Lupinus angustifolius Protein Hydrolysates Reduce Abdominal Adiposity and Ameliorate Metabolic Associated Fatty Liver Disease (MAFLD) in Western Diet Fed-ApoE−/− Mice

Metabolic-associated fatty liver disease (MAFLD) is the most important cause of liver disease worldwide. It is characterized by the accumulation of fat in the liver and is closely associated with abdominal obesity. In addition, oxidative stress and inflammation are significant features involved in MAFLD. Recently, our group demonstrated that lupin protein hydrolysates (LPHs) had lipid lowering, antioxidant, and anti-inflammatory effects. Sixty male mice fed with a Western diet were intragastrically treated with LPHs (or vehicle) for 12 weeks. Liver and adipose tissue lipid accumulation and hepatic inflammatory and oxidant status were evaluated. A significant decrease in steatosis was observed in LPHs-treated mice, which presented a decreased gene expression of CD36 and LDL-R, crucial markers in MAFLD. In addition, LPHs increased the hepatic total antioxidant capacity and reduced the hepatic inflammatory status. Moreover, LPHs-treated mice showed a significant reduction in abdominal adiposity. This is the first study to show that the supplementation with LPHs markedly ameliorates the generation of the steatotic liver caused by the intake of a Western diet and reduces abdominal obesity in ApoE−/− mice. Future clinical trials should shed light on the effects of LPHs on MAFLD.Ministerio de Economía y Competitividad [AGL2012-40247-C02-01 and AGL2012-40247-C02-02]Andalusian Government Ministry of Health [PC-0111-2016-0111]PAIDI Program from the Andalusian Government [CTS160]Spanish Ministerio de Educación, Cultura y Deporte [FPU16/02339] and [FPU13/01210]The National Net RETICEF for Aging Studies (RD12/0043/0012 from the Instituto de Salud Carlos III, Spanish Ministerio de Ciencia e Innovación)European Social Fund and Spanish Ministerio de Empleo y Seguridad Social [EJ-086]Andalusian Government Ministry of Health [PI-0136-2019]VI Program of Inner Initiative for Research and Transfer of University of Seville (VI PPIT-US

Anxiolytic-like effects of Lupinus angustifolious protein hydrolysates in alzheimer model mice

Alzheimer’s disease (AD), which is characterized by a devastating and progressive loss of memory, is the principal neurodegenerative disease in the elderly population worldwide. As a consequence, AD patients present neuropsychiatric symptoms, such as anxiety, causing sleeping difficulty, irritability, agitation, or aggressiveness. Previous studies have demonstrated that a high-fat diet, in addition to exacerbating AD, aggravates anxiety. We have demonstrated that Lupinus angustifolius protein hydrolysates (LPHs) have anti-inflammatory and antioxidant effects, key factors for AD and anxiety. Thus, this study aimed to evaluate the potential effects of LPHs on spatial memory and anxiety of a preclinical model of AD. ApoE−/− mice fed with a western diet were intragastrically treated with LPHs (or a vehicle) for 14 weeks. Spatial memory and anxiety were then assessed through the Morris water maze and elevated plus maze, respectively. The results did not show significant differences in spatial memory between groups. However, a significant increase (p < 0.05) in time in open arms, center time, the number of crossings, and a reduction of anxiety behavior were observed in LPH-treated mice. This is the first study showing that an LPH treatment causes anxiolytic effects, pointing to LPHs as a potential component of future nutritional therapies.Ministerio de Economía y Competitividad AGL2012-40247-C02-01Ministerio de Economía y Competitividad AGL2012-40247-C02-02Ministerio de Salud del Gobierno Andaluz PC-0111-2016-0111Gobierno Andaluz CTS160Ministerio de Educación, Cultura y Deporte FPU16/02339Ministerio de Educación, Cultura y Deporte FPU13/01210Universidad de Sevilla VI PPIT-U

Bioactive Peptides from Lupin (Lupinus angustifolius) Prevent the Early Stages of Atherosclerosis in Western Diet-Fed ApoE–/– Mice

We have previously reported the in vitro hypocholesterolemic, anti-inflammatory, and antioxidant effects of Alcalase-generated lupin protein hydrolysate (LPH). Given that lipoprotein deposition, oxidative stress, and inflammation are the main components of atherogenesis, we characterized the LPH composition, in silico identified LPH–peptides with activities related to atherosclerosis, and evaluated the in vivo LPH effects on atherosclerosis risk factors in a mouse model of atherosclerosis. After 15 min of Alcalase hydrolysis, peptides smaller than 8 kDa were obtained, and 259 peptides out of 278 peptides found showed biological activities related to atherosclerosis risk factors. Furthermore, LPH administration for 12 weeks reduced the plasma lipids, as well as the cardiovascular and atherogenic risk indexes. LPH also increased the total antioxidant capacity, decreased endothelial permeability, inflammatory response, and atherogenic markers. Therefore, this study describes for the first time that LPH prevents the early stages of atherosclerosis

