion of palmitate. We explore which of these metabolite changes triggers adipose browning by generating eight adipose-selective KO mouse models with loss of ATP-citrate lyase (ACLY), acetyl-CoA carboxylase 1 (ACC1), ACC2, malonyl-CoA decarboxylase (MCD) or FASN, or dual KOs ACLY/FASN, ACC1/FASN, and ACC2/FASN. Preventing elevation of acetyl-CoA and malonyl-CoA by depletion of adipocyte ACLY or ACC1 in combination with FASN KO does not block the browning of iWAT. Conversely, elevating malonyl-CoA levels in MCD KO mice does not induce browning. Strikingly, adipose ACC1 KO induces a strong iWAT thermogenic response similar to FASN KO while also blocking malonyl-CoA and palmitate synthesis. Thus, ACC1 and FASN are strong suppressors of adipocyte thermogenesis through promoting lipid synthesis rather than modulating the DNL intermediates acetyl-CoA or malonyl-CoA., Competing Interests: Declaration of interests The authors declare no competing interests., (Copyright © 2023 The Author(s). Published by Elsevier Inc. All rights reserved.)

Adipocytes robustly synthesize fatty acids (FA) from carbohydrate through the de novo lipogenesis (DNL) pathway, yet surprisingly DNL contributes little to their abundant triglyceride stored in lipid droplets. This conundrum raises the hypothesis that adipocyte DNL instead enables membrane expansions to occur in processes like autophagy, which requires an abundant supply of phospholipids. We report here that adipocyte Fasn deficiency in vitro and in vivo markedly impairs autophagy, evident by autophagosome accumulation and severely compromised degradation of the autophagic substrate p62. Our data indicate the impairment occurs at the level of autophagosome-lysosome fusion, and indeed, loss of Fasn decreases certain membrane phosphoinositides necessary for autophagosome and lysosome maturation and fusion. Autophagy dependence on FA produced by Fasn is not fully alleviated by exogenous FA in cultured adipocytes, and interestingly, imaging studies reveal that Fasn colocalizes with nascent autophagosomes. Together, our studies identify DNL as a critical source of FAs to fuel autophagosome and lysosome maturation and fusion in adipocytes., (© 2023. The Author(s).)

The purpose of this study was to compare the clinical outcomes of 13 patients with optic disc pit maculopathy (ODP-M) - progressive visual loss, serous macular detachment, and/or intraretinal fluid - who underwent different surgical approaches. This was a retrospective study including a consecutive sample of 13 patients aged 13-74 years (mean 35.38 ± 19.66 years) diagnosed with ODP-M and submitted to vitreoretinal surgery between 2005 and 2021. All patients underwent pars plana vitrectomy, posterior hyaloid detachment, and gas tamponade. Endolaser photocoagulation was applied to the temporal margin of the optic disc in 8 cases; internal limiting membrane (ILM) peeling was performed in 9 cases; and ILM inverted flap technique in 5 cases. Stuffing of the pit with an ILM flap was performed in 3 cases. Mean best-corrected visual acuity improved from 20/200 (1.04 ± 0.56 LogMAR) to 20/50 (0.43 ± 0.54 LogMAR) within 4-36 months. Central retinal thickness decreased from 587.5 ± 158.01 μm to 253.9 ± 33.55 μm, and 7 out of 10 patients had complete resolution of intraretinal fluid. All patients had complete retinal reattachment; however, a few years after surgery, 4 patients had recurrence of serous retinal detachment. The only adjunctive technique associated with greater visual improvement was endolaser ( p = 0.033) and not performing peeling of the ILM was also associated with better visual results ( p = 0.013), independently of preoperative visual acuity or age at the time of surgery. None of the adjunctive procedures was a significant predictor of better anatomical outcomes. In conclusion, all of these approaches for the surgical management of ODP-M were safe and effective. In this study, vitrectomy with endolaser was a good option for management of ODP-M., Competing Interests: The authors have no relevant financial or non-financial interests to disclose., (Copyright © 2022 by The Author(s). Published by S. Karger AG, Basel.)

Background: This study aimed to use the Charlson Comorbidity Index (CCI) to assess the prevalence of medical comorbidities among hospitalization episodes with a primary Bipolar Disorder (BD) diagnosis, and to analyze its association with hospitalization outcomes., Methods: A population-based observational retrospective study was conducted using a Portuguese administrative database containing all mainland public hospitalizations. From 2008-2015, hospitalization episodes with a primary diagnosis of BD were analysed. Outcomes included: length of stay (LoS), in-hospital mortality and discharge destination., Results: Overall, 20807 hospitalization episodes were analysed. Mean±standard deviation age at admission was 47.9±14.3 years, and these episodes mostly refer to women's admissions (66.6%). Median (1st quartile; 3rd quartile) LoS was 16.0 (9.0; 25.0) days. A total of 2145 (10.3%) episodes had ≥1 CCI comorbidities registered, being diabetes the most prevalent. LoS was significantly higher for episodes with secondary diagnoses of congestive heart failure, cerebrovascular disease, dementia, diabetes, renal disease and malignancy (all p<0.05). Episodes with a registry of myocardial infarction, peripheral vascular disease, malignancy and renal disease diagnoses had higher in-hospital mortality., Limitations: Limitations include the use of data registered for administrative reasons rather than research purposes, and the analysis of hospitalization episodes, instead of patients., Conclusions: In this Portuguese nationwide study, greater comorbidity had a measurable impact on BD hospitalization outcomes. During the study period the prevalence of CCI comorbidities rose from 8.1% to 17.4%, which may reflect the overall increasing quality of hospital-coded data in Portugal throughout the years. The detection and timely management of medical comorbid conditions will likely prevent the high BD medical burden., (Copyright © 2021. Published by Elsevier B.V.)

