Although controversial, cancer stem cells (CSCs) are thought to be one tumor component, being characterized by their strong self-renewal and survival properties. Cancer cells, CSCs included, are thought to rely mostly on glycolysis, even in the presence of oxygen, which confers them adaptive advantages. Adenine nucleotide translocator 2 (ANT2), responsible for the exchange of ADP and ATP in the mitochondrial inner membrane, has been correlated with a higher glycolytic metabolism and is known to be overexpressed in cancer cells. Using P19 embryonal carcinoma stem cells, we inhibited ANT2 translation by using siRNA. ANT2 protein levels were shown to be overexpressed in P19 undifferentiated cells (P19SCs) when compared to their differentiated counterparts (P19dCs). Furthermore, we showed here that the OXPHOS machinery and mitochondrial membrane potential are compromised after ANT2 depletion, leading to a metabolic adaptation towards a less oxidative phenotype. Interestingly, hexokinase II levels were downregulated, which was also accompanied by decreased cell growth, and reduced ability to form spheroids. Our findings underscore ANT2 as a key regulator of metabolic remodeling and cell survival of cancer stem-like cells, suggesting its potential as a therapeutic target for controlling CSC-driven tumor progression.

Diosgenin is a phytosteroid sapogenin with reported antitumoral activity. Despite the evidence indicating a lower incidence of prostate cancer (PCa) associated with a higher consumption of phytosteroids and the beneficial role of these compounds, only a few studies have investigated the effects of diosgenin in PCa, and its mechanisms of action remain to be disclosed. The present study investigated the effect of diosgenin in modulating PCa cell fate and glycolytic metabolism and explored its potential interplay with G protein-coupled oestrogen receptor (GPER). Non-neoplastic (PNT1A) and neoplastic (LNCaP, DU145, and PC3) human prostate cell lines were stimulated with diosgenin in the presence or absence of the GPER agonist G1 and upon GPER knockdown. Diosgenin decreased the cell viability, as indicated by the MTT assay results, which also demonstrated that castrate-resistant PCa cells were the most sensitive to treatment (PC3 > DU145 > LNCaP > PNT1A; IC50 values of 14.02, 23.21, 56.12, and 66.10 µM, respectively). Apoptosis was enhanced in diosgenin-treated cells, based on the increased caspase-3-like activity, underpinned by the altered expression of apoptosis regulators evaluated by Western blot analysis, which indicated the activation of the extrinsic pathway. Exposure to diosgenin also altered glucose metabolism. Overall, the effects of diosgenin were potentiated in the presence of G1. Moreover, diosgenin treatment augmented GPER expression, and the knockdown of the GPER gene suppressed the proapoptotic effects of diosgenin in PC3 cells. Our results support the antitumorigenic role of diosgenin and its interest in PCa therapy, alone or in combination with G1, mainly targeting the more aggressive stages of the disease.

Cancer stem cells (CSCs) are amongst the group of cells constituting tumors, being characterized by their strong self-renewal and survival properties. Cancer cells, CSCs included, are thought to rely mostly on glycolysis, even in the presence of oxygen, which confers them adaptative advantages. Adenine nucleotide translocator 2 (ANT2), responsible for the exchange of ADP and ATP in the mitochondrial inner membrane, has been correlated with a higher glycolytic metabolism and is known to be overexpressed in cancer cells. Using P19 embryonal carcinoma stem cells (P19SCs) as a CSCs model, we inhibit ANT2 translation by using siRNA. ANT2 protein levels were shown to be overexpressed in P19SC when compared to their differentiated counterparts. Furthermore, we showed here that the OXPHOS machinery and mitochondrial membrane potential are compromised after ANT2 depletion, exhibiting a metabolic adaptation towards a less oxidative phenotype. Interestingly, hexokinase II levels were downregulated, which was also accompanied by decreased cell growth, and decreased ability to form spheroids. Our findings underscore ANT2 as a key regulator of metabolic remodeling and cell survival of CSCs, suggesting its potential as a therapeutic target for controlling CSC-driven tumor progression. Highlights ### Competing Interest Statement The authors have declared no competing interest.