Anti-obesogenic effect of lupin-derived protein hydrolysate through modulation of adiposopathy, insulin resistance and gut dysbiosis in a diet-induced obese mouse

The prevalence of obesity is increasingly widespread, resembling a global epidemic. Lifestyle changes, such as consumption of high-energy-dense diets and physical inactivity, are major contributors to obesity. Common features of this metabolic pathology involve an imbalance in lipid and glucose homeostasis including dyslipidemia, insulin resistance and adipose tissue dysfunction. Moreover, the importance of the gut microbiota in the development and susceptibility to obesity has recently been highlighted. In recent years, new strategies based on the use of functional foods, in particular bioactive peptides, have been proposed to counteract obesity outcomes. In this context, the present study examines the effects of a lupin protein hydrolysate (LPH) on obesity, dyslipidemia and gut dysbiosis in mice fed a high-fat diet (HFD). After 12 weeks of LPH treatment, mice gained less weight and showed decreased adipose dysfunction compared to the HFD-fed group. HFD-induced dyslipidemia (increased triglycerides, cholesterol and LDL concentration) and insulin resistance were both counteracted by LPH consumption. Discriminant analysis differentially distributed LPH-treated mice compared to non-treated mice. HFD reduced gut ecological parameters, promoted the blooming of deleterious taxa and reduced the abundance of commensal members. Some of these changes were corrected in the LPH group. Finally, correlation analysis suggested that changes in this microbial population could be responsible for the improvement in obesity outcomes. In conclusion, this is the first study to show the effect of LPH on improving weight gain, adiposopathy and gut dysbiosis in the context of diet-induced obesity, pointing to the therapeutic potential of bioactive peptides in metabolic diseases

Melatonin diminishes oxidative damage in sperm cells, improving assisted reproductive techniques

Los procedimientos de preparación de esperma son un generador potencial de daño en el ADN inducido por el estrés oxidativo, lo que conduce a una caída dramática en la fertilidad. Un número creciente de estudios sugiere que la melatonina reduce el estrés oxidativo inducido por la manipulación. Sin embargo, se sabe muy poco sobre el papel conservador de la melatonina en el medio de preparación de esperma durante los procedimientos de reproducción asistida. Para lograr este objetivo, el semen se dividió en dos fracciones y se preincubó con y sin melatonina 1 mM. Posteriormente, ambas fracciones se dividieron en dos subfracciones para realizar un baño en presencia y ausencia de melatonina 1 mM. El marcado con anti-CD46 y antipactivo caspasa-3 permitió el monitoreo de la reacción del acrosoma y la apoptosis por citometría de flujo. La fragmentación y la compactación del ADN espermático se analizaron mediante tinción con yoduro de propidio. Las muestras de normozoospermia y oligozoospermia que se preincubaron con melatonina experimentaron un aumento significativo en la proporción de espermatozoides adecuados y una reducción de la activación de caspasa-3. Además, la preincubación con melatonina mejoró la migración de espermatozoides con ADN compactado en muestras oligozoospérmicas (P <0.05) y evitó la fragmentación del ADN en muestras normozoospérmicas (P <0.05). A la luz de los resultados actuales, la capacidad citoprotectora y la inocuidad de la melatonina lo convierten en un gran candidato para ser aplicado en técnicas de reproducción asistida para prevenir el daño oxidativo iatrogénico.Sperm preparation procedures are a potential generator of oxidative stress-induced DNA damage, which leads to a dramatic drop in fertility. An increasing number of studies suggest that melatonin reduces the oxidative stress induced by manipulation. However, very little is known about the preservative role of melatonin in sperm preparation medium during assisted reproduction procedures. For this aim to be achieved, semen was divided into two fractions and preincubated with and without 1 mM melatonin. Afterwards, both fractions were divided into two subfractions to perform swim-up in the presence and absence of 1 mM melatonin. Labeling with anti-CD46 and antiactive caspase-3 allowed the monitoring of acrosome reaction and apoptosis by flow cytometry. Sperm DNA fragmentation and compaction were analyzed through propidium iodide staining. The normozoospermic and oligozoospermic samples that were preincubated with melatonin underwent a significant increase in the ratio of adequate spermatozoa and a reduction of caspase-3 activation. Additionally, preincubation with melatonin enhanced the migration of sperm cells with compacted DNA in oligozoospermic samples (P < 0.05) and prevented DNA fragmentation in normozoospermic samples (P < 0.05). In light of the current results, the cytoprotective capacity and innocuousness of melatonin make it a great candidate to be applied in assisted reproduction techniques in order to prevent iatrogenic oxidative damage.• Angelini Farmacéutica S.A. Ayuda • Junta de Extremadura y Fondo Social Europeo. Ayuda GR15051/BB021 • Gobierno de Extremadura y Fondo Social Europeo. Ayuda. Beca postdoctoral P01 401, para Javier Espino Palma • Fundación Tatiana Pérez de Guzmán el Bueno. Beca predoctoral para Ana María Marchena LópezpeerReviewe

