Effects of Tryptophan and Physical Exercise on the Modulation of Mechanical Hypersensitivity in a Fibromyalgia-like Model in Female Rats.

Though the mechanisms are not fully understood, tryptophan (Trp) and physical exercise seem to regulate mechanical hypersensitivity in fibromyalgia. Here, we tested the impact of Trp supplementation and continuous low-intensity aerobic exercise on the modulation of mechanical hypersensitivity in a fibromyalgia-like model induced by acid saline in female rats. Twelve-month-old female Wistar rats were randomly divided into groups: [control (n = 6); acid saline (n = 6); acid saline + exercise (n = 6); acid saline + Trp (n = 6); and acid saline + exercise + Trp (n = 6)]. Hypersensitivity was caused using two intramuscular jabs of acid saline (20 µL; pH 4.0; right gastrocnemius), 3 days apart. The tryptophan-supplemented diet contained 7.6 g/hg of Trp. The three-week exercise consisted of progressive (30-45 min) treadmill running at 50 to 60% intensity, five times (Monday to Friday) per week. We found that acid saline induced contralateral mechanical hypersensitivity without changing the levels of Trp, serotonin (5-HT), and kynurenine (KYN) in the brain. Hypersensitivity was reduced by exercise (~150%), Trp (~67%), and its combination (~160%). The Trp supplementation increased the levels of Trp and KYN in the brain, and the activity of indoleamine 2,3-dioxygenase (IDO), and decreased the ratio 5-HT:KYN. Exercise did not impact the assessed metabolites. Combining the treatments reduced neither hypersensitivity nor the levels of serotonin and Trp in the brain. In conclusion, mechanical hypersensitivity induced by acid saline in a fibromyalgia-like model in female rats is modulated by Trp supplementation, which increases IDO activity and leads to improved Trp metabolism via the KYN pathway. In contrast, physical exercise does not affect mechanical hypersensitivity through brain Trp metabolism via either the KYN or serotonin pathways. Because this is a short study, generalizing its findings warrants caution.

Association of the Serotonin and Kynurenine Pathways as Possible Therapeutic Targets to Modulate Pain in Patients with Fibromyalgia.

Fibromyalgia (FM) is a disorder characterized by widespread chronic pain, significant depression, and various neural abnormalities. Recent research suggests a reciprocal exacerbation mechanism between chronic pain and depression. In patients with FM, dysregulation of tryptophan (Trp) metabolism has been identified. Trp, an essential amino acid, serves as a precursor to serotonin (5-HT), a neuromodulator that influences mood, appetite, sleep, and pain perception through the receptors 5-HT1, 5-HT2, and 5-HT3. Additionally, Trp is involved in the kynurenine pathway, a critical route in the immune response, inflammation, and production of neuroactive substances and nicotinamide adenine dinucleotide (NAD+). The activation of this pathway by pro-inflammatory cytokines, such as tumor necrosis factor α (TNF-α) and interferon gamma (IFN-γ), leads to the production of kynurenic acid (KYNA), which has neuroprotective properties, and quinolinic acid (QA), which is neurotoxic. These findings underscore the crucial balance between Trp metabolism, 5-HT, and kynurenine, where an imbalance can contribute to the dual burden of pain and depression in patients with FM. This review proposes a novel therapeutic approach for FM pain management, focusing on inhibiting QA synthesis while co-administering selective serotonin reuptake inhibitors to potentially increase KYNA levels, thus dampening pain perception and improving patient outcomes.

Effect of Chronic Tibolone Administration on Memory and Choline Acetyltransferase and Tryptophan Hydroxylase Content in Aging Mice.

Gonadal steroids exert different effects on the central nervous system (CNS), such as preserving neuronal function and promoting neuronal survival. Estradiol, progesterone, and testosterone reduce neuronal loss in the CNS in animal models of neurodegeneration. However, hormone replacement therapy has been associated with higher rates of endometrial, prostate, and breast cancer. Tibolone (TIB), the metabolites of which show estrogenic and progestogenic effects, is an alternative to reduce this risk. However, the impact of TIB on memory and learning, as well as on choline acetyltransferase (ChAT) and tryptophan hydroxylase (TPH) levels in the hippocampus of aging males, is unknown. We administered TIB to aged C57BL/6J male mice at different doses (0.01 or 1.0 mg/kg per day for 12 weeks) and evaluated its effects on memory and learning and the content of ChAT and TPH. We assessed memory and learning with object recognition and elevated T-maze tasks. Additionally, we determined ChAT and TPH protein levels in the hippocampus by Western blotting. TIB administration increased the percentage of time spent on the novel object in the object recognition task. In addition, the latency of leaving the enclosed arm increased in both TIB groups, suggesting an improvement in fear-based learning. We also observed decreased ChAT content in the group treated with the 0.01 mg/kg TIB dose. In the case of TPH, no changes were observed with either TIB dose. These results show that long-term TIB administration improves memory without affecting locomotor activity and modulates cholinergic but not serotonergic systems in the hippocampus of aged male mice.

Size matters: lower body weight pigs have a different response to immune challenge and amino acids supplementation above the estimated requirement compared to heavy pigs.

