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Objectives Antihypertensives reduce the risk of myocardial infarction and stroke. Restrictions during the COVID-19 pandemic limited access to healthcare, which may have had a negative impact on drug prescribing. This study aimed to assess the effect of the COVID-19 pandemic on the initiation of antihypertensive drugs.Design Interrupted time series study using a segmented linear regression model.Setting Swedish population assessed through linked national healthcare registers.Participants 720 300 new users of antihypertensives.Intervention March 2020, COVID-19 pandemic onset.Main outcomes measures The change in the initiation of antihypertensives expressed as monthly cumulative incidence, stratified by age and sex. Data on dispensed prescriptions of diuretics, beta-blockers, calcium channel blockers, ACE inhibitors (ACEi) and angiotensin receptor blockers were extracted from the Swedish Prescribed Drug Register, from March 2018 to November 2021. Initiation (new use) was defined as having no previous dispensations before March 2019. Monthly cumulative incidence in March 2019–November 2021 was calculated as the number of patients initiating each drug class in each month divided by the population.Results The start of the pandemic was associated with an immediate drop in the initiation of any antihypertensive, but no sustained effects were observed, as the incidence continued to increase in the postinterruption period by +0.02% each month in both sexes. The immediate drop was statistically significant for ACEi in both sexes and all antihypertensive classes except diuretics in patients >65 years. А significant postintervention trend change was observed for initiation of diuretics (+0.013% overall), driven mainly by a significant increase in patients >65 years. Similar findings were also observed for diuretics in females (+0.02%) and ACEi (+0.03%) in patients >65 years.Conclusions The pandemic had an immediate negative short-term effect, but we found no major long-term negative influence of the COVID-19 pandemic on initiation of any type of antihypertensive drugs.

One Health, as a multi-disciplinary approach, considers animal, human, and environmental health. Serbia continuously records high rates of antimicrobial resistance, with a knowledge gap on practices regarding veterinary antibiotic use, as no standardized data regarding veterinary consumption of antibiotics that would allow for temporal or spatial comparison are available in Serbia. This study aimed to describe the sales of veterinary antibiotics in Serbia. Aggregated wholesale data on veterinary drug sales were retrieved from the annual reports available from the Medicines and Medical Devices Agency of the Republic of Serbia for a period between 2017 and 2020. Veterinary use was analyzed using standard European Medicines Agency proposed methodology and expressed in milligrams of active substance per population correction unit (mg/PCU). Overall, there has been a 13% increase in aggregated sales, in mg/PCU, of antibiotic veterinary medicinal products (VMPs) for food-producing animals in Serbia between 2017 and 2020. With sales of around 110 mg/PCU in 2020, Serbia would have ranked ninth in Europe. In 2020, the overall highest-selling antibiotic classes were penicillins (27.62 mg/PCU), tetracyclines (27.54 mg/PCU), and aminoglycosides (12.8 mg/PCU), accounting for 61.9% of the total sales of antibiotic VMPs for food-producing animals, in mg/PCU. An increase in the use of macrolides and lincosamides and a decrease in the use of penicillins and the group classified as “other antibacterials” was noted. The current study identified an increase in the sales of veterinary antibiotics in Serbia and pointed out several potential problem areas.

Textile based sensors have gained tremendous attention in moisture sensing. Moisture monitoring is crucial in wound healing. To promote the healing process, it is essential to maintain an optimal level of moisture while limiting unnecessary dressing changes. The objective of this research was to test how well a textile moisture sensor can detect moisture from the body and wound fluid when attached to the dressing. To this end, a 3D interdigitated capacitive structure was embroidered with silver conductive threads on the textile substrate, and a bandage was placed in the centre of the multilayer structure. When compared to traditional planar interdigitated structures, the main innovation of the proposed 3D capacitive structure lies in a larger surface area for interaction with the surrounding environment, leading to enhanced sensitivity to changes in capacitance with the respect to the moisture. The 3D structure of the bandage increased the ratio between electrodes and surface area to impact the surface charge sensitivity towards the adsorbed charges of the wound and body fluid. To observe the performance of the sensor, the bandage was exposed to simulated body fluid and wound fluid. Between dry and wet conditions, the sensor can detect capacitance differences of several orders of magnitude. The threshold volume for the 3D bandage was 30 μL to 50 μL, depending on the type of biofluids. The capacitive bandage integration with the inductor was tested at high frequencies (1–400 MHz). The shift in impedance was observed for the tested fluids. Finally, to compare sensing properties of the proposed structure against the previously reported designs, a bandage sensitivity was checked for three different configurations: (a) the proposed 3D bandage, (b) a 2D bandage, composed of textile substrate with both electrodes embroidered on top of bandage, and (c) classical interdigital structure without bandage composed of textile with embroidered electrodes on the top of it. The sensitivity of the 3D bandage for simulated body fluid and wound fluid is >6.6% and 7.4%, respectively, higher than the other two structures. The integration of the proposed textile-based sensor into a bandage could facilitate wound care and have a significant impact on efficacy for patients.

Introduction: Surgical training traditionally adheres to the apprenticeship paradigm, potentially exposing trainees to an increased risk of complications stemming from their limited experience. To mitigate this risk, augmented and virtual reality have been considered, though their effectiveness is difficult to assess. Research question: The PASSION study seeks to investigate the improvement of manual dexterity following intensive training with neurosurgical simulators and to discern how surgeons' psychometric characteristics may influence their learning process and surgical performance. Material and methods: Seventy-two residents were randomized into the simulation group (SG) and control group (CG). The course spanned five days, commencing with assessment of technical skills in basic procedures within a wet-lab setting on day 1. Over the subsequent core days, the SG engaged in simulated procedures, while the CG carried out routine activities in an OR. On day 5, all residents' technical competencies were evaluated. Psychometric measures of all participants were subjected to analysis. Results: The SG demonstrated superior performance (p

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Dyslipidemia and obesity are recognized as two of the major global health issues and main risk factors for coronary heart disease and cerebrovascular disease. In recent years, carob has shown certain antioxidant and anti-dyslipidemic potential. In this study, Wistar rats were fed with a standard and cholesterol-enriched diet and treated orally with carob extract and simvastatin for four weeks. After sacrifice, blood samples were collected for biochemical analysis, and liver tissue was taken for histological and immunohistochemical assessment. Weight gain was significantly higher in groups fed with cholesterol-fortified granules; total cholesterol was found to be significantly lower in the hypercholesterolemic groups treated with simvastatin and simvastatin/carob combined regimens compared with hypercholesterolemic animals treated with saline (p < 0.05). The same was true for low-density lipoprotein cholesterol and the LDL/HDL ratio (p < 0.05). Adiponectin was remarkably higher in animals treated with simvastatin compared to all other groups (p < 0.05). Leptin was significantly lower in groups treated with carob and simvastatin compared to the hypercholesterolemic group treated with saline (p < 0.05). Carob/simvastatin co-administration reduced hepatocyte damage and improved liver morphology. A study confirmed the anti-dyslipidemic, anti-obesity, and hepatoprotective potential of carob pulp alone or in combination with simvastatin in the treatment of high-fat diet-fed rats.

