Effects of Virgin Olive Oil on Blood Pressure and Renal Aminopeptidase Activities in Male Wistar Rats

High saturated fat diets have been associated with the development of obesity and hypertension, along with other pathologies related to the metabolic syndrome. In contrast, the Mediterranean diet, characterized by its high content of monounsaturated fatty acids, has been proposed as a dietary factor capable of positively regulating cardiovascular function. These effects have been linked to changes in the local renal renin angiotensin system (RAS) and the activity of the sympathetic nervous system. The main goal of this study was to analyze the role of two dietary fat sources on aminopeptidases activities involved in local kidney RAS. Male Wistar rats (six months old) were fed during 24 weeks with three different diets: the standard diet (S), the standard diet supplemented with virgin olive oil (20%) (VOO), or the standard diet enriched with butter (20%) plus cholesterol (0.1%) (Bch). Kidney samples were separated in medulla and cortex for aminopeptidase activities (AP) assay. Urine samples were collected for routine analysis by chemical tests. Aminopeptidase activities were determined by fluorometric methods in soluble (sol) and membrane-bound (mb) fractions of renal tissue, using arylamide derivatives as substrates. After the experimental period, the systolic blood pressure (SBP) values were similar in standard and VOO animals, and significantly lower than in the Bch group. At the same time, a significant increase in GluAP and IRAP activities were found in renal medulla of Bch animals. However, in VOO group the increase of GluAP activity in renal medulla was lower, while AspAP activity decreased in the renal cortex. Furthermore, the VOO diet also affected other aminopeptidase activities, such as TyrAP and pGluAP, related to the regulation of the sympathetic nervous system and the metabolic rate. These results support the beneficial effect of VOO in the regulation of SBP through changes in local AP activities of the kidney.ACCIÓN 1 PAIUJA 2019 2020: BIO22

The Role of High Fat Diets and Liver Peptidase Activity in the Development of Obesity and Insulin Resistance in Wistar Rats

Technical and human support provided by CICT and CEPA of Universidad de Jaén (UJA, MINECO, Junta de Andalucía; FEDER) is gratefully acknowledged.High-fat diets (HFD) have been widely associated with an increased risk of metabolic disorders and overweight. However, a high intake of sources that are rich in monounsaturated fatty acids has been suggested as a dietary agent that is able to positively influence energy metabolism and vascular function. The main objective of this study was to analyze the role of dietary fats on hepatic peptidases activities and metabolic disorders. Three diets: standard (S), HFD supplemented with virgin olive oil (VOO), and HFD supplemented with butter plus cholesterol (Bch), were administered over six months to male Wistar rats. Plasma and liver samples were collected for clinical biochemistry and aminopeptidase activities (AP) analysis. The expression of inducible nitric oxide synthase (iNOS) was also determined by Western blot in liver samples. The diet supplement with VOO did not induce obesity, in contrast to the Bch group. Though the VOO diet increased the time that was needed to return to the basal levels of plasma glucose, the fasting insulin/glucose ratio and HOMA2-%B index (a homeostasis model index of insulin secretion and valuation of β-cell usefulness (% β-cell secretion)) were improved. An increase of hepatic membrane-bound dipeptidyl-peptidase 4 (DPP4) activity was found only in VOO rats, even if no differences in fasting plasma glucagon-like peptide 1 (GLP-1) were obtained. Both HFDs induced changes in hepatic pyroglutamyl-AP in the soluble fraction, but only the Bch diet increased the soluble tyrosyl-AP. Angiotensinase activities that are implicated in the metabolism of angiotensin II (AngII) to AngIV increased in the VOO diet, which was in agreement with the higher activity of insulin-regulated-AP (IRAP) in this group. Otherwise, the diet that was enriched with butter increased soluble gamma-glutamyl transferase (GGT) and Leucyl-AP, iNOS expression in the liver, and plasma NO. In summary, VOO increased the hepatic activity of AP that were related to glucose metabolism (DPP4, angiotensinases, and IRAP). However, the Bch diet increased activities that are implicated in the control of food intake (Tyrosine-AP), the index of hepatic damage (Leucine-AP and GGT), and the expression of hepatic iNOS and plasma NO. Taken together, these results support that the source of fat in the diet affects several peptidases activities in the liver, which could be related to alterations in feeding behavior and glucose metabolism.This work was supported by grant code: ACCIÓN 1 PAIUJA 2019 2020: BIO221