Introduction: To report the first case of a serpiginous choroiditis presenting after SARS-CoV-2 infection in a previously healthy young woman., Case Description: A 41-year-old woman reported blurry vision OS 1 month after a mild SARS-CoV-2 infection. Left eye fundus examination revealed multiple peripapillary atrophic lesions, adjacent to a larger diffuse, ill-defined, yellow-whitish deep amoeboid-like patch, involving the peripapillary region and extending temporally to the fovea. Multimodal imaging including fluorescein angiography, indocyanine-green angiography, fundus autofluorescence and optical coherence tomography was consistent with serpiginous choroiditis. A complete systemic work-up was performed to exclude potential infectious or inflammatory etiologies. The active choroidal lesions responded to high dose corticosteroids, with functional improvement. Immunomodulatory therapy with methotrexate was initiated for long-term management., Conclusion: Serpiginous choroiditis is a rare but important sight-threatening condition that has been previously associated to viral infections, which seem to have a role in the induction and/or perpetuation of choroidal inflammation. SARS-CoV-2 infection appears to have played a role as a possible trigger for intraocular inflammation in this case. Therefore, COVID-19 patients reporting visual symptoms should be carefully evaluated in order to obtain adequate ophthalmological management to avoid irreversible visual damage.

The emergence of deep learning has considerably advanced the state-of-the-art in cardiac magnetic resonance (CMR) segmentation. Many techniques have been proposed over the last few years, bringing the accuracy of automated segmentation close to human performance. However, these models have been all too often trained and validated using cardiac imaging samples from single clinical centres or homogeneous imaging protocols. This has prevented the development and validation of models that are generalizable across different clinical centres, imaging conditions or scanner vendors. To promote further research and scientific benchmarking in the field of generalizable deep learning for cardiac segmentation, this paper presents the results of the Multi-Centre, Multi-Vendor and Multi-Disease Cardiac Segmentation (M&Ms) Challenge, which was recently organized as part of the MICCAI 2020 Conference. A total of 14 teams submitted different solutions to the problem, combining various baseline models, data augmentation strategies, and domain adaptation techniques. The obtained results indicate the importance of intensity-driven data augmentation, as well as the need for further research to improve generalizability towards unseen scanner vendors or new imaging protocols. Furthermore, we present a new resource of 375 heterogeneous CMR datasets acquired by using four different scanner vendors in six hospitals and three different countries (Spain, Canada and Germany), which we provide as open-access for the community to enable future research in the field.

Obesity and type 2 diabetes are associated with disturbances in insulin-regulated glucose and lipid fluxes and severe comorbidities including cardiovascular disease and steatohepatitis. Whole body metabolism is regulated by lipid-storing white adipocytes as well as "brown" and "brite/beige" adipocytes that express thermogenic uncoupling protein 1 (UCP1) and secrete factors favorable to metabolic health. Implantation of brown fat into obese mice improves glucose tolerance, but translation to humans has been stymied by low abundance of primary human beige adipocytes. Here we apply methods to greatly expand human adipocyte progenitors from small samples of human subcutaneous adipose tissue and then disrupt the thermogenic suppressor gene NRIP1 by CRISPR. Ribonucleoprotein consisting of Cas9 and sgRNA delivered ex vivo are fully degraded by the human cells following high efficiency NRIP1 depletion without detectable off-target editing. Implantation of such CRISPR-enhanced human or mouse brown-like adipocytes into high fat diet fed mice decreases adiposity and liver triglycerides while enhancing glucose tolerance compared to implantation with unmodified adipocytes. These findings advance a therapeutic strategy to improve metabolic homeostasis through CRISPR-based genetic enhancement of human adipocytes without exposing the recipient to immunogenic Cas9 or delivery vectors., (© 2021. The Author(s).)

Aims: Artificial intelligence (A.I) driven voice-based assistants may facilitate data capture in clinical care and trials; however, the feasibility and accuracy of using such devices in a healthcare environment are unknown. We explored the feasibility of using the Amazon Alexa ('Alexa') A.I. voice-assistant to screen for risk factors or symptoms relating to SARS-CoV-2 exposure in quaternary care cardiovascular clinics., Methods and Results: We enrolled participants to be screened for signs and symptoms of SARS-CoV-2 exposure by a healthcare provider and then subsequently by the Alexa. Our primary outcome was interrater reliability of Alexa to healthcare provider screening using Cohen's Kappa statistic. Participants rated the Alexa in a post-study survey (scale of 1 to 5 with 5 reflecting strongly agree). This study was approved by the McGill University Health Centre ethics board. We prospectively enrolled 215 participants. The mean age was 46 years [17.7 years standard deviation (SD)], 55% were female, and 31% were French speakers (others were English). In total, 645 screening questions were delivered by Alexa. The Alexa mis-identified one response. The simple and weighted Cohen's kappa statistic between Alexa and healthcare provider screening was 0.989 [95% confidence interval (CI) 0.982-0.997] and 0.992 (955 CI 0.985-0.999), respectively. The participants gave an overall mean rating of 4.4 (out of 5, 0.9 SD)., Conclusion: Our study demonstrates the feasibility of an A.I. driven multilingual voice-based assistant to collect data in the context of SARS-CoV-2 exposure screening. Future studies integrating such devices in cardiovascular healthcare delivery and clinical trials are warranted., Registration: https://clinicaltrials.gov/ct2/show/NCT04508972., (© The Author(s) 2021. Published by Oxford University Press on behalf of the European Society of Cardiology.)

