Antiangiogenic properties of BthMP, a P-I metalloproteinase from Bothrops moojeni snake venom by VEGF pathway in endothelial cells.

Angiogenesis is a process that is controlled by a delicate combination of proangiogenic and antiangiogenic molecules and can be disrupted in various illnesses, including cancer. Non-cancerous diseases can also have an abnormal or insufficient vascular growth, inflammation and hypoxia, which exacerbate angiogenesis. These conditions include atherosclerosis, psoriasis, endometriosis, asthma, obesity and AIDS. Based on that, the present work assessed the in vitro and ex vivo antiangiogenic properties stemming from BthMP, a P-I metalloproteinase from Bothrops moojeni snake venom, via the VEGF pathway. BthMP at a concentration of 5 and 40 µg/mL showed no toxicity to endothelial cells (HUVEC) in the MTT assay and was not able to induce necrosis and colony proliferation. Interestingly, BthMP inhibited adhesion, migration and invasion of HUVECs in Matrigel and arrested in vitro angiogenesis by reducing the average number of nodules in toxin-treated cells by 9.6 and 17.32 at 5 and 40 µg/mL, respectively, and the number of tubules by 15.9 at 5 µg/mL and 21.6 at 40 µg/mL in a VEGF-dependent way, an essential proangiogenic property. Furthermore, BthMP inhibited the occurrence of the angiogenic process in an ex vivo aortic ring test by decreasing new vessel formation by 52% at 5 µg/mL and by 66% at 40 µg/mL and by increasing the expression of an antiangiogenic gene, SFLT-1, and decreasing the expression of the proangiogenic genes VEGFA and ANGPT-1. Finally, this toxin reduces the production of nitric oxide, a marker that promotes angiogenesis and VEGF modulation, and decreases the protein expression of VEGFA in the supernatant of the HUVEC culture by about 30 %. These results suggest that BthMP has a promising antiangiogenic property and proves to be a biotechnological mechanism for understanding the antiangiogenic responses induced by snake venom metalloproteinases, which could be applied to a variety of diseases that exhibit an imbalance of angiogenesis mechanisms.

Regulation of GABAergic neurotransmission by purinergic receptors in brain physiology and disease.

Purinergic receptors regulate the processing of neural information in the hippocampus and cerebral cortex, structures related to cognitive functions. These receptors are activated when astrocytic and neuronal populations release adenosine triphosphate (ATP) in an autocrine and paracrine manner, following sustained patterns of neuronal activity. The modulation by these receptors of GABAergic transmission has only recently been studied. Through their ramifications, astrocytes and GABAergic interneurons reach large groups of excitatory pyramidal neurons. Their inhibitory effect establishes different synchronization patterns that determine gamma frequency rhythms, which characterize neural activities related to cognitive processes. During early life, GABAergic-mediated synchronization of excitatory signals directs the experience-driven maturation of cognitive development, and dysfunctions concerning this process have been associated with neurological and neuropsychiatric diseases. Purinergic receptors timely modulate GABAergic control over ongoing neural activity and deeply affect neural processing in the hippocampal and neocortical circuitry. Stimulation of A2 receptors increases GABA release from presynaptic terminals, leading to a considerable reduction in neuronal firing of pyramidal neurons. A1 receptors inhibit GABAergic activity but only act in the early postnatal period when GABA produces excitatory signals. P2X and P2Y receptors expressed in pyramidal neurons reduce the inhibitory tone by blocking GABAA receptors. Finally, P2Y receptor activation elicits depolarization of GABAergic neurons and increases GABA release, thus favoring the emergence of gamma oscillations. The present review provides an overall picture of purinergic influence on GABAergic transmission and its consequences on neural processing, extending the discussion to receptor subtypes and their involvement in the onset of brain disorders, including epilepsy and Alzheimer's disease.

The role of clinical and demographic predictors for understanding the cognitive impairment in Spinal Cord Injury (SCI) patients.

