Participation of Ca2+-calmodulin dependent protein kinase II in the antidepressant like effects of melatonin.

Melatonin (N-acetyl-5 methoxytriptamine) is an indolamine secreted by the pineal gland during the dark phase of the photoperiod. Its main function is the synchronization of different body rhythms with the dark-light cycle. Research on melatonin has significantly advanced since its discovery and we now know that it has considerable significance in various physiological processes, including immunity, aging, and reproduction. Moreover, in recent years evidence of the pharmacological possibilities of melatonin has increased. The indolamine, on the other hand, has antidepressant-like effects in rodents, which may be mediated by the activation of CaMKII and are also related to the regulation of neuroplasticity processes, including neurogenesis, synaptic maintenance, and long-term potentiation. Remarkably, patients with major depression show decreased levels of circulating melatonin in plasma. This review presents evidence of the antidepressant-like effects of melatonin in preclinical models, and the participation of CaMKII in these actions. CaMKII's role in cognition and memory processes, which are altered in depressive states, are part of the review and the effects of melatonin in these processes are also reviewed. Furthermore, participation of CaMKII on structural and synaptic plasticity and the effects of melatonin are also described. Finally, the advantages of using melatonin in combination with other antidepressants such as ketamine for neuroplasticity are described. Evidence supports that CaMKII is activated by melatonin, downstream melatonin receptors and may be the common effector in the synergistic effects of melatonin with other antidepressants. Significance Statement This review compiled evidence in support that melatonin causes antidepressant-like effects in mice through calmodulin kinase II stimulation downstream melatonin receptors, as well as the participation of this enzyme in neuroplasticity, memory, and cognition. Finally, we describe evidence about the effectiveness of antidepressant like effects of melatonin in combination with ketamine.

Neuroinflammation, Microbiota-Gut-Brain Axis, and Depression: The Vicious Circle.

Depression is the leading cause of disability worldwide, contributing to the global disease burden. From above, it is a priority to investigate models that fully explain its physiopathology to develop new treatments. In the last decade, many studies have shown that gut microbiota (GM) dysbiosis influences brain functions and participate, in association with immunity, in the pathogenesis of depression. Thereby, GM modulation could be a novel therapeutic target for depression. This review aims to evidence how the GM and the immune system influence mental illness, particularly depression. Here, we focus on the communication mechanisms between the intestine and the brain and the impact on the development of neuroinflammation contributing to the development of Major Depressive Disorder (MDD). However, most of the current findings are in animal models, suggesting the need for studies in humans. In addition, more analysis of metabolites and cytokines are needed to identify new pathophysiological mechanisms improving anti-depression treatments.

Contribution of Trimethylamine N-Oxide (TMAO) to Chronic Inflammatory and Degenerative Diseases.

Trimethylamine N-oxide (TMAO) is a metabolite produced by the gut microbiota and has been mainly associated with an increased incidence of cardiovascular diseases (CVDs) in humans. There are factors that affect one's TMAO level, such as diet, drugs, age, and hormones, among others. Gut dysbiosis in the host has been studied recently as a new approach to understanding chronic inflammatory and degenerative diseases, including cardiovascular diseases, metabolic diseases, and Alzheimer's disease. These disease types as well as COVID-19 are known to modulate host immunity. Diabetic and obese patients have been observed to have an increase in their level of TMAO, which has a direct correlation with CVDs. This metabolite is attributed to enhancing the inflammatory pathways through cholesterol and bile acid dysregulation, promoting foam cell formation. Additionally, TMAO activates the transcription factor NF-κB, which, in turn, triggers cytokine production. The result can be an exaggerated inflammatory response capable of inducing endoplasmic reticulum stress, which is responsible for various diseases. Due to the deleterious effects that this metabolite causes in its host, it is important to search for new therapeutic agents that allow a reduction in the TMAO levels of patients and that, thus, allow patients to be able to avoid a severe cardiovascular event. The present review discussed the synthesis of TMAO and its contribution to the pathogenesis of various inflammatory diseases.

Antidepressant Low Doses of Ketamine and Melatonin in Combination Produce Additive Neurogenesis in Human Olfactory Neuronal Precursors.