BACKGROUND:Amyotrophic lateral sclerosis (ALS) is a fatal and rapidly progressing neurodegenerative disease that affects motor neurons. This disease is associated with oxidative stress especially in mutant superoxide dismutase 1 (mutSOD1) patients. However, less is known for the most prevalent sporadic ALS form, due to a lack of disease models. Here, we studied oxidative stress profiles in lymphoblasts from ALS patients with mutSOD1 or unknown (undSOD1) mutations. METHODS:mutSOD1 and undSOD1 lymphoblasts, as well as sex/age-matched controls (3/group) were obtained from Coriell and divided into 46 years-old-men (C1), 46 years-old-women (C2) or 26/27 years-old-men (C3) cohorts. Growth curves were performed, and several parameters associated with redox homeostasis were evaluated, including SOD activity and expression, general oxidative stress levels, lipid peroxidation, response to oxidative stimulus, glutathione redox cycle, catalase expression, and activity, and Nrf2 transcripts. Pooled (all cohorts) and paired (intra-cohort) statistical analyses were performed, followed by clustering and principal component analyses (PCA). RESULTS:Although a high heterogeneity among lymphoblast redox profiles was found between cohorts, clustering analysis based on 7 parameters with high chi-square ranking (total SOD activity, oxidative stress levels, catalase transcripts, SOD1 protein levels, metabolic response to mM concentrations of tert-butyl hydroperoxide, glutathione reductase activity, and Nrf2 transcript levels) provided a perfect cluster segregation between samples from healthy controls and ALS (undSOD1 and mutSOD1), also visualized in the PCA. CONCLUSIONS:Our results show distinct redox signatures in lymphoblasts from mutSOD1, undSOD1 and healthy controls that can be used as therapeutic targets for ALS drug development.

Amyotrophic lateral sclerosis (ALS) is a fatal and rapidly progressing neurodegenerative disease that affects motor neurons. This disease is associated with oxidative stress especially in mutant superoxide dismutase 1 (mutSOD1) patients. However, less is known for the most prevalent sporadic ALS due to a lack of disease models. Here, we studied oxidative the stress profiles in lymphoblasts from ALS patients with mutSOD1 or unknown (undSOD1) mutations.mutSOD1 and undSOD1 lymphoblasts, as well as sex/age-matched controls (3/group) were obtained from Coriell and divided in 46 years-old-men (C1), 46 years-old-women (C2) or 26/27 years-old-men (C3) cohorts. Growth curves were performed, and several parameters associated with redox homeostasis were evaluated, including SOD activity and expression, general oxidative stress levels, lipid peroxidation, response to oxidative stimulus, glutathione redox cycle, catalase expression, and activity, and Nrf2 transcripts. Pooled (all cohorts) and paired (intra-cohort) statistical analyses were performed, followed by clustering and principal component analyses (PCA).Although a high heterogeneity among lymphoblast redox profiles was found between cohorts, clustering analysis based on 7 parameters with high chi-square ranking (total SOD activity, oxidative stress levels, catalase transcripts, SOD1 protein levels, metabolic response to mM concentrations of tert-butyl hydroperoxide, glutathione reductase activity, and Nrf2 transcript levels) provided a perfect separation between samples from healthy controls and ALS (undSOD1 and mutSOD1), also visualized in the PCA analysis.Our results show distinct redox signatures in lymphoblasts from mutSOD1, undSOD1, and healthy controls that can be used as therapeutic targets for ALS drug development.Highlights ### Competing Interest StatementThe authors have declared no competing interest.* ALS : Amyotrophic Lateral Sclerosis ARE : Antioxidant Response Element BCA : Bicinchoninic Acid CSF : Cerebrospinal Fluid DTT : Dithiothreitol ECL : Enhanced Chemiluminescence reagent EDTA : Ethylenediaminetetraacetic Acid EGTA : Ethylene Glycol-bis (2-aminoethylether) N, N,’N, ’N-tetraacetic Acid fALS : Familial ALS GPx : Glutathione Peroxidase GR : Glutathione Reductase GSH : Reduced Glutathione GSSG : Oxidized Glutathione H2DCFDA : 2’,7’-Dichlorodihydrofluorescein Diacetate HPLC : High-Performance Liquid Chromatograph HRP : Horseradish Peroxidase MDA : Malondialdehyde mutSOD1 : Mutant SOD1 NAM : Nicotinamide Nrf2 : Nuclear Factor Erythroid-2-related Factor PBMCs : Peripheral Blood Mononuclear Cells PBS : Phosphate-Buffered Saline PCA : Principal Component Analysis PMSF : Phenylmethylsulfonyl Fluoride PVDF : Polyvinylidene Difluoride sALS : Sporadic ALS SDS : Sodium Dodecyl Sulfate SOD1 : Superoxide Dismutase 1 Ta : Annealing Temperature TBA : Thiobarbituric Acid t-BHP : tert-Butyl Hydroperoxide undSOD1 : Undetermined SOD1 Mutations vit E : Vitamin E