Waterhouse-Friderichsen syndrome (WFS), defined as severe adrenal insufficiency due to bilateral adrenal gland haemorrhagic necrosis, occurred in a 59-year-old woman. An underlying serogroup Y Neisseria meningitidis (NM) infection was diagnosed, with a rapid progression to purpura fulminans, disseminated intravascular coagulation and WFS. Intensive treatment including fluid resuscitation, broad-spectrum antibiotic therapy, ventilatory support, platelet and factor replacement were administered. The meningococcaemia in the presence of WFS had a fulminant progression, leading to a fatal outcome within 24 hours of symptom onset. This case details the diagnosis and management challenges of the WFS, a rare complication of NM septicaemia, and describes the identification of a NM serogroup that is rare in Portugal in middle-aged patients., Competing Interests: Competing interests: None declared., (© BMJ Publishing Group Limited 2021. No commercial re-use. See rights and permissions. Published by BMJ.)

Adipocytes deficient in fatty acid synthase (iAdFASNKO) emit signals that mimic cold exposure to enhance the appearance of thermogenic beige adipocytes in mouse inguinal white adipose tissues (iWATs). Both cold exposure and iAdFASNKO upregulate the sympathetic nerve fiber (SNF) modulator Neuregulin 4 (Nrg4), activate SNFs, and require adipocyte cyclic AMP/protein kinase A (cAMP/PKA) signaling for beige adipocyte appearance, as it is blocked by adipocyte Gsα deficiency. Surprisingly, however, in contrast to cold-exposed mice, neither iWAT denervation nor Nrg4 loss attenuated adipocyte browning in iAdFASNKO mice. Single-cell transcriptomic analysis of iWAT stromal cells revealed increased macrophages displaying gene expression signatures of the alternately activated type in iAdFASNKO mice, and their depletion abrogated iWAT beiging. Altogether, these findings reveal that divergent cellular pathways are sufficient to cause adipocyte browning. Importantly, adipocyte signaling to enhance alternatively activated macrophages in iAdFASNKO mice is associated with enhanced adipose thermogenesis independent of the sympathetic neuron involvement this process requires in the cold., Competing Interests: Declaration of Interests The authors declare no competing interests., (Copyright © 2020 The Author(s). Published by Elsevier Inc. All rights reserved.)

Aerobic exercise training (AER) may promote several adaptations in white adipose tissue (WAT), including a phenotypic change known as browning. The present study aimed at assessing if resistance exercise training (RES) would be as efficient as AER in inducing a brown-like adipocyte reprogramming in WAT. Thirty Swiss male mice were randomly divided into 3 groups with 10 animals each: 1 ) sedentary (SED), 2 ) AER, and 3 ) RES. After the adaptation training, an incremental test was performed at the beginning of each week to adjust training load. Mice were submitted to 8 wk of AER or RES. After the experimental period, inguinal and retroperitoneal WAT (iWAT and rpWAT) and brown adipose tissue (BAT) were collected. The prescription of AER and RES was effective in increasing the performance of both groups. Also, RES presented a lower body weight than AER/SED. AER and RES reduced the area of iWAT and rpWAT adipocytes and the lipid area of BAT, induced an increase of vascular endothelial growth factor (VEGF) and cluster of differentiation 31 (CD31) and uncoupling protein 1 (UCP-1), and increased the expression of selective genes of brown and beige phenotype in adipocytes after 8 wk. In general, we demonstrated here that AER and RES training similarly induced the browning of iWAT and rpWAT. NEW & NOTEWORTHY Aerobic exercise training (AER) induces the browning of white adipose tissue, turning adipocytes multilocular, highly vascularized and expressing uncoupling protein 1 (UCP-1). The current study compared the efficiency of resistance to aerobic exercise training to promote a brown-like phenotype. Our results suggest that both types of training similarly induce subcutaneous and visceral adipose tissue browning.

Here, we show that β adrenergic signaling coordinately upregulates de novo lipogenesis (DNL) and thermogenesis in subcutaneous white adipose tissue (sWAT), and both effects are blocked in mice lacking the cAMP-generating G protein-coupled receptor Gs (Adipo-GsαKO) in adipocytes. However, UCP1 expression but not DNL activation requires rapamycin-sensitive mTORC1. Furthermore, β3-adrenergic agonist CL316243 readily upregulates thermogenic but not lipogenic genes in cultured adipocytes, indicating that additional regulators must operate on DNL in sWAT in vivo. We identify one such factor as thyroid hormone T3, which is elevated locally by adrenergic signaling. T3 administration to wild-type mice enhances both thermogenesis and DNL in sWAT. Mechanistically, T3 action on UCP1 expression in sWAT depends upon cAMP and is blocked in Adipo-GsαKO mice even as elevated DNL persists. Thus, T3 enhances sWAT thermogenesis by amplifying cAMP signaling, while its control of adipocyte DNL can be mediated independently of both cAMP and rapamycin-sensitive mTORC1., Competing Interests: Declaration of Interests The authors declare no competing interests., (Copyright © 2020 The Authors. Published by Elsevier Inc. All rights reserved.)