Asymmetric Pattern of Correlations of Leucine Aminopeptidase Activity between Left or Right Frontal Cortex versus Diverse Left or Right Regions of Rat Brains

Previous studies demonstrated an asymmetry of left predominance for mean values of soluble leucine aminopeptidase (LeuAP) activity in the frontal cortex (FC) and hypothalamus of adult male rats, fluorimetrically analyzed by the hydrolysis of Leu- -naphthylamide as a substrate. No asymmetries were observed in nine other left (L) and right (R) regions obtained from rostrocaudally sectioned coronal slices. Neither had inter-hemispheric differences observed for lactate dehydrogenase (LDH), analyzed simultaneously in the same brain regions (L and R) of the same animals. However, the level of intra-hemispheric or inter-hemispheric correlation of LeuAP or LDH between such brain regions has not been analyzed. In order to obtain additional suggestions on the functional heterogeneity between regions of LeuAP and LDH, in the present investigation, the level of intra-hemispheric and inter-hemispheric correlations of the frontal cortex with the rest of the regions studied is described: (A) between the left frontal cortex (LFC) and the rest of the left regions; (B) between the right frontal cortex (RFC) and the rest of the right regions; (C) between the left frontal cortex and all of the right regions; and (D) between the right frontal cortex and all of the left regions. All of the correlations obtained were positive. The intra-hemispheric analysis showed a greater heterogeneity of values in the correlations observed between RFC and the rest of the right regions than between LFC and the rest of the left regions. Greater heterogeneity is observed when comparing RFC correlations with left regions than when comparing LFC correlations with right regions. In conclusion, the greatest heterogeneity (suggesting a greater functional variability) was observed in the right intra-hemispheric analysis and in the inter-hemispheric analysis between the RFC and the left hemisphere. The results for LDH showed a great homogeneity between regions both in the intra- and inter-hemispheric studies.University of Jaén “Neuroendocrinology and Nutrition” BIO-22

Brain Asymmetry: Towards an Asymmetrical Neurovisceral Integration

This review was supported by the group of the University of Jaen "Neuroendocrinology and Nutrition" BIO-221.Despite the ancestral evidence of an asymmetry in motor predominance, going through the inspiring discoveries of Broca and Wernicke on the localization of language processing, continuing with the subsequent noise coinciding with the study of brain function in commissurotomized patients—and the subsequent avalanche of data on the asymmetric distribution of multiple types of neurotransmitters in physiological and pathological conditions—even today, the functional significance of brain asymmetry is still unknown. Currently, multiple evidence suggests that functional asymmetries must have a neurochemical substrate and that brain asymmetry is not a static concept but rather a dynamic one, with intra- and inter-hemispheric interactions between its various processes, and that it is modifiable depending on changing endogenous and environmental conditions. Furthermore, based on the concept of neurovisceral integration in the overall functioning of an organism, some evidence has emerged suggesting that this integration could be organized asymmetrically, using the autonomic nervous system as a bidirectional communication pathway, whose performance would also be asymmetric. However, the functional significance of this distribution, as well as the evolutionary advantage of an asymmetric nervous organization, is still unknown.University of Jaen "Neuroendocrinology and Nutrition" BIO-22

High-Fat Diets Modify the Proteolytic Activities of Dipeptidyl-Peptidase IV and the Regulatory Enzymes of the Renin–Angiotensin System in Cardiovascular Tissues of Adult Wistar Rats