Melatonin (MEL), an indolamine with diverse functions in the brain, has been shown to produce antidepressant-like effects, presumably through stimulating neurogenesis. We recently showed that the combination of MEL with ketamine (KET), an NMDA receptor antagonist, has robust antidepressant-like effects in mice, at doses that, by themselves, are non-effective and have no adverse effects. Here, we show that the KET/MEL combination increases neurogenesis in a clone derived from human olfactory neuronal precursors, a translational pre-clinical model for effects in the human CNS. Neurogenesis was assessed by the formation of cell clusters > 50 µm in diameter, positively stained for nestin, doublecortin, BrdU and Ki67, markers of progenitor cells, neurogenesis, and proliferation. FGF, EGF and BDNF growth factors increased the number of cell clusters in cultured, cloned ONPs. Similarly, KET or MEL increased the number of clusters in a dose-dependent manner. The KET/MEL combination further increased the formation of clusters, with a maximal effect obtained after a triple administration schedule. Our results show that the combination of KET/MEL, at subeffective doses that do not produce adverse effects, stimulate neurogenesis in human neuronal precursors. Moreover, the mechanism by which the combination elicits neurogenesis is meditated by melatonin receptors, CaM Kinase II and CaM antagonism. This could have clinical advantages for the fast treatment of depression.

Further Evidence of the Melatonin Calmodulin Interaction: Effect on CaMKII Activity.

Melatonin (MEL) is a pleiotropic indolamine that reaches multiple intracellular targets. Among these, MEL binds to calmodulin (CaM) with high affinity. In presence of Ca2+, CaM binds to CaM-dependent kinase II (CaMKII). The Ca2+-CaM/CaMKII pathway regulates a myriad of brain functions in different cellular compartments. Evidence showing the regulation of this cellular pathway by MEL is scarce. Thus, our main objective was to study the interaction of MEL with CaM and its effects on CaMKII activity in two microenvironments (aqueous and lipidic) naturally occurring within the cell. In addition, colocalization of MEL with CaM in vivo was explored in mice brain hippocampus. In vitro CaM-MEL interaction and the structural conformations of CaM in the presence of this indoleamine were assessed through electrophoretic mobility and isoelectric point. The functional consequence of this interaction was evaluated by measuring CaMKII activity. Ca2+-CaM-MEL increased the activity of CaMKII in aqueous buffer but reduced the kinase activity in lipid buffer. Importantly, MEL colocalizes in vivo with Ca2+-CaM in the hippocampus. Our evidence suggests that MEL regulates the key cellular Ca2+-CaM/CaMKII pathway and might explain why physiological MEL concentrations reduce CaMKII activity in some experimental conditions, while in others it drives biological processes through activation of this kinase.

Low Doses of Ketamine and Melatonin in Combination Produce Additive Antidepressant-like Effects in Mice.

Major depressive disorder is a disabling disease with the number of affected individuals increasing each year. Current antidepressant treatments take between three to six weeks to be effective with forty percent of patients being resistant to treatment, making it necessary to search for new antidepressant treatments. Ketamine, a phencyclidine hydrochloride derivative, given intravenously, induces a rapid antidepressant effect in humans. In mice, it causes increased neurogenesis and antidepressant-like effects. However, it also produces psychomimetic effects in humans and in rodents increases the locomotor activity. In contrast, melatonin, a hormone secreted by the pineal gland and synthesized in extrapineal sites, increases new neuron formation and causes antidepressant-like effects in adult rodents with no collateral effects. Here, we assessed the effects of a non-effective dose of ketamine in combination with melatonin (KET/MEL), both on neurogenesis as well as on the antidepressant-like effect in mice. Our results showed that KET/MEL combination increased neurogenesis and produced antidepressant-like effects without altering locomotor activity after both single and triple administration protocols. Our data strongly suggest that KET/MEL combination could be used to simultaneously promote neurogenesis, reverting neuronal atrophy and inducing antidepressant-like effects.

EhRab21 associates with the Golgi apparatus in Entamoeba histolytica.

Rab proteins constitute the largest group of small GTPases and act as molecular switches in a wide variety of cellular processes, including proliferation, cytoskeleton assembly, and membrane trafficking in all eukaryotic cells. Rab21 has been reported in several eukaryotic cells, and our results suggest that in Entamoeba histolytica, Rab21 is involved in the vesicular traffic associated with the Golgi apparatus, where its function appears to be important to maintain the structure of this organelle. In addition, proteins such as Rab1A and Sec24, identified in this work associated with EhRab21, participate in the traffic of COPII vesicles from the endoplasmic reticulum to the Golgi apparatus and are necessary to maintain the latter's structure in human cells. In addition, EhRab21 probably affects the lysosome biogenesis, as indicated by an increase in the number of lysosomes as a result of the increase in EhRab21 activity. The participation of EhRab21 in the pathogenesis of amebiasis was verified on the amoebic liver abscess formation model using hamsters (Mesocricetus auratus), in which the overexpression of EhRab21Q64L (positive dominant mutant protein) decreased the number of liver abscesses formed.