The brown alga Lobophora (Dictyotales, Phaeophyceae) is an important macroalga in the North-eastern Atlantic archipelagos (i.e., Macaronesia). Notably in the Canaries it can dominate benthic assemblages. While the genus has been the subject of several ecological studies in the Canaries, no study has yet been conducted to assess species-level diversity of Lobophora in Macaronesia. We reassessed the diversity of Lobophora in Macaronesia, reporting the presence of seven species (L. caboverdeana sp. nov., L. canariensis, L. dagamae sp. nov., L. delicata, L. dispersa, L. littlerorum, and L. schneideri). Lobophora spp. from Macaronesia are morphologically and ecologically distinguishable. In the Canaries, L. schneideri dominates the photophilic assemblages from the intertidal to 20-30 m depth. Lobophora dagamae sp. nov. grows in less illuminated shallow habitats, and replaces L. schneideri from 30 to ~80 m. Lobophora canariensis also has a wide vertical distribution, from the intertidal to deep waters, while L. delicata, L. dispersa and L. littlerorum grow in shallow waters. The dominance of species with an upright habit versus prostrate or crustose species may be mediated by the pressure of herbivores. Four species have an amphi-Atlantic distribution: L. littlerorum, L. canariensis, L. delicata, and L. schneideri. Lobophora schneideri and L. delicata are furthermore distributed in the Mediterranean Sea. By sampling a pivotal region in the Atlantic, this study significantly improves our knowledge of Lobophora biogeography in the Atlantic Ocean. Macaronesia constitutes a species-poor region for Lobophora where no diversification events occurred, and a region of overlap between the Greater Caribbean and the Indo-Pacific., (© 2019 Phycological Society of America.)

Adipose tissue comprises adipocytes and many other cell types that engage in dynamic crosstalk in a highly innervated and vascularized tissue matrix. Although adipose tissue has been studied for decades, it has been appreciated only in the past 5 years that extensive arborization of nerve fibres has a dominant role in regulating the function of adipose tissue. This Review summarizes the latest literature, which suggests that adipocytes signal to local sensory nerve fibres in response to perturbations in lipolysis and lipogenesis. Such adipocyte signalling to the central nervous system causes sympathetic output to distant adipose depots and potentially other metabolic tissues to regulate systemic glucose homeostasis. Paracrine factors identified in the past few years that mediate such adipocyte-neuron crosstalk are also reviewed. Similarly, immune cells and endothelial cells within adipose tissue communicate with local nerve fibres to modulate neurotransmitter tone, blood flow, adipocyte differentiation and energy expenditure, including adipose browning to produce heat. This understudied field of neurometabolism related to adipose tissue biology has great potential to reveal new mechanistic insights and potential therapeutic strategies for obesity and type 2 diabetes mellitus.

We consider a Wireless Sensor Network (WSN) monitoring environmental data. Compressive Sensing (CS) is explored to reduce the number of coefficients to transmit and consequently save the energy of sensor nodes. Each sensor node collects N samples of environmental data, these are CS coded to transmit M < N values to a sink node. The M CS coefficients are uniformly quantized and entropy coded. We investigate the rate-distortion performance of this approach even under CS coefficient losses. The results show the robustness of the CS coding framework against packet loss. We devise a simple strategy to successively approximate/quantize CS coefficients, allowing for an efficient incremental transmission of CS coded data. Tests show that the proposed successive approximation scheme provides rate allocation adaptivity and flexibility with a minimum rate-distortion performance penalty.

Cancer-induced cachexia, characterized by systemic inflammation, body weight loss, adipose tissue (AT) remodeling and muscle wasting, is a malignant metabolic syndrome with undefined etiology. Here, we show that both genetic ablation and pharmacological inhibition of TLR4 were able to attenuate the main clinical markers of cachexia in mice bearing Lewis lung carcinoma (LLC). AT remodelling was not found in LLC tumor-bearing (TB) TLR4 -/- mice due to reduced macrophage infiltration and adipocyte atrophy. TLR4 -/- mice were also resistant to cold-induced browning of subcutaneous AT (scAT). Importantly, pharmacological inhibition of TLR4 (Atorvastatin) reproduced the main protective effect against AT remodeling found in TLR4 -/- TB mice. Moreover, the treatment was effective in prolonging survival and attenuating tumor mass growth when compared to non-treated-TB animals. Furthermore, tumor-induced elevation of circulating pro-inflammatory cytokines was similarly abolished in both genetic ablation and pharmacological inhibition of TLR4. These data suggest that TLR4 is a critical mediator and a promising target for novel anti-cachexia therapies.

RNA-guided, engineered nucleases derived from the prokaryotic adaptive immune system CRISPR-Cas represent a powerful platform for gene deletion and editing. When used as a therapeutic approach, direct delivery of Cas9 protein and single-guide RNA (sgRNA) could circumvent the safety issues associated with plasmid delivery and therefore represents an attractive tool for precision genome engineering. Gene deletion or editing in adipose tissue to enhance its energy expenditure, fatty acid oxidation, and secretion of bioactive factors through a "browning" process presents a potential therapeutic strategy to alleviate metabolic disease. Here, we developed "CRISPR-delivery particles," denoted CriPs, composed of nano-size complexes of Cas9 protein and sgRNA that are coated with an amphipathic peptide called Endo-Porter that mediates entry into cells. Efficient CRISPR-Cas9-mediated gene deletion of ectopically expressed GFP by CriPs was achieved in multiple cell types, including a macrophage cell line, primary macrophages, and primary pre-adipocytes. Significant GFP loss was also observed in peritoneal exudate cells with minimum systemic toxicity in GFP-expressing mice following intraperitoneal injection of CriPs containing Gfp -targeting sgRNA. Furthermore, disruption of a nuclear co-repressor of catabolism, the Nrip1 gene, in white adipocytes by CriPs enhanced adipocyte browning with a marked increase of uncoupling protein 1 (UCP1) expression. Of note, the CriP-mediated Nrip1 deletion did not produce detectable off-target effects. We conclude that CriPs offer an effective Cas9 and sgRNA delivery system for ablating targeted gene products in cultured cells and in vivo , providing a potential therapeutic strategy for metabolic disease., (© 2018 Shen et al.)

