Platinum iodido drugs show potential anti-tumor activity, affecting cancer cell metabolism and inducing ROS and senescence in gastrointestinal cancer cells.

Cisplatin-based chemotherapy has associated clinical disadvantages, such as high toxicity and resistance. Thus, the development of new antitumor metallodrugs able to overcome different clinical barriers is a public healthcare priority. Here, we studied the mechanism of action of the isomers trans and cis-[PtI2(isopropylamine)2] (I5 and I6, respectively) against gastrointestinal cancer cells. We demonstrate that I5 and I6 modulate mitochondrial metabolism, decreasing OXPHOS activity and negatively affecting ATP-linked oxygen consumption rate. Consequently, I5 and I6 generated Reactive Oxygen Species (ROS), provoking oxidative damage and eventually the induction of senescence. Thus, herein we propose a loop with three interconnected processes modulated by these iodido agents: (i) mitochondrial dysfunction and metabolic disruptions; (ii) ROS generation and oxidative damage; and (iii) cellular senescence. Functionally, I5 reduces cancer cell clonogenicity and tumor growth in a pancreatic xenograft model without systemic toxicity, highlighting a potential anticancer complex that warrants additional pre-clinical studies.

Uncovering microplastics contamination in canned seafood.

There is limited research on the occurrence of microplastics (MPs) in canned seafood. All types of canned seafood investigated in the present study were contaminated. After sample digestion in 30 % hydrogen peroxide, a total of 40 MPs were recovered. Fibers were the most common type, blue was the dominant colour, and Fourier Transform Infrared Spectroscopy (FTIR) identified polyester as the most common polymer. Considering all samples, an average of 3.5 ± 5.2 MPs/can was obtained, with octopus in tomato sauce and tuna in olive oil presenting the highest contamination (5.2 ± 7.5 MPs/can and 5.2 ± 5.1 MPs/can, respectively). Also, significant differences between the number of MPs in the seafood tissues and immersion liquids were verified. The present study demonstrates MPs occurrence in canned seafood, a potential contamination pathway for humans. More research on the different stages of the canning processing is vital for understanding MPs contamination in cans.

Quantifying the role of saltmarsh as a vulnerable carbon sink: A case study from Northern Portugal.

Saltmarshes play a crucial role in carbon sequestration and storage, although they are increasingly threatened by climate change-induced sea level rise (SLR). This study assessed the potential variation in Blue Carbon stocks across regional and local scales, and estimated their economic value and potential habitat loss due to SLR based on the IPCC AR6 scenarios for 2050 and 2100 in three estuarine saltmarshes in northern Portugal, the saltmarshes of the Minho, Lima and Cávado estuaries. The combined carbon stock of these saltmarshes was 38,798 ± 2880 t of organic carbon, valued at 3.96 ± 0.38 M€. Local and regional differences in carbon stocks were observed between common species, with the cordgrass Spartina patens and the reed Phragmites australis consistently showing higher values in the Lima saltmarsh in some of the parameters. Overall, the Lima saltmarsh had the highest total carbon per species cover, with S. patens showing the highest values among common species. Bolboschoenus maritimus had the highest values in the Minho saltmarsh, while the other species presented a similar carbon storage capacity. Potential habitat loss due to SLR was most evident in the Cávado saltmarsh over shorter timescales, with a significant risk of inundation even for median values of SLR, while the Lima saltmarsh was shown to be more resistant and resilient. If habitat loss directly equates to carbon loss within these saltmarshes, projected CO2 emissions may range from 22,000 to 43,449 t by 2050 and 33,000 to 130,000 t by 2100 (under the IPCC SSP5-8.5 scenario). The study shows the importance of Blue Carbon site-specific estimates, acknowledging the potential future repercussions from habitat loss due to SLR. It emphasizes the need to consider local and regional variability in Blue Carbon stocks assessments and highlights the critical importance of preserving and rehabilitating these ecosystems to ensure their continued efficacy as vital carbon sinks, thereby contributing to climate change mitigation efforts.