The immune response varies between pigs, as not all pigs have the same response to a stressor. This variation may exist between individuals due to body weight (BW) or body composition, which may impact the capacity for coping with an immune challenge (IC). Tryptophan (Trp), threonine (Thr), and methionine (Met) requirements might also play a considerable part in supporting immune system activation while reducing variation between pigs; however, the latter has yet to be reported. This exploratory study investigated the effect of initial BW (light vs. heavy-weight) and supplementation of Trp, Thr, and Met above National Research Council (NRC) requirements on feeding behavior and the coping capacity of growing pigs under an IC. Eighty gilts were categorized into 2 groups according to BW: light-weight (LW, 22.5 kg) and heavy-weight pigs (HW, 28.5 kg). Both BW groups were group-housed for a 28-d trial in a good or poor sanitary condition (SC). Pigs within a poor SC were orally inoculated with 2 × 109 colony units of Salmonella Typhimurium, and fresh manure from a pig farm was spread on the floor. Pigs within good SC were not inoculated, nor was manure spread. Two diets were provided within each SC: control (CN) or supplemented (AA+) with Trp, Thr, and Met at 120% of NRC recommended levels. A principal component analysis was performed in R, and a feeding behavior index was calculated in SAS. Results showed that LW and HW pigs were clustered separately on day 0, where LW pigs had a positive correlation with body lipid percentage (r = 0.83), and HW pigs had a positive correlation with body protein percentage (r = 0.75). After the IC, the cluster configuration changed, with diets influencing LW more than HW pigs within poor SC. On day 14, LW fed AA + diet in poor SC was clustered separately from LW pigs fed CN diet, whereas LW fed AA + and CN diets in good SC were clustered together. For feeding behavior, in both analyzed periods (period 1: days 7 to 14; period 2: days 21 to 28), LW had lower total feed intake and shorter meals than HW pigs (P < 0.10), independent of the SC. Furthermore, LW pigs fed AA + diet had a more regular feed intake pattern than those fed CN diet, while a more irregular pattern was observed for HW pigs fed AA + diet than CN diet at period 2. These findings suggest that supplementing Trp, Thr, and Met above requirements may be a nutritional strategy for LW pigs under IC by improving feed intake regularity and reducing the probability of being susceptible to IC.

An immune challenge impacts pig welfare and may decrease growth and protein deposition. These may happen due to the different nutrient requirements of immune-challenged pigs compared to non-challenged. Dietary supplementation of tryptophan, threonine, and methionine has been proven to be a strategy to mitigate performance losses by supporting immune system functioning, maintaining gut barrier integrity, and reducing oxidative status. However, individuals within a population with similar age and genetics have distinct responses to dietary strategies due to different coping abilities to an immune challenge, which may be due to body weight (BW)/body composition and feeding behavior patterns. In this context, this study investigated the effect of BW (light-weight vs. heavy-weight) and tryptophan, threonine, and methionine supplementation on feeding behavior and the coping capacity of growing pigs under an immune challenge. Heavy-weight pigs had greater feed intake regularity and coping abilities over time when compared to light-weight pigs. However, increasing the amino acid level in the diet improved feed intake regularity in light-weight pigs. The amino acid supplementation may be a potential precision nutrition strategy for light-weight pigs by improving feed intake regularity over time, reducing susceptibility to an immune challenge.

Evaluation of real-time fluorescence sensors and benchtop fluorescence for tracking and predicting sewage contamination in the Tijuana River Estuary at the US-Mexico border.

Cross-border flow of untreated sewage from Mexico into the USA via the Tijuana River is public health issue with negative consequences for coastal communities. Here we evaluate the potential application of fluorescence-based, submersible tryptophan-like (TRP) and humic-like (CDOM) fluorescence sensors for real-time tracking of wastewater pollution in an estuarine environment. Sonde fluorescence measurements were compared with benchtop fluorescence, fecal indicator bacteria (FIB) concentrations, and real-time specific conductivity measurements in the Tijuana River Estuary during dry and wet weather conditions, and with and without cross-border flow. TRP and CDOM fluorescence concentrations were low during times without cross-border flow and two-three orders of magnitude higher during storm events and after cross-border sewage flow events. Major deterioration in water quality, including hypoxic conditions, was observed after consistent, long-term cross-border sewage flow. Real-time TRP and CDOM fluorescence concentrations had a significant linear relationship with fecal indicator bacteria (FIB) concentrations during dry weather periods with cross-border flow (p < 0.001) but were poorly correlated during stormflow and during less polluted periods with no cross-border flow. TRP and CDOM fluorescence acquired on discrete samples using a benchtop fluorometer correlated significantly (p < 0.001) with FIB concentrations under all cross-border flow conditions. Based on relationships between benchtop TRP fluorescence and percent wastewater, the greatest amount of untreated wastewater in the estuary's surface layer during cross-border flow events was estimated at >80 % and occurred during neap tides, when concentrated, sewage-laden freshwater flowed over dense saline seawater due to stratification and lack of mixing in the estuary. These results are important because exposure to untreated sewage poses severe health risks for residents and visitors to adjacent coastal areas. While benchtop fluorescence was more effective for estimating the degree of wastewater pollution, submersible TRP and CDOM sensors provided a real-time alert of sewage contamination, which can be utilized in other sewage impacted estuarine environments.