Objective: Crosstalk between adipocytes and local neurons may be an important regulatory mechanism to control energy homeostasis. We previously reported that perturbation of adipocyte de novo lipogenesis (DNL) by deletion of fatty acid synthase (FASN) expands sympathetic neurons within white adipose tissue (WAT) and stimulates the appearance of "beige" adipocytes. Here we tested whether WAT DNL activity can also influence neuronal regulation and thermogenesis in brown adipose tissue (BAT)., Methods and Results: Induced deletion of FASN in all adipocytes in mature mice (iAdFASNKO) enhanced sympathetic innervation and neuronal activity as well as UCP1 expression in both WAT and BAT. This increased sympathetic innervation could be observed at both 22 °C and 30 °C, indicating it is not a response to heat loss but rather adipocyte signaling. In contrast, selective ablation of FASN in brown adipocytes of mice (iUCP1FASNKO) failed to modulate sympathetic innervation and the thermogenic program in BAT. Surprisingly, DNL in brown adipocytes was also dispensable in maintaining euthermia when UCP1FASNKO mice were cold-exposed., Conclusion: These results indicate that DNL in white adipocytes influences long distance signaling to BAT, which can modify BAT sympathetic innervation and expression of genes involved in thermogenesis., (Copyright © 2018 The Authors. Published by Elsevier GmbH.. All rights reserved.)

Cancer cachexia (CC) is a multifactorial syndrome with an unknown etiology. The primary symptom is the progressive reduction of the body weight. Recently, down-regulation of adipogenic and lipogenic genes were demonstrated to be early affected during cachexia progression in adipose tissue (AT), resulting in AT remodeling. Thus, this study aimed to evaluate in a co-culture system the influence of the Lewis Lung Carcinoma (LLC) tumor cells (c/c-LLC) in an established pre-adipocyte cell line 3T3-L1 adipogenic capacity. c/c-LLC in the presence of 3T3-L1 caused a reduction in lipids accumulation, suggesting that secretory tumor cells products may affect adipogenesis. Interestingly, a very early (day 2) down-regulation of proliferator-activated receptor gamma (PPARγ) and CCAAT/enhancer-binding protein alpha (C/EBPα), followed by late genes (day 4 and 8), adiponectin, perilipin, and fatty acid-binding protein 4 (FABP4). Caspase-3 expression was increased on the last day of cell differentiation; it occurred in the expression of pro-inflammatory cytokines interleukin-6 (IL-6) and tumor necrosis factor alpha (TNF-α). Overall, our results suggest that LLC secretory products impair adipocyte differentiation in a co-culture system and increased apoptosis. In summary, our study has shown the inhibition of the adipogenic process in the 3T3-L1 co-culture system with LLC cells.

Pigmented purpuric dermatosis is a chronic benign skin disorder of unknown aetiology. Although there are several other potential cofactors, drugs are the most frequent cause. This paper describes the case of a 71-year-old woman who was admitted in the emergency department with skin lesions on the lower extremities, characteristics of Schamberg's disease. After a medical study and treatment, it was concluded that the lesions were caused by amlodipine administration. To the authors' knowledge, only one previous case describing an association between this disease and amlodipine administration has been reported in the medical literature., Learning Points: Schamberg's disease is the most common form of pigmented purpuric dermatosis.The condition can be caused by amlodipine administration.This is a rare but benign skin disorder., Competing Interests: Conflicts of Interests: The Authors declare that there are no competing interests.

Purpose . The aim was to evaluate the efficacy of aflibercept in patients with diabetic macular edema (DME) unresponsive to prior anti-VEGF therapy. Methods . Retrospective review of DME unresponsive to previous anti-VEGF switched to aflibercept with 3 months of follow-up. Changes in best correct visual acuity (BCVA), central retinal thickness (CRT), and frequency of injections were analyzed. The percentage of subjects who had ≥20/40 (logMAR equivalent 0.3) and ≤20/200 (logMAR equivalent 1) was evaluated. Results . A total of 32 eyes from 26 patients were included. Mean age was 65 ± 10 years old. The mean number of previous anti-VEGF injections was 5.34 ± 2.38, and the mean number of aflibercept injections at the end of the study was 2.00 ± 0.00. The CRT at baseline was 501.47 ± 150.51  μ m and 367.97 ± 124.61  μ m at 3 months of follow-up ( P < 0.001). The logMAR BCVA at baseline was 0.71 ± 0.36 and 0.65 ± 0.33 at the end of the follow-up ( P = 0.037). At baseline, 12.5% of patients had ≥20/40 compared with 25% at the end of follow-up. At baseline, 28.13% of patients had 20/200 or inferior vision compared with 15.63% at the end of the follow-up. Conclusions . DME patients unresponsive to previous multiple ranibizumab injections demonstrate a significant anatomical and functional improvement with the switch to aflibercept.

In this work, a non-commercial triaxial geometry energy dispersive X-ray Fluorescence (EDXRF) setup and a benchtop µ-XRF system were used to identify postmortem contamination in buried bones. For two of the individuals, unusually high concentrations of Cu and Pb, but also Zn (in one individual) were observed. The pigments of the burial shroud coverings have been identified as the source of contamination. Accurate and precise quantitative results were obtained by nondestructive process using fundamental parameters method taking into account the matrix absorption effects. A total of 30 bones from 13 individuals, buried between the mid-XVIIIth to early XIXth centuries, were analyzed to study the elemental composition and elemental distribution. The bones were collected from a church in Almada (Portugal), called Ermida do Espírito Santo, located near the Tagus River and at the sea neighbourhood. The triaxial geometry setup was used to quantify Ca, Fe, Cu, Zn, Br, Sr and Pb of powder pressed bone pellets (n=9 for each bone). Cluster analysis was performed considering the elemental concentrations for the different bones. There was a clear association between some bones regarding Fe, Cu, Zn, Br and Pb content but not a categorization between cortical and trabecular bones. The elemental distribution of Cu, Zn and Pb were assessed by the benchtop μ-analysis, the M4 Tornado, based on a polycapillary system which provides multi-elemental 2D maps. The results showed that contamination was mostly on the surface of the bone confirming that it was related to the burial shroud covering the individuals., (Copyright © 2016 Elsevier B.V. All rights reserved.)