Urbanization, self-harm, and suicidal ideation in left-behind children and adolescents in China: a systematic review and meta-analysis.

PURPOSE OF REVIEW: Economic development and urbanisation have prompted many Chinese parents to move from rural to urban regions for better job opportunities. Their children, who remain behind in rural regions, become left-behind children (LBC). With absent parents, children and adolescents are unable to maintain the secure attachment required for healthy social and emotional development, increasing the risk of mental illness. This study aimed to compare risk of self-harm and suicidal ideation in LBC and non-LBC in China. RECENT FINDINGS: Greater risks for poor mental health outcomes including worse depression, loneliness and anxiety have been identified in LBC in cross-sectional studies. Previous studies have also identified higher prevalence of bullying victimization, poorer school performance and worse school attendance amongst LBC. SUMMARY: Findings indicate that prolonged separation from parents put LBC at greater risks of poor mental health. Policy changes to allow children to migrate with their parents and policies to reduce inequalities in job opportunities between urban and rural regions are needed.

Screening of native Saccharomyces cerevisiae strains from Chile for beer production.

Introduction: Beer is one of the most consumed alcoholic drinks in the world, and this industry is a growing market that demands different properties to satisfy new consumers. The yeasts are used in different fermented beverages to contribute to new flavors. However, yeast strains used in the beer industry are limited so far, thus the diversity of flavors is very restricted. Therefore, the use of native yeast strains has been taking more importance with the purpose of conferring differentiated organoleptic properties to the product. Based on this observation the potentiality of native Saccharomyces cerevisiae strains obtained from different localities in Chile was researched. Methods: In this work was selected those strains that produced the highest ethanol concentration (nearly 6% v/v), consumed the highest amounts of sugars, and produced the lowest amounts of organic acids in the resulting beers. Finally, we did a beer tasting to select those strains that added different flavors to the final beer compared with a commercial strain used. Results and discussion: In this study, two native strains that produced fruity descriptors are described, which could be used in the future in brewing, craft or industrial production.

The efficacy of chemotherapy is limited by intratumoral senescent cells expressing PD-L2.

Chemotherapy often generates intratumoral senescent cancer cells that strongly modify the tumor microenvironment, favoring immunosuppression and tumor growth. We discovered, through an unbiased proteomics screen, that the immune checkpoint inhibitor programmed cell death 1 ligand 2 (PD-L2) is highly upregulated upon induction of senescence in different types of cancer cells. PD-L2 is not required for cells to undergo senescence, but it is critical for senescent cells to evade the immune system and persist intratumorally. Indeed, after chemotherapy, PD-L2-deficient senescent cancer cells are rapidly eliminated and tumors do not produce the senescence-associated chemokines CXCL1 and CXCL2. Accordingly, PD-L2-deficient pancreatic tumors fail to recruit myeloid-derived suppressor cells and undergo regression driven by CD8 T cells after chemotherapy. Finally, antibody-mediated blockade of PD-L2 strongly synergizes with chemotherapy causing remission of mammary tumors in mice. The combination of chemotherapy with anti-PD-L2 provides a therapeutic strategy that exploits vulnerabilities arising from therapy-induced senescence.

Targeting cancer stem cell OXPHOS with tailored ruthenium complexes as a new anti-cancer strategy.

BACKGROUND: Previous studies by our group have shown that oxidative phosphorylation (OXPHOS) is the main pathway by which pancreatic cancer stem cells (CSCs) meet their energetic requirements; therefore, OXPHOS represents an Achille's heel of these highly tumorigenic cells. Unfortunately, therapies that target OXPHOS in CSCs are lacking. METHODS: The safety and anti-CSC activity of a ruthenium complex featuring bipyridine and terpyridine ligands and one coordination labile position (Ru1) were evaluated across primary pancreatic cancer cultures and in vivo, using 8 patient-derived xenografts (PDXs). RNAseq analysis followed by mitochondria-specific molecular assays were used to determine the mechanism of action. RESULTS: We show that Ru1 is capable of inhibiting CSC OXPHOS function in vitro, and more importantly, it presents excellent anti-cancer activity, with low toxicity, across a large panel of human pancreatic PDXs, as well as in colorectal cancer and osteosarcoma PDXs. Mechanistic studies suggest that this activity stems from Ru1 binding to the D-loop region of the mitochondrial DNA of CSCs, inhibiting OXPHOS complex-associated transcription, leading to reduced mitochondrial oxygen consumption, membrane potential, and ATP production, all of which are necessary for CSCs, which heavily depend on mitochondrial respiration. CONCLUSIONS: Overall, the coordination complex Ru1 represents not only an exciting new anti-cancer agent, but also a molecular tool to dissect the role of OXPHOS in CSCs. Results indicating that the compound is safe, non-toxic and highly effective in vivo are extremely exciting, and have allowed us to uncover unprecedented mechanistic possibilities to fight different cancer types based on targeting CSC OXPHOS.