Tryptophan intrinsic fluorescence from wound healing correlates with re-epithelialization in a rabbit model.

BACKGROUND: Wound healing monitoring and timely decision-making are critical for wound classification. Tryptophan (Tr) intrinsic fluorescence, detected at 295/340 nm, provides a noninvasive approach for wound assessment. Our previous work demonstrated that this autofluorescence is associated with keratinocytes in a highly proliferative state in vitro. OBJECTIVE: We investigated the correlation between Tr fluorescence and key wound healing parameters, including re-epithelialization, fibrosis, neovascularization, and acute and chronic inflammation, using a rabbit model. METHODS: Seven rabbits underwent wound healing assessment over a 15-day period. We employed histological analysis from central and marginal biopsies, and UV fluorescence imaging captured by a monochromatic near-UV sensitive camera equipped with a passband optical filter (340 nm/12 nm). Excitation was achieved using a 295 nm LEDs ring lamp. Normalized fluorescence values were correlated with histological measurements using Pearson correlation. RESULTS: The UV fluorescence strongly exhibited a strong correlation with re-epithelization (r = 0.8) at the wound edge, with peak intensity observed between the sixth and ninth days. Notably, wound-healing dynamics differed between the wound center and edge, primarily attributed to variations in re-epithelialization, neovascularization, and chronic inflammation. CONCLUSION: Our findings highlight the presence of autofluorescence at 295/340 nm during wound healing, demonstrating a robust association with re-epithelialization. This excitation/emission signal holds promise as a valuable noninvasive strategy for monitoring wound closure, re-epithelialization, and other biological processes where Tr plays a pivotal role.

Oral Administration of Efavirenz Dysregulates the Tph2 Gene in Brain Serotonergic Areas and Alters Weight and Mood in Mice.

Most HIV-antiretroviral drugs have adverse effects. Efavirenz (EFV) is an example of a drug with neuropsychiatric effects, such as anxiety, depression, and suicidal thoughts, in people living with HIV (PLWH). The mechanisms by which EFV causes neuropsychiatric alterations in PLWH are complex, multifactorial, and not fully understood, although several studies in animals have reported changes in brain energy metabolism, alterations in monoamine turnover, GABA, and glutamate levels, and changes in 5-HT receptors. In this report, we studied the effects of EFV on the serotonergic system in healthy mice, specifically, whether EFV results in alterations in the levels of the tryptophan hydroxylase 2 (Tph2) gene in the brain. EFV (10 mg/kg) and distilled water (1.5 µL/kg) (control group) were orally administered to the mice for 36 days. At the end of the treatment, Tph2 expression levels in mouse brains were measured, and mood was evaluated by three trials: the forced swim test, elevated plus maze, and open field test. Our results revealed dysregulation of Tph2 expression in the brainstem, amygdala, and hypothalamus in the EFV group, and 5-HT levels increased in the amygdala in the EFV group. In the behavioral tests, mice given EFV exhibited a passive avoidance response in the forced swim test and anxiety-like behavior in the elevated plus maze, and they lost weight. Herein, for the first time, we showed that EFV triggered dysregulation of the Tph2 gene in the three serotonergic areas studied; and 5-HT levels increased in the amygdala using the ELISA method. However, further studies will be necessary to clarify the increase of 5-HT in the amygdala as well as understand the paradoxical decrease in body weight with the simultaneous increase in food consumption. It will also be necessary to measure 5-HT by other techniques different from ELISA, such as HPLC.

Amelioration of Neurochemical Alteration and Memory and Depressive Behavior in Sepsis by Allopurinol, a Tryptophan 2,3-Dioxygenase Inhibitor.

BACKGROUND: In response to inflammation and other stressors, tryptophan is catalyzed by Tryptophan 2,3-Dioxygenase (TDO), which leads to activation of the kynurenine pathway. Sepsis is a serious condition in which the body responds improperly to an infection, and the brain is the inflammation target in this condition. OBJECTIVE: This study aimed to determine if the induction of TDO contributes to the permeability of the Blood-Brain Barrier (BBB), mortality, neuroinflammation, oxidative stress, and mitochondrial dysfunction, besides long-term behavioral alterations in a preclinical model of sepsis. METHODS: Male Wistar rats with two months of age were submitted to the sepsis model using Cecal Ligation and Perforation (CLP). The rats received allopurinol (Allo, 20 mg/kg, gavage), a TDO inhibitor, or a vehicle once a day for seven days. RESULTS: Sepsis induction increased BBB permeability, IL-6 level, neutrophil infiltrate, nitric oxide formation, and oxidative stress, resulting in energy impairment in 24h after CLP and Allo administration restored these parameters. Regarding memory, Allo restored short-term memory impairment and decreased depressive behavior. However, no change in survival rate was verified. CONCLUSION: In summary, TDO inhibition effectively prevented depressive behavior and memory impairment 10 days after CLP by reducing acute BBB permeability, neuroinflammation, oxidative stress, and mitochondrial alteration.

