Geriatric screening and comprehensive geriatric assessment during initial oncology appointments.

INTRODUCTION: Geriatric oncology underscores the significance of assessing functional age in guiding medical decisions, endeavouring to delineate practical and efficacious methodologies for evaluating functionality, adapting therapeutic regimens and attenuating the risks of treatment-related deterioration. OBJECTIVES AND METHODS: In this prospective study, we aimed to delineate the characteristics of older patients presenting for their initial oncology appointment by using geriatric screening (G8 score) and comprehensive geriatric assessment (CGA), while also assessing the feasibility of these evaluations. Secondary objectives included comparing the initial Eastern Cooperative Oncology Group (ECOG) performance status and any deviations from standard therapeutic strategies against the identified frailty in geriatric assessment. RESULTS: Most patients exhibited a G8 score ≤14 and underwent comprehensive geriatric assessment. While oncologists typically perceive patients' general conditions, CGA enables a systematic assessment, providing a comprehensive characterisation of elderly patients to inform therapeutic decisions and address identified fragilities. The CGA highlighted vulnerabilities across all primary domains. Notably, even among patients with ECOG scores of 0 and 1, the application of G8 score and CGA revealed numerous fragilities. Consistent with existing literature, these scales offered additional insights beyond ECOG evaluation alone, suggesting their potential to guide therapeutic adaptations for this demographic. CONCLUSION: Ongoing research and continuous evaluation are imperative to refine and broaden the implementation of geriatric-focused interventions.

LPS-Induced Mortality in Zebrafish: Preliminary Characterisation of Common Fish Pathogens.

Disease outbreaks are a common problem in aquaculture, with serious economic consequences to the sector. Some of the most important bacterial diseases affecting aquaculture are caused by Gram-negative bacteria including Vibrio spp. (vibriosis), Photobacterium damselae (photobacteriosis), Aeromonas spp. (furunculosis; haemorrhagic septicaemia) or Tenacibaculum maritimum (tenacibaculosis). Lipopolysaccharides (LPS) are important components of the outer membrane of Gram-negative bacteria and have been linked to strong immunogenic responses in terrestrial vertebrates, playing a role in disease development. To evaluate LPS effects in fish, we used a hot-phenol procedure to extract LPS from common fish pathogens. A. hydrophila, V. harveyi, T. maritimum and P. damselae purified LPS were tested at different concentrations (50, 100, 250 and 500 µg mL-1) at 3 days post-fertilisation (dpf) Danio rerio larvae, for 5 days. While P. damselae LPS did not cause any mortality under all concentrations tested, A. hydrophila LPS induced 15.5% and V. harveyi LPS induced 58.3% of zebrafish larvae mortality at 500 µg mL-1. LPS from T. maritimum was revealed to be the deadliest, with a zebrafish larvae mortality percentage of 80.6%. Analysis of LPS separated by gel electrophoresis revealed differences in the overall LPS structure between the bacterial species analysed that might be the basis for the different mortalities observed.

Feeding Yellow Worms to Meagre: Effects on Whole-Body Fatty Acid Profile and Hepatic and Intestine Oxidative Status.

This study aimed to determine the effects of dietary inclusion of Tenebrio molitor larvae (yellow worms) meal (TM) on meagre fish (Argyrosomus regius) whole-body fatty acids (FA) profile and hepatic and intestine oxidative status. For that purpose, fish were fed for 9 weeks a fishmeal-based diet (control) or diets including 10%, 20%, or 30% TM. With the increase in dietary TM level, whole-body oleic acid, linoleic acid, monounsaturated FA, and n-6 polyunsaturated FA (PUFA) increased while saturated FA (SFA), n-3 PUFA, n-3 long chain-PUFA, SFA:PUFA ratio, n3:n6 ratio, and FA retention decreased. Hepatic superoxide dismutase (SOD), glucose-6-phosphate dehydrogenase (G6PDH), and glutathione reductase (GR) activities increased and catalase (CAT) and glutathione peroxidase (GPX) activities decreased with dietary TM inclusion. Hepatic total and reduced glutathione were lower in fish fed 20% TM. Intestinal CAT activity and oxidized glutathione increased and GPX activity decreased with dietary TM inclusion. Intestine SOD, G6PDH, and GR activities increased and malondialdehyde concentration decreased in fish fed the diets with lower TM inclusion levels. Liver and intestine oxidative stress index and liver malondialdehyde concentration were unaffected by dietary TM. In conclusion, to avoid major whole-body FA changes or antioxidant status imbalances, it is recommended to limit TM to 10% inclusion in meagre diets.