Self-reading is a process in which the consumer is responsible for measuring his own energy consumption, which can be done through digital platforms, such as websites or mobile applications. The Equatorial Energy group's electric utilities have been working on developing a chatbot application through which consumers can send an image of their energy meter to a server that runs a method based on image processing and deep learning for the automatic recognition of consumption reading. However, these methods in a solution available to the public should consider factors such as response time and accuracy, so that it presents a satisfactory response time when it needs to handle a large number of simultaneous requests. Therefore, this paper presents a comparative study between approaches developed for the automatic recognition of consumption readings in images of electric meters sent to the server. Response time performances are analyzed through stress tests that simulate the real application scenario. The mean average precision (mAP) and the accuracy metrics of the methods are also analyzed in order to evaluate the generalization of the used convolutional neural networks.

To adapt to tumoral environment conditions or even to escape chemotherapy, cells rapidly reprogram their metabolism to handle adversities and survive. Given the rapid rise of studies uncovering novel insights and therapeutic opportunities based on the role of mitochondria in tumor metabolic programing and therapeutics, this review summarizes most significant developments in the field. Taking in mind the key role of mitochondria on carcinogenesis and tumor progression due to their involvement on tumor plasticity, metabolic remodeling, and signaling re-wiring, those organelles are also potential therapeutic targets. Among other topics, we address the recent data intersecting mitochondria as of prognostic value and staging in cancer, by mitochondrial DNA (mtDNA) determination, and current inhibitors developments targeting mtDNA, OXPHOS machinery and metabolic pathways. We contribute for a holistic view of the role of mitochondria metabolism and directed therapeutics to understand tumor metabolism, to circumvent therapy resistance, and to control tumor development.

Abstract Background Metastatic breast cancer is a major cause of cancer-related deaths in woman. Brain metastasis is a common and devastating site of relapse for several breast cancer molecular subtypes, including oestrogen receptor-positive disease, with life expectancy of less than a year. While efforts have been devoted to developing therapeutics for extra-cranial metastasis, drug penetration of blood–brain barrier (BBB) remains a major clinical challenge. Defining molecular alterations in breast cancer brain metastasis enables the identification of novel actionable targets. Methods Global transcriptomic analysis of matched primary and metastatic patient tumours (n = 35 patients, 70 tumour samples) identified a putative new actionable target for advanced breast cancer which was further validated in vivo and in breast cancer patient tumour tissue (n = 843 patients). A peptide mimetic of the target’s natural ligand was designed in silico and its efficacy assessed in in vitro, ex vivo and in vivo models of breast cancer metastasis. Results Bioinformatic analysis of over-represented pathways in metastatic breast cancer identified ADAM22 as a top ranked member of the ECM-related druggable genome specific to brain metastases. ADAM22 was validated as an actionable target in in vitro, ex vivo and in patient tumour tissue (n = 843 patients). A peptide mimetic of the ADAM22 ligand LGI1, LGI1MIM, was designed in silico. The efficacy of LGI1MIM and its ability to penetrate the BBB were assessed in vitro, ex vivo and in brain metastasis BBB 3D biometric biohybrid models, respectively. Treatment with LGI1MIM in vivo inhibited disease progression, in particular the development of brain metastasis. Conclusion ADAM22 expression in advanced breast cancer supports development of breast cancer brain metastasis. Targeting ADAM22 with a peptide mimetic LGI1MIM represents a new therapeutic option to treat metastatic brain disease.

Brazilian Electricity Regulatory Agency (ANEEL) defines as non-technical losses the problems that happen during the energy consumption measurement process. A feasible alternative to reduce those losses is the energy consumption reading process performed by customers, called self-reading. This process might be executed across digital platforms such as mobile applications and the customers would register and send consumption information. In this context, the consumption reading using voice is a simple way for the public that has lower affinity for technology to perform self-reading. Therefore, this work proposes an end-to-end speech recognition method applied to the energy consumption measurement that uses a neural network architecture based on Recurrent Neural Network Transducer (RNN- T). The proposed method uses a modified architecture called Residual Recurrent Neural Network Transducer (RRNN-T) which contains a Residual Skip that make its behavior similar to a network shallower than RNN - T by skipping some layers in the training process. That strategy saves computational cost of the inference process and disk space. A spelled sequence of digits is the input of the proposed method that recognizes those digits and outputs the electrical energy consumption reading measured in kilowatts per hour. The dataset used in this work contains 111,224 samples of spelled sequences divided into 70% for training and 30% for test. The proposed method obtains 0.05 word error rate (WER) and inference time less than 3 seconds on mobile devices.