Is It Possible to Mitigate Fear of Fireworks in Dogs? A Study on the Behavioural and Physiological Effects of a Psychoactive Supplement.

Canine fear of fireworks is a common problem worldwide, with serious implications for the welfare of both dogs and their owners. Therapies for the problem are available, and herbal and nutraceutical agents are increasingly suggested by professionals; nonetheless, studies on their real efficacy in reducing firework fear are lacking. In a randomised, double-blinded, placebo-controlled study, 44 dogs (25 in the "supplement" group and 19 in the "placebo" group) completed a long-term continuous treatment with either a supplement made of tryptophan, valerian, and passiflora or a placebo, including two real exposures to fireworks (on 2020 Christmas and 2021 New Years' Eve, after 42 and 48 days of treatment, respectively). Owners of both groups received the same general environmental management and food/toy offering recommendations for trying with their dogs on those nights. Behavioural (measured by LSSS-Lincoln Sound Sensitivity Scale and PANAS-Positive and Negative Activation scale, as rated by the owners) and stress (measured via salivary cortisol measures) reactions were evaluated. Significantly greater fear decrease (LSSS) was recorded in the "supplement" dogs, as compared to the "placebo" group. Cortisol dosages on New Year's Eve ("noisy" night) were in line with behavioural results; "supplement" dogs showed a smaller increase in the stress response from 22:30 to 00:30 h on New Year's Eve and a greater decrease in their stress response from 02:30 h to 10:30 h on New Year's Day compared to "placebo" dogs. Smaller cortisol levels were also shown by "supplement" dogs than "placebo" dogs on a controlled "quiet night" (27th December). Owners' rates on PANAS remained stable during the whole period of therapy for both groups. The evaluated supplement, a combination of tryptophan, valerian, and passiflora, showed satisfactory results and rare side effects when treating dogs fearful of fireworks.

Genetic regulation of L-tryptophan metabolism in Psilocybe mexicana supports psilocybin biosynthesis.

BACKGROUND: Although Basidiomycota produce pharmaceutically and ecologically relevant natural products, knowledge of how they coordinate their primary and secondary metabolism is virtually non-existent. Upon transition from vegetative mycelium to carpophore formation, mushrooms of the genus Psilocybe use L-tryptophan to supply the biosynthesis of the psychedelic tryptamine alkaloid psilocybin with the scaffold, leading to a strongly increased demand for this particular amino acid as this alkaloid may account for up to 2% of the dry mass. Using Psilocybe mexicana as our model and relying on genetic, transcriptomic, and biochemical methods, this study investigated if L-tryptophan biosynthesis and degradation in P. mexicana correlate with natural product formation. RESULTS: A comparative transcriptomic approach of gene expression in P. mexicana psilocybin non-producing vegetative mycelium versus producing carpophores identified the upregulation of L-tryptophan biosynthesis genes. The shikimate pathway genes trpE1, trpD, and trpB (encoding anthranilate synthase, anthranilate phosphoribosyltransferase, and L-tryptophan synthase, respectively) were upregulated in carpophores. In contrast, genes idoA and iasA, encoding indole-2,3-dioxygenase and indole-3-acetaldehyde synthase, i.e., gateway enzymes for L-tryptophan-consuming pathways, were massively downregulated. Subsequently, IasA was heterologously produced in Escherichia coli and biochemically characterized in vitro. This enzyme represents the first characterized microbial L-tryptophan-preferring acetaldehyde synthase. A comparison of transcriptomic data collected in this study with prior data of Psilocybe cubensis showed species-specific differences in how L-tryptophan metabolism genes are regulated, despite the close taxonomic relationship. CONCLUSIONS: The upregulated L-tryptophan biosynthesis genes and, oppositely, the concomitant downregulated genes encoding L-tryptophan-consuming enzymes reflect a well-adjusted cellular system to route this amino acid toward psilocybin production. Our study has pilot character beyond the genus Psilocybe and provides, for the first time, insight in the coordination of mushroom primary and secondary metabolism.

Exploring the interplay between running exercises, microbial diversity, and tryptophan metabolism along the microbiota-gut-brain axis.

The emergent recognition of the gut-brain axis connection has shed light on the role of the microbiota in modulating the gut-brain axis's functions. Several microbial metabolites, such as serotonin, kynurenine, tryptamine, indole, and their derivatives originating from tryptophan metabolism have been implicated in influencing this axis. In our study, we aimed to investigate the impact of running exercises on microbial tryptophan metabolism using a mouse model. We conducted a multi-omics analysis to obtain a comprehensive insight into the changes in tryptophan metabolism along the microbiota-gut-brain axis induced by running exercises. The analyses integrated multiple components, such as tryptophan changes and metabolite levels in the gut, blood, hippocampus, and brainstem. Fecal microbiota analysis aimed to examine the composition and diversity of the gut microbiota, and taxon-function analysis explored the associations between specific microbial taxa and functional activities in tryptophan metabolism. Our findings revealed significant alterations in tryptophan metabolism across multiple sites, including the gut, blood, hippocampus, and brainstem. The outcomes indicate a shift in microbiota diversity and tryptophan metabolizing capabilities within the running group, linked to increased tryptophan transportation to the hippocampus and brainstem through circulation. Moreover, the symbiotic association between Romboutsia and A. muciniphila indicated their potential contribution to modifying the gut microenvironment and influencing tryptophan transport to the hippocampus and brainstem. These findings have potential applications for developing microbiota-based approaches in the context of exercise for neurological diseases, especially on mental health and overall well-being.