Children from a rural region in The Chiapas Highlands, Mexico, show an increased risk of stunting and intestinal parasitoses when compared with urban children / Alto riesgo de desmedro y parasitosis intestinal en niños de una región rural de los Altos de Chiapas, México, en comparación con niños de una región urbana

Abstract Background: The state of Chiapas has held the first place of extreme poverty in Mexico. The majority of Chiapas' municipalities are inhabited by marginalized, indigenous populations, who usually present diarrhea of unknown etiology. We evaluated the nutritional status, intestinal parasites, and common bacterial pathogens, including DEC (diarrheagenic Escherichia coli) strains, in 178 children under five years of age with a high (rural) and a moderate (urban) degree of marginalization. Methods: Z-scores for anthropometric indexes from the children were obtained, whereas intestinal parasites were investigated by using a direct coproparasitoscopic analysis and a concentration method. DEC strains were detected by polymerase chain reaction (PCR). Results: The stunting prevalence in children from the rural and urban regions was 79.8 and 7.5%, respectively. Only children from rural municipalities were parasitized (72.6%), being Ascaris lumbricoides and Entamoeba histolytica/Entamoeba dispar the most prevalent parasites (57.1 and 38.1%, respectively). More than half of the children presented moderated ascariasis. Besides Giardia intestinalis, these parasites were associated with stunting. The prevalence of DEC strains was similar in both regions. Conclusions: Only children from the Chiapas Highlands (rural zone) exhibited high prevalences of stunting and intestinal parasites. A reevaluation of social development programs should be in place to address stunting and intestinal parasitoses, mainly in rural regions of Chiapas, to avoid adverse functional consequences on these children.

Resumen Introducción: El estado de Chiapas ha ostentado el primer lugar de pobreza extrema en México. La mayor parte de la población de los municipios de Chiapas es indígena, vive en condiciones de marginación y padece de diarrea de etiología desconocida. Este trabajo evaluó el estado nutricional, la presencia de parásitos intestinales y patógenos bacterianos comunes, además de cepas DEC (Escherichia coli diarreogénica) en 178 niños menores de cinco años, provenientes de una localidad con alto grado de marginación (rural) y de una con moderada marginación (urbana). Métodos: Se obtuvieron los puntajes Z de los índices antropométricos de los niños. Los parásitos intestinales se investigaron con el método coproparasitoscópico directo y un método de concentración. Las cepas DEC se detectaron mediante reacción en cadena de la polimerasa. Resultados: La prevalencia de desmedro en niños de la zona rural y urbana fue de 79.8 y 7.5%, respectivamente. Únicamente los niños de la zona rural estuvieron parasitados (72.6%), y los más prevalentes fueron Ascaris lumbricoides y Entamoeba histolytica/Entamoeba dispar (57.1 y 38.1%, respectivamente). Más de la mitad de los infantes exhibieron ascariasis moderada. Estos parásitos, además de Giardia intestinalis, se asociaron con el desmedro. En ambas regiones, la prevalencia de DEC fue similar. Conclusiones: Solo los niños de los Altos de Chiapas (zona rural) exhibieron alta prevalencia de desmedro y parásitos intestinales. Para evitar las consecuencias adversas entre los infantes, es necesario reevaluar los programas de desarrollo social para combatir el desmedro y la parasitosis intestinal, principalmente en las regiones rurales de Chiapas.

Children from a rural region in The Chiapas Highlands, Mexico, show an increased risk of stunting and intestinal parasitoses when compared with urban children.

Background: The state of Chiapas has held the first place of extreme poverty in Mexico. The majority of Chiapas' municipalities are inhabited by marginalized, indigenous populations, who usually present diarrhea of unknown etiology. We evaluated the nutritional status, intestinal parasites, and common bacterial pathogens, including DEC (diarrheagenic Escherichia coli) strains, in 178 children under five years of age with a high (rural) and a moderate (urban) degree of marginalization. Methods: Z-scores for anthropometric indexes from the children were obtained, whereas intestinal parasites were investigated by using a direct coproparasitoscopic analysis and a concentration method. DEC strains were detected by polymerase chain reaction (PCR). Results: The stunting prevalence in children from the rural and urban regions was 79.8 and 7.5%, respectively. Only children from rural municipalities were parasitized (72.6%), being Ascaris lumbricoides and Entamoeba histolytica/Entamoeba dispar the most prevalent parasites (57.1 and 38.1%, respectively). More than half of the children presented moderated ascariasis. Besides Giardia intestinalis, these parasites were associated with stunting. The prevalence of DEC strains was similar in both regions. Conclusions: Only children from the Chiapas Highlands (rural zone) exhibited high prevalences of stunting and intestinal parasites. A reevaluation of social development programs should be in place to address stunting and intestinal parasitoses, mainly in rural regions of Chiapas, to avoid adverse functional consequences on these children.