Intermittent normobaric hypoxia alters substrate partitioning and muscle oxygenation in individuals with obesity: implications for fat burning.

This single-blind, crossover study aimed to measure and evaluate the short-term metabolic responses to continuous and intermittent hypoxic patterns in individuals with obesity. Indirect calorimetry was used to quantify changes in resting metabolic rate (RMR), carbohydrate (CHOox, %CHO), and fat oxidation (FATox, %FAT) in nine individuals with obesity pre and post: 1) breathing normoxic air [normoxic sham control (NS-control)], 2) breathing continuous hypoxia (CH), or 3) breathing intermittent hypoxia (IH). A mean peripheral oxygen saturation ([Formula: see text]) of 80-85% was achieved over a total of 45 min of hypoxia. Throughout each intervention, pulmonary gas exchanges, oxygen consumption (V̇o2) carbon dioxide production (V̇co2), and deoxyhemoglobin concentration (Δ[HHb]) in the vastus lateralis were measured. Both RMR and CHOox measured pre- and postinterventions were unchanged following each treatment: NS-control, CH, or IH (all P > 0.05). Conversely, a significant increase in FATox was evident between pre- and post-IH (+44%, P = 0.048). Although the mean Δ[HHb] values significantly increased during both IH and CH (P < 0.05), the greatest zenith of Δ[HHb] was achieved in IH compared with CH (P = 0.002). Furthermore, there was a positive correlation between Δ[HHb] and the shift in FATox measured pre- and postintervention. It is suggested that during IH, the increased bouts of muscle hypoxia, revealed by elevated Δ[HHb], coupled with cyclic periods of excess posthypoxia oxygen consumption (EPHOC, inherent to the intermittent pattern) played a significant role in driving the increase in FATox post-IH.

Unusual Trisomy X Phenotype Associated with a Concurrent Heterozygous 16p11.2 Deletion: Importance of an Integral Approach for Proper Diagnosis.

Trisomy X is the most frequent sex chromosome anomaly in women, but it is often underdiagnosed postnatally because most patients do not show any clinical manifestation. It is estimated that only 10% of patients with trisomy X are diagnosed by clinical findings. Thus, it has been proposed that the clinical spectrum is not yet fully delimited, and additional uncommon or atypical clinical manifestations could be related to this entity. The present report describes a female carrying trisomy X but presenting atypical manifestations, including severe intellectual disability, short stature, thymus hypoplasia, and congenital hypothyroidism (CH). These clinical findings were initially attributed to trisomy X. However, chromosome microarray analysis (CMA) subsequently revealed that the patient also bears a heterozygous 304-kb deletion at 16p11.2. This pathogenic copy-number variant (CNV) encompasses 13 genes, including TUFM. Some authors recommend that when a phenotype differs from that described for an identified microdeletion, the presence of pathogenic variants in the non-deleted allele should be considered to assess for an autosomal recessive disorder; thus, we used a panel of 697 genes to rule out a pathogenic variant in the non-deleted TUFM allele. We discuss the possible phenotypic modifications that might be related to an additional CNV in individuals with sex chromosome aneuploidy (SCA), as seen in our patient. The presence of karyotype-demonstrated trisomy X and CMA-identified 16p11.2 deletion highlights the importance of always correlating a patient's clinical phenotype with the results of genetic studies. When the phenotype includes unusual manifestations and/or exhibits discrepancies with that described in the literature, as exemplified by our patient, a more extensive analysis should be undertaken to enable a correct diagnosis that will support proper management, genetic counseling, and medical follow-up.