Fungi Tryptophan Synthases: What Is the Role of the Linker Connecting the α and ß Structural Domains in Hemileia vastatrix TRPS? A Molecular Dynamics Investigation.

Tryptophan synthase (TRPS) is a complex enzyme responsible for tryptophan biosynthesis. It occurs in bacteria, plants, and fungi as an αßßα heterotetramer. Although encoded by independent genes in bacteria and plants, in fungi, TRPS is generated by a single gene that concurrently expresses the α and ß entities, which are linked by an elongated peculiar segment. We conducted 1 µs all-atom molecular dynamics simulations on Hemileia vastatrix TRPS to address two questions: (i) the role of the linker segment and (ii) the comparative mode of action. Since there is not an experimental structure, we started our simulations with homology modeling. Based on the results, it seems that TRPS makes use of an already-existing tunnel that can spontaneously move the indole moiety from the α catalytic pocket to the ß one. Such behavior was completely disrupted in the simulation without the linker. In light of these results and the αß dimer's low stability, the full-working TRPS single genes might be the result of a particular evolution. Considering the significant losses that Hemileia vastatrix causes to coffee plantations, our next course of action will be to use the TRPS to look for substances that can block tryptophan production and therefore control the disease.

Can supplementation of tryptophan in parenteral nutrition increase melatonin and alleviate inflammatory response?

SUMMARY OBJECTIVE: Endogenous melatonin is produced from tryptophan which is an essential amino acid. Besides its role in the regulation of sleep patterns, melatonin has anti-inflammatory effects. In this case-control study, we aimed to compare tryptophan and melatonin levels and their relationship with the inflammatory response, specifically serum interleukin-1, interleukin-6, and c-reactive protein levels following major abdominal surgery in patients with food restriction and who receive parenteral nutritional therapy. METHODS: We enrolled 40 patients between the ages of 18 and 65 years in the study. We collected blood and urine samples 48 h before the operation and on postoperative days 1, 3, and 5. RESULTS AND CONCLUSION: The tryptophan levels in the experimental group were higher than in the control group but failed to reach any statistical difference. Melatonin levels were increased in both groups following the surgery compared with preoperative levels. The increase in the experimental group was statistically different 3 days after the surgery. The difference in the level of interleukin-1 between the control and the experimental groups was greatest on postoperative day 3. On postoperative day 3, the interleukin-6 level in the treatment group was slightly higher than in the control group. We did not find any difference in the levels of c-reactive protein between the groups. As a result, the levels of tryptophan and melatonin were increased in the parenteral nutrition group, irrespective of the postoperative inflammatory response.

Análise do transcriptoma de diferentes tipos de células únicas do sangue periférico de pacientes infectados com o vírus da Dengue para detalhamento da massiva geração de células secretoras de anticorpos / Evaluation of the tryptophan metabolism in the massive generation of antibody-secreting cells in Dengue patients through the transcriptome of single blood cells

A Dengue induz uma resposta exacerbada e transitória das células secretoras de anticorpos (ASCs) no sangue de pacientes cerca de sete dias após o início dos sintomas. A frequência dessas ASCs chega a representar mais de 50% de todas as células B circulantes neste período. No entanto, ainda é desconhecido se a magnitude dessa resposta tem relação com a gravidade da Dengue. Nosso grupo de pesquisa já mostrou que a cultura de células nucleares do sangue periférico (PBMCs) de indivíduos saudáveis com partículas do vírus da Dengue (DENV), por 7 dias, levava a diferenciação de células B em ASCs em magnitude similar àquelas estimuladas por mitógenos. Essas culturas apresentavam um consumo significativamente maior de triptofano (TRP), associado à maior expressão das enzimas IDO1 e IDO2, e, consequentemente, maior síntese de quinurenina (KYN) em relação ao estímulo por mitógenos. Considerando que as concentrações de TRP e KYN detectadas nos sobrenadantes dessas culturas eram diretamente proporcionais ao aumento de ASCs, decidimos investigar o papel desse metabolismo do TRP e de seus respectivos metabólitos na diferenciação das ASCs. Para isso, análises do transcriptoma público de células únicas do sangue periférico de pacientes com Dengue (estudo E-MTAB- 9467) foram realizadas para inferir a real participação do metabolismo do TRP na geração de ASCs. Com o programa R foram executadas análises de Downstream. Identificamos um aumento massivo das ASCs nas amostras dos pacientes infectados com Dengue. No entanto, os principais genes desencadeadores da ativação do metabolismo do TRP (IDO1 e IDO2) não foram expressos nas subpopulações de células B, mas sim em células dendríticas e monócitos CD14+ respectivamente. Isso sugeriria que esta via não seria ativada nos linfócitos B. Por outro lado, genes codificadores de outros participantes da via do TRP (HSD17B10, ECHS1 e SIRT3) foram detectados em células B e podem estar relacionados com a proliferação das ASCs. Além disso, a análise de enriquecimento mostrou uma aumentada expressão de genes associados com moléculas de MHC de classe II em plasmablastos e plasmócitos de pacientes com Dengue. Porém, com a expressão aumentada de ENTPD1 nessas células durante a fase sintomatológica, nossos dados sugerem também que um eventual papel de plasmablastos e plasmócitos como apresentadoras de antígenos na Dengue poderia induzir uma resposta supressora de células T