Background and Objectives: This study aimed to examine the efficacy of tapentadol immediate release (IR) and morphine hydrochloride in the treatment of acute postoperative pain after total abdominal hysterectomy, as well as to examine the frequency of opioid-related side effects in observed patients. Materials and Methods: The prospective observational study was conducted over five months, and it included a total number of 100 patients. The two cohorts had different types of postoperative analgesia, and the effects were observed for 24 h postoperatively, by following the pain scores on NRS (Numerical Pain Scale), contentment with analgesia, and opioid-related side effects. Results: Statistical significance was found when assessing pain 24 h after surgery while coughing, where patients in the tapentadol IR group had significantly higher mean pain scores (p < 0.01). The subjective feeling of satisfaction with postoperative analgesia was statistically significant in the tapentadol IR group (p = 0.005). Vertigo appeared significantly more in patients from the morphine group (p = 0.03). Conclusions: Tapentadol IR (immediate release) and morphine hydrochloride are both effective analgesics used in the first 24 h after total transabdominal hysterectomy. Overall satisfaction of patients with analgesia was good. The frequency of side effects was higher in the morphine group, with statistical significance regarding the vertigo.

IntroductionThere is limited data on the awareness and use of synthetic cannabinoids (SCs) in high-risk population in Serbia, despite SCs becoming more and more common at illicit drug market.AimThis pilot study aimed to examine the awareness and prevalence of use of SCs in patients with an opioid-use disorder and to identify patient characteristics and other factors associated with SCs use.Patients and methodsThis cross-sectional study was conducted at the Clinic for Psychiatry, Clinical Center Vojvodina, Serbia, the largest tertiary health care institution in this region of the country. All patients hospitalized due to the treatment of opioid dependence during November and December 2017 were included (response rate 100%), and filled-out an anonymous questionnaire specifically developed for the purpose of this study. Differences between patients reporting SCs use and those who did not were compared using chi-square test with values of p

Objectives To determine rate of blood pressure (BP) control and to analyse patient-related, medication-related and healthcare system-related factors associated with poor BP control in outpatients with hypertension (HT).Design Cross-sectional study.Setting Two study sites with different levels of healthcare (primary healthcare (PHC) and secondary level of healthcare (SHC)) in Vojvodina, Northern Serbia.Participants A total of 581 patients (response rate 96.8%) visiting their primary care physician between July 2019 and June 2020 filled out a pretested semistructured questionnaire and had a BP reading during their regular appointments.Primary and secondary outcome measures Data on demographics, medication, BP control (target systolic BP≤140 mm Hg and∕ or diastolic BP≤90 mm Hg) and knowledge on HT was collected. Based on the median of knowledge score, patients were classified as having poor, average and adequate knowledge.Results Majority of the respondents (74.9%) had poorly controlled BP and had HT longer than 10 years. Larger number of patients at PHC site was managed with monotherapy while at the SHC majority received three or more antihypertensive drugs. Respondents from SHC showed a significantly lower knowledge score (9, 2–15) compared with the respondents from PHC (11, 4–15, p=0.001). The share of respondents with adequate knowledge on HT was significantly higher in the group with good BP control (26% and 9.2%, respectively). In a multivariate regression analysis, factors associated with poor BP control were knowledge (B=−1.091; p

IntroductionOne of the most common oral diseases affecting children is early childhood caries (ECC). The link between oxidative stress and ECC has been proven in numerous clinical studies. Technical and biological variability were so high in most of the studies that none of the markers have yet been proven suitable for routine clinical use. This study aimed to evaluate the antioxidant status and the levels of leptin and adiponectin in saliva of children with severe early childhood caries (S-ECC).MethodsMorning unstimulated saliva samples were collected from children (n = 40, 0–6 years old) for the evaluation of oxidative stress which were measured by total antioxidant capacity (TAC), and by the ferric reducing antioxidant power (FRAP) assays, as well as to assess the salivary levels of leptin and adiponectin. FRAP, TAC, leptin and adiponectin concentrations were evaluated in S-ECC group (n = 31) and caries free group CF (n = 9). All results were analyzed based on age and sex.ResultsOverall median salivary leptin and adiponectin levels were 5.59 pg/mL and 24.86 ng/mL, respectively. Significantly lower leptin levels were observed in saliva of caries free children (4.66 pg/mL) than in the S-ECC group (6.64 pg/mL, p < 0.01). No significant difference was observed for adiponectin levels (S-ECC and CF, 25.31 and 23.2 ng/mL, respectively, p = 0.961). TAC and FRAP values of saliva had similar values in children with S-ECC and caries free children. TAC and FRAP values also remained stable with the age of the children, without significant differences with respect to sex.ConclusionThe increased concentrations of leptin in saliva of children with S-ECC suggests that leptin may play a role in inflammatory and immune responses in the development of early childhood caries.

The global prevalence of dyslipidemia (elevated plasma levels of total cholesterol, LDL-Cholesterol, triglycerides, and lower plasma levels of HDL-Cholesterol) is constantly on the rise. Lately, carob pulp has been recognized as an effective natural product for the treatment of dyslipidemia. The two main components of the carob pulp, polyphenols, and insoluble fiber are believed to have beneficial effects on lipid metabolism. Studies on humans and animals confirmed its lipid-lowering effects. Several mechanisms have been proposed to explain this phenomenon, namely by affecting three organ systems: 1) gastrointestinal tract, 2) liver and 3) adipose tissue. Also, carob products have antioxidative, anti-inflammatory, and vascular-protective activity.

Objective: Considering that adherence level affects diabetes treatment success and maintenance of glycemic control greatly, the aim of this study was to examine diabetes patients’ adherence to oral hypoglycemic agents (OHAs) and knowledge about their mechanism of action, dosing regimen, and side effects. Material and Methods: This cross-sectional study was conducted on a sample of 100 patients with type 2 diabetes in order to assess their knowledge of OHAs using anonymous questionnaires. Results: Most patients had used OHAs between 2 and 5 years (61.0%), where 78.0% were treated with metformin, and the remaining 22.0% were prescribed sulfonylurea derivatives. Besides drugs used for the treatment of type 2 diabetes, 58.0% of patients took another 1-3 drugs daily for the treatment of other conditions. Although 75.0% achieved a score of 5-6 out of the maximum score of 8, only 2.0% of respondents listed at least 2 side effects of the OHA they used, and none of them could explain its mechanism of action. Most of the information about OHAs was given to patients by endocrinologists (53.0%). Conclusion: More than half of participants considered their knowledge of OHAs insufficient. Results clearly indicate that the respondents were not sufficiently familiar with the mechanism of action and possible side effects of such medications. Information about OHAs given in written form as well as via community pharmacists would contribute to educating type 2 diabetes patients significantly.

The framework of this study was a comprehensive investigation of Morus nigra L. extracts, with the aim to establish the correlation between chemical composition and antioxidant/hepatoprotective activity of a series of black mulberry extracts obtained from aerial parts of the plant. Black mulberry leaf (MLEE), bark (MBEE), juice (MJ) and fresh fruit (MFEE) extracts were obtained using the conventional Soxhlet extraction, while the supercritical CO2 extraction procedure was employed for preparation of the seed oil (MSO). Analysis of the chemical composition was performed using spectrophotometric, HPLC and GC methods. For the evaluation of antioxidant activity, in vitro FRAP and DPPH assays were applied. In Haan strain NMRI mice with streptozotocin-induced oxidative stress, in vivo antioxidant activity and liver tissue integrity were examined. The content of polyphenolic compounds was the highest in MBEE (68.3 ± 0.7 mgGAE/g) with the most abundant compounds being polyphenolic acids, followed by MLEE (23.4 ± 0.5 mgGAE/g) with the flavonoids isoquercetin and rutin being present in a significant amount. An analysis of MSO revealed a high content of γ-linoleic acid. The highest antioxidant activity in vitro (FRAP and DPPH) was observed for MLEE, MBEE and MSO. Beneficial effects were confirmed in vivo, with lower values of hepatosomatic index, potentiation of the activity of the enzymes superoxide dismutase and catalase, a lower rate of lipid peroxidation and reduced positivity for the P450 enzyme in animals treated with MLEE, MBEE and MSO. Black mulberry leaf and bark extracts as well as seed oil exhibited significant antioxidant activity. Apart from the confirmed biological properties of the fruit and leaf extracts, the observed activities of black mulberry seed oil and bark extract imply its importance as a sustainable source of phytochemicals.