Polysomnographic Phenotypes of Obstructive Sleep Apnea in a Real-Life Cohort: A Pathophysiological Approach

Introduction: Obstructive sleep apnea (OSA) is heterogeneous and complex, but its severity is still based on the apnea–hypoapnea index (AHI). The present study explores using cluster analysis (CA), the additional information provided from routine polysomnography (PSG) to optimize OSA categorization. Methods: Cross-sectional study of OSA subjects diagnosed by PSG in a tertiary hospital sleep unit during 2016–2020. PSG, demographical, clinical variables, and comorbidities were recorded. Phenotypes were constructed from PSG variables using CA. Results are shown as median (interquartile range). Results: 981 subjects were studied: 41% females, age 56 years (45–66), overall AHI 23events/h (13–42) and body mass index (BMI) 30kg/m2 (27–34). Three PSG clusters were identified: Cluster 1: “Supine and obstructive apnea predominance” (433 patients, 44%). Cluster 2: “Central, REM and shorter-hypopnea predominance” (374 patients, 38%). Cluster 3: “Severe hypoxemic burden and higher wake after sleep onset” (174 patients, 18%). Based on classical OSA severity classification, subjects are distributed among the PSG clusters as severe OSA patients (AHI≥30events/h): 46% in cluster 1, 17% in cluster 2 and 36% in cluster 3; moderate OSA (15≤AHI<30events/h): 57% in cluster 1, 34% in cluster 2 and 9% in cluster 3; mild OSA (5≤AHI<15events/h): 28% in cluster 1, 68% in cluster 2 and 4% in cluster 3. Conclusions: The CA identifies three specific PSG phenotypes that do not completely agree with classical OSA severity classification. This emphasized that using a simplistic AHI approach, the OSA severity is assessed by an incorrect or incomplete analysis of the heterogeneity of the disorder. (AU)

Polysomnographic Phenotypes of Obstructive Sleep Apnea in a Real-Life Cohort: A Pathophysiological Approach.

INTRODUCTION: Obstructive sleep apnea (OSA) is heterogeneous and complex, but its severity is still based on the apnea-hypoapnea index (AHI). The present study explores using cluster analysis (CA), the additional information provided from routine polysomnography (PSG) to optimize OSA categorization. METHODS: Cross-sectional study of OSA subjects diagnosed by PSG in a tertiary hospital sleep unit during 2016-2020. PSG, demographical, clinical variables, and comorbidities were recorded. Phenotypes were constructed from PSG variables using CA. Results are shown as median (interquartile range). RESULTS: 981 subjects were studied: 41% females, age 56 years (45-66), overall AHI 23events/h (13-42) and body mass index (BMI) 30kg/m2 (27-34). Three PSG clusters were identified: Cluster 1: "Supine and obstructive apnea predominance" (433 patients, 44%). Cluster 2: "Central, REM and shorter-hypopnea predominance" (374 patients, 38%). Cluster 3: "Severe hypoxemic burden and higher wake after sleep onset" (174 patients, 18%). Based on classical OSA severity classification, subjects are distributed among the PSG clusters as severe OSA patients (AHI≥30events/h): 46% in cluster 1, 17% in cluster 2 and 36% in cluster 3; moderate OSA (15≤AHI<30events/h): 57% in cluster 1, 34% in cluster 2 and 9% in cluster 3; mild OSA (5≤AHI<15events/h): 28% in cluster 1, 68% in cluster 2 and 4% in cluster 3. CONCLUSIONS: The CA identifies three specific PSG phenotypes that do not completely agree with classical OSA severity classification. This emphasized that using a simplistic AHI approach, the OSA severity is assessed by an incorrect or incomplete analysis of the heterogeneity of the disorder.