Dengue can cause an exacerbated and transient antibody-secreting cell (ASC) response in the blood of patients nearly 7 days of symptomatology. The ASC frequency reaches more than 50% of all circulating B cells during that period. However, it is still unknown whether the magnitude of this response may be directly related to the severity of Dengue. Our research group has already shown that the culture of peripheral blood mononuclear cells (PBMCs) from healthy individuals with Dengue virus (DENV) particles for 7 days led to a differentiation of B cells into ASCs to a magnitude similar to those stimulated by mitogens. These cultures showed significantly higher consumption of tryptophan (TRP), associated with higher expression of enzymes IDO1 and IDO2, and consequently, higher synthesis of quinurenine (KYN) compared to mitogen stimulation. The concentrations of TRP and KYN detected in the supernatants of these cultures were directly proportional to ASC frequency increase. Thus, we have decided to investigate the role of TRP metabolism and its respective metabolites in ASC differentiation. For this, we performed an analysis of single-cell transcriptome with peripheral blood from Dengue patients (dataset from E-MTAB-9467 study). Downstream analyses were performed with R software. Corroborating with literature, we identified a massive increase in ASC frequency of Dengue infected patients. However, the main genes triggering TRP metabolism activation (IDO1 and IDO2) were not expressed in B-cell subsets, but in dendritic cells and CD14+ monocytes, respectively. This would suggest that this pathway would not be activated in B lymphocytes. Nevertheless, genes encoding other participants in the TRP pathway (HSD17B10, ECHS1, and SIRT3) were detected in B cells and may be related to ASC proliferation. Futhermore, a Gene Ontology analysis showed an increased expression of genes associated with MHC class II molecules in plasmablasts and plasma cells of Dengue patients. As these cells also presented an increased expression of ENTPD1 during the symptomatic phase, our data suggest a potential role of plasmablasts and plasma cells as antigen-presenting cells associated with a suppressive T cell response in Dengue

Confirmation of the presence of serotonergic cells in the fetal cerebral cortex in cultures and in situ.

BACKGROUND: Early appearance of serotonin in the fetal brain and its effects on brain morphogenesis support its neurotrophic role. OBJECTIVE: To determine the presence of serotonergic cells and the expression of tryptophan-5-hydroxylase (TPH), 5-hydroxytryptamine (5-HT), serotonin transporter (SERT), 5-HT1A receptor and Pet-1 during the development of the cerebral cortex, both in situ and in tissue cultures. MATERIAL AND METHODS: A descriptive, observational study was carried out in pregnant Wistar rats. The presence of the plug was regarded as the beginning of gestation. On days 13, 16 and 17, cesarean sections were performed to obtain the fetuses, and the brains were then immediately dissected to identify the presence of serotonergic cells, TPH, 5-HT, SERT, 5-HT1A and Pet-1 in tissue cultures and in situ by immunostaining detected on a confocal microscope. RESULTS: Serotonergic cells and terminals were observed in the midbrain on day 17 of gestation, and in neopallium cocultures on days 13 and 16. TPH, 5-HT, SERT and Pet-1 immunopositive cells were also observed in the neopallium on day 12 of culture. CONCLUSIONS: The presence of serotonergic cells and other elements of the serotonergic system in the early cerebral cortex was confirmed, which may be transient and participate in cortical maturation processes during brain development.

ANTECEDENTES: La aparición temprana de serotonina en el cerebro fetal y sus efectos en la morfogénesis cerebral apoyan su papel neurotrófico. OBJETIVO: Determinar la presencia de células serotoninérgicas y la expresión de triptófano-5-hidroxilasa (TPH), 5-hidroxitriptamina (5-HT), transportador de serotonina (SERT), receptor 5-HT1A y Pet-1 durante el desarrollo de la corteza cerebral, tanto in situ como en cultivo de tejidos. MATERIAL Y MÉTODOS: Se realizó estudio observacional descriptivo en ratas Wistar preñadas. La presencia del tapón se consideró el inicio de la gestación; en los días 13, 16 y 17 se practicaron cesáreas para obtener los fetos e inmediatamente se disecaron los cerebros para identificar células serotoninérgicas, TPH, 5-HT, SERT, 5-HT1A y Pet-1 en cultivo de tejido e in situ mediante inmunomarcaje detectado en un microscopio confocal. RESULTADOS: Células y terminales serotoninérgicas fueron observadas en el mesencéfalo el día 17 de gestación y en cocultivos de neopalio los días 13 y 16. También se observaron células inmunopositivas a TPH, 5-HT, SERT y Pet-1 en el neopalio en el día 12 del cultivo. CONCLUSIONES: Se confirmó la presencia de células serotoninérgicas y otros elementos del sistema serotoninérgico en la corteza cerebral temprana, la cual puede ser transitoria y participar en los procesos de maduración cortical durante el desarrollo cerebral.