STUDY DESIGN: Using a cross-sectional design, we extracted sociodemographic and clinical data from 488 Spinal Cord Injury (SCI) patients during their initial assessment before receiving intensive rehabilitation treatment. OBJECTIVES: The primary objectives of this study were to ascertain the prevalence of cognitive impairment in the study sample and specify the key clinical and demographic predictors of cognitive functioning in SCI patients. SETTING: Lucy Montoro Rehabilitation Institute (LMRI), University of Sao Paulo, Sao Paulo, Brazil. METHODS: We utilized independent univariate and multivariate regression models with the Montreal Cognitive Assessment (MoCA) scale, adapted for individuals with visual impairment. Moreover, we consider scores from the execution tasks (visuospatial/executive) as the dependent variable. RESULTS: Our findings demonstrate that approximately 80% of the evaluated study sample exhibited cognitive impairment. Through the multivariate regression models, we show that several factors, including age, education, depression levels, and the use of analgesics and/or opioids, are significant predictors of total cognitive scores. These factors are independent of the clinical features associated with SCI, such as age, sex, education, and time since the injury. CONCLUSIONS: The results indicate a high prevalence of significant cognitive impairment within the sample, with age, education, depression levels, and the use of analgesics and/or opioids emerging as the primary predictors of total cognitive scores, independent of the clinical features correlated to SCI. These findings hold significant implications for both clinical research and practice, offering valuable guidance for comprehensive management throughout hospitalization and rehabilitation.

Seizure Induced by Single-Pulse TMS Delivered to Lesioned Hemisphere in Stroke.

ABSTRACT: Transcranial magnetic stimulation (TMS) is used for therapeutic and research purposes, but it is still important to establish safety guidelines and recommendations mainly related to serious adverse effect (SAE). As part of this, safety reports need to be published. Our report highlights a case of a male patient who was a 30-year-old with a history of hemorrhagic stroke following an accidental seizure episode during a single-pulse TMS.

NEOS: An odour-induced affect scale for use in the cosmetic industry.

This work proposes an odour-induced affect scale for use in the cosmetic industry that relies on the approach that produced the UniGEOS, a universal odour-related emotional scale from the Swiss Center for Affective Sciences. The Natura Emotion and Odor Scale (NEOS) was built on experiments conducted with a larger set of participants (491) and a set of 35 scents that combine seven commercial perfumes from Natura & Co cosmetic company with 28 odours from different olfactory classes important for the cosmetic industry. The results showed the stability of 60 Emotion-Related terms in Brazilian Portuguese split into five emotion-related dimensions: Romance, Attention, Energy, Well-being and Negative feelings. The association of the scents evoking these five dimensions has direct implications in the design of new products.

Ce travail propose une échelle d'affect induite par des odeurs passible d'être utilisée dans l'industrie cosmétique. Cette échelle s'appuie sur l'approche qui a produit l'UniGEOS, une échelle affective universelle liée aux odeurs du Swiss Center for Affective Sciences. L'échelle Natura Emotion and Odor Scale (NEOS) a été construite sur la base d'expériences menées avec un plus grand nombre de participants (491) et un ensemble de 35 arômes combinant sept parfums commerciaux de la compagnie Natura & Co avec 28 odeurs de différentes classes olfactives importantes pour l'industrie cosmétique. Les résultats ont montré la stabilité de 60 termes liés aux émotions en portugais brésilien, répartis en cinq dimensions liées aux émotions : Romance, Attention, Énergie, Bien-être et Sentiments négatifs. L'association des arômes évoquant ces cinq dimensions a des implications directes dans la conception de nouveaux produits.

Does it matter who harmed whom? A cross-cultural study of moral judgments about harm by and to insiders and outsiders.