Melatonin diminishes oxidative damage in sperm cells,improving assisted reproductive techniques

Los procedimientos de preparación de esperma son un generador potencial de daño en el ADN inducido por el estrés oxidativo, lo que conduce a una caída dramática en la fertilidad. Un número creciente de estudios sugiere que la melatonina reduce el estrés oxidativo inducido por la manipulación. Sin embargo, se sabe muy poco sobre el papel conservador de la melatonina en el medio de preparación de esperma durante los procedimientos de reproducción asistida. Para lograr este objetivo, el semen se dividió en dos fracciones y se preincubó con y sin melatonina 1 mM. Posteriormente, ambas fracciones se dividieron en dos subfracciones para realizar un baño en presencia y ausencia de melatonina 1 mM. El marcado con anti-CD46 y antipactivo caspasa-3 permitió el monitoreo de la reacción del acrosoma y la apoptosis por citometría de flujo. La fragmentación y la compactación del ADN espermático se analizaron mediante tinción con yoduro de propidio. Las muestras de normozoospermia y oligozoospermia que se preincubaron con melatonina experimentaron un aumento significativo en la proporción de espermatozoides adecuados y una reducción de la activación de caspasa-3. Además, la preincubación con melatonina mejoró la migración de espermatozoides con ADN compactado en muestras oligozoospérmicas (P <0.05) y evitó la fragmentación del ADN en muestras normozoospérmicas (P <0.05). A la luz de los resultados actuales, la capacidad citoprotectora y la inocuidad de la melatonina lo convierten en un gran candidato para ser aplicado en técnicas de reproducción asistida para prevenir el daño oxidativo iatrogénico.Sperm preparation procedures are a potential generator of oxidative stress-induced DNA damage, which leads to a dramatic drop in fertility. An increasing number of studies suggest that melatonin reduces the oxidative stress induced by manipulation. However, very little is known about the preservative role of melatonin in sperm preparation medium during assisted reproduction procedures. For this aim to be achieved, semen was divided into two fractions and preincubated with and without 1 mM melatonin. Afterwards, both fractions were divided into two subfractions to perform swim-up in the presence and absence of 1 mM melatonin. Labeling with anti-CD46 and antiactive caspase-3 allowed the monitoring of acrosome reaction and apoptosis by flow cytometry. Sperm DNA fragmentation and compaction were analyzed through propidium iodide staining. The normozoospermic and oligozoospermic samples that were preincubated with melatonin underwent a significant increase in the ratio of adequate spermatozoa and a reduction of caspase-3 activation. Additionally, preincubation with melatonin enhanced the migration of sperm cells with compacted DNA in oligozoospermic samples (P < 0.05) and prevented DNA fragmentation in normozoospermic samples (P < 0.05). In light of the current results, the cytoprotective capacity and innocuousness of melatonin make it a great candidate to be applied in assisted reproduction techniques in order to prevent iatrogenic oxidative damage.• Angelini Farmacéutica S.A. Ayuda • Junta de Extremadura y Fondo Social Europeo. Ayuda GR15051/BB021 • Gobierno de Extremadura y Fondo Social Europeo. Ayuda. Beca postdoctoral P01 401, para Javier Espino Palma • Fundación Tatiana Pérez de Guzmán el Bueno. Beca predoctoral para Ana María Marchena LópezpeerReviewe

Autophagy-related proteins are functionally active in human spermatozoa and may be involved in the regulation of cell survival and motility