The aim of this systematic review is to determine the diagnostic accuracy of clinical feeding evaluation (CFE) compared to instrumental assessments in detecting oropharyngeal aspiration (OPA) in children. This is important to support clinical decision-making and to provide safe, cost-effective, higher quality care. All published and unpublished studies in all languages assessing the diagnostic accuracy of CFE compared to videofluoroscopic swallowing study (VFSS) and/or fibre-optic endoscopic examination of swallowing (FEES) in detecting OPA in paediatric populations were sought. Databases were searched from inception to April 2015. Grey literature, citations, and references were also searched. Two independent reviewers extracted and analysed data. Accuracy estimates were calculated. Research reports were translated into English as required. Six studies examining the diagnostic accuracy of CFE using VFSS and/or FEES were eligible for inclusion. Sample sizes, populations studied, and CFE characteristics varied widely. The overall methodological quality of the studies, assessed with QUADAS-2, was considered 'low'. Results suggested that CFEs trialling liquid consistencies might provide better accuracy estimates than CFEs trialling solids exclusively. This systematic review highlights the critical lack of evidence on the accuracy of CFE in detecting OPA in children. Larger well-designed primary diagnostic test accuracy studies in this area are needed to inform dysphagia assessment in paediatrics., (© 2016 Mac Keith Press.)

Aim: Evaluate the real-life experience with ocriplasmin on vitreomacular traction (VMT) release and full-thickness macular hole (FTMH) closure in Portugal., Methods: Multicentric, retrospective study of 83 eyes of 78 patients who were treated with intravitreal ocriplasmin for VMT with and without FTMH. Primary outcomes were VMT release and FTMH closure. Secondary outcomes included visual acuity changes and structural features on spectral-domain ocular coherence tomography., Results: VMT resolved in 47 of the 83 eyes (56.6%) and 6 of the 12 FTMH were closed (50.0%). Mean best-corrected visual acuity (BCVA) improved from 65.1 at baseline to 70.8 ETDRS letters at the end of follow-up (p < 0.0001) with a mean follow-up of 138.8 days. Improvement in BCVA was significantly better in eyes with VMT release (p = 0.021). Approximately 73% of patients had normal ellipsoid zone integrity at the end of follow-up, 87% had no neurosensorial detachment and 40% had no intra- or subretinal fluid., Conclusion: VMT release and FTMH closure were achieved in more than half of the treated eyes and were correlated with significant BCVA improvements and favorable baseline characteristics. In fact, if a careful patient selection is carried out, VMT resolution with ocriplasmin can be optimized, tailoring the best approach to each patient., (© 2016 S. Karger AG, Basel.)

Purpose: To perform a comprehensive and systematic review regarding ophthalmic adverse drug reactions (ADRs) to systemic drugs to: (i) systematically summarize existing evidence, (ii) identify areas, ophthalmic ADRs or drugs that lacked systematization or assessment (namely drugs with original studies characterizing specific ophthalmic ADRs but without causality assessment nor without meta-analysis)., Methods: Systematic review of several electronic databases (last search 1/7/2012): Medline, SCOPUS, ISI web of knowledge, ISI Conference Proceedings, International Pharmaceutical Abstracts and Google scholar. Search query included: eye, ocular, ophthalmic, ophthalmology, adverse and reaction. Inclusion criteria were: (i) Primary purpose was to assess an ophthalmic ADR to a systemic medication; (ii) Patient evaluation performed by an ophthalmologist; (iii) Studies that specified diagnostic criteria for an ocular ADR. Different types of studies were included and analyzed separately. Two independent reviewers assessed eligibility criteria, extracted data and evaluated risk of bias., Results: From 562 studies found, 32 were included (1 systematic review to sildenafil, 11 narrative reviews, 1 trial, 1 prospective study, 6 transversal studies, 6 spontaneous reports and 6 case series). Drugs frequently involved included amiodarone, sildenafil, hydroxychloroquine and biphosphonates. Frequent ophthalmic ADRs included: keratopathy, dry eye and retinopathy., Conclusions: To increase evidence about ophthalmic ADRs, there is a need for performing specific systematic reviews, applying strictly the World Health Organization's (WHO) definition of ADR and WHO causality assessment of ADRs. Some ophthalmic ADRs may be frequent, but require ophthalmological examination; therefore, ophthalmologists' education and protocols of collaboration between other specialties whenever they prescribe high-risk drugs are suggestions for the future., (Copyright © 2014 John Wiley & Sons, Ltd.)

Purpose: To report the clinical course of 2 pediatric ocular rosacea cases with a significant delay until diagnosis., Methods: We report 2 interventional case reports. Case 1 is a 10-year-old boy with 2 years of recurrent bilateral blepharitis, repetition chalazion, conjunctival hyperemia, and corneal ulcers, without response to topical antibiotics or topical and systemic steroids. Case 2 is a 9-year-old girl with keratoconjunctivitis and repetition chalazion since she was 2 years old, without improvement after consulting several ophthalmologists and performing several treatments throughout those years., Results: Rapid response to systemic erythromycin with marked improvement of both cases within a few weeks., Conclusions: Ocular rosacea is frequently misdiagnosed, particularly in the pediatric population. To our knowledge, this report demonstrates a case with the longest history before diagnosis (7 years) and another case in which a conjunctival biopsy was performed.

