Daniel Salete-Granado

Background: MicroRNAs (miRNAs) are a subclass of small non-coding RNAs that regulate gene expression and play important roles in infectious diseases. Echinococcosis is a globally distributed disease caused by various species of Echinococcus . The disease can be asymptomatic in the early stages, but if left untreated, it can lead to severe complications. Like other helminths, Echinococcus spp. exhibit immunomodulatory capacities that favour infection establishment and progression. Currently, there is no fully accurate, non-invasive method for early detection of echinococcosis. This systematic review provides a comprehensive analysis of the existing literature and discusses recent advances in the study of miRNAs during host– Echinococcus spp. interactions, emphasizing their intriguing role in immunomodulation and their potential as biomarkers for detection, disease activity, prognosis, and therapeutic monitoring, as well as their possible relevance for future therapeutic research. Methods: This systematic review was conducted in accordance with the PRISMA 2020 guidelines. Searches were performed in PubMed, Web of Science, and Scopus without date restrictions to identify studies assessing the diagnostic potential of miRNAs in cystic echinococcosis (CE) and alveolar echinococcosis (AE). Fourteen original research articles were included in the final analysis. Results: Parasite-derived miRNAs have been detected in plasma, serum, and liver tissue, with egr-let-7-5p and egr-miR-71-5p emerging as candidates for CE detection and monitoring. In CE, dysregulation of host miRNAs was observed, displaying cyst activity-dependent patterns associated with inflammation, immunomodulation, and fibrosis, including a consistent upregulation of hsa-miR-146a-5p in plasma and serum samples. In AE, several host miRNAs exhibited altered expression in liver tissue and plasma, with hsa-miR-483-3p consistently upregulated. Comparative analyses revealed partially distinct but overlapping miRNA profiles in CE and AE, reflecting host-driven regulatory responses related to immunomodulation and tissue remodeling with stage-dependent patterns. Most included studies were based on case–control designs, with small sample sizes and limited validation, suggesting high overall risk of bias. Conclusions: Current evidence suggests that miRNAs have translational potential as complementary biomarkers for diagnosis, patient stratification, prognostic assessment, and disease monitoring in echinococcosis and may support future therapeutic research. Nonetheless, methodological heterogeneity and limited validation may contribute to overestimating diagnostic performance and hinder clinical translation.

Alcohol use disorder (AUD) is influenced by genetic factors that affect key neurobiological systems, including dopaminergic and GABAergic pathways, which regulate neurobehavioral functions and are modulated by brain-derived neurotrophic factor (BDNF). Variations in these genes contribute to individual vulnerability to AUD. In this study, we investigated single-nucleotide polymorphisms (SNPs) and haplotypic associations in GABRA1, GABRA2, and GABRA6, along with the dopaminergic pathway genes DRD2/ANKK1 and BDNF, in a Spanish cohort. Peripheral blood-derived genomic DNA was genotyped, and haplotype analyses were conducted. Individual SNPs in GABRA1, GABRA2, BDNF, and DRD2/ANKK1 showed no significant associations with AUD. In GABRA6, the rs3219151 T allele was more frequent in AUD patients than in controls (57.9% vs. 49.3%; p = 0.03), while the C allele appeared to show a potential protective association. In addition, the GAC haplotype of GABRA6 (rs2197414, rs1992647, rs3219151) was less frequent in AUD than in controls (0.071 vs. 0.122) and showed a protective association (OR = 0.58; 95% CI = 0.34–0.99; p = 0.045). Our findings provide exploratory evidence suggesting that specific genetic variants and haplotypes may contribute to AUD susceptibility and support the relevance of multigenic and haplotypic approaches for exploring the neurobiological mechanisms underlying AUD.