The dose-response relationship of sildenafil effects on cardiac function is not completely elucidated. The aim of this study was to assess the effects of different doses of sildenafil on coronary flow and oxidative stress in isolated rat hearts. Coronary flow and markers of oxidative stress, including nitrite outflow, and superoxide anion production in coronary effluent, were determined for isolated rat hearts. The experiments were performed during control conditions and in the presence of sildenafil (10, 20, 50, 200 nM) alone or with Nω-nitro-L-arginine monomethyl ester (L-NAME) (30 μM). Sildenafil was shown to result in a significant increase in coronary flow at lower coronary perfusion pressure (CPP) values at all administered doses, whereas, with an increase in CPP, a reduction in coronary flow was observed. An increase in nitric oxide (NO) was most pronounced in the group treated with the lowest dose of sildenafil at the highest CPP value. After the inhibition of the NO-cyclic guanosine monophosphate (cGMP) signaling (NOS) system by L-NAME, only a dose of 200 nM sildenafil was high enough to overcome the inhibition and to boost release of O2−. That effect was CPP-dependent, with statistical significance reached at 80, 100 and 120 mmHg. Our findings indicate that sildenafil causes changes in heart vasculature in a dose-dependent manner, with a shift from a vasodilatation effect to vasoconstriction with a pressure increase. The highest dose administered is capable of producing superoxide anion radicals in terms of NOS system inhibition.

This research aimed to examine the effects of pre-treatment with basil extract, prepared as an infusion, on acetaminophen-induced acute liver injury in rats. In vitro analysis of total phenolic, flavonoid content, and antioxidant activity was performed by spectrophotometric methods. Effects of basil extract on oxidative stress parameters, biochemical markers of liver injury, and liver morphology were determined in an in vivo model of acetaminophen-induced liver injury in Wistar rats. Total phenolic content was 52.61 ± 1.35 mg GAE/g of DE, and flavonoid content was 0.5 ± 0.2 mg QE/g of DE. IC50 values ranged from 0.22 to 45.76 μg/ml for DPPH radical, OH radical, H2O2 and lipid peroxidation. Ascorbic acid was used as a positive control. The extract lowered the intensity of lipid peroxidation and potentiated the activity of antioxidant enzymes. In addition, serum levels of aspartate aminotransferase and alanine aminotransferase were significantly lower in basil extract-treated animals in comparison to respective controls.

Resveratrol was recognized as the major factor responsible for the beneficial properties of red wine. Several resveratrol-based dietary supplements are available, but their efficacy has not been sufficiently tested. This study was designed to examine the effect of resveratrol supplementation, using a commercially available product, on the metabolic status of experimental animals with induced hyperlipidemia or type 2 diabetes mellitus (T2DM). Hyperlipidemia was induced by feeding the rats a standard pellet diet supplemented with cholesterol. T2DM was induced by adding 10% fructose to drinking water and streptozotocin. Treatment with resveratrol-based supplement improved glycemic control in diabetic animals and significantly decreased serum low-density-lipoprotein (LDL) and triglyceride levels, concurrently increasing the high-density-lipoprotein (HDL) levels in animals with hyperlipidemia. Resveratrol-treated animals had improved tolerance to glucose loading. Supplementation did not induce alterations in parameters of liver and renal function. Findings indicate that commercial resveratrol supplement improves metabolic control in rats with induced hyperlipidemia and T2DM. Keywords: Resveratrol, Diabetes, Hyperlipidemia, Metabolic parameters

The complex issue of antibacterial resistance (ABR) requires actions taken with the One Health approach, involving both human and veterinarian medicine. It can spread from animals to humans through the food chain or through direct contact. Health profession students, as the future antibiotic providers, can greatly impact antibiotic-related issues in the future. The study was conducted to evaluate knowledge, attitudes and practice of future antibiotic prescribers in relation to judicious use of antibiotics. This cross-sectional, questionnaire-based study was performed on 400 students of health professions who were allowed to prescribe antibiotics of the University of Novi Sad, Serbia. Students of medicine and students of dentistry showed a significantly higher knowledge score compared to students of veterinary medicine (p = 0.001). Multivariate regression identified predictors of adequate antibiotic knowledge: being a female student (B = 0.571; p = 0.020), higher grade average (B = 1.204; p = 0.001), students of medicine (B = 0.802; p = 0.006) and dentistry (B = 0.769; p = 0.026), and students who used a complete package of antibiotics during the last infection (B = 0.974; p = 0.001) or for the period recommended by the doctor (B = 1.964; p = 0.001). Out of the total sample, self-medication was reported among 42.8% of students. The identified predictors of self-medication were: more frequent (B = 0.587; p = 0.001) and irregular (B = 0.719; p = 0.007) antibiotic use, taking antibiotics until symptoms disappeared (B = 2.142; p = 0.001) or until the bottle was finished (B = 1.010; p = 0.001) during the last infection. It seems prudent to reevaluate the educational curricula regarding antibiotic use and ABR of prospective prescribers in Serbia.

This study investigated the chemical and nutritional profile and antioxidative properties of cultivated Coprinus comatus. Proximate analysis revealed that C. comatus is rich in carbohydrates, dietary fibres and proteins, and could also be a valuable source of phenolics. Additionally, fat content is low, consisting mainly of polyunsaturated and omega-3 fatty acids. Furthermore, the safety profile of C. comatus is satisfactory, with all elements of toxicological importance within the proposed limits. Oral treatment with C. comatus for 42 days improved the antioxidant capabilities and ameliorated carbon tetrachloride-induced liver damage in rats, marked by decreased serum aminotransferase levels and lipid peroxidation intensity. Glutathione concentrations increased in a dose-dependent manner. Histological morphometric and immunohistochemical analysis confirmed antioxidative and hepatoprotective potential. These findings imply that cultivated C. comatus could be considered a nutraceutical, having beneficial nutrient and therapeutic properties.

This study aimed to determine and describe the prevalence of combination antibiotics dispensed in outpatients with health insurance in Syria. Data on all dispensed medicines between June 2018 and May 2019 for 81,314 adults were obtained, and medicines belonging to the J01 group of the World Health Organization (WHO) anatomical therapeutic classification (ATC) were included in the analysis. Prescriptions were stratified according to the number of antibiotics, age, and sex. Antibiotic utilization was expressed as the number of prescriptions per 1000 persons per year. Out of 59,404 prescriptions for antibiotics, 14.98% contained antibiotic combinations, distributed to 22.49% of the patients. The prevalence of dispensing antibiotic combinations was higher in female patients (23.00%), and the youngest (18–30 years, 26.19%) and oldest age groups (>70 years, 25.19%). The antibiotics most commonly combined were co-amoxiclav, second- and third-generation cephalosporins, and macrolides. Over 60% of the combinations contained ceftriaxone alone or in combination with sulbactam. The present study shows an alarmingly widespread prescription of antibiotic combinations, posing a risk to global health by promoting resistance development.

