Salivary alpha-amylase stress reactivity in advanced-aged marmosets (Callithrix jacchus): Impacts of cognitive function and oral health status.

Salivary alpha-amylase (sAA) is an enzyme found in saliva and is considered a noninvasive biomarker for sympathetic nervous system activity. While a wide range of sAA activity in response to stress has been reported in nonhuman primates, the effects of stress on sAA activity in common marmosets are still unknown. We tested the hypothesis that advanced age and cognitive function may have an impact on stress-related sAA reactivity in marmosets. Thirteen marmosets (nine males and five females) had saliva samples collected during a stressful condition (manual restraint stress) at two different time points, with a 60-min interval. On the next day, the animals underwent the object recognition test (ORT, a type of cognitive test), and then oral examinations. The animals were categorized into two age groups: old (10-13 years), and very old (15-22 years). Irrespective of age, sAA levels showed a significant difference between T1 (mean 2.07 ± 0.86 U/mL) and T2 samples (mean 1.03 ± 0.67 U/mL), with higher values observed at T1 (p < 0.001). The intra-assay coefficients of variation (CV) for low and high sAA concentrations were 10.79% and 8.17%, respectively, while the interassay CVs for low and high sAA concentrations were 6.39% and 4.38%, respectively. Oral health issues were common but did not significantly impact sAA levels. The ORT indicated that the animals could recognize an object placed in the cage 6 h after familiarization. In conclusion, all marmosets showed a higher sAA concentration in the first saliva sample as compared to the second saliva sample collected 1 h later, indicating adaptation to stress. No significant differences in sAA levels were observed between sexes, ORT performance, or oral health. Our results indicate that autonomic responsivity and cognitive (memory) functions were preserved even in very old marmosets.

Validation and measurement invariance of the Compulsive Exercise Test among Brazilian and American young adults.

PURPOSE: To evaluate compulsive exercise, researchers often rely on the widely used Compulsive Exercise Test (CET). However, the measure has shown unstable factor structure in several validation studies and is not available in Portuguese for use in Brazil. We aimed to describe the translation and cultural adaptation of the CET to Brazilian Portuguese, to test several factor structures among Brazilian and US samples of men and women, to test measurement invariance across countries, and to evaluate its internal consistency. Furthermore, we sought to evaluate convergent validity, correlating the CET with a measure of eating disorder symptoms, and to compare compulsive exercise symptoms between countries. METHODS: Four models of the latent structure of the CET were tested using confirmatory factor analyses (CFAs), three-factor structure with 15 items, three-factor structure with 18 items, four-factor structure with 21 items, and the original five-factor structure with 24 items, in a sample of 1,531 young adults (601 Brazilians and 930 Americans), aged 18-35 years. RESULTS: A series of CFAs demonstrated that the three-factor structure with 15 items showed a better fit to the data. This model demonstrated good convergent validity and internal consistency. Results from the CET multigroup CFA showed evidence for the invariance at the configural, metric, and scalar levels across Brazilians and Americans. Furthermore, significant differences were found between Brazilians and Americans, with Brazilians demonstrating higher scores on the Avoidance and rule-driven behavior and Mood improvement subscales, whereas US participants scored higher on the Weight control exercise subscale. CONCLUSIONS: Results support the three-factor structure with 15 items to be used as a measure of compulsive exercise among Brazilians and Americans, allowing cross-cultural comparisons between these countries. LEVEL OF EVIDENCE: Level V, Cross-sectional, Psychometric study.

Multidisciplinary approach to multiple dental anomalies in pediatric patients: a case report with 4-year follow-up.

The objective of this article is to report the clinical case and 4-year follow-up of a 5-year-old child with multiple dental anomalies, emphasizing the importance of early diagnosis and use of combined pediatric surgery and orthodontic approaches. A 5-year-old boy, accompanied by his mother, sought dental care for dental caries and tooth pain. Clinical and radiographic examinations revealed active caries, a supernumerary primary tooth in the region of the mandibular right second premolar, and severe ankylosis of the primary mandibular right second molar. The treatment plan involved extraction of the supernumerary tooth as well as sectioning and extraction of the ankylosed molar. When the patient was 6 years old, the permanent mandibular right first molar showed signs of an altered eruptive process, and orthodontic treatment was initiated. A unilateral band-and-loop space maintainer with coil springs designed to move the permanent first molar was placed on the primary first molar. A new panoramic radiograph, obtained when the patient was aged 7 years, suggested the presence of an odontoma in the apical region of the primary maxillary right canine. Surgical removal and histopathologic examination of the lesion confirmed that it was a developing odontoma. After surgery, due to occlusal anomalies that included transverse maxillary deficiency, deep overbite, and midline deviation, the patient underwent rapid maxillary expansion therapy with a Haas-type appliance. When the patient was 8 years old, orthodontic treatment continued with a removable palatal Hawley expander and a orthodontic mandibular lingual arch. Currently, at the age of 9 years, the child is still undergoing fixed orthodontic treatment after surgical exposure of the impacted permanent maxillary right canine and bonding of an orthodontic attachment to enable traction. A multidisciplinary approach to the management of dental anomalies promotes a favorable prognosis and ensures comprehensive treatment of young patients.