Chitinases, a group of glycosyl hydrolases (GHs), catalyze the degradation of chitin by releasing N-acetylglucosamine subunits. GHs can be found in all three domains of life. Among the three GH families (GH18, GH19, and GH20), GH18 chitinases are the most conserved and extensively studied. These enzymes have been implicated in nutrition, immune modulation, invasion, and virulence across diverse pathogens. In some parasitic protists, GH18 chitinases are essential for transmission. However, in kinetoplastids, including Leishmania spp., these enzymes remain poorly characterized. This study aimed to identify and characterize chitinase across kinetoplastids, with a particular focus on GH18 chitinase in Leishmania braziliensis , the causative agent of muco-cutaneous leishmaniasis. A bioinformatic pipeline was implemented to retrieve and annotate chitinase genes from multiple databases. Catalytic domains and subcellular localization were identified using dedicated computational tools. Phylogenetic relationships were reconstructed in MEGA12 using maximum likelihood and neighbor-joining methods. RNA-seq data were analyzed to evaluate stage-specific expression. Structural models of L. braziliensis GH18 chitinase (Lbr_ChGH18) were created with AlphaFold and subsequently refined. We identified GH18 chitinases genes across Leishmania species and other kinetoplastids, together with GH19 and GH20 chitinase genes. Lbr_ChGH18 was detected across all life-cycle stages, with peak levels in amastigotes. Docking analyses identified closantel, argifin, and argadin as potential inhibitors of Lbr_ChGH18. Epitope prediction revealed conserved B- and T-cell epitopes in GH18 chitinases from Old and New World Leishmania , capable of binding multiple HLA class I and II molecules, highlighting their potential as diagnostic and vaccine candidates. The discovery of GH20 chitinase-like genes in Trypanosoma species offers new insights into the evolution and functional diversification of GHs in kinetoplastids. The high conservation and expression of GH18 chitinases underscore their potential as promising targets for drug and vaccine development against leishmaniasis.

Alcohol exposure affects immune regulation and tissue homeostasis. Antimicrobial peptides (AMPs) are essential components of innate immunity, not only defending against pathogens but also modulating processes such as inflammation. However, their tissue-specific regulation in response to alcohol remains poorly characterized, particularly in humans after acute intoxication. We evaluated the expression of AMPs in the peripheral blood of patients with alcohol use disorder (AUD, n = 9), individuals with acute alcohol consumption (AAC, n = 9), and controls using quantitative polymerase chain reaction (qPCR). Additionally, we analyzed AMP expression in selected tissues of mice exposed to chronic ethanol feeding (National Institute on Alcohol Abuse and Alcoholism model for 5 days) and performed a systematic review of AMP regulation in alcohol-related disorders (2005–2025; n = 36 studies, reflecting a limited and heterogeneous body of available evidence). Human cathelicidin antimicrobial peptide (LL-37), lipopolysaccharide-binding protein (LBP), and bactericidal/permeability-increasing protein (BPI) were significantly upregulated in patients with AUD, whereas LL-37 and LBP were significantly upregulated in AAC. In the livers of ethanol-fed mice, LEP2, LCN2, and LBP levels were markedly increased, whereas LL-37 and LEP1 were downregulated. Duodenal tissue exhibited upregulation of DEFB1. In adipose tissue, DEFA2 was significantly increased in peripheral depots, whereas only LCN2 was upregulated in brain tissue. The systematic review demonstrated complex, heterogeneous, and organ-dependent AMP regulation and also highlighted the paucity of human data on AAC, a gap that our study partially addresses. Our results are consistent with the hypothesis that selected AMPs may serve as candidate markers of organ damage or microbial translocation and as possible therapeutic targets, a hypothesis that requires confirmation in larger, adequately powered studies.

Glycolysis is primarily involved in ATP production but also modulates oxidative stress. Chronic alcohol consumption is correlated with an increased incidence of multiple diseases, including cancer and neurodegenerative diseases (NDDs), though the underlying mechanisms remain unclear. Guided by a literature review and bioinformatics analysis, we evaluated the expression of 22 genes encoding various isoforms of seven glycolytic enzymes (GEs) in the peripheral blood of patients with alcohol use disorder (AUD), individuals with acute alcohol consumption (AAC), and their respective control groups using qPCR. In parallel, we evaluated the expression of selected genes coding for GEs linked to NDDs, as well as astrocytic markers in primary mouse astrocyte cultures exposed to ethanol. Thirteen GE-related genes, including non-canonical isoforms, were significantly dysregulated in AUD patients; notably, eight of these genes showed similar alterations in individuals with AAC. Several enzymes encoded by these genes are known to be regulated by oxidative stress. Ethanol-exposed astrocytes also showed altered expression of glycolytic genes associated with NDDs and astrocyte function. These findings indicate that glycolytic dysregulation is driven by ethanol intake, regardless of exposure duration or organic damage, highlighting a link between ethanol-driven redox imbalance and glycolytic remodeling, which could contribute to organ damage.