Oxidative Stress Response of Meagre to Dietary Black Soldier Fly Meal.

This study aimed to assess the effect of Hermetia illucens meal (HM) dietary inclusion on meagre oxidative status. Thus, fish were fed a fishmeal-based diet (CTR diet) and three other diets with increasing levels of HM inclusion, namely 10%, 20%, and 30% (diets HM10, HM20, and HM30, respectively). At the end of the trial, hepatic and intestine superoxide dismutase, catalase, and glucose-6-phosphate dehydrogenase activities and malondialdehyde concentration were unaffected by the diet composition. Liver glutathione peroxidase activity was higher in the fish fed the HM20 diet than in the fish fed the CTR and HM30 diets, and glutathione reductase activity linearly increased with the dietary HM level. The hepatic total glutathione and reduced glutathione contents were significantly lower in fish fed the HM20 diet than in fish fed the CTR and HM10 diets. In the intestine, the oxidized glutathione (GSSG) content and oxidative stress index linearly increased with the increase in dietary HM level, with the GSSG content of fish fed the HM20 diet being significantly higher than of fish fed the CTR diet. In conclusion, 30% HM might be included in meagre diets without negatively affecting hepatic and intestine oxidative status.

Oral vaccination of fish against vibriosis using spore-display technology.

Oral vaccines are highly demanded by the aquaculture sector, to allow mass delivery of antigens without using the expensive and labor-intensive injectable vaccines. These later require individual handling of fish, provoking stress-related mortalities. One possible strategy to create injection-free vaccine delivery vehicles is the use of bacterial spores, extremely resistant structures with wide biotechnological applications, including as probiotics, display systems, or adjuvants. Bacterial spores, in particular those of Bacillus subtilis, have been shown to behave as mucosal vaccine adjuvants in mice models. However, such technology has not been extensively explored against fish bacterial disease. In this study, we used a laboratory strain of B. subtilis, for which a variety of genetic manipulation tools are available, to display at its spores surface either a Vibrio antigenic protein, OmpK, or the green fluorescence protein, GFP. When previously vaccinated by immersion with the OmpK- carrying spores, zebrafish survival upon a bacterial challenge with V. anguillarum and V. parahaemolyticus, increased up to 50 - 90% depending on the pathogen targeted. Further, we were able to detect anti-GFP-antibodies in the serum of European seabass juveniles fed diets containing the GFP-carrying spores and anti-V. anguillarum antibodies in the serum of European seabass juveniles fed the OmpK-carrying spores containing diet. More important, seabass survival was increased from 60 to 86% when previously orally vaccinated with in-feed OmpK- carrying spores. Our results indicate that B. subtilis spores can effectively be used as antigen-carriers for oral vaccine delivery in fish.

Novel protein carrier system based on cyanobacterial nano-sized extracellular vesicles for application in fish.

Aquaculture has been one of the fastest-growing food industry sectors, expanding at the pace of consumers' demands. To promote safe and effective fish growth performance strategies, and to stimulate environmentally friendly solutions to protect fish against disease outbreaks, new approaches are needed to safeguard fish welfare, as well as farmers and consumers interests. Here, we tested the use of cyanobacterial extracellular vesicles (EVs) as a novel nanocarrier system of heterologous proteins for applications in fish. We started by incubating zebrafish larvae with Synechocystis sp. PCC6803 EVs, isolated from selected mutant strains with different cell envelope characteristics. Results show that Synechocystis EVs are biocompatible with fish larvae, regardless of their structural composition, as EVs neither induced fish mortality nor triggered significant inflammatory responses. We establish also that cyanobacteria are amenable to engineering heterologous protein expression and loading into EVs, for which we used the reporter sfGFP. Moreover, upon immersion treatment, we successfully demonstrate that sfGFP-loaded Synechocystis EVs accumulate in the gastrointestinal tract of zebrafish larvae. This work opens the possibility of using cyanobacterial EVs as a novel biotechnological tool in fish, with prospective applications in carrying proteins/enzymes, for example for modulating their nutritional status or stimulating specific adaptive immune responses.