Introducción: El estado de Chiapas ha ostentado el primer lugar de pobreza extrema en México. La mayor parte de la población de los municipios de Chiapas es indígena, vive en condiciones de marginación y padece de diarrea de etiología desconocida. Este trabajo evaluó el estado nutricional, la presencia de parásitos intestinales y patógenos bacterianos comunes, además de cepas DEC (Escherichia coli diarreogénica) en 178 niños menores de cinco años, provenientes de una localidad con alto grado de marginación (rural) y de una con moderada marginación (urbana). Métodos: Se obtuvieron los puntajes Z de los índices antropométricos de los niños. Los parásitos intestinales se investigaron con el método coproparasitoscópico directo y un método de concentración. Las cepas DEC se detectaron mediante reacción en cadena de la polimerasa. Resultados: La prevalencia de desmedro en niños de la zona rural y urbana fue de 79.8 y 7.5%, respectivamente. Únicamente los niños de la zona rural estuvieron parasitados (72.6%), y los más prevalentes fueron Ascaris lumbricoides y Entamoeba histolytica/Entamoeba dispar (57.1 y 38.1%, respectivamente). Más de la mitad de los infantes exhibieron ascariasis moderada. Estos parásitos, además de Giardia intestinalis, se asociaron con el desmedro. En ambas regiones, la prevalencia de DEC fue similar. Conclusiones: Solo los niños de los Altos de Chiapas (zona rural) exhibieron alta prevalencia de desmedro y parásitos intestinales. Para evitar las consecuencias adversas entre los infantes, es necesario reevaluar los programas de desarrollo social para combatir el desmedro y la parasitosis intestinal, principalmente en las regiones rurales de Chiapas.

EhRab21 mobilization during erythrophagocytosis in Entamoeba histolytica.

Rab proteins are present in all eukaryotic lineages and regulate vesicular trafficking. Entamoeba histolytica has approximately 100 genes encoding Rab proteins, among which 16 have homology with human Rab proteins. Human Rab21 participates in integrin recycling, and thus amoebic Rab21 was believed to regulate the mobilization of Ehß1FNR (integrin-like fibronectin receptor related with human integrin ß1). We analyzed the distribution of EhRab21 using a polyclonal antibody produced with a specific peptide against the amoebic Rab protein, using confocal microscopy and specific probes for different organelles. EhRab21 was not associated with Ehß1FNR in fibronectin-stimulated trophozoites. However, EhRab21 was relocalized to lysosomes in erythrophagocytosis assays and was also found in Golgi-positive structures and the nuclear periphery. These results suggest that EhRab21, unlike its human homologue, is not present in the recycling pathway. However, according to the results, EhRab21 may regulate the trafficking between lysosomes and the Golgi apparatus.

Dengue virus induced changes in Ca2+ homeostasis in human hepatic cells that favor the viral replicative cycle.

The role of Ca2+ during dengue virus (DENV) replication is unknown; thus, changes in Ca2+ homeostasis in DENV infected human hepatic HepG2 and Huh-7 cells were analyzed. Infected HepG2 cells, but not Huh-7 cells, showed a significant increase in plasma membrane permeability to Ca2+, while both cell lines showed marked reduced levels of Ca2+ stored in the endoplasmic reticulum. While the expression levels of STIM1 and ORAI1 showed no changes, STIM1 and ORAI1 were shown to co-localized in infected cells, indicating activation of the store-operated Ca2+ entry (SOCE) pathway. Finally, manipulation in the infected cells of the intra and extracellular Ca2+ levels by chelators (BAPTA-AM and EGTA), SOC inhibitor (SKF96365), IP3 Receptor antagonist (2APB) or increase of extracellular [Ca2+], significantly reduced DENV yield, but not vesicular stomatitis virus yield, used as a control. These results show that DENV infection alters cell Ca2+ homeostasis and that such changes favor viral replication.