This cross-cultural study compared judgments of moral wrongness for physical and emotional harm with varying combinations of in-group vs. out-group agents and victims across six countries: the United States of America (N = 937), the United Kingdom (N = 995), Romania (N = 782), Brazil (N = 856), South Korea (N = 1776), and China (N = 1008). Consistent with our hypothesis we found evidence of an insider agent effect, where moral violations committed by outsider agents are generally considered more morally wrong than the same violations done by insider agents. We also found support for an insider victim effect where moral violations that were committed against an insider victim generally were seen as more morally wrong than when the same violations were committed against an outsider, and this effect held across all countries. These findings provide evidence that the insider versus outsider status of agents and victims does affect moral judgments. However, the interactions of these identities with collectivism, psychological closeness, and type of harm (emotional or physical) are more complex than what is suggested by previous literature. Supplementary information: The online version contains supplementary material available at 10.1007/s12144-023-04986-3.

Prevalence of Neisseria gonorrhoeae and Chlamydia trachomatis infections among adolescent men who have sex with men and transgender women in Salvador, Northeast Brazil.

Adolescent men who have sex with men (AMSM) and transgender women (ATGW) enrolled as part of the PrEP1519 study between April 2019 and February 2021 in Salvador were tested for Neisseria gonorrhoeae (NG) and Chlamydia trachomatis (CT) infections.We performed real-time polymerase chain reaction using oropharyngeal, anal, and urethral swabs; assessed factors associated with NG and CT infections using multivariable Poisson regression analysis with robust variance; and estimated the prevalence ratios (PRs) and 95% confidence intervals (95% CIs). In total, 246 participants were included in the analyses (median age: 18.8; IQR: 18.2-19.4 years). The overall oropharyngeal, anal, and urethral prevalence rates of NG were 17.9%, 9.4%, 7.6%, and 1.9%, respectively. For CT, the overall, oropharyngeal, anal, and urethral prevalence rates were 5.9%, 1.2%, 2.4%, and 1.9%, respectively. A low level of education, clinical suspicion of STI (and coinfection with Mycoplasma hominis were associated with NG infection. The prevalence of NG and CT, especially extragenital infections, was high in AMSM and ATGW. These findings highlight the need for testing samples from multiple anatomical sites among adolescents at a higher risk of STI acquisition, implementation of school-based strategies, provision of sexual health education, and reduction in barriers to care.

Prevalence of Mollicutes among men who have sex with men and transgender women aged 15 to 19 years in Salvador, North-eastern Brazil.

BACKGROUND: Some species of Mollicutes have been associated with different pathologies of the urogenital tract in humans, with a high prevalence among adult men who have sex with men (MSM) and transgender women (TGW). However, few studies have been performed to investigate its prevalence among adolescents. In this study, we estimated the initial prevalence of Mycoplasma genitalium (MG), Mycoplasma hominis (MH), Ureaplasma urealyticum (UU), and Ureaplasma parvum (UP); the rate of misdiagnosis at different anatomical sites; and the associated factors with positive tests for Mollicutes among MSM and TGW aged 15 to 19 years enrolled in the PrEP1519 study. METHODS: PrEP-1519 is the first study to investigate the effectiveness of pre-exposure prophylaxis for human immunodeficiency virus among adolescent MSM and TGW aged 15 to 19 in Latin America. Oral, anal, and urethral swabs were taken from 246 adolescents upon enrolment in the study to detect MG, MH, UU, and UP by quantitative polymerase chain reaction (qPCR). Bivariate and multivariate analyses were conducted by Poisson regression and 95% confidence intervals (95% CI) were estimated. RESULTS: The prevalence of Mollicutes was 32.1%. UU was the most prevalent species (20.7%), followed by MH (13.4%), MG (5.7%), and UP (3.2%); 67.3% of the positive samples would have been missed if only urethral samples had been taken. Receptive anal sex (prevalence ratio [PR] = 1.79; 95% CI = 1.07-3.01) and clinical suspicion of sexually transmitted infection (PR = 1.62; 95% CI = 1.01-2.61) were factors associated with the detection of Mollicutes in general. Group sex (PR = 1.98; 95% CI = 1.12-3.50) and receptive anal sex (PR = 2.36; 95% CI = 0.95-5.86) were associated with the detection of Mycoplasma spp. No sociodemographic, clinical, or behavioural variable was significantly associated with the detection of Ureaplasma spp. CONCLUSIONS: A high prevalence of Mollicutes was observed among adolescent MSM and TGW, especially at extragenital sites. Further research is required to understand the epidemiological profile of high-risk adolescents in different regions and contexts, and to investigate the pathogenesis of Mollicutes in the oral and anal mucosa before routine screening can be recommended in clinical practice.