La macroautofagia (en lo sucesivo denominado autofagia) es un proceso conservador de las células que participa alta y evolutivamente en el mantenimiento de la homeostasis intracelular a través de la degradación de proteínas más perdurables y de los orgánulos. La autofagia participa en algunos eventos reproductivos; sin embargo, no hay información sobre la función de la autofagia en la regulación de la fisiología del espermatozoide. Por lo tanto, el objetivo de este estudio es investigar si las proteínas en relación con la autofagía están presentes y son funcionalmente activas en espermatozoides humanos. Las proteínas relacionadas con la autofagia/proceso mitofágico (LC3, ATG5, ATG16, Beclin 1, p62, m-TOR, AMPKα 1/2 y PINK1) están presentes en espermatozoides humanos. El LC3 está colocalizado con la p62 en la parte central de los espermatozoides. La activación de la autofagia induce un aumento significativo de la motilidad y una disminución en el PINK1, en la proteína Tom20 y la activación de caspasa 3/7. Sin embargo, la inhibición de la autofagia causó una disminución de la motilidad, viabilidad, ATP y la concentración de calcio intracelular mientras PINK1, proteína Tom20, fosforilación AMPK y activación de la caspasa 3/7 aumentaron considerablemente. En conclusión nuestros resultados demuestran que la autofagia en relación con proteínas y reguladores upstream están presentes y activos en espermatozoides humanos. La modificación de proteínas mitocondriales después de la autofagia activación/inhibición puede estar indicando una forma especializada de la autofagia denominado mitofagía pueden estar regulando la función espermática como motilidad, su viabilidad y pueden estar cooperando con la apoptosis.Macroautophagy (hereafter autophagy) is an evolutionarily highly conserved cellular process that participates in the maintenance of intracellular homeostasis through the degradation of most longlived proteins and entire organelles. Autophagy participates in some reproductive events; however, there are not reports regarding the role of autophagy in the regulation of sperm physiology. Hence, the aim of this study was to investigate whether autophagy-related proteins are present and functionally active in human spermatozoa. Proteins related to autophagy/mitophagy process (LC3, Atg5, Atg16, Beclin 1, p62, m-TOR, AMPKα 1/2, and PINK1) were present in human spermatozoa. LC3 colocalized with p62 in the middle piece of the spermatozoa. Autophagy activation induced a significant increase in motility and a decrease in PINK1, TOM20 expression and caspase 3/7 activation. In contrast, autophagy inhibition resulted in decreased motility, viability, ATP and intracellular calcium concentration whereas PINK1, TOM20 expression, AMPK phosphorylation and caspase 3/7 activation were significantly increased. In conclusion our results show that autophagy related proteins and upstream regulators are present and functional in human spermatozoa. Modification of mitochondrial proteins expression after autophagy activation/inhibition may be indicating that a specialized form of autophagy named mitophagy may be regulating sperm function such as motility and viability and may be cooperating with apoptosis.• Gobierno de Extremadura y Fondos FEDER. Ayudas DE12006, GR10010 y PCE1002 • Ministerio de Ciencia e Innovación-FEDER. Proyectos BFU2011-30261 y AGL2013-43211-R (I+D+i)peerReviewe

Colombian surgical outcomes study insights on perioperative mortality rate, a main indicator of the lancet commission on global surgery – a prospective cohort studyResearch in context

Summary: Background: Surgical care holds significant importance in healthcare, especially in low and middle-income countries, as at least 50% of the 4.2 million deaths within the initial 30 days following surgery take place in these countries. The Lancet Commission on Global Surgery proposed six indicators to enhance surgical care. In Colombia, studies have been made using secondary data. However, strategies to reduce perioperative mortality have not been implemented. This study aims to describe the fourth indicator, perioperative mortality rate (POMR), with primary data in Colombia. Methods: A multicentre prospective cohort study was conducted across 54 centres (hospitals) in Colombia. Each centre selected a 7-day recruitment period between 05/2022 and 01/2023. Inclusion criteria involved patients over 18 years of age undergoing surgical procedures in operating rooms. Data quality was ensured through a verification guideline and statistical analysis using mixed-effects multilevel modelling with a case mix analysis of mortality by procedure-related, patient-related, and hospital-related conditions. Findings: 3807 patients were included with a median age of 48 (IQR 32–64), 80.3% were classified as ASA I or II, and 27% of the procedures had a low-surgical complexity. Leading procedures were Orthopedics (19.2%) and Gynaecology/Obstetrics (17.7%). According to the Clavien–Dindo scale, postoperative complications were distributed in major complications (11.7%, 10.68–12.76) and any complication (31.6%, 30.09–33.07). POMR stood at 1.9% (1.48–2.37), with elective and emergency surgery mortalities at 0.7% (0.40–1.23) and 3% (2.3–3.89) respectively. Interpretation: The POMR was higher than the ratio reported in previous national studies, even when patients had a low–risk profile and low-complexity procedures. The present research represents significant public health progress with valuable insights for national decision-makers to improve the quality of surgical care. Funding: This work was supported by Universidad del Rosario and Fundación Cardioinfantil-Instituto de Cardiología grant number CTO-057-2021, project-ID IV-FGV017