(1) Background: The replacement of diets high in saturated fat (SAFA) with monounsaturated fatty acids (MUFA) is associated with better cardiovascular function and is related to the modulation of the activity of the local renin–angiotensin system (RAS) and the collagenase activity of dipeptidyl peptidase IV (DPP-IV). The objective of the work was to verify the capacity of different types of dietary fat on the regulatory activities of RAS and DPP-IV. (2) Methods: Male Wistar rats were fed for 24 weeks with three different diets: the standard diet (S), the standard diet supplemented with virgin olive oil (20%) (VOO), or with butter (20%) plus cholesterol (0.1%) (Bch). The proteolytic activities were determined by fluorometric methods in the soluble (sol) and membrane-bound (mb) fractions of the left ventricle and atrium, aorta, and plasma samples. (3) Results: With the VOO diet, angiotensinase values were significantly lower than with the Bch diet in the aorta (GluAP and ArgAP (mb)), ventricle (ArgAP (mb)) and atrium (CysAP (sol)). Significant decreases in DPP-IV (mb) activity occurred with the Bch diet in the atrium and aorta. The VOO diet significantly reduced the activity of the cardiac damage marker LeuAP (mb) in the ventricle and aorta, except for LeuAP (sol) in the ventricle, which was reduced with the Bch diet. (4) Conclusions: The introduction into the diet of a source rich in MUFA would have a beneficial cardiovascular effect on RAS homeostasis and cardiovascular functional stability.ACCIÓN 1 PAIUJA 2019 2020: BIO221

Correlational Study of Aminopeptidase Activities between Left or Right Frontal Cortex versus the Hypothalamus, Pituitary, Adrenal Axis of Spontaneously Hypertensive Rats Treated with Hypotensive or Hypertensive Agents

Supplementary Materials: The supporting information can be downloaded at: https://www.mdpi. com/article/10.3390/ijms242116007/s1It has been suggested that the neuro-visceral integration works asymmetrically and that this asymmetry is dynamic and modifiable by physio-pathological influences. Aminopeptidases of the renin–angiotensin system (angiotensinases) have been shown to be modifiable under such conditions. This article analyzes the interactions of these angiotensinases between the left or right frontal cortex (FC) and the same enzymes in the hypothalamus (HT), pituitary (PT), adrenal (AD) axis (HPA) in control spontaneously hypertensive rats (SHR), in SHR treated with a hypotensive agent in the form of captopril (an angiotensin-converting enzyme inhibitor), and in SHR treated with a hypertensive agent in the form of the L-Arginine hypertensive analogue L-NG-Nitroarginine Methyl Ester (L-NAME). In the control SHR, there were significant negative correlations between the right FC with HPA and positive correlations between the left FC and HPA. In the captopril group, the predominance of negative correlations between the right FC and HPA and positive correlations between the HPA and left FC was maintained. In the L-NAME group, a radical change in all types of interactions was observed; particularly, there was an inversion in the predominance of negative correlations between the HPA and left FC. These results indicated a better balance of neuro-visceral interactions after captopril treatment and an increase in these interactions in the hypertensive animals, especially in those treated with L-NAME.Ministry of Science and Innovation SAF 2008 04685 C02 0

Asymmetric Interaction of Neuropeptidase Activities between Cortico-Limbic Structures, Plasma and Cardiovascular Function after Unilateral Dopamine Depletions of the Nigrostriatal System