Neuroplasticity changes in knee osteoarthritis (KOA) indexed by event-related desynchronization/synchronization during a motor inhibition task.

PURPOSE: Event-related desynchronisation (ERD) and event-related synchronisation (ERS) reflect pain perception and integration of the nociceptive sensory inputs. This may contribute to the understanding of how neurophysiological markers of Knee Osteoarthritis (KOA) patients can differ from control individuals, which would improve aspects such as prediction and prognosis. We performed a cross-sectional analysis of our cohort study (DEFINE cohort), KOA arm, with 71 patients, compared with 65 control participants. The study aimed to examine possible differences between ERD and ERS in control participants compared to Knee Osteoarthritis (KOA) patients when adjusting for important covariates. MATERIALS AND METHODS: We performed independent multivariate regression models considering as dependent variables the power value related to ERD and ERS for four different sensorimotor tasks (Motor Execution, Motor Imagery, Active Observation and Passive Observation) and four sensorimotor oscillations (Alpha, Beta, Low Beta, and High Beta), each model, controlled by age and sex. RESULTS: We demonstrate that the differences between KOA and healthy subjects are frequency specific, as most differences are in the beta bandwidth range. Also, we observed that subjects in the KOA group had significantly higher ERD and ERS. This may be correlated to the amount of lack of brain organisation and a subsequent attempt at compensation induced by KOA. CONCLUSIONS: Our findings strengthen the notion that subjects with KOA have a higher degree of brain plasticity changes that are also likely correlated to the degree of compensation and behavioural dysfunction.

Integrative Analysis of the Ethanol Tolerance of Saccharomyces cerevisiae.

Ethanol (EtOH) alters many cellular processes in yeast. An integrated view of different EtOH-tolerant phenotypes and their long noncoding RNAs (lncRNAs) is not yet available. Here, large-scale data integration showed the core EtOH-responsive pathways, lncRNAs, and triggers of higher (HT) and lower (LT) EtOH-tolerant phenotypes. LncRNAs act in a strain-specific manner in the EtOH stress response. Network and omics analyses revealed that cells prepare for stress relief by favoring activation of life-essential systems. Therefore, longevity, peroxisomal, energy, lipid, and RNA/protein metabolisms are the core processes that drive EtOH tolerance. By integrating omics, network analysis, and several other experiments, we showed how the HT and LT phenotypes may arise: (1) the divergence occurs after cell signaling reaches the longevity and peroxisomal pathways, with CTA1 and ROS playing key roles; (2) signals reaching essential ribosomal and RNA pathways via SUI2 enhance the divergence; (3) specific lipid metabolism pathways also act on phenotype-specific profiles; (4) HTs take greater advantage of degradation and membraneless structures to cope with EtOH stress; and (5) our EtOH stress-buffering model suggests that diauxic shift drives EtOH buffering through an energy burst, mainly in HTs. Finally, critical genes, pathways, and the first models including lncRNAs to describe nuances of EtOH tolerance are reported here.

Ureaplasma diversum-Ureaplasma sp. alone or concomitantly with Mycoplasma hyopneumoniae in pig lungs with and without pneumonia: A descriptive exploratory study in abattoirs.