Effectiveness of Pediatric Competency-Based Orientation (PCBO) on Knowledge, Confidence, and Manifestation of Early Recognition of Nursing Expertise Among PACU Nurses.

PURPOSE: To evaluate the effectiveness of the American Society of Perianesthesia Nurses (ASPAN) pediatric competency-based orientation (PCBO) education program on knowledge, confidence, and early recognition of nursing expertise among perianesthesia nurses in an acute care setting. DESIGN: A quasi-experimental pre/post survey-intervention design. METHODS: Sixty perianesthesia nurses with experience ranging from less than 5 years to more than 20 years were included. A chapter review survey was completed to assess knowledge before reviewing the chapters and again after reviewing the ASPAN PCBO materials. A presurvey assessing confidence levels, decision making abilities, and early detection of knowledge regarding pediatric patients' expertise were obtained at the beginning of the study. At the end of the study, a post-study survey was completed to evaluate the effectiveness of the intervention. Random codes were assigned to each participant to ensure that the participants' information was blinded. FINDINGS: Perianesthesia nurses' knowledge increased from pre to post intervention with one of the three sets of chapters (Set 2) statistically significant. Perianesthesia nurses' confidence and recognition of nursing expertise scores increased statistically significant from pre to post intervention. Both confidence (with 33 items [P value ≤ .001]) and recognition of nursing expertise (with 16 items [P value ≤ 0.001]) were statistically significant. CONCLUSIONS: The ASPAN PCBO was shown to be statistically effective at increasing knowledge, building expertise, promoting confidence, and improving decision making skills. The plan is for the ASPAN PCBO to be incorporated into the new-hire perianesthesia orientation didactic and the competency plan.

Care Robotics: An Assessment of Professional Perception in the Face of the COVID-19 Pandemic.

The COVID-19 crisis accelerated the adoption of technologies. Technological advancement is also expected in robotics applied to any sector, including in healthcare. The aim is to assess the professional perception of care robotics facing COVID-19. This study aimed to (1) select a tool for assessing different aspects of healthcare, (2) analyse the professional perception about the development, usefulness and helpfulness of technologies and robotics in the field of healthcare and (3) evaluate the correlation between the perceived helpfulness of care robotics and the selected tool. We implement five validated clinical tests which integrate 80 items about a person and their clinical situation. From the sample of 46 professionals, 95.65% affirmed that technology was moderately to completely useful for professional performance in the context of the pandemic, lowering to 67.39% when asked only about robotics; 93.48% stated that the inclusion of robotics in at least one health area affected by COVID-19 would have helped them. Finally, the variables extracted from clinical tests corresponded to the most relevant health areas as identified by the professionals. This research shows the potential of care robotics oriented towards healthcare from a care paradigm.

Microplastic contamination in large migratory fishes collected in the open Atlantic Ocean.

Fishes are one of the most important components of the oceans and are exposed to several anthropogenic pressures, namely microplastic (MP), contaminants that are now ubiquitous worldwide. Taking advantage of the 2020 Circumnavigation Expedition carried by the NRP Sagres tall ship of the Portuguese Navy, fish samples from the southern Atlantic ocean were collected to evaluate possible MP contamination. In a total of 14 weeks of campaign, seven large migratory fishes of commercial interest were collected across the middle Atlantic Ocean and along the South American Atlantic coast. All individuals were contaminated with MPs, with an average of 18 ± 11 MPs/fish. In all fish sampled, both the gastrointestinal tract and gills presented MPs, indicating different contamination pathways including via their preys and from surrounding water, respectively. A total of 124 MPs were observed, where 72 % were fibers and 28 % particles, mostly of blue color (85 %), and with rayon and nylon as the most abundant polymers. This study is an important contribution to increase the scientific knowledge of MP contamination in mesopelagic fishes used for human consumption and collected in the open waters, reinforcing the need for further research regarding MP contamination in top predatory species from high trophic levels.

Therapy-Induced Senescence Enhances the Efficacy of HER2-Targeted Antibody-Drug Conjugates in Breast Cancer.