The aim of this study was to determine and describe trends in antibiotics utilization in Serbia over a ten-year period. Data were retrieved from publicly available annual reports (2010–2019). The results were expressed as Defined Daily Dose (DDD) per 1000 inhabitants per day (DID). All calculations were performed using the DDD values for the 2020 Anatomical Therapeutic Chemical/Defined Daily Dose (ATC/DDD) version for each year of the study, to account for the DDD changes during the study period. Antibiotics were classified using the WHO Access, Watch, Reserve (AWaRe) classification. Total utilization of antibacterials for systemic use increased from 17.25 DID in 2010 to 28.65 DID in 2019. A statistically significant increasing trend in the use of the Watch category antibiotics was observed. A tendency towards use of broad-spectrum antibiotics, apparent by a statistically significant increase in the rate of utilization of broad-spectrum macrolides, quinolones and third-generation cephalosporins vs. narrow-spectrum ones, as well as a significant increasing trend in the use of quinolones was identified. Total antibiotic utilization was found to be well above the European average. Several specific problem areas were identified, which requires further efforts to improve antibiotic prescribing. The present study provides the information needed to facilitate antibiotic stewardship in Serbia further and proposes specific interventions to optimize antibiotic use in Serbia.

In this work the effect of partial or total replacement of fishmeal by plant protein sources and krill and squid meal on growth performance and shelf-life of gilthead sea bream was evaluated. Plant protein dietswith 50 g kg−1 of krill and 100 g kg−1 of squid were supplemented with synthetic amino acidsand at the end of the growing period weight showed no significant differences. The spoilage process of the fish was followed by physicochemical and microbiological measurements together with a colorimetric sensor array (CSA) specially designed for that purpose. The changes in the physicochemical parameters and microbial growth showed that shelf-life of samples were in all cases lower than ninedays. The CSA was not able to show significant differences between both diets, confirming the physicochemical and microbiological results. The fact that the type of feed had no effect on the freshness parameters studied demonstrates that total fishmeal replacement with plant protein blends in the proportions used in this work could be an excellent alternative for feed formulation in aquaculture.

There is little published information about antibiotic utilization in outpatients in Serbia. The objective of this study was to determine the amount and structure of outpatient antibiotic use in South Backa District (SBD) in Serbia, to assess prescibing quality of antibiotics and to compare with results from Scandinavian countries. Data on the antibiotic use were collected from all private and state-owned pharmacies from January through March 2008 in SBD. Results were expressed as the number of defined daily doses/1,000 inhabitants/day. The drug utilization 90% method was also used. Penicillins were the most frequently used antibiotic subgroup in SBD (35.20%), followed by cephalosporins (19.16%) and macrolides (13.18%). Thirteen drugs accounted for 90% of total antibiotics consumption (DU90% segment). The average cost/DDD within the DU90% segment was 0.95 euros, whereas the average cost/DDD beyond the DU90% segment was 1.89 euros, indicating that less expensive antibiotics were more frequently used. High use of ampicillin, third-generation cefalosporins, co-trimoxazole, and gentamicin, will aggravate the alarming problem of resistance in Serbia. Differences in the amount and structure of antibiotic consumption between SBD and Scandinavian countries indicate the need of updated national guidelines for rational antimicrobial drug use in Serbia.

Ciprofloxacin is often used in treatment of complicated urinary tract infections in areas with high rates of resistance to first line agents. The aim of this study was to evaluate efficacy of ciprofloxacin in standard dosing regimens in treatment of complicated urinary tract infections. Plasma concentration curves were simulated and minimum inhibitory concentration (MIC) and post-antibiotic effect were determined. Ciprofloxacin MIC ranged from 0.0156 for Gram-negative and to 0.125-0.5 µg/mL for Gram-positive bacteria. Both dosing regimens were suitable for eradication of Gram-negative bacteria, with slight supremacy of 750 mg/12 hours over 500 mg/12 hours dosing regimen. Even though all strains were fully susceptible to ciprofloxacin, pharmaco-kinetic/pharmacodynamic parameters did not meet target thresholds for pathogens with MIC over 0.1-0.2 µg/mL regardless of the administered dose. Ciprofloxacin remains an excellent choice for treatment of complicated urinary tract infections caused by Gram-negative bacteria, but in infection caused by Gram-positive strains, deeper analysis is necessary in order to achieve optimal results.

La creciente preocupación por la contaminación generada por los plásticos derivados del petróleo representa uno de los desafíos ambientales más apremiantes a nivel global en la actualidad. El uso generalizado de plásticos en una amplia gama de productos, combinado con una gestión inadecuada de los residuos, ha dado lugar a una acumulación masiva de estos materiales en entornos naturales como océanos, ríos, suelos y diversos ecosistemas. Esta problemática no solo afecta la biodiversidad y la calidad de vida de las comunidades, sino que también plantea interrogantes éticos y de sostenibilidad para las generaciones presentes y futuras. En este contexto de urgencia ambiental, es necesario explorar y promover soluciones innovadoras y sostenibles que aborden de manera integral la problemática de los plásticos y fomenten una cultura de reducción, reutilización y reciclaje. Dentro de este marco, el arte emerge como una poderosa herramienta capaz de generar conciencia, transformación social y ambiental, así como de inspirar cambios significativos en las prácticas individuales y colectivas. El artículo se centra en el desarrollo de un modelo de evaluación para el Arte Público Participativo que utiliza desechos plásticos como material artístico. Este enfoque, conocido como Upcycling o Supra-reciclaje, tiene como objetivo principal la reutilización creativa de los plásticos en la creación de obras artísticas. Además de su función estética, estas obras tienen la intención de empatizar y transmitir emociones que aporten concienciación sobre sostenibilidad, consumo responsable y gestión de residuos. Este estudio analiza cómo el arte puede ser un instrumento efectivo para abordar desafíos ambientales globales al transformar desechos en recursos artísticos y educativos. Para fundamentar y enriquecer este estudio, se recurre a diversas corrientes teóricas y disciplinas, que van desde la sociología ambiental hasta la psicología social y la ecología cultural. Entre las teorías y consideraciones relevantes se encuentran la "Teoría del valor, las normas y las creencias hacia el medioambiente" de Stern et al. (2000), que proporciona un marco conceptual para comprender cómo se construyen y transforman las actitudes y comportamientos hacia el entorno natural. Asimismo, se considera la "Psicología social de los objetos" de Mead (1932), que explora la relación entre los individuos, los objetos y la identidad personal en el contexto de la sociedad contemporánea. Además, se aborda la "Teoría de la Basura" de Thompson (1979), que analiza la evolución del valor social de los objetos, incluyendo los desechos, en el contexto de la cultura material y el consumo. La metodología propuesta para la evaluación de estos proyectos artísticos participativos se sustenta en un conjunto de variables fundamentales que abarcan aspectos como el grado de participación comunitaria en la creación de las obras, la selección y uso innovador de materiales plásticos, el impacto ambiental de las instalaciones artísticas, la integración con el entorno urbano o natural, así como el mensaje temático y simbólico que transmiten las obras a la audiencia. Este modelo de evaluación no solo busca valorar la calidad estética y conceptual de las obras, sino también su impacto social, educativo y medioambiental. Desde una perspectiva más amplia, esta focalización de Arte Público Participativo orientado en la reutilización creativa de desechos plásticos representa una oportunidad única para transformar la percepción y las prácticas relacionadas con los residuos y el consumo en nuestra sociedad. En resumen, este artículo aborda el papel crucial que desempeña el arte, especialmente cuando se integra en proyectos participativos que incluyen la reutilización creativa de desechos plásticos. Se examina cómo esta guía puede ser fundamental en la sensibilización, educación y movilización de la sociedad hacia prácticas más sostenibles y conscientes. Además de su impacto en el ámbito artístico, este enfoque contribuye significativamente a la construcción de una cultura de respeto y cuidado hacia el medio ambiente y los recursos naturales