Occurrence of Ureaplasma diversum (U. diversum) has been associated with reproductive failures in cattle and detected in pigs with and without pneumonia. However, its role in the porcine respiratory disease complex (PRDC) is unclear. A cross-sectional study was conducted in abattoirs, inspecting 280 pig lungs from eight herds. All the lungs were inspected, processed and classified according to the histopathological analysis. Moreover, bronchoalveolar lavage (BAL) specimens were collected and processed by PCR for detection of U. diversum and Mycoplasma hyopneumoniae (M. hyopneumoniae). Ureaplasma sp.-U. diversum and M. hyopneumoniae were detected in 17.1% and 29.3% of the analyzed BAL specimens, respectively. The concomitant presence of both microorganisms was detected in 12.5% of the inspected lungs. Both agents were found in lungs with and without pneumonia. M. hyopneumoniae was detected in 31.8% of pig lungs with enzootic pneumonia-like lesions, while Ureaplasma sp.-U. diversum was detected in 27.5% of lungs with these lesions. This descriptive exploratory study provides information for future experimental and field-based studies to better define the pathogenic role of this organism within the PRDC.

Host cell interactions of novel antigenic membrane proteins of Mycoplasma agalactiae.

BACKGROUND: Mycoplasma agalactiae is the main etiological agent of Contagious Agalactia syndrome of small ruminants notifiable to the World Organization for Animal Health. Despite serious economic losses, successful vaccines are unavailable, largely because its colonization and invasion factors are not well understood. This study evaluates the role of two recently identified antigenic proteins (MAG_1560, MAG_6130) and the cytadhesin P40 in pathogenicity related phenotypes. RESULTS: Adhesion to HeLa and sheep primary mammary stromal cells (MSC) was evaluated using ELISA, as well as in vitro adhesion assays on monolayer cell cultures. The results demonstrated MAG_6130 as a novel adhesin of M. agalactiae whose capacity to adhere to eukaryotic cells was significantly reduced by specific antiserum. Additionally, these proteins exhibited significant binding to plasminogen and extracellular matrix (ECM) proteins like lactoferrin, fibrinogen and fibronectin, a feature that could potentially support the pathogen in host colonization, tissue migration and immune evasion. Furthermore, these proteins played a detrimental role on the host cell proliferation and viability and were observed to activate pro-apoptotic genes indicating their involvement in cell death when eukaryotic cells were infected with M. agalactiae. CONCLUSIONS: To summarize, the hypothetical protein corresponding to MAG_6130 has not only been assigned novel adhesion functions but together with P40 it is demonstrated for the first time to bind to lactoferrin and ECM proteins thereby playing important roles in host colonization and pathogenicity.

Clinical and demographic predictors of symptoms of depression and anxiety in patients with spinal cord injury.

Spinal Cord Injury (SCI) is a condition whose consequences can impact the physical, emotional, and social aspects of patient's life, including Depression and Anxiety disorders. STUDY DESIGN: Using a cross-sectional design, sociodemographic and clinical data were extracted from 556 SCI patients at the time of initial assessment, prior to intensive rehabilitation treatment at the local rehabilitation institute. OBJECTIVES: Identify the predictive and multivariate relationship between different sociodemographic and clinical variables of Depression and Anxiety symptoms in SCI patients. SETTING: Lucy Montoro Rehabilitation Institute (LMRI), University of Sao Paulo, Sao Paulo, Brazil. METHODS: We performed independent univariate and multivariate regression models using the Hospital Anxiety and Depression Scale (HADS) as dependent variable. RESULTS: Symptoms of depression and anxiety present in SCI patients negatively correlate with the level of independence for locomotion, personal hygiene, bowel control, social interaction measured by the Functional Independence Scale (FIM), type of medication in use and the Language subtest of the MoCA Scale. Unlike previous studies, we did not find a relationship with the use of alcohol and illicit drugs, injury levels, etiological diagnosis or duration of injury. For the anxiety models, the main predictor was Depression symptoms, with SCI-related aspects not being significant. It was found that characteristics of the FIM Scale and cognitive aspects of the MoCA Scale were the main predictors of symptoms of Depression. Characteristics of the injury and motor deficit were not statistically significant. CONCLUSIONS: These findings can potentially be used to guide clinical practices to identify patients at higher risk of experiencing symptoms of Depression and Anxiety.

Lactiplantibacillus plantarum strains isolated from spontaneously fermented cocoa exhibit potential probiotic properties against Gardnerella vaginalis and Neisseria gonorrhoeae.