Functional Feeds to Tackle Meagre (Argyrosomus regius) Stress: Physiological Responses under Acute Stressful Handling Conditions.

Marine algae are recognised sources of bioactive compounds that have attracted great interest as nutritional supplements for aquaculture fish. Intensive rearing conditions often expose fish to husbandry-related stressors, rendering fish more susceptible to disease and reducing production yields. The present work evaluated the potential of two marine algae extracts (Fucus vesiculosus and Nannochloropsis gaditana) as nutritional supplements to mitigate stress effects in meagre (Argyrosomus regius) exposed to an acute handling stress (AS). A plant-based diet was used as a control, and three other diets were prepared, which were similar to the control diet but supplemented with 1% of each algal extract or a combination of the two extracts (0.5% each). The effects of supplemented diets on stress biomarkers, antioxidant enzyme activities, and immune response were analysed in fish exposed to AS after 4 weeks of feeding. Supplemented diets did not affect growth performance but the inclusion of F. vesiculosus promoted higher feed efficiency, as compared to the control group. Dietary algal extracts supplementation reduced plasma glucose levels, increased white blood cell counts, and reduced the expression of pro-inflammatory genes when compared with the control. N. gaditana supplementation led to a reduction in hepatic antioxidant enzyme activity and glutathione levels, while F. vesiculosus supplementation increased muscle glutathione reductase activity and reduced lipid peroxidation. These findings support the potential of algal extracts as nutraceuticals in aquafeeds to enhance the ability of fish to cope with husbandry-related stressful conditions and ultimately improve fish health and welfare.

Chlorella sp. and Nannochloropsis sp. Inclusion in Plant-Based Diets Modulate the Intestine and Liver Antioxidant Mechanisms of European Sea Bass Juveniles.

This study aimed to evaluate the effects of including microalgae Chlorella sp. or Nannochloropsis sp. in plant-based diets on antioxidant mechanisms of European sea bass (Dicentrarchus labrax) juveniles. For this purpose, three isoproteic (50%) and isolipidic (19%) diets were formulated: a practical diet, containing 15% fish meal (FM) and plant ingredients as the protein source and a mixture of fish oil and vegetable oils (40: 60) as lipid source (control diet); and two diets identical to the control but with the FM replaced by Nannochloropsis sp. or Chlorella sp. (diets Nanno and Chlo, respectively). The diets were offered to quadruplicate groups of 25 fish (initial body weight: 24 ± 1 g) for 11 weeks and then enzymatic and non-enzymatic antioxidant mechanisms and lipid oxidative biomarkers were assessed in the liver and intestine of these fish. Results showed that the antioxidant response was tissue-dependent, with the liver exhibiting lower glutathione peroxidase and glucose-6-phosphate dehydrogenase (only in Chlo group) activities, and intestine lower superoxide dismutase activity with the diets including microalgae compared to control diet. An increase of oxidized glutathione content was also observed in the intestine of fish fed the microalgae diets. Catalase and glutathione reductase activities, oxidative stress index, and total and reduced glutathione, were unaffected by dietary treatments in both tissues. Overall, the lipid peroxidation status was not compromised by the replacement of FM by microalgae.

Nutritional history does not modulate hepatic oxidative status of European sea bass (Dicentrarchus labrax) submitted to handling stress.