Successful metastatic spreading relies on cancer cells with stem-like properties, glycolytic metabolism and increased antioxidant protection, allowing them to escape anoikis and to survive in circulation. The expression of P-cadherin, a poor prognostic factor in breast cancer, is associated with hypoxic, glycolytic and acidosis biomarkers. In agreement, P-cadherin-enriched breast cancer cell populations presents a glycolytic and an acid-resistance phenotype. Our aim was to evaluate whether P-cadherin expression controls the glycolytic and oxidative phosphorylation fluxes of matrix-detached breast cancer cells, acting as an antioxidant and enhancing their survival in anchorage-independent conditions. By using matrix-detached breast cancer cells, we concluded that P-cadherin increases glucose-6-phosphate dehydrogenase expression, up-regulating the carbon flux through the pentose phosphate pathway, while inhibiting pyruvate oxidation to acetyl-coA via pyruvate dehydrogenase kinase-4 (PDK-4) activation. Accordingly, P-cadherin expression conferred increased sensitivity to dichloroacetate (DCA), a PDK inhibitor. P-cadherin expression also regulates oxidative stress in matrix-detached breast cancer cells, through the control of antioxidant systems, such as catalase and superoxide dismutases (SOD)1 and 2, providing these cells with an increased resistance to doxorubicin-induced anoikis. Importantly, this association was validated in primary invasive breast carcinomas, where an enrichment of SOD2 was found in P-cadherin-overexpressing breast carcinomas. In conclusion, we propose that P-cadherin up-regulates carbon flux through the pentose phosphate pathway and decreases oxidative stress in matrix-detached breast cancer cells. These metabolic remodeling and antioxidant roles of P-cadherin can promote the survival of breast cancer cells in circulation and in metastatic sites, being a possible player in breast cancer therapeutic resistance to pro-oxidant-based interventions.

A 40-year-old man presented to the emergency department with dyspnoea and fatigue after bupropion and popper consumption. Clinical examination was remarkable for central cyanosis not responding to supplementary oxygen. Arterial blood gas analysis showed a methaemoglobin value of 30.3%. Methaemoglobinemia was diagnosed and the patient was treated with methylene blue. However, during methylene blue administration, the patient developed a generalized tonic-clonic seizure that was successfully managed with diazepam. Combined intoxications can be a critical problem in the emergency department. Early recognition and treatment of poisoning are key for good patient outcome.LEARNING POINTS:Distinguishing toxidromes is critical for adequate treatment of patients with drug intoxication; the most common side effect of bupropion consumption is dose-dependent seizures.The diagnosis of methaemoglobinemia requires a high index of suspicion, particularly in a patient presenting with central cyanosis not responding to supplementary oxygen.Treatment with methylene blue is recommended when the percentage of methaemoglobin is above 30% or when the patient has symptoms related to methaemoglobinemia.

Environmental exposures associated with host immune system factors can trigger and worse several forms of allergic diseases. Second-hand smoke (SHS) is a known factor that exacerbates numerous diseases in children, such as asthma, respiratory tract infections and sudden infant death syndrome. During childhood the main source of exposure to SHS is the children own home. This study aimed to compare the prevalence of asthma and its impact on life quality of children exposed or not to SHS. Children with ages between 7 to 17 years old answered an ISAAC (International Study of Asthma and Allergies in Childhood) questionnaire to diagnose asthma. Subsequently they were questioned about the presence of smokers at home. Those classified as asthmatics answered a PAQLQ-A (Pediatric Asthma Quality of Life Questionnaire-Adapted). Five hundred and forty questionnaires were distributed and 257 were correctly filled, the mean age of the volunteers was (9,61±1,73 years). According to our data, ISAAC global score pointed out to a 21.01% prevalence of asthma. Among the asthmatic children 66.67% were exposed to SHS at home and 37.44% of non-asthmatic children were also exposed to SHS at home. The association between asthma symptoms and SHS at home was observed with the odds ratio test (p <0.001). Regarding the quality of life (QL) evaluation we observed a significant worsening (p <0.05) in all domains of the PAQLQ-A (i.e. symptoms, activity limitation and emotional function) in children exposed to SHS at home compared to those non-exposed, indicating a comprehensive worsening in the children’s quality of life. Therefore, we conclude that SHS may contribute to trigger asthma in children and it also significantly interferes in the QL, especially for those with asthma.