Antibody-drug conjugates (ADC) are antineoplastic agents recently introduced into the antitumor arsenal. T-DM1, a trastuzumab-based ADC that relies on lysosomal processing to release the payload, is approved for HER2-positive breast cancer. Next-generation ADCs targeting HER2, such as [vic-]trastuzumab duocarmazine (SYD985), bear linkers cleavable by lysosomal proteases and membrane-permeable drugs, mediating a bystander effect by which neighboring antigen-negative cells are eliminated. Many antitumor therapies, like DNA-damaging agents or CDK4/6 inhibitors, can induce senescence, a cellular state characterized by stable cell-cycle arrest. Another hallmark of cellular senescence is the enlargement of the lysosomal compartment. Given the relevance of the lysosome to the mechanism of action of ADCs, we hypothesized that therapies that induce senescence would potentiate the efficacy of HER2-targeting ADCs. Treatment with the DNA-damaging agent doxorubicin and CDK4/6 inhibitor induced lysosomal enlargement and senescence in several breast cancer cell lines. While senescence-inducing drugs did not increase the cytotoxic effect of ADCs on target cells, the bystander effect was enhanced when HER2-negative cells were cocultured with HER2-low cells. Knockdown experiments demonstrated the importance of cathepsin B in the enhanced bystander effect, suggesting that cathepsin B mediates linker cleavage. In breast cancer patient-derived xenografts, a combination treatment of CDK4/6 inhibitor and SYD985 showed improved antitumor effects over either treatment alone. These data support the strategy of combining next-generation ADCs targeting HER2 with senescence-inducing therapies for tumors with heterogenous and low HER2 expression. SIGNIFICANCE: Combining ADCs against HER2-positive breast cancers with therapies that induce cellular senescence may improve their therapeutic efficacy by facilitating a bystander effect against antigen-negative tumor cells.

Optimization of an Analytical Protocol for the Extraction of Microplastics from Seafood Samples with Different Levels of Fat.

Marine organisms are affected by the ubiquitous occurrence of microplastics (MPs) in the environment. Several protocols have been described to extract and quantify MPs in seafood, although their complex matrices, with high level of fat, can compromise the efficiency of MPs extraction. To solve this issue, the present study aimed to develop a detailed methodology suitable to process seafood samples with different levels of fat, namely fish and molluscs, from fresh and canned sources, including the immersive liquids from the cans. Sample digestion was tested using different solutions (10% KOH, 30% H2O2), temperatures (40 °C, 65 °C) and incubation times (24, 48, 72 h). For fat removal, three detergents (two laboratory surfactants and a commercial dish detergent) and 96% ethanol were tested, as well as the manual separation of fat. The methodology optimized in this study combined a digestion with 30% H2O2 at 65 °C, during 24 to 48 h, with a manual separation of the fat remaining after the digestion. All steps from the present methodology were tested in six types of polymers (PE-LD, PET, PE, AC, PS, and lycra), to investigate if these procedures altered the integrity of MPs. Results showed that the optimized methodology will allow for the efficient processing of complex seafood samples with different fat levels, without compromising MPs integrity (recoveries rate higher than 89% for all the polymers tested).

What Leads to a Patient Refusal for Ambulatory Surgery? A Logistic Regression Prediction Model Based on a 5-year Retrospective Analysis of Patients with Abdominal Wall Hernia.