The aim of the present study was to assess the impact of an acute handling stress on hepatic oxidative status of European sea bass (Dicentrarchus labrax) juveniles fed diets differing in lipid so urce and carbohydrate content. For that purpose, four diets were formulated with fish oil (FO) and vegetable oils (VO) as lipid source and with 20 or 0% gelatinized starch as carbohydrate source. Triplicate groups of fish with 74 g were fed each diet during 13 weeks and then subjected to an acute handling stress. Stress exposure decreased hematocrit (Ht) and hemoglobin (Hb) levels. Independent of dietary treatment, stress exposure increased hepatic lipid peroxidation (LPO). Stressed fish exhibited lower glucose 6-phosphate dehydrogenase (G6PD), catalase (CAT), and superoxide dismutase (SOD) activities, independent of previous nutritional history. In the VO groups, stress exposure increased glutathione peroxidase (GPX) activity. Diet composition had no effect on Ht and Hb levels. In contrast, dietary carbohydrate decreased hepatic LPO and CAT activity and increased glutathione reductase (GR) and G6PD activities. Dietary lipids had no effect on LPO. Fish fed the VO diets exhibited higher G6PD activity than fish fed the FO diets. In conclusion, dietary carbohydrates contributed to the reduction of oxidative stress in fish. However, under the imposed handling stress conditions, liver enzymatic antioxidant mechanisms were not enhanced, which may explain the overall increased oxidative stress.

Merkel cell carcinoma and the challenge in its approach: a review based on a clinical context of immunosuppression.

Merkel cell carcinoma (MCC) is a rare and aggressive neuroendocrine cancer with high rate to local relapse and metastasis. Its connection to immunosuppression is well known, with reported association to human immunodeficiency virus (HIV). The authors present an 87-year-old woman, infected by HIV type 2 at advanced stage of the disease, whom presented a painless papule on left cheek in 2011. After its total excision, the histopathology confirmed MCC "in situ," with no regional or distant metastases. Simultaneously, she revealed a viral load of 2220 copies/mL and 224 CD4/mm3. Five months later, the patient presented a local and distance relapse with an aggressive behavior and positive regional lymph node. Stage IV disease was confirmed due to presence of liver metastases. Concurrently to the relapse, it was detected low CD4 levels. In our multidisciplinary team decision meeting, it has been decided conservative treatment due to low Karnofsky status, comorbidities, and stage of disease.

An Overview of Teaching Communication of Bad News in Medical School: Should a Lecture be Adequate to Address the Topic?

INTRODUCTION: Delivering bad news is very common in medical daily practice. Several studies have shown a lack of effective communication skills amongst medical students, particularly concerning how to deliver bad news. The SPIKES protocol allows communicating bad news in a 6-step method. The aim of this study is to investigate the perspective of students related to this subject. MATERIAL AND METHODS: A 45 minute lecture "Breaking Bad News" was given to 160 students in the fifth and sixth years of the Medicine course, using the SPIKES' protocol training. After the lecture, an online survey was given to all students, and a cross-sectional and descriptive analysis of data extracted from survey was undertaken. RESULTS: Fifty-four students (21% of overall) answered the online survey. Eighty three percent said that theme should have an important role in their further daily medical practice, and most of students rated the physicians' role as challenging. Sixty percent of students expressed that communicating bad news was an integral part of the medical course curriculum. Regarding the SPIKES´ protocol, 48% felt that the first step would be the easiest to put in practice, and 40% felt that the fifth step related to "Emotions" would be the most difficult. DISCUSSION: In general, the students would like to gain competencies in breaking bad news using a practical approach Conclusions: Students highly valued theoretical and practical approaches in teaching of communication of bad news. Therefore, we encourage a combination approach in pre-graduate medical education.

Introdução: A comunicação de más notícias é muito comum na prática médica diária. Vários estudos têm demonstrado um défice de competências por parte dos estudantes de medicina quanto à forma de dar más notícias. Com a aplicação do protocolo SPIKES torna-se possível adquirir treino de competências relacionadas com tal questão em seis etapas. O objetivo deste estudo é investigar a perspetiva dos estudantes sobre o problema. Material e Métodos: Uma palestra de 45 minutos sobre "Comunicação de Más Notícias" visou os 160 estudantes do quinto e sexto ano de Medicina, usando o treino segundo protocolo de SPIKES. Após a palestra, foi implementado um inquérito online a todos os alunos, e foi realizada uma análise transversal e descritiva dos dados extraídos da pesquisa. Resultados: Cinquenta e quatro alunos (21% do total) responderam ao questionário online. Oitenta e três por cento afirmaram que o tema deverá ter um papel importante na sua futura prática médica diária, e a maioria dos estudantes avaliou o papel dos médicos como desafiante. Sessenta por cento dos estudantes expressaram que comunicar más notícias fez parte integrante do currículo do curso de medicina. Em relação ao protocolo de SPIKES, 48% sentiram que o primeiro passo seria o mais fácil de colocar em prática, e 40% sentiram que o quinto passo relacionado com as "Emoções" seria o mais difícil. Discussão: Em geral, os alunos gostariam de se sentir capacitados para obter competências nas más notícias usando uma abordagem prática. Conclusões: Os estudantes valorizam as abordagens teóricas e práticas no ensino da comunicação de más notícias. Deste modo, recomenda-se um modelo pedagógico combinado na educação médica pré-graduada.