Fishmeal and fish oil substitution in aquafeeds might have adverse effects on fish growth and health, mainly in carnivorous species, such as Mediterranean yellowtail (Seriola dumerili). Mediterranean yellowtail shows great potential as an alternative aquaculture species due to its fast growth and high price on the market, but the need for high-quality protein and fatty acid content in its diets is limiting its production. In order to improve the sustainability of its production, this study was conducted with 360 fish of 35 g to evaluate the effects on fish growth and health. Six diets were used: one control diet without replacement, three with FM replacement (FM66, FM33, and FM0) (33%, 66%, and 100% FM replacement), and two with FO replacement (FO50 and FO0) (50% and 100% FO replacement). The substitution of FM was with vegetable (VM) (corn gluten) and animal (AM) (krill and meat meal) meals. The reductions in FM and FO of up to 33 and 0%, respectively, did not affect the growth and survival of S. dumerili at the intestinal morphology level, except for the anterior intestine regarding the lower villi length and width and the posterior intestine regarding the lower width of the lamina propria. On the other hand, the substitution of fish ingredients in the diet affects liver morphology, indicating alterations in the major diameter of hepatocytes or their nuclei. Finally, diet did not affect the gut microbiota with respect to the control, but significant differences were found in alpha and beta diversity when FO and FM microbiota were compared. A 66% FM replacement and total FO replacement would be possible without causing major alterations in the fish.

Two novel conjugate molecules were designed: pyrene and phenanthridine-amino acid units with a different linker length between the aromatic fragments. Molecular modelling combined with spectrophotometric experiments revealed that in neutral and acidic buffered water solutions conjugates predominantly exist in intramolecularly stacked conformations because of the π–π stacking interaction between pyrene and phenanthridine moieties. The investigated systems exhibited a pH-dependent excimer formation that is significantly red-shifted relative to the pyrene and phenanthridine fluorescence. While the conjugate with a short linker showed negligible spectrophotometric changes due to the polynucleotide addition, the conjugate with a longer and more flexible linker exhibited a micromolar and submicromolar binding affinity for ds-polynucleotides and inactivated a mutant of dipeptidyl peptidase enzyme E451A. Confocal microscopy revealed that the conjugate with the longer linker entered the HeLa cell membranes and blue fluorescence was visualized as the dye accumulated in the cell membrane.

A reunificação familiar é um processo complexo que se inicia com o acolhimento residencial da criança e/ou jovem na instituição e continua após o regresso a casa, podendo contribuir para consolidação e sucesso das medidas de proteção. O presente estudo teve como objetivo analisar as dinâmicas geradoras dos processos de resiliência em famílias após implementação do Programa “Caminhar em Família” que visa a reunificação familiar por meio de promoção de competências parentais durante o acolhimento. Foi desenvolvido um estudo de caso qualitativo de caráter descritivo e retrospectivo. A amostra não probabilística foi composta por 4 famílias, sendo 5 progenitores e 6 crianças/jovens que saíram do acolhimento no ano de 2019 e estavam aptas à reunificação. A participação das famílias no programa permitiu confirmar o fortalecimento da resiliência no sistema relacional de pais e filhos graças à compreensão, capacitação e empoderamento suscitados nas diferentes etapas da medida de proteção. A associação conceitual de dois modelos teóricos da resiliência familiar das autoras Froma Walsh e Lietz e Strenght possibilitou identificar diferentes forças e recursos individuais e familiares na gestão da crise e respostas específicas às necessidades das famílias nos diferentes estádios do acolhimento e reunificação. Os resultados reforçam a importância de programas socioeducativos que privilegiam o fortalecimento das dimensões promotoras de aspectos saudáveis que compõe a resiliência em famílias. Estes elementos devem ser os norteadores das intervenções para uma reunificação familiar segura e estável.

The use of organic ingredients as a source of protein in aquaculture diets has gained significant attention due to the growing demand for organic seafood products. This study aimed to evaluate the potential for the use of organic ingredients as protein sources in the diet of juvenile organic seabass (Dicentrarchus labrax). A total of 486 juvenile seabass with an average weight of 90 g were fed six diets containing varied organic proteins. The control group (CON) was fed a diet with conventional fishmeal from sustainable fisheries as the primary protein source. The other five groups were fed diets with different compositions: organic Iberian pig meal byproduct (IB diet), a combination of organic Iberian pig meal byproduct and insect meal (IB-IN diet), a mix of organic Iberian pig meal byproduct and organic rainbow trout meal byproduct (IB-TR diet), a blend of organic rainbow trout meal byproduct and insect meal (TR-IN), and a mixed diet containing all of these protein sources (MIX diet). Over a 125-day feeding trial, growth performance, feed utilisation, feed digestibility, and histological parameters were assessed. The results showed that the fish fed the control diet had the highest final weight and specific growth rate, followed by the fish fed the TR-IN and IB-TR diets. The IB-TR diet had the highest apparent digestibility coefficients (ADCs) for protein, while the TR-IN diet had the lowest. Histological analysis revealed that fish fed the control diet had the largest nucleus diameter and hepatocyte diameter. Use of IN seems to penalise performance in several ways. Fish fed diets containing insect meal grew less, and those diets had lower digestibility. Fish fed the TR and IB diets grew at rates near that of the control, and the feed had acceptable digestibility.

A 120-day experiment investigated the new organic raw materials for Gilthead seabream (Sparus aurata) feeding on growth, nutritional parameters, digestibility, and histology. An organic control diet (CON) and three diets with 100% organic raw materials—the rest of rainbow trout, visceral Iberian pig, and insects as a protein source (TRO, IBE, and INS)—were tested. After the experiment, growth, nutritional parameters, digestibility, and histology were measured. The CON diet-fed seabream obtained the highest weight; there was no difference between the experimental diets. The crude protein content was the highest in seabream fed the TRO and INS diets. Crude fat was the highest value observed in the CON diet. High digestibility was observed in seabream-fed IBE and INS diets. Except for EAA methionine (Met), there were no static differences in retention efficiency. The diet with the highest hydrolysis rate was IBE. Diet differences were significant but had the typical healthy liver morphology of seabream. Seabream fed on the TRO and INS diets had shorter distal gut measurements. In conclusion, the full substitution of fishmeal with organic raw materials, including rainbow trout remains, Iberian pig viscera, and insects, offers several benefits in terms of digestibility, histology, and growth performance and may help improve sustainable and healthy aquaculture practices.