BACKGROUND: Probiotics are important tools in therapies against vaginal infections and can assist traditional antibiotic therapies in restoring healthy microbiota. Recent research has shown that microorganisms belonging to the genus Lactobacillus have probiotic potential. Thus, this study evaluated the potential in vitro probiotic properties of three strains of Lactiplantibacillus plantarum, isolated during the fermentation of high-quality cocoa, against Gardnerella vaginalis and Neisseria gonorrhoeae. Strains were evaluated for their physiological, safety, and antimicrobial characteristics. RESULTS: The hydrophobicity of L. plantarum strains varied from 26.67 to 91.67%, and their autoaggregation varied from 18.10 to 30.64%. The co-aggregation of L. plantarum strains with G. vaginalis ranged from 14.73 to 16.31%, and from 29.14 to 45.76% with N. gonorrhoeae. All L. plantarum strains could moderately or strongly produce biofilms. L. plantarum strains did not show haemolytic activity and were generally sensitive to the tested antimicrobials. All lactobacillus strains were tolerant to heat and pH resistance tests. All three strains of L. plantarum showed antimicrobial activity against the tested pathogens. The coincubation of L. plantarum strains with pathogens showed that the culture pH remained below 4.5 after 24 h. All cell-free culture supernatants (CFCS) demonstrated activity against the two pathogens tested, and all L. plantarum strains produced hydrogen peroxide. CFCS characterisation in conjunction with gas chromatography revealed that organic acids, especially lactic acid, were responsible for the antimicrobial activity against the pathogens evaluated. CONCLUSION: The three strains of L. plantarum presented significant probiotic characteristics against the two pathogens of clinical importance. In vitro screening identified strong probiotic candidates for in vivo studies for the treatment of vaginal infections.

Ureaplasma diversum clearance in lung mice infection is mediated by neutrophils.

Pneumonia in cattle is one of the causes of morbidity rates and economic loss. The host response to lung infections caused by Ureaplasma diversum in bovines is virtually unknown. Here in the immune response was evaluated in a murine model for an experimental pulmonary infection by U. diversum. Therefore, AJ, BALB/C and C57BL/6 mice received intratracheal inoculation of U. diversum and were evaluated after 1, 2, 3, 7 and 14 days and the clinical specimens were collected. In bronchoalveolar lavages (BAL) an increase of inflammatory cells was observed. Neutrophils were the main cells recruited to the site of infection and the infiltration was coincided with the production of pro-inflammatory cytokines. We found a large amount of neutrophil in this initial period, followed by a decrease 7 and 14 days post infection, accompanied by bacterial clearance. Our results evidenced the presence of U. diversum within the neutrophil that suggests a phagocytic role of this cell in the elimination of the infection. The immune response features reported here are the initial evidence that healthy immune systems may control these microorganisms. This may be the first step to design new strategies immune based to control the infections in naturally infected hosts.

Antimicrobial activity of Lactobacillus fermentum TcUESC01 against Streptococcus mutans UA159.

Dental caries is a multifactorial chronic-infection disease, which starts with a bacterial biofilm formation caused mainly by Streptococcus mutans. The use of probiotics has shown numerous health benefits, including in the fight against oral diseases. Strains of Lactobacillus fermentum have already shown probiotic potential against S. mutans, but there are still few studies. Thus, the aim of our study was to evaluate the antimicrobial activity of the inoculum and metabolites produced by L. fermentum TcUESC01 against S. mutans UA159. For this, a growth curve of L. fermentum was performed and both the inoculum and the metabolites formed in the fermentation were tested against the growth of S. mutans UA159 in agar diffusion tests, and only its metabolites were tested to determine the minimum inhibitory concentration, minimal bactericidal concentration and inhibition of cell adhesion. Inhibition of biofilm formation, pH drop and proton permeability were also tested with the metabolites. The zone of inhibition began to be formed at 14 h and continued until 16 h. The inoculum containing L. fermentum also showed zone of inhibition. The MIC for the metabolites was 1280 mg/mL and the MBC was obtained with a concentration higher than the MIC equal to 5120 mg/mL. Half of the MIC concentration (640 mg/mL) was required to inhibit S. mutans adhesion to the surface of the microplates. In the biofilm analyzes, the treatment with the metabolites in the tested concentration was not able to reduce biomass, insoluble glucans and alkali soluble compared to the control biofilm (p > 0.05). The metabolites also did not affect acid production and acid tolerance of S. mutans cells in biofilms compared to saline group (p > 0.05). Lactic acid (50.38%) was the most abundant organic acid produced by L. fermentum. This is the first report showing that the metabolites produced by the Lactobacillus fermentum TcUESC01 have a potential to be used as an antimicrobial agent against S. mutans, showing anti-adherence and bactericidal activity against planktonic cells of S. mutans. Thus, further studies should be carried out in order to better understand the antimicrobial activity of metabolites of L. fermentum TCUESC01.