INTRODUCTION: Ambulatory surgery has proven benefits in patient wellbeing and cost reduction in healthcare systems. However, some patients referred for ambulatory surgery are refused and directed instead towards inpatient care, which generates several drawbacks. The reasons for this refusal have not been yet studied. The aim of this study is to identify, retrospectively, significant variables associated with patient refusal for ambulatory surgery and develop a mathematical tool able to predict with strong accuracy those who will be rejected. MATERIAL AND METHODS: Over a 5-year period (2014 - 2018), all patients that underwent abdominal hernia repair in our hospital in an inpatient setting, and that had been previously refused for ambulatory surgery, were analysed for a total of 94 variables. A multivariate logistic regression model was developed to identify risk factors associated with refusal using data from 136 patients (65 refused vs 71 accepted). A prediction index for refusal in ambulatory surgery - IRAS - was derived and tested (n = 62 patients). RESULTS: The risk index included five significant risk factors: type 2 diabetes mellitus [OR 14.669 (2.982; 72.154)], physical status [OR 49.155 (15.532; 155.555)], prior malignancy [OR 14.518 (2.653; 79.441)], prior abdominal surgery [OR 3.455 (1.006; 11.866)] and usage of antiplatelet agents [OR 25.600 (4.309; 152.066)]. All risk factors were associated with a high risk of refusal (OR between 3.455 for history of prior abdominal surgery and 49.155 according to the American Society of Anaesthesiologists physical status classification). Defining five points as the maximum IRAS score that predicts suitability for ambulatory surgery resulted in a positive predictive value of 93.55% and negative predictive value of 87.10%. DISCUSSION: Significant patient variables for refusal of an ambulatory procedure were determined and an easy to use risk index - IRAS - was built that is able to predict with good accuracy which patients will be refused. CONCLUSION: IRAS is a useful tool that can contribute to reduce time to surgery and improve patients' quality of life.

Introdução: A cirurgia de ambulatório tem benefícios comprovados no bem-estar dos doentes e na redução de custos dos sistemas de saúde. Porém, alguns doentes referenciados para cirurgia de ambulatório são recusados e encaminhados para internamento. Os motivos desta recusa ainda não foram estudados. Neste trabalho identificámos, retrospectivamente, variáveis significativas na recusa dos doentes e fornecemos uma ferramenta matemática capaz de prever de forma precisa aqueles que serão rejeitados. Material e Métodos: Ao longo de cinco anos (2014 - 2018), todos os doentes submetidos a correção cirúrgica de hérnia abdominal em regime de internamento no nosso centro hospitalar previamente recusados para cirurgia de ambulatório foram analisados para um total de 94 variáveis. Um modelo de regressão logística multivariada foi desenvolvido para identificar os fatores de risco para recusa usando dados de 136 doentes (65 recusados vs 71 aceites). Um índice preditivo para recusa de cirurgia em ambulatório, IRAS, foi criado e testado (n = 62 doentes). Resultados: O IRAS incluiu cinco fatores de risco significativos: diabetes mellitus tipo 2 [OR 14,669 (2,982; 72,154)], estado físico [OR 49,155 (15,532; 155,555)], neoplasia maligna prévia [OR 14,518 (2,653; 79,441)], cirurgia abdominal prévia [OR 3,455 (1,006; 11,866)] e uso de agentes antiplaquetários [OR 25,600 (4,309; 152,066)]. Todos os fatores de risco foram associados a elevado risco de recusa (OR entre 3,455 para história de cirurgia abdominal prévia e 49,155 de acordo com a classificação do estado físico segundo a American Society of Anaesthesiologists). A definição de cinco pontos como a pontuação máxima do IRAS que prevê adequação para cirurgia de ambulatório resultou num valor preditivo positivo de 93,55% e um valor preditivo negativo de 87,10%. Discussão: Foram determinadas variáveis significativas para recusa de um doente para cirurgia de ambulatório e construído um índice de risco de fácil utilização, IRAS, capaz de predizer que doentes serão recusados com boa precisão. Conclusão: O índice IRAS é uma ferramenta útil que pode contribuir para a redução dos tempos de espera e melhorar a qualidade de vida dos doentes.

Dysfunctional mitochondrial translation and combined oxidative phosphorylation deficiency in a mouse model of hepatoencephalopathy due to Gfm1 mutations.