Skin cancers are among the most common cancers worldwide and are increasingly prevalent. Cutaneous melanoma (CM) is characterized by the malignant transformation of melanocytes in the epidermis. Although CM shows lower incidence than other skin cancers, it is the most aggressive and responsible for the vast majority of skin cancer-related deaths. Indeed, 75% of patients present with invasive or metastatic tumors, even after surgical excision. In CM, the photoprotective pigment melanin, which is produced by melanocytes, plays a central role in the pathology of the disease. Melanin absorbs ultraviolet radiation and scavenges reactive oxygen/nitrogen species (ROS/RNS) resulting from the radiation exposure. However, the scavenged ROS/RNS modify melanin and lead to the induction of signature DNA damage in CM cells, namely cyclobutane pyrimidine dimers, which are known to promote CM immortalization and carcinogenesis. Despite triggering the malignant transformation of melanocytes and promoting initial tumor growth, the presence of melanin inside CM cells is described to negatively regulate their invasiveness by increasing cell stiffness and reducing elasticity. Emerging evidence also indicates that melanin secreted from CM cells is required for the immunomodulation of tumor microenvironment. Indeed, melanin transforms dermal fibroblasts in cancer-associated fibroblasts, suppresses the immune system and promotes tumor angiogenesis, thus sustaining CM progression and metastasis. Here, we review the current knowledge on the role of melanin secretion in CM aggressiveness and the molecular machinery involved, as well as the impact in tumor microenvironment and immune responses. A better understanding of this role and the molecular players involved could enable the modulation of melanin secretion to become a therapeutic strategy to impair CM invasion and metastasis and, hence, reduce the burden of CM-associated deaths.

Introduction: PIK3CA is one of the most mutated oncogenes in solid tumors. In breast cancer (ER-positive, HER2-negative), these events represent a predictive biomarker of response to alpelisib. In glioblastomas (GBM), PIK3CA mutations were described as early constitutive events. Here, we investigated PIK3CA mutational profile across glioma molecular subgroups and its relevance during glioma recurrence. Furthermore, PIK3CA mutations’ effect in PI3K pathway, prognosis, and response to therapy was also explored. Material and Methods: Exons 10 and 21 of PIK3CA mutations were evaluated in 394 gliomas and 19 glioma recurrences from Instituto Português de Oncologia Lisboa Francisco Gentil (IPOLFG) and compared with The Cancer Genome Atlas (TCGA) data. TIMER2.0 and NetMHCpan4.1 were used to assess the immune-microenvironment contribution. Results: PIK3CA mutations were identified among all glioma subgroups, although with no impact on their stratification or prognosis. In both cohorts (IPOLFG and TCGA), PIK3CA mutation frequencies in IDH -mutant and IDH -wild-type GBM were similar (IPOLFG: 9% and 3%; TCGA: 8% and 2%). These mutations were not mutually exclusive with PTEN deletion and EGFR amplification. Despite their reduced frequency, we discovered PIK3CA mutations were maintained during glioma recurrence regardless of administered therapies. The immune microenvironment might not contribute to this phenotype as PIK3CA mutations did not influence immune cell infiltration. Conclusions: Despite the absence of a predominant effect in glioma stratification, PIK3CA mutations were maintained during glioma recurrence, possibly contributing to glioma cell survival, representing promising therapeutic targets in recurrent glioma. Nevertheless, understanding the potential synergistic effects between PIK3CA mutations, PTEN deletion, and EGFR amplification is pivotal to targeted therapies’ efficiency.

The fishmeal replacement by vegetable meals or other alternative sources, without affecting fish performance and productivity, is one of the principal challenges in aquaculture. The use of hydrolyzed porcine mucosa (HPM) and nucleotide (NT) concentrates, as feed additives in gilthead seabream (Sparus aurata L.) non-fishmeal diets was assessed in order to determine the possible effects on growth, feed efficiency, protein digestion, and gut histology when these were included in a plant-based diet (HPM 1% and 2%, P1 and P2; NT 250 and 500 ppm, N250 and N500), in comparison with two control diets, AA0 (100% plant-protein-based diet) and FM100 (100% fishmeal-protein-based diet). Diets were assayed in triplicate and the growth assay lasted 134 days. Results showed a significant improvement in all groups in terms of final weight and specific growth rate in comparison with the AA0 group. An improvement in the feed conversion ratio and the protein efficiency ratio was also observed when the additives were included in lower percentages (P1 and N250) compared to the FM100 group. Significant differences were found in hepatosomatic index, villi thickness, and goblet cells. Thus, the inclusion of NT and HPM was tested as beneficial for the improvement of efficiency of plant feed in seabream.

The binding interactions of six ligands, neutral and monocationic asymmetric monomethine cyanine dyes comprising benzoselenazolyl moiety with duplex DNA and RNA and G-quadruplex structures were evaluated using fluorescence, UV/Vis (thermal melting) and circular dichroism (CD) spectroscopy. The main objective was to assess the impact of different substituents (methyl vs. sulfopropyl vs. thiopropyl/thioethyl) on the nitrogen atom of the benzothiazolyl chromophore on various nucleic acid structures. The monomethine cyanine dyes with methyl substituents showed a 100-fold selectivity for G-quadruplex versus duplex DNA. Study results indicate that cyanines bind with G-quadruplex via end π-π stacking interactions and possible additional interactions with nucleobases/phosphate backbone of grooves or loop bases. Cyanine with thioethyl substituent distinguishes duplex DNA and RNA and G-quadruplex structures by distinctly varying ICD signals. Furthermore, cell viability assay reveals the submicromolar activity of cyanines with methyl substituents against all tested human cancer cell lines. Confocal microscopy analysis shows preferential accumulation of cyanines with sulfopropyl and thioethyl substituents in mitochondria and indicates localization of cyanines with methyl in nucleus, particularly nucleolus. This confirms the potential of examined cyanines as theranostic agents, possessing both fluorescent properties and cell viability inhibitory effect.

O objetivo deste texto é identificar o espaço urbano da cidade de Lisboa entre os princípios do século XIX e os princípios do século XXI recorrendo a uma metodologia que possa ser aplicada de forma homogénea ao longo do todo o período considerado. Para essa abordagem, tomam-se como base os dados dos recenseamentos da população realizados ao longo desses séculos e informações sobre a rede de transportes da região de Lisboa no mesmo período. Identifica-se a área com população de densidade urbana em torno do núcleo central constituído pela cidade na sua definição legal e administrativa; a área onde predominam atividades económicas de natureza urbana, isto é, dos setores secundário e terciário, em torno do mesmo núcleo central; e a área abrangida pelo sistema de transportes públicos da região de Lisboa. Conclui-se que a área urbana de Lisboa viveu um período de estagnação durante a primeira metade do século XIX, um período de expansão moderada e intensiva entre meados do século XIX e meados do século XX e um período de expansão acelerada e extensiva desde meados do século XX, alterando-se significativamente a distribuição no interior desse espaço.

Triploid, sterile Atlantic salmon (Salmo salar) could make a contribution to the development of the farming industry, but uncertainties about the performance and welfare of triploids have limited their adoption by farmers. In this study, we compared the ontogeny of digestive tract morphology and enzyme activities (pepsin, trypsin, chymotrypsin, alkaline phosphatase and aminopeptidase) of diploid and triploid Atlantic salmon. Fish were fed diets based on fishmeal (STD) or a mix of fishmeal and hydrolysed fish proteins (HFM) whilst being reared at low temperature from start-feeding to completion of the parr-smolt transformation. Fish weights for each ploidy and feed combination were used to calculate thermal growth coefficients (TGCs) that spanned this developmental period, and the data were used to examine possible relationships between enzyme activities and growth. At the end of the experiment, faeces were collected and analyzed to determine the apparent digestibility coefficients (ADCs) of the dietary amino acids (AAs). Digestive tract histo-morphology did not differ substantially between ploidies and generally reflected organ maturation and functionality. There were no consistent differences in proteolytic enzyme activities resulting from the inclusion of HFM in the diet, nor was there improved digestibility and AA bioavailability of the HFM feed in either diploid or triploid fish. The triploid salmon had lower ADCs than diploids for most essential and non-essential AAs in both diets (STD and HFM), but without there being any indication of lower intestinal protease activity in triploid fish. When trypsin-to-chymotrypsin activity and trypsin and alkaline phosphatase (ALP) ratios (T:C and T:ALP, respectively) were considered in combination with growth data (TGC) low T:C and T:ALP values coincided with times of reduced fish growth, and vice versa, suggesting that T:C and T:ALP may be used to predict recent growth history and possible growth potential.