Confirmación de la presencia de células serotoninérgicas en la corteza cerebral fetal en cultivos e in situ / Confirmation of the presence of serotonergic cells in the fetal cerebral cortex in cultures and in situ

Resumen Antecedentes: La aparición temprana de serotonina en el cerebro fetal y sus efectos en la morfogénesis cerebral apoyan su papel neurotrófico. Objetivo: Determinar la presencia de células serotoninérgicas y la expresión de triptófano-5-hidroxilasa (TPH), 5-hidroxitriptamina (5-HT), transportador de serotonina (SERT), receptor 5-HT1A y Pet-1 durante el desarrollo de la corteza cerebral, tanto in situ como en cultivo de tejidos. Material y métodos: Se realizó estudio observacional descriptivo en ratas Wistar preñadas. La presencia del tapón se consideró el inicio de la gestación; en los días 13, 16 y 17 se practicaron cesáreas para obtener los fetos e inmediatamente se disecaron los cerebros para identificar células serotoninérgicas, TPH, 5-HT, SERT, 5-HT1A y Pet-1 en cultivo de tejido e in situ mediante inmunomarcaje detectado en un microscopio confocal. Resultados: Células y terminales serotoninérgicas fueron observadas en el mesencéfalo el día 17 de gestación y en cocultivos de neopalio los días 13 y 16. También se observaron células inmunopositivas a TPH, 5-HT, SERT y Pet-1 en el neopalio en el día 12 del cultivo. Conclusiones: Se confirmó la presencia de células serotoninérgicas y otros elementos del sistema serotoninérgico en la corteza cerebral temprana, la cual puede ser transitoria y participar en los procesos de maduración cortical durante el desarrollo cerebral.

Abstract Background: Early appearance of serotonin in the fetal brain and its effects on brain morphogenesis support its neurotrophic role. Objective: To determine the presence of serotonergic cells and the expression of tryptophan-5-hydroxylase (TPH), 5-hydroxytryptamine (5-HT), serotonin transporter (SERT), 5-HT1A receptor and Pet-1 during the development of the cerebral cortex, both in situ and in tissue cultures. Material and methods: A descriptive, observational study was carried out in pregnant Wistar rats. The presence of the plug was regarded as the beginning of gestation. On days 13, 16 and 17, cesarean sections were performed to obtain the fetuses, and the brains were then immediately dissected to identify the presence of serotonergic cells, TPH, 5-HT, SERT, 5-HT1A and Pet-1 in tissue cultures and in situ by immunostaining detected on a confocal microscope. Results: Serotonergic cells and terminals were observed in the midbrain on day 17 of gestation, and in neopallium cocultures on days 13 and 16. TPH, 5-HT, SERT and Pet-1 immunopositive cells were also observed in the neopallium on day 12 of culture. Conclusions: The presence of serotonergic cells and other elements of the serotonergic system in the early cerebral cortex was confirmed, which may be transient and participate in cortical maturation processes during brain development.

A Minimal Kynurenine Pathway Was Preserved for Rhodoquinone but Not for De Novo NAD+ Biosynthesis in Parasitic Worms: The Essential Role of NAD+ Rescue Pathways.

Aims: To determine the role of the kynurenine (KYN) pathway in rhodoquinone (RQ) and de novo NAD+ biosynthesis and whether NAD+ rescue pathways are essential in parasitic worms (helminths). Results: We demonstrate that RQ, the key electron transporter used by helminths under hypoxia, derives from the tryptophan (Trp) catabolism even in the presence of a minimal KYN pathway. We show that of the KYN pathway genes only the kynureninase and tryptophan/indoleamine dioxygenases are essential for RQ biosynthesis. Metabolic labeling with Trp revealed that the lack of the formamidase and kynurenine monooxygenase genes did not preclude RQ biosynthesis in the flatworm Mesocestoides corti. In contrast, a minimal KYN pathway prevented de novo NAD+ biosynthesis, as revealed by metabolic labeling in M. corti, which also lacks the 3-hydroxyanthranilate 3,4-dioxygenase gene. Our results indicate that most helminths depend solely on NAD+ rescue pathways, and some lineages rely exclusively on the nicotinamide salvage pathway. Importantly, the inhibition of the NAD+ recycling enzyme nicotinamide phosphoribosyltransferase with FK866 led cultured M. corti to death. Innovation: We use comparative genomics of more than 100 hundred helminth genomes, metabolic labeling, HPLC-mass spectrometry targeted metabolomics, and enzyme inhibitors to define pathways that lead to RQ and NAD+ biosynthesis in helminths. We identified the essential enzymes of these pathways in helminth lineages, revealing new potential pharmacological targets for helminthiasis. Conclusion: Our results demonstrate that a minimal KYN pathway was evolutionary maintained for RQ and not for de novo NAD+ biosynthesis in helminths and shed light on the essentiality of NAD+ rescue pathways in helminths.