Molecular basis of carrageenan-induced cytokines production in macrophages.

BACKGROUND: Low molecular weight carrageenan (Cg) is a seaweed-derived sulfated polysaccharide widely used as inflammatory stimulus in preclinical studies. However, the molecular mechanisms of Cg-induced inflammation are not fully elucidated. The present study aimed to investigate the molecular basis involved in Cg-induced macrophages activation and cytokines production. METHODS: Primary culture of mouse peritoneal macrophages were stimulated with Kappa Cg. The supernatant and cell lysate were used for ELISA, western blotting, immunofluorescence. Cg-induced mouse colitis was also developed. RESULTS: Here we show that Cg activates peritoneal macrophages to produce pro-inflammatory cytokines such as TNF and IL-1ß. While Cg-induced TNF production/secretion depends on TLR4/MyD88 signaling, the production of pro-IL-1ß relies on TLR4/TRIF/SYK/reactive oxygen species (ROS) signaling pathway. The maturation of pro-IL1ß into IL-1ß is dependent on canonical NLRP3 inflammasome activation via Pannexin-1/P2X7/K+ efflux signaling. In vivo, Cg-induced colitis was reduced in mice in the absence of NLRP3 inflammasome components. CONCLUSIONS: In conclusion, we unravel a critical role of the NLRP3 inflammasome in Cg-induced pro-inflammatory cytokines production and colitis, which is an important discovery on the pro-inflammatory properties of this sulfated polysaccharide for pre-clinical studies. Video abstract Carrageenan (Cg) is one the most used flogistic stimulus in preclinical studies. Nevertheless, the molecular basis of Cg-induced inflammation is not totally elucidated. Herein, Lopes et al. unraveled the molecular basis for Cg-induced macrophages production of biological active IL-1ß. The Cg-stimulated macrophages produces pro-IL-1ß depends on TLR4/TRIF/Syk/ROS, whereas its processing into mature IL-1ß is dependent on the canonical NLRP3 inflammasome.

Characterization of 3-aminospirostane alkaloids from roots of Solanum paniculatum L. with hepatoprotective activity.

RATIONALE: Solanum paniculatum L., popularly known as jurubeba, has traditionally been used in Brazilian folk medicine for liver diseases. However, there is a lack of knowledge about the chemical characterization of 3-aminospirostane alkaloids, an important class related to pharmacological activities. This work aimed to characterize the alkaloids using liquid chromatography with tandem mass spectrometry (LC/MS/MS) supported by molecular networking and theoretical calculations as well as to evaluate the contribution to hepatoprotective activity. METHODS: S. paniculatum roots were collected and macerated with MeOH/H2 O (8:2) obtaining the crude extract (SP-CE). From this, partition using EtOAc with pH variation yielded the alkaloidic fraction (SP-AF). Both were evaluated in an acute liver injury model (100 and 200 mg/kg), after intraperitoneal administration of carbon tetrachloride (CCl4 ) in mice. AST (aspartate transaminase) and ALT (alanine transaminase) serum levels were investigated, as well as the histopathological characteristics. The SP-CE and SP-AF were analyzed by LC/MS/MS, using quadrupole/time-of-flight and ion-trap systems. The alkaloids annotated by the GNPS molecular network had their structures defined using gas-phase ionization and fragmentation reaction supported by theoretical calculations. RESULTS: The SP-CE and SP-AF decreased the ALT serum levels compared with the negative control. The group treated with the SP-CE (at the highest dose) demonstrated a significant decrease of ALT. Hepatic cell degeneration decrease was observed mainly at the highest dose of the treatment. Detailed electrospray ionization MS/MS data allowed us to identify alkaloids not previously reported, to propose their gas-phase reactions and to redefine the initial open ring fragmentation mechanism of the steroidal alkaloids with the jurubidine moiety. CONCLUSIONS: The results allowed us to identify seven steroidal alkaloids from jurubeba and redefine the initial mechanism of fragmentation. A significant hepatoprotective effect was also demonstrated, corroborating its traditional use.