Abstract Purpose Studying the effect of citrus pulp silage in the growth performance, amino acid retention of sharpsnout sea bream juveniles. Methods Citrus pulp is an energy and protein source that has been used to replace wheat meal in Diplodus puntazzo fingerlings; 96 fish of 21.5 ± 0.67 g were nourished with two experimental diets for 105 days. The control diet (CON) contained 421.6 g/kg of protein and 22.6 MJ/kg of energy. The citrus pulp diet (CPD) included 442.7 g/kg of protein and 22.02 MJ/kg of energy. Results At the end of this trial, the CPD did not affect the growth parameters, but the feed efficiency values exhibited statistically significant differences, with those fish fed the CPD having higher values, possibly due to the presence of limonene in the diet. The CPD also resulted in a reduction in amino acid (AA) content due to the protein being bound to fibre; this consequence did not affect the final fish AA profile. For AA retention, only arginine displayed significant differences, although the rest of the AAs also showed effects of AA reduction from the CPD. Conclusion Citrus pulp silage can be used in D. puntazzo feeding, without adverse in the growth performance and the amino acid profile, although it did cause a reduction in feed efficiency.

Abstract Background In order to ensure sustainability of aquaculture production of carnivourous fish species such as the gilthead seabream (Sparus aurata, L.), the impact of the inclusion of alternative protein sources to fishmeal, including plants, has been assessed. With the aim of evaluating long-term effects of vegetable diets on growth and intestinal status of the on-growing gilthead seabream (initial weight = 129 g), three experimental diets were tested: a strict plant protein-based diet (VM), a fishmeal based diet (FM) and a plant protein-based diet with 15% of marine ingredients (squid and krill meal) alternative to fishmeal (VM+). Intestines were sampled after 154 days. Besides studying growth parameters and survival, the gene expression related to inflammatory response, immune system, epithelia integrity and digestive process was analysed in the foregut and hindgut sections, as well as different histological parameters in the foregut. Results There were no differences in growth performance (p = 0.2703) and feed utilization (p = 0.1536), although a greater fish mortality was recorded in the VM group (p = 0.0141). In addition, this group reported a lower expression in genes related to pro-inflammatory response, as Interleukine-1β (il1β, p = 0.0415), Interleukine-6 (il6, p = 0.0347) and cyclooxigenase-2 (cox2, p = 0.0014), immune-related genes as immunoglobulin M (igm, p = 0.0002) or bacterial defence genes as alkaline phosphatase (alp, p = 0.0069). In contrast, the VM+ group yielded similar survival rate to FM (p = 0.0141) and the gene expression patterns indicated a greater induction of the inflammatory and immune markers (il1β, cox2 and igm). However, major histological changes in gut were not detected. Conclusions Using plants as the unique source of protein on a long term basis, replacing fishmeal in aqua feeds for gilthead seabream, may have been the reason of a decrease in the level of different pro-inflammatory mediators (il1 β, il6 and cox2) and immune-related molecules (igm and alp), which reflects a possible lack of local immune response at the intestinal mucosa, explaining the higher mortality observed. Krill and squid meal inclusion in vegetable diets, even at low concentrations, provided an improvement in nutrition and survival parameters compared to strictly plant protein based diets as VM, maybe explained by the maintenance of an effective immune response throughout the assay.

Mia40/CHCHD4 and Erv1/ALR are essential for oxidative protein folding in the mitochondrial intermembrane space of yeast and mammals. In contrast, many protists, including important apicomplexan and kinetoplastid parasites, lack Mia40. Furthermore, the Erv homolog of the model parasite Leishmania tarentolae (LtErv) was shown to be incompatible with Saccharomyces cerevisiae Mia40 (ScMia40). Here we addressed structure-function relationships of ScErv1 and LtErv as well as their compatibility with the oxidative protein folding system in yeast using chimeric, truncated, and mutant Erv constructs. Chimeras between the N-terminal arm of ScErv1 and a variety of truncated LtErv constructs were able to rescue yeast cells that lack ScErv1. Yeast cells were also viable when only a single cysteine residue was replaced in LtErvC17S. Thus, the presence and position of the C-terminal arm and the kinetoplastida-specific second (KISS) domain of LtErv did not interfere with its functionality in the yeast system, whereas a relatively conserved cysteine residue before the flavodomain rendered LtErv incompatible with ScMia40. The question whether parasite Erv homologs might also exert the function of Mia40 was addressed in another set of complementation assays. However, neither the KISS domain nor other truncated or mutant LtErv constructs were able to rescue yeast cells that lack ScMia40. The general relevance of Erv and its candidate substrate small Tim1 was analyzed for the related parasite L. infantum. Repeated unsuccessful knockout attempts suggest that both genes are essential in this human pathogen and underline the potential of mitochondrial protein import pathways for future intervention strategies.

The CRISPR/Cas9 prokaryotic adaptive immune system and its swift repurposing for genome editing enables modification of any prespecified genomic sequence with unprecedented accuracy and efficiency, including targeted gene repair. We used the CRISPR/Cas9 system for targeted repair of patient-specific point mutations in the Cytochrome b-245 heavy chain gene (CYBB), whose inactivation causes chronic granulomatous disease (XCGD)—a life-threatening immunodeficiency disorder characterized by the inability of neutrophils and macrophages to produce microbicidal reactive oxygen species (ROS). We show that frameshift mutations can be effectively repaired in hematopoietic cells by non-integrating lentiviral vectors carrying RNA-guided Cas9 endonucleases (RGNs). Because about 25% of most inherited blood disorders are caused by frameshift mutations, our results suggest that up to a quarter of all patients suffering from monogenic blood disorders could benefit from gene therapy employing personalized, donor template-free RGNs.

The aquaculture of carnivorous fish is in continuous expansion, which leads to the need to reduce the dependence on fishmeal (FM). Plant proteins (PP) represent a suitable protein alternative to FM and are increasingly used in fish feed. However, PP may lead to stunted growth and enteritis. In the current study, the effect of high FM substitution by PP sources on the growth, mortality and intestinal health of rainbow trout (Oncorhynchus mykiss) was evaluated in terms of the histological intestine parameters and expression of genes related to inflammation (IL-1β, IL-8 and TGF-β) and immune responses (Transferrin, IgT and IFN-γ). The results show that a total substitution registered lower growth and survival rates, probably due to a disruption to the animal’s health. Confirming this hypothesis, fish fed FM0 showed histological changes in the intestine and gene changes related to inflammatory responses, which in the long-term could have triggered an immunosuppression. The FM10 diet presented not only a similar expression to FM20 (control diet), but also similar growth and survival. Therefore, 90% of FM substitution was demonstrated as being feasible in this species using a PP blend of wheat gluten (WG) and soybean meal (SBM) as a protein source.