This work was supported by Junta de Andalucia through the research groups PAI CVI-221 (Peptides and Peptidases) and CTS 438 (Group for Neurological Diseases Research in Southern Spain).In emotional processing, dopamine (DA) plays an essential role, and its deterioration involves important consequences. Under physiological conditions, dopamine exhibits brain asymmetry and coexists with various neuropeptides that can coordinate the processing of brain functions. Brain asymmetry can extend into a broader concept of asymmetric neurovisceral integration, including behavior. The study of the activity of neuropeptide regulatory enzymes (neuropeptidases, NPs) is illustrative. We have observed that the left and right brain areas interact intra- and interhemispherically, as well as with peripheral tissues or with physiological parameters such as blood pressure or with behaviors such as turning preference. To obtain data that reflect this integrative behavior, we simultaneously analyzed the impact of left or right brain DA depletion on the activity of various NPs in corticolimbic regions of the left and right hemispheres, such as the medial prefrontal cortex, amygdala and hippocampus, as well as on the plasma activity of the same aminopeptidase activities (APs) and on systolic blood pressure (SBP). Intra- and inter-hemispheric interactions as well as the interactions of NPs from the left or right hemispheres were analyzed with the same plasma APs and the SBP obtained from sham and from left or right lesioned rats. The results demonstrate a complex profile depending on the hemisphere considered. They definitively confirm an asymmetric neurovisceral integration and reveal a higher level of inter-hemispheric corticolimbic interactions including with SBP after left dopamine depletion.Junta de Andalucia PAI CVI-221 CTS 43

Aminopeptidase Activities Interact Asymmetrically between Brain, Plasma and Systolic Blood Pressure in Hypertensive Rats Unilaterally Depleted of Dopamine

Brain dopamine, in relation to the limbic system, is involved in cognition and emotion. These functions are asymmetrically processed. Hypertension not only alters such functions but also their asymmetric brain pattern as well as their bilateral pattern of neurovisceral integration. The central and peripheral renin-angiotensin systems, particularly the aminopeptidases involved in its enzymatic cascade, play an important role in blood pressure control. In the present study, we report how these aminopeptidases from left and right cortico-limbic locations, plasma and systolic blood pressure interact among them in spontaneously hypertensive rats (SHR) unilaterally depleted of dopamine. The study comprises left and right sham and left and right lesioned (dopaminedepleted) rats as research groups. Results revealed important differences in the bilateral behavior comparing sham left versus sham right, lesioned left versus lesioned right, and sham versus lesioned animals. Results also suggest an important role for the asymmetrical functioning of the amygdala in cardiovascular control and an asymmetrical behavior in the interaction between the medial prefrontal cortex, hippocampus and amygdala with plasma, depending on the left or right depletion of dopamine. Compared with previous results of a similar study inWistar-Kyoto (WKY) normotensive rats, the asymmetrical behaviors differ significantly between both WKY and SHR strains.Junta de Andalucia PAI CVI-221 CTS 43

Systematic Review and Meta-Analyses of Aminopeptidases as Prognostic Biomarkers in Amyotrophic Lateral Sclerosis

Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disease characterized by the progressive loss of motor neurons in the spinal cord, brain stem, and cerebral cortex. Biomarkers for ALS are essential for disease detection and to provide information on potential therapeutic targets. Aminopeptidases catalyze the cleavage of amino acids from the amino terminus of protein or substrates such as neuropeptides. Since certain aminopeptidases are known to increase the risk of neurodegeneration, such mechanisms may reveal new targets to determine their association with ALS risk and their interest as a diagnostic biomarker. The authors performed a systematic review and meta-analyses of genome-wide association studies (GWASs) to identify reported aminopeptidases genetic loci associated with the risk of ALS. PubMed, Scopus, CINAHL, ISI Web of Science, ProQuest, LILACS, and Cochrane databases were searched to retrieve eligible studies in English or Spanish, published up to 27 January 2023. A total of 16 studies were included in this systematic review, where a series of aminopeptidases could be related to ALS and could be promising biomarkers (DPP1, DPP2, DPP4, LeuAP, pGluAP, and PSA/NPEPPS). The literature reported the association of singlenucleotide polymorphisms (SNPs: rs10260404 and rs17174381) with the risk of ALS. The genetic variation rs10260404 in the DPP6 gene was identified to be highly associated with ALS susceptibility, but meta-analyses of genotypes in five studies in a matched cohort of different ancestry (1873 cases and 1861 control subjects) showed no ALS risk association. Meta-analyses of eight studies for minor allele frequency (MAF) also found no ALS association for the “C” allele. The systematic review identified aminopeptidases as possible biomarkers. However, the meta-analyses for rs1060404 of DPP6 do not show a risk associated with ALS.PPJIA2022.09, “Plan Propio de Investigación y Transferencia de la Universidad de Granada2022: Programa 20Proyectos de Investigación Precompetitivos para Jóvenes Investigadores. Modalidad 20.aProyectos para jóvenes doctores” corresponding to the year 2022 with reference no. 11 (File no. 1586950)University of Granada (Spain). Accepted: 23 December 202