Electrophysiological indexes of ToM and non-ToM humor in healthy adults.

The cognitive processes involved in humor comprehension were analyzed by directly comparing the time course of brain activity associated with the perception of slapstick humor and that associated with the comprehension of humor requiring theory of mind (ToM). Four different comic strips (strips containing humorous scenes that required ToM, non-ToM humorous strips, non-humorous semantically coherent strips and non-humorous semantically incoherent strips) were presented to participants, while their EEG response was recorded. Results showed that both of the humorous comic strips and the semantically incongruent strip elicited an N400 effect, suggesting similar cognitive mechanisms underlying the processing of incongruent and humorous comic strips. The results also showed that the humorous ToM strips elicited a frontal late positive (LP) response, possibly reflecting the active deployment of ToM abilities such as perspective-taking and empathy that allow for the resolution and interpretation of apparently incongruent situations. In addition, the LP response was positively correlated with ratings of perceived amusement as well as individual empathy scores, suggesting that the increased LP response to ToM humorous strips reflects the combined activation of neural mechanisms involved in the experience of amusement and ToM abilities. Overall, humor comprehension appears to demand distinct cognitive steps such as the detection of incongruent semantic components, the construction of semantic coherence, and the appreciation of humoristic elements such as maladaptive emotional reactions. Our results show that the deployment of these distinct cognitive steps is at least partially dependent on individual empathic abilities.

Multilocus sequence typing characterizes diversity of Ureaplasma diversum strains, and intra-species variability induces different immune response profiles.

BACKGROUND: Ureaplasma diversum is a pathogen found in the genital tract of cattle and associated with genital disorders such as infertility, placentitis, abortion, birth of weak calves, low sperm motility, seminal vesiculitis and epididymitis. There are few studies evaluating the genetic diversity of U. diversum strains and their influence on the immune response in cattle. Therefore, to better understand genetic relationships of the pathogenicity of U. diversum, a multilocus sequence typing (MLST) scheme was performed to characterize the ATCC 49782 strain and another 40 isolates recovered from different Brazilian states. RESULTS: Primers were designed for housekeeping genes ftsH, polC, rpL22, rpoB, valS and ureA and for virulence genes, phospholipase D (pld), triacylglycerol lipase (tgl), hemolysin (hlyA), MIB-MIP system (mib,mip), MBA (mba), VsA (VsA) and ribose transporter (tABC). PCRs were performed and the targeted gene products were purified and sequenced. Sequence types (STs), and clonal complexes (CCs) were assigned and the phylogenetic relationship was also evaluated. Thus, a total of 19 STs and 4 CCs were studied. Following the molecular analysis, six isolates of U. diversum were selected, inoculated into bovine monocyte/macrophage culture and evaluated for gene expression of the cytokines TNF-α, IL-1, IL-6, IL-10 and IL-17. Differences were detected in the induction of cytokines, especially between isolates 198 and BA78, promoted inflammatory and anti-inflammatory profiles, respectively, and they also differed in virulence factors. CONCLUSION: It was observed that intra-species variability between isolates of U. diversum can induce variations of virulent determinants and, consequently, modulate the expression of the triggered immune response.