The authors report the case of a 45-years-old female patient, with progressive hemiatrophy of the left face and tongue. The laboratorial and image studies were normal, including routine blood exams, cerebral-spinal fluid, hepatic function tests, renal and rheumatic; besides X ray, computerized tomography and magnetic resonance of skull. The electroneuromyography showed peripheral neurogenic alterations of motor trigeminal and hypoglossal nerves, markedly on the left side. Those findings are compatible with Romberg's facial hemiatrophy diagnosis, motivating discussion of several aspects of this rare disease according to the account and literature.

A prospective study was designed to follow patients with syndromes of brain stem during 12 months (in 1991-1992). The aim was to correlate clinical and radiologic findings and comparing them with the classical descriptions of brain stem syndromes. Twenty one consecutive patients were admitted at the Hospital de Base do Distrito Federal. The diagnosis and the follow up were carried out by neurological examination and neuroradiologic images: CT (100%), angiography (24%, 5) and MRI (5%, 1). Hypertension and old age were the most important risk factors and hemorrhage at pons with extension to midbrain was the most frequent finding. Coma at admission was associated to a poor outcome. The outcome was defined by the Glasgow Outcome Scale: 28% rated 5, 24% rated 4, 5% rated 3, 28% rated 2 and 14% rated 1. Our results show that the clinical presentation of ischemic strokes and hemorrhages of the brain stem very frequently follow a mixed pattern, which do not conciliate with the classical of these syndromes.

The authors report a case of symptomatic basilar impression and Arnold-Chiari malformation being presented as the first symptom of hemifacial spasm. The surgical treatment of the malformation resulted in improvement of the clinical manifestation with reduction of the hemifacial spasm. The need for the aetiological therapy for the hemifacial spasm is emphasized, before symptomatic treatment with botulinum toxin is tried.

Recent claims of metal contamination in the lower reaches of the Sado River, in the Alcácer do Sal region, Portugal, a major rice-producing area were investigated by carrying out metal surveys in the area. The elements Fe, Mn, Zn, Cu and Pb were measured in the soil and in rice plant parts--roots, shoots and grain--as well as in some weeds growing in the Sado banks, near the paddy fields. Results showed that the metal contents of paddy soils were similar to background concentrations, with the exception of Zn and Cu, which were above those concentrations and reached their highest levels at Vale de Guizo, the monitored station located furthest upstream in the Sado River. At some sites, plant roots accumulated relatively large amounts of Fe, Mn, Zn and Cu, but the shoot levels of these metals were within the normal range for rice plants. It is possible that varying, but significant, amounts of Fe associated with the roots were in the form of ferric hydroxide plaque covering their surfaces. Copper levels in the shoots of rice were below the normal contents cited for this plant in the literature. Metal levels of river sediments collected near Vale de Guizo seem to corroborate the possibility of some metal contamination in the Sado River, most probably derived from pyrites mining activity in the upper zone of the Sado basin.

The authors report a case of Raeder's paratrigeminal syndrome and make some comments about the diagnosis: a dissecant aneurysm of internal carotid artery.

A comparative study of peptide composition and freeze-fracture morphology of chloroplast membranes from a chlorophyll b-less mutant and a normal barley plant (Hordeum vulgare L.) is reported in this work. Using a high resolution, discontinuous sodium dodecyl sulfate-acrylamide gel electrophoretic system, we show that the mutant chloroplast membranes not only completely lack the 25-kilodalton peak, which accounts for about 18% of the chloroplast membrane protein in the normal plant, but also exhibit gross reduction in other components at 27.5- and 20-kilodalton regions. Despite such extensive deletions in the peptide composition of the mutant chloroplast lamellae, no alteration could be detected in either density or size of the intramembranous particles, visualized by freeze-fracturing.

Park and co-workers (Annu. Rev. Plant Physiol. 22: 395-430) have suggested that stroma lamellae, which perform only photosystem I contain a developing photosystem II which becomes functional upon the folding of these membranes to form grana stacks.The present study was undertaken to test this hypothesis. The lipid and protein constituents of chloroplast membranes were pulse-labeled radioactively with (14)C, and the specific activity of stroma and grana components were studied in a pulse-chase experiment. The components of both grana and stroma lamellae become labeled and decay at similar rates which suggests that the Park and Sane hypothesis is not correct. The results also show large differences in the turnover rates of some chloroplast membrane components supporting the multistep model for membrane synthesis and maintenance in this organelle.

The case of a 17-year-old boy who had been well until the age of four when progressive stiffness of lower limbs developed is reported. Soon walking became difficult and on on his thighs, buttocks and calves one could notice the appearance of myokymia. Painful and intermittent spasms were often present specially after physical strain. This was followed by profuse sweating. Strength was normal but muscle relaxing was slow mainly in the quadriceps. The familiar history for neuromuscular pathology was negative. Muscle enzymes, thyroid function tests were within normal limits and muscular biopsy showed no abnormalities. The EMG disclosed a permanent spontaneous activity, more evident in the quadriceps, which was normal in shape, amplitude and duration. In association to this activity multiplets discharges and low motor units were also observed; in interferencial pattern appeared at maximum contraction but the multiplets and low motor units potentials kept on showing even though voluntary muscular activity ceased. The therapeutic essay with carbamazepin had a dramatic effect upon the course of the patient's conditions. He showed a great improvement with this drug and today leads a normal life being able even to play soccer and ride a bicycle. An interesting remark which should be done is the fact that the flexion of the neck inhibits the quadriceps myokymia whereas the extension makes them to appear and even worsen. Unfortunately is was not possible to carry out a more accurate investigation concerning electropharmacology and hystopathology as the patient was uncooperative and refused to undergo such examinations. However the authors believe that this clinical picture can be considered as a case of "continuous muscle-fibre activity" due to the great similarity it bears to the cases related by Isaacs in 1961. An extensive review concerning the pertinent literature has been done.