The kynurenine pathway in major depressive disorder, bipolar disorder, and schizophrenia: a systematic review and meta-analysis of cerebrospinal fluid studies

Objectives: The kynurenine (KYN) pathway has been attracting attention as a relevant pathway in schizophrenia (SZ), bipolar disorder (BD), and major depressive disorder (MDD). We conducted a systematic review and meta-analysis of studies examining KYN pathway metabolites from cerebrospinal fluid (CSF) samples in SZ, BD, and MDD. Methods: The PubMed and Scopus databases were systematically searched to identify peer-reviewed case-control studies published until April 2022 that assessed KYN metabolites, namely, tryptophan (TRP), KYN, kynurenic acid (KA), quinolinic acid (QA), and 3-hydroxykynurenine (3-HK), in subjects with SZ, BD, or MDD compared with healthy controls (HC). The Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines were followed. The random effects model method was selected for comparison of standardized mean differences (SMD) between two groups. Results: Twenty-three articles met the inclusion criteria (k = 8, k = 8, k = 11, for SZ, BD, and MDD, respectively). In SZ, KA levels were increased (SMD = 2.64, confidence interval [CI] = 1.16 to 4.13, p = 0.0005, I2 = 96%, k = 6, n=384). TRP (k = 5) and KYN (k = 4) did not differ significantly. In BD, TRP levels (k = 7) did not differ significantly. The level of KA was increased in MDD (k = 2), but the small number of studies precluded evaluation of statistical significance. Finally, in MDD, although some studies tended to show an increased level of KYN in those with remission vs. decreased levels in those with current depression, no significant difference was found in any KYN metabolite levels. Similarly, an increased level of QA was found, but the number of studies (k = 2) was small. Conclusion: KA, which has possibly neuroprotective effects, is increased in SZ. QA, which has neurotoxic effects, may be increased in MDD. There were no alterations in BD. Alterations in the KYN pathway may occur based on population characteristics and mood states. Future studies should explore the utility of these metabolites as biomarkers.

Biogenic amines and stable isotopes in the quality and authenticity of honeys from Brazil.

The identification of biogenic amines and some precursor amino acids and the adulteration through stable isotopes was carried out in 114 honey from different geographic regions in Brazil (states of São Paulo (SP) and Santa Catarina (SC)) as support for evaluating quality control and food safety. Serotonin was detected in all samples, while melatonin was quantified in 92.2% of honey from SP and in 94% of SC. l-Dopa, dopamine and histamine appeared at higher levels in honey from SP. Cadaverine, putrescine, spermidine and spermine, varied little according to botanical source. Three honey from the metropolitan region of SP were considered adulterated (C4SUGARS > 7%), 92 were authentic samples (C4SUGARS - 7 to 7%) and 19 unadulterated (C4SUGARS less than - 7%), with isotopic values of δ13CH and δ13CP > 7%. The data were important for differentiating quality as a function of biogenic amines and stable isotope technique was important in detecting honey adulteration.

The kynurenine pathway in major depressive disorder, bipolar disorder, and schizophrenia: a systematic review and meta-analysis of cerebrospinal fluid studies.

INTRODUCTION: The kynurenine pathway has been attracting attention as a relevant pathway in schizophrenia (SZ), bipolar disorder (BD), and major depressive disorder (MDD). We conducted a systematic review and meta-analysis of the kynurenine pathway metabolites from cerebrospinal fluid (CSF) samples in SZ, BD, and MDD. METHODS: PubMed and Scopus databases were systematically searched to identify peer-reviewed case-control studies until April 2022 that assessed kynurenine metabolites, namely, tryptophan (TRP), kynurenine (KYN), kynurenic acid (KA), quinolinic acid (QA), and 3- hydroxykynurenine (3-HK) in SZ, BD, or MDD subjects compared with healthy controls (HC). The Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines were followed. The random effects model method was selected when comparing the standardized mean differences (SMD) between two groups. RESULTS: There were 23 articles that met the inclusion criteria (k=8, k=8, k=11, for SZ, BD, and MDD, respectively). In SZ, KA levels were increased [SMD=2.64, confidence interval (CI) =1.16 to 4.13, p=0.0005, I2=96%, k=6, n=384 subjects]. TRP (k=5) and KYN (k=4) did not differ significantly. In BD, TRP levels (k=7) did not differ significantly. The level of KA was increased in MDD (k=2), but the small number of studies made not possible for statistical significance evaluation. Finally, in MDD, although some studies tended to have an increased level of KYN in those with remission versus decreased levels in those with current depression, no significant difference was found in any of the kynurenine metabolite levels. Similarly, there was an increased level of QA (k=2) but the number of studies (k= 2) was small. CONCLUSION: KA, which has possibly neuroprotective effects, is increased in SZ. QA, which has neurotoxic effects, may be increased in MDD. There were no alterations in BD. There may be alterations in this pathway based on population characteristics and mood states. Future studies should explore the utility of these metabolites as biomarkers.