Glycolysis, present in most organisms, is evolutionarily one of the oldest metabolic pathways. It has great relevance at a physiological level because it is responsible for generating ATP in the cell through the conversion of glucose into pyruvate and reducing nicotinamide adenine dinucleotide (NADH) (that may be fed into the electron chain in the mitochondria to produce additional ATP by oxidative phosphorylation), as well as for producing intermediates that can serve as substrates for other metabolic processes. Glycolysis takes place through 10 consecutive chemical reactions, each of which is catalysed by a specific enzyme. Although energy transduction by glucose metabolism is the main function of this pathway, involvement in virulence, growth, pathogen–host interactions, immunomodulation and adaptation to environmental conditions are other functions attributed to this metabolic pathway. In humans, where glycolysis occurs mainly in the cytosol, the mislocalization of some glycolytic enzymes in various other subcellular locations, as well as alterations in their expression and regulation, has been associated with the development and progression of various diseases. In this review, we describe the role of glycolytic enzymes in the pathogenesis of diseases of clinical interest. In addition, the potential role of these enzymes as targets for drug development and their potential for use as diagnostic and prognostic markers of some pathologies are also discussed.

El grupo de Alcohol, Metabolismo y Sistema Inmune es un grupo multidisciplinar ubicado en el Departamento de Medicina de la Facultad de Medicina y en el Servicio de Medicina Interna del Hospital Universitario de Salamanca. El investigador principal es el doctor Miguel Marcos y tenemos colaboraciones con grupos pertenecientes a centros relevantes en el ámbito nacional e internacional como CNIO, IBFG y CIC.

Excessive alcohol consumption impairs the immune system, induces oxidative stress, and triggers the activation of peripheral blood (PB) monocytes, thereby contributing to alcoholic liver disease (ALD). We analyzed the M1/M2 phenotypes of circulating classical monocytes and macrophage-derived monocytes (MDMs) in excessive alcohol drinkers (EADs). PB samples from 20 EADs and 22 healthy controls were collected for isolation of CD14+ monocytes and short-term culture with LPS/IFNγ, IL4/IL13, or without stimulation. These conditions were also used to polarize MDMs into M1, M2, or M0 phenotypes. Cytokine production was assessed in the blood and culture supernatants. M1/M2-related markers were analyzed using mRNA expression and surface marker detection. Additionally, the miRNA profile of CD14+ monocytes was analyzed. PB samples from EADs exhibited increased levels of pro-inflammatory cytokines. Following short-term culture, unstimulated blood samples from EADs showed higher levels of soluble TNF-α and IL-8, whereas monocytes expressed increased levels of surface TNF-α and elevated mRNA expression of pro-inflammatory cytokines and inducible nitric oxide synthase. MDMs from EADs showed higher levels of TNF-α and CD206 surface markers and increased IL-10 production. LPS/IFNγ induced higher mRNA expression of Nrf2 only in the controls. miRNA analysis revealed a distinctive miRNA profile that is potentially associated with liver carcinogenesis and ALD through inflammation and oxidative stress. This study confirms the predominantly pro-inflammatory profile of PB monocytes among EADs and suggests immune exhaustion features in MDMs.

Ethanol consumption triggers oxidative stress by generating reactive oxygen species (ROS) through its metabolites. This process leads to steatosis and liver inflammation, which are critical for the development of alcoholic liver disease (ALD). Autophagy is a regulated dynamic process that sequesters damaged and excess cytoplasmic organelles for lysosomal degradation and may counteract the harmful effects of ROS-induced oxidative stress. These effects include hepatotoxicity, mitochondrial damage, steatosis, endoplasmic reticulum stress, inflammation, and iron overload. In liver diseases, particularly ALD, macroautophagy has been implicated as a protective mechanism in hepatocytes, although it does not appear to play the same role in stellate cells. Beyond the liver, autophagy may also mitigate the harmful effects of alcohol on other organs, thereby providing an additional layer of protection against ALD. This protective potential is further supported by studies showing that drugs that interact with autophagy, such as rapamycin, can prevent ALD development in animal models. This systematic review presents a comprehensive analysis of the literature, focusing on the role of autophagy in oxidative stress regulation, its involvement in organ–organ crosstalk relevant to ALD, and the potential of autophagy-targeting therapeutic strategies.