Respiratory therapies for Amyotrophic Lateral Sclerosis: A state of the art review.

Amyotrophic lateral sclerosis (ALS) is a neurodegenerative condition noteworthy for upper and lower motor neuron death. Involvement of respiratory motor neuron pools leads to progressive pathology. These impairments include decreases in neural activation and muscle coordination, progressive airway obstruction, weakened airway defenses, restrictive lung disease, increased risk of pulmonary infections, and weakness and atrophy of respiratory muscles. These neural, airway, pulmonary, and neuromuscular changes deteriorate integrated respiratory-related functions including sleep, cough, swallowing, and breathing. Ultimately, respiratory complications account for a large portion of morbidity and mortality in ALS. This state-of-the-art review highlights applications of respiratory therapies for ALS, including lung volume recruitment, mechanical insufflation-exsufflation, non-invasive ventilation, and respiratory strength training. Therapeutic acute intermittent hypoxia, an emerging therapeutic tool for inducing respiratory plasticity will also be introduced. A focus on emerging evidence and future work underscores the common goal to continue to improve survival for patients living with ALS.

Galectin-3 regulates proinflammatory cytokine function and favours Brucella abortus chronic replication in macrophages and mice.

In this study, we provide evidence that galectin-3 (Gal-3) plays an important role in Brucella abortus infection. Our results showed increased Gal-3 expression and secretion in B. abortus infected macrophages and mice. Additionally, our findings indicate that Gal-3 is dispensable for Brucella-containing vacuoles disruption, inflammasome activation and pyroptosis. On the other hand, we observed that Brucella-induced Gal-3 expression is crucial for induction of molecules associated to type I IFN signalling pathway, such as IFN-ß: Interferon beta (IFN-ß), C-X-C motif chemokine ligand 10 (CXCL10) and guanylate-binding proteins. Gal-3 KO macrophages showed reduced bacterial numbers compared to wild-type cells, suggesting that Gal-3 facilitates bacterial replication in vitro. Moreover, priming Gal-3 KO cells with IFN-ß favoured B. abortus survival in macrophages. Additionally, we also observed that Gal-3 KO mice are more resistant to B. abortus infection and these animals showed elevated production of proinflammatory cytokines when compared to control mice. Finally, we observed an increased recruitment of macrophages, dendritic cells and neutrophils in spleens of Gal-3 KO mice compared to wild-type animals. In conclusion, this study demonstrated that Brucella-induced Gal-3 is detrimental to host and this molecule is implicated in inhibition of recruitment and activation of immune cells, which promotes B. abortus spread and aggravates the infection. TAKE AWAYS: Brucella abortus infection upregulates galectin-3 expression Galectin-3 regulates guanylate-binding proteins expression but is not required for Brucella-containing vacuole disruption Galectin-3 modulates proinflammatory cytokine production during bacterial infection Galectin-3 favours Brucella replication.

Non-Alcoholic Fatty Liver Disease-Related Hepatocellular Carcinoma: Immunohistochemical Assessment of Markers of Cancer Cell Metabolism.

INTRODUCTION: Hepatocellular carcinoma (HCC) has been associated to non-alcoholic fatty liver disease (NAFLD). We sought to investigate the immunoexpression of several glycolytic metabolism-associated markers in patients with HCC associated to NAFLD and associate these factors to their clinical-pathological characteristics. METHODS: We evaluated 35 HCC specimens from 21 patients diagnosed with non-alcoholic steatohepatitis (NASH) undergoing liver resection (12 patients), liver transplantation (8 patients), or both (1 patient). Histological features, clinical aspects, demographic and biochemical data, as well as the immunohistochemical reactivity for monocarboxylate transporters 1, 2, and 4; their chaperone CD147; carbonic anhydrase IX; and glucose transporter-1 (GLUT1) were assessed. RESULTS: Metabolic-associated cirrhosis was present in 12 of the 21 patients (8 child A and 4 child B scores). From 9 patients without cirrhosis, 3 presented NASH F3 and 6 NASH F2. Sixteen (76%) had diabetes mellitus, 17 (81%) arterial hypertension, and 19 (90%) body mass index above 25 kg/m2; 8 (38%) had dyslipidemia. From 35 nodules, steatosis was found in 26, ballooning in 31 nodules, 25 of them diagnosed as steatohepatitic subtype of HCC. MCT4 immunoexpression was