Seabass is one of the leading aquaculture species in Europe. Sensory analysis is essential for new product development. This research focused on establishing and differentiating the opinion of consumers about seabass quality obtained with organic feeding. Fish were fed for 196 days with four treatments (a control diet with 30% fishmeal and three diets with different levels of fishmeal supplemented with organic vegetable ingredients: 25%, 30% and 35%). Experimental diets were compared with commercial samples from the retail industry that were considered as “adequate quality for fish”. Two sensory analyses were carried out, check-all-that-apply (CATA) to obtain feedback on consumers’ characterization towards a different type of fish evaluated and projective mapping (PM) to measure the similarity among a set of products and establish a comparison between results provided by both methods. According to the CATA results, white color, softness, meaty taste and juicy texture were considered relevant attributes, also showing a good relationship with an adequate cooked fish description. A penalty analysis confirmed that the previous characteristics were considered essential while fibrous was an undesirable attribute. The projective mapping showed a similar sensory configuration to the CATA, corroborating these findings that showed that commercial fish were placed in a position away from the rest of the treatments, and the organic diet with a higher level of fishmeal (35%) was the most distant from the control diet.

This study was conducted to evaluate the apparent availability and P and N excretion in rainbow trout (Oncorhynchus mykiss) using different inorganic phosphorus sources. With this goal, fish (153 ± 14.1 g) fed four inorganic P sources were assayed: monoammonium phosphate (MAP, NH4H2PO4), monosodium/monocalcium phosphate (SCP-2%, AQphos+, NaH2PO4/Ca(H2PO4)2·H2O in proportion 12/88), monosodium/monocalcium phosphate (SCP-5%, NaH2PO4/Ca(H2PO4)2·H2O in proportion 30/70) and monocalcium phosphate (MCP, Ca(H2PO4)2·H2O). Phosphorus (P) digestibility, in diets that included MAP and SCP-2% as inorganic phosphorus sources, were significantly higher than for SCP-5% and MCP sources. In relation to the P excretion pattern, independent of the diet, a peak at 6 h after feeding was registered, but at different levels depending on inorganic P sources. Fish fed an MAP diet excreted a higher amount of dissolved P in comparison with the rest of the inorganic P sources, although the total P losses were lower in MAP and SCP-2% (33.02% and 28.13, respectively) than in SCP-5% and MCP sources (43.35% and 47.83, respectively). Nitrogen (N) excretion was also studied, and the fish fed an SCP-5% diet provided lower values (15.8%) than MAP (28.0%). When N total wastes were calculated, SCP-2% and SCP-5% showed the lowest values (31.54 and 28.25%, respectively). In conclusion, based on P and N digestibility and excretion, the SCP-2% diet showed the best results from a nutritional and environmental point of view.

The meagre is a carnivorous species and might be a suitable candidate species for the diversification of aquaculture in the Mediterranean region. This is based on its high growth and flesh quality. Nevertheless, there is little information available about its growth rates and nutrient requirements. The objective of this study was to determine the protein and energy requirements of juvenile meagre (Argyrosomus regius). Two trials for different weights of 53 and 188 g were conducted with rations from starvation to apparent satiation with the scope of studying its nutritional needs. In the first trial, the initial mean body weight of the fish was 53 g, and they were fed at feeding rates, measured as a percentage of the body weight, of 0, 0.75, 1.5, 2.5, 3.5, and 4.5%, with two replicates per treatment. In a second trial, another group with approximately 188 g of initial body weight was fed at feeding rates of 0, 0.5, 1.5, and 2.5%, with two replicates per treatment. The optimum thermal growth coefficient was obtained with a feed intake of 2.2% day−1 in trial A and 1.73% day−1 in trial B. The digestible protein (DP) intake for maintenance was determined as 0.57 g kg−0.7 day−1, the DP intake for maximum growth was 6.0 g kg−0.7 day−1, and the point for maximum efficiency in protein retention was 1.8 g kg−0.7 day−1. The requirement for digestible energy (DE) intake for maintenance was recorded at 25.4 kJ kg−0.82 day−1, the DE intake to maximize growth was 365 kJ kg−0.82 day−1, and the point for maximum efficiency in energy retention occurs with a digestible energy intake of 93 kJ kg−0.82 day−1. The requirements and retention efficiency of protein and energy in Argyrosomus regius tend to be within the range other fish species. The maintenance needs are in agreement with species with low voluntary activity and growth requirements in agreement with fast-growth species.

Cutaneous melanoma is the deadliest type of skin cancer, characterized by a high molecular and metabolic heterogeneity which contributes to therapy resistance. Despite advances in treatment, more efficient therapies are needed. Olive oil compounds have been described as having anti-cancer properties. Here, we clarified the cytotoxic potential of oleic acid, homovanillyl alcohol, and hydroxytyrosol on melanoma cells. Metabolic viability was determined 48 h post treatment of A375 and MNT1 cells. Metabolic gene expression was assessed by qRT-PCR and Mitogen-Activated Protein Kinase (MAPK) activation by Western blot. Hydroxytyrosol treatment (100 and 200 µM) significantly reduced A375 cell viability (p = 0.0249; p < 0.0001) which, based on the expression analysis performed, is more compatible with a predominant glycolytic profile and c-Jun N-terminal kinase (JNK) activation. By contrast, hydroxytyrosol had no effect on MNT1 cell viability, which demonstrates an enhanced oxidative metabolism and extracellular signal-regulated kinase (ERK) activation. This compound triggered cell detoxification and the use of alternative energy sources in A375 cells, inhibiting JNK and ERK pathways. Despite oleic acid and homovanillyl alcohol demonstrating no effect on melanoma cell viability, they influenced the MNT1 glycolytic rate and A375 detoxification mechanisms, respectively. Both compounds suppressed ERK activation in MNT1 cells. The distinct cell responses to olive oil compounds depend on the metabolic and molecular mechanisms preferentially activated. Hydroxytyrosol may have a cytotoxic potential in melanoma cells with predominant glycolytic metabolism and JNK activation.

The effect of different weekly feeding frequencies on Nile tilapia fingerlings of 2.02 g, was determined during 12 weeks. This was done by feeding the fish 7 days/week, 6 days/week or 5 days/week. After this restriction feeding period, all fish were fed as the control group (7 days/week) during 26 days to study the capability of the fish to compensate the growth during this re-feeding period. At the end of the feeding restriction period, there were significant differences in weights among the different treatments, although the significance was detected only at 7 days/week level, which presented the highest final body weight compared with the other 2 treatments. The daily feed intake and the feed conversion and protein efficiency ratios did not present significant differences. Crude protein efficiency (CPE) and gross energy efficiency (GEE) were affected by the feeding frequency, presenting high values in fish fed 7 days/week. Growth results obtained during this re-feeding period indicate that weight gain (WG) and specific growth rate (SGR) presented a linear increase from 7 to 5 days/week, i.e. with increasing feed deprivation period the fish could compensate the growth effectively, trying to reach to the weight as those of the control group.

The study aimed to evaluate how replacing different proportions of fish oil (FO) with vegetable oils (VO) in the diet of Mediterranean yellowtail, Seriola dumerili (Risso, 1810), affects the fatty acids (FA) signature, i.e.; overall FA profile, in different tissues. A total of 225 Mediterranean yellowtail juveniles (initial live weight: 176 ± 3.62 g) were fed for 109 days with one of three diets: A control diet (FO 100), with FO as the only lipid source, or diets with 75% and 100% of FO replaced with a VO mixture. At the end of the feeding trial, the brains, muscles, livers, and visceral fat were sampled in four fish per tank (12 per treatment), and their fat were extracted and used for FA analysis. The FA signatures of red and white muscle, liver, and visceral fat tissues changed when the dietary FA source changed, whereas FA signatures in the brain were rather robust to such dietary changes. These new insights might help evaluate whether key physiological functions are preserved when fish are fed diets with low FO levels, as well as define the dietary FA requirements of Mediterranean yellowtail to improve the sustainability of the production and welfare of the fish.