Hepatoencephalopathy due to combined oxidative phosphorylation deficiency type 1 (COXPD1) is a recessive mitochondrial translation disorder caused by mutations in GFM1, a nuclear gene encoding mitochondrial elongation factor G1 (EFG1). Patients with COXPD1 typically present hepatoencephalopathy early after birth with rapid disease progression, and usually die within the first few weeks or years of life. We have generated two different mouse models: a Gfm1 knock-in (KI) harboring the p.R671C missense mutation, found in at least 10 patients who survived more than 1 year, and a Gfm1 knock-out (KO) model. Homozygous KO mice (Gfm1-/- ) were embryonically lethal, whereas homozygous KI (Gfm1R671C/R671C ) mice were viable and showed normal growth. R671C mutation in Gfm1 caused drastic reductions in the mitochondrial EFG1 protein content in different organs. Six- to eight-week-old Gfm1R671C/R671C mice showed partial reductions of in organello mitochondrial translation and respiratory complex IV enzyme activity in the liver. Compound heterozygous Gfm1R671C/- showed a more pronounced decrease of EFG1 protein in liver and brain mitochondria, as compared with Gfm1R671C/R671C mice. At 8 weeks of age, their mitochondrial translation rates were significantly reduced in both tissues. Additionally, Gfm1R671C/- mice showed combined oxidative phosphorylation deficiency (reduced complex I and IV enzyme activities in liver and brain), and blue native polyacrylamide gel electrophoresis analysis revealed lower amounts of both affected complexes. We conclude that the compound heterozygous Gfm1R671C/- mouse presents a clear dysfunctional molecular phenotype, showing impaired mitochondrial translation and combined respiratory chain dysfunction, making it a suitable animal model for the study of COXPD1.

Synonymous mutations in the phosphoglycerate kinase 1 gene induce an altered response to protein misfolding in Schizosaccharomyces pombe.

Background: Proteostasis refers to the processes that regulate the biogenesis, folding, trafficking, and degradation of proteins. Any alteration in these processes can lead to cell malfunction. Protein synthesis, a key proteostatic process, is highly-regulated at multiple levels to ensure adequate adaptation to environmental and physiological challenges such as different stressors, proteotoxic conditions and aging, among other factors. Because alterations in protein translation can lead to protein misfolding, examining how protein translation is regulated may also help to elucidate in part how proteostasis is controlled. Codon usage bias has been implicated in the fine-tuning of translation rate, as more-frequent codons might be read faster than their less-frequent counterparts. Thus, alterations in codon usage due to synonymous mutations may alter translation kinetics and thereby affect the folding of the nascent polypeptide, without altering its primary structure. To date, it has been difficult to predict the effect of synonymous mutations on protein folding and cellular fitness due to a scarcity of relevant data. Thus, the purpose of this work was to assess the effect of synonymous mutations in discrete regions of the gene that encodes the highly-expressed enzyme 3-phosphoglycerate kinase 1 (pgk1) in the fission yeast Schizosaccharomyces pombe. Results: By means of systematic replacement of synonymous codons along pgk1, we found slightly-altered protein folding and activity in a region-specific manner. However, alterations in protein aggregation, heat stress as well as changes in proteasome activity occurred independently of the mutated region. Concomitantly, reduced mRNA levels of the chaperones Hsp9 and Hsp16 were observed. Conclusion: Taken together, these data suggest that codon usage bias of the gene encoding this highly-expressed protein is an important regulator of protein function and proteostasis.

Bioremediation of Petroleum Hydrocarbons in Seawater: Prospects of Using Lyophilized Native Hydrocarbon-Degrading Bacteria.

This work aimed to develop a bioremediation product of lyophilized native bacteria to respond to marine oil spills. Three oil-degrading bacterial strains (two strains of Rhodococcus erythropolis and one Pseudomonas sp.), isolated from the NW Portuguese coast, were selected for lyophilization after biomass growth optimization (tested with alternative carbon sources). Results indicated that the bacterial strains remained viable after the lyophilization process, without losing their biodegradation potential. The biomass/petroleum ratio was optimized, and the bioremediation efficiency of the lyophilized bacterial consortium was tested in microcosms with natural seawater and petroleum. An acceleration of the natural oil degradation process was observed, with an increased abundance of oil-degraders after 24 h, an emulsion of the oil/water layer after 7 days, and an increased removal of total petroleum hydrocarbons (47%) after 15 days. This study provides an insight into the formulation and optimization of lyophilized bacterial agents for application in autochthonous oil bioremediation.