Metabolic responses to dietary protein/carbohydrate ratios in zebra sea bream (Diplodus cervinus, Lowe, 1838) juveniles.

This study aims to evaluate the effect of diets with different protein to carbohydrate ratios (P:C) on the omnivorous zebra sea bream (Diplodus cervinus) juveniles growth performance, feed efficiency, N excretion and metabolic response of intermediary metabolism enzymes. Four isoenergetic and isolipidic diets were formulated to contain increasing protein levels (25, 35, 45 and 55%) at the expense of carbohydrates (43, 32, 21 and 9%): diets P25C43, P35C32, P45C21 and P55C9. Growth performance, feed efficiency (FE), N intake [(g kg(-1) average body weight (ABW) day(-1))], N retention (g kg(-1) ABW day(-1)) and energy retention (kJ kg(-1) ABW day(-1)) increased with the increase of P:C ratio. The best growth performance and FE were achieved with diet P45C21. Ammonia excretion (mg NH4­N kg(-1) ABW day(-1)) increased as dietary protein level increased. Alanine aminotransferase and glutamate dehydrogenase activities increased with the increase of dietary P:C ratio. The opposite was observed for malic enzyme activity. Aspartate aminotransferase, hexokinase, glucokinase, fructose-1, 6-bisphosphatase and fatty acid synthetase activities were unaffected by dietary treatments. Response of key amino acid catabolic enzymes and N excretion levels to dietary P:C ratio supports the metabolic adaptability of this species to dietary protein inclusion levels. Overall, zebra sea bream seems capable of better utilize dietary protein rather than dietary carbohydrates as energy source which may be an obstacle for using more economically diets and thus for reducing environmental N loads in semi-intensive aquaculture of this species.

Dietary carbohydrate and lipid sources affect differently the oxidative status of European sea bass (Dicentrarchus labrax) juveniles.

This study aimed to evaluate the effects of dietary lipid source and carbohydrate content on the oxidative status of European sea bass (Dicentrarchus labrax) juveniles. For that purpose, four diets were formulated with fish oil (FO) and vegetable oils (VO) as the lipid source and with 20 or 0 % gelatinised starch as the carbohydrate source, in a 2×2 factorial design. Liver and intestine antioxidant enzyme activities (catalase (CAT), superoxide dismutase (SOD), glutathione peroxidase (GPX), glutathione reductase (GR), glucose-6-phosphate dehydrogenase (G6PD)), hepatic and intestinal lipid peroxidation (LPO), as well as hepatic oxidative stress index (OSI), were measured in fish fed the experimental diets for 73 d (n 9 fish/diet). Carbohydrate-rich diets promoted a decrease in hepatic LPO and OSI, whereas the lipid source induced no changes. Inversely, dietary lipid source, but not dietary carbohydrate concentration, affected LPO in the intestine. Lower intestinal LPO was observed in VO groups. Enzymes responsive to dietary treatments were GR, G6PD and CAT in the liver and GR and GPX in the intestine. Dietary carbohydrate induced GR and G6PD activities and depressed CAT activity in the liver. GPX and GR activities were increased in the intestine of fish fed VO diets. Overall, effects of diet composition on oxidative status were tissue-related: the liver and intestine were strongly responsive to dietary carbohydrates and lipid sources, respectively. Furthermore, different metabolic routes were more active to deal with the oxidative stress in the two organs studied.