Técnicas basadas en las tecnologías de Bioréplicas impresas en 3D y Realidad Aumentada de alteraciones anatómicas y disfunciones fisiológicas para la mejora docente en las asignaturas de Anatomía y Fisiología

Memoria final de la CONVOCATORIA DE PROYECTOS DE INNOVACIÓN Y BUENAS PRÁCTICAS DOCENTES (PIBD Fase I), 2020/2021. Este proyecto tiene como objetivos revelar los factores que ayudan a integrar el binomio enseñanza-aprendizaje empleando las tecnologías de realidad aumentada (RA) y el modelaje de bioréplicas por renderización (a través de segmentación, fotogrametría o diseño asistido por ordenador), preferiblemente a través de imágenes digitales de pacientes reales o colaboradores, y/o deposición fundida (FDM). Los entornos de aprendizaje elegidos fueron las asignaturas del departamento de fisiología en el Campus de Ceuta ("Anatomía Humana" y "Fisiología I"), impartidas en el grado de enfermería de la Universidad de Granada. Junto con las clases tradicionales, en prácticas y seminarios se complementó la docencia tradicional con la tecnología emergente para comprobar los resultados que podrían tener en el aprendizaje del discente, además de comprobar su apreciación ante la utilidad percibida y satisfacción. Las encuestas utilizadas fueron de tipo Likert, con valoraciones del 1 al 7, siendo el valor de 1 el más desfavorable. El modelo de aceptación tecnológica (TAM) se pasó a los discentes antes y después de la experiencia. Solo la encuesta de motivación de materiales de instrucción (IMMS) se pasó al finalizar el curso. Además, se valoró el conocimiento mediante una prueba de evaluación al finalizar las asignaturas. Los resultados mostraron que ambas tecnologías eran igual de eficientes, mejorando el aprendizaje de las materias. Además, estas técnicas permite que los discentes de primer año de grado puedan correlacionar los conceptos teóricos con modelos visuales reales. De esa manera profundizan en conceptos difíciles de explicar de manera tradicional, al no permitir su visualización y creación de una imagen mental, y por otro lado porque no tendrían acceso hasta último año de grado, situación donde ocurre un acercamiento real a los pacientes del hospital. ----- This project aims to reveal the factors that help to integrate the teaching-learning binomial using augmented reality (AR) technologies and the modeling of bioreplicas by rendering (through segmentation, photogrammetry or computer-aided design), preferably through of digital images of real patients or collaborators, and/or fused deposition (FDM). The chosen learning environments were the subjects of the physiology department at the Ceuta Campus ("Human Anatomy" and "Physiology I"), taught in the nursing degree at the University of Granada. Along with traditional classes, practices and seminars, traditional teaching was complemented with emerging technology to verify the results that they could have in the student's learning, in addition to checking their appreciation of the perceived usefulness and satisfaction. The surveys used were of the Likert type, with ratings from 1 to 7, with a value of 1 being the most unfavorable. The technology acceptance model (TAM) was passed to the students before and after the experience. Only the Instructional Materials Motivation Survey (IMMS) was passed at the end of the course. In addition, knowledge was assessed through an evaluation test at the end of the subjects. The results showed that both technologies were equally efficient, improving the learning of the subjects. In addition, these techniques allow first-year students to correlate theoretical concepts with real visual models. In this way, they delve into concepts that are difficult to explain in a traditional way, by not allowing their visualization and creation of a mental image, and on the other hand because they would not have access until the last year of degree, a situation where a real approach to hospital patients occurs.Departamento de Fisiología