The study of the ciliospinal reflex in twenty-three patients is reported. The patients were divided into two groups: the first with spinal cord lesions and the second one with brain stem lesions. The authors discuss the pathways and the mechanisms of the ciliospinal reflex in man and emphasize their value on the diagnosis of brain stem lesions.

In the present study we extend previous work from this laboratory on the polypeptide composition of photosynthetic lamellae. Using a high resolution sodium dodecyl sulfate gel electrophoresis technique, we show that both grana and stroma lamellae have qualitatively very similar polypeptide compositions although some clear quantitative differences are demonstrated.

The authors report a case of bilateral ptosis with miosis, paralysis of upward gaze and of smooth pursuit movements to the right, as well as a slowly regressive state of lassitude. This syndrome followed a haemorrhagic brain stem infarction. After a review of the pathophysiology of ptosis and of vertical eye movements, together with analysis of data provided by computerized tomography studies--since the case was not fatal--explanations are offered for each clinical sign and it is proposed that the lesion, a paramedian, slightly deviated to the right haemorrhagic focus in the midbrain, with a diameter of 15 mm, might have involved several structures, namely the rostral interstitial nucleus of the medial longitudinal fasciculus, the mesencephalic reticular formation, as well as cortico-nuclear and sympathetic pathways.

A comparative study of polypeptide composition and freeze-fracture morphology of chloroplast membranes isolated from different, but photosynthetically active, regions of a romaine lettuce shoot is presented. Chloroplasts prepared from outer dark-green leaves possess a fully developed light-harvesting chlorophyll-protein complex, have low chlorophyll a:b ratios and display fracture faces similar to those found in other higher plant chloroplast membranes; chloroplasts from leaves more to the interior of the shoot, have a much lower content of chlorophyll, show high chlorophyll a:b ratios, are depleted in components of the light-harvesting chlorophyll-protein complex, and exhibit extensive modifications of their fracture faces. We have also re-examined the freeze-fracture morphology of chloroplast membranes from a barley b-deficient mutant that lacks the light-harvesting chlorophyll-protein complex. A tentative interpretation of our findings suggests correlating the assembly of the light-harvesting chlorophyll-protein complex into the chloroplast membranes with the appearance of large freeze-fracture B face particles in the stacked interior-membrane region, and differentiation of typical fracture faces.

Sodium dodecyl sulfate-solubilized chloroplast internal membranes of higher plants (cowpea [Vigna unguiculata L. Walp], chinese cabbage [Brassica chinensi L.], and tobacco [Nicotiana tabacum L.]) are resolved by polyacrylamide gel electrophoresis into two chlorophyll a- and three chlorophyll a,b-proteins. A small portion (about 15%) of the membrane chlorophyll migrates as a component of high electrophoretic mobility and presumably consists of detergent-complexed, protein-free pigment.One of the chlorophyll a-proteins is qualitatively similar to the P(700) chlorophyll a-protein but contains a much larger proportion of total chlorophyll (about 30%) than previously reported. The second chlorophyll a-protein is a recently discovered component of the membrane and accounts for about 7% of the total chlorophyll. The absorption and fluorescence emission spectra of these two chlorophyll a-proteins differ.The three chlorophyll a,b-proteins are components of the chloroplast membrane chlorophyll a,b-light-harvesting complex which was previously resolved as a single chlorophyll-protein band. The two additional chlorophyll a,b-proteins observed in our work probably represent larger aggregates contained within that membrane complex which are preserved under the solubilization and electrophoretic conditions used here.

Extraction of an aqueous suspension of spinach chloroplast lamellae with a chloroform/methanol mixture leads to solubilization of about 1/3 of the total membrane protein. Amino acid analysis of the chloroform/methanol-soluble protein shows that this fraction is largely enriched in the hydrophobic residues proline, leucine, alanine and phenylalanine and considerably depleted in polar amino acids, namely lysine and arginine. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of the solubilized material reveals the presence of a variety of low molecular weight polypeptides (molecular weight less than or equal to 25 000), with more than 50% of the total fraction being contributed by a 25 000 dalton band. This band, which accounts for about 25% of the total chloroplast lamellar protein, has recently been identified as the main component of the light-harvesting chlorophyll-protein complex. The physiological role of most of the chloroform/methanol-soluble protein fraction is not known at present. From its chemical properties and apparent biological inertness, we propose that it plays mainly a structural role in situ, interacting with the lipid moiety of the chloroplast membrane. The material insoluble in the aqueous chloroform/methanol mixture is largely enriched in manganese, iron, cytochrome and water-soluble proteins, such as chloroplast coupling factor and ribulose diphosphate carboxylase.

The protein moiety of the two major chlorophyll-protein complexes associated with chloroplast membranes of outer, dark green leaves of a romaine lettuce shoot (Lactuca sativa L. var. Romana) has been analyzed by discontinuous sodium dodecyl sulfate-polyacrylamide disc gel electrophoresis. Complex II, also termed light-harvesting chlorophyll-protein complex, is shown to consist of a major polypeptide of 25 kilodaltons (kD) and two minor ones of 27.5 and 23 kD. The 25 kD subunit is the single largest polypeptide component of the chloroplast membranes, accounting for about 25% of their total protein. Complex I contains only high molecular weight subunits, the major one being at 67 kD, these subunits representing only a small percentage of the chloroplast membrane total protein.These data, suggesting an oligomeric nature for the apoprotein of these two chlorophyll-protein complexes, are difficult to reconcile with the estimated molecular weights of the native complexes and raise some intriguing questions as to the types of interactions among the components of these major lipoproteins of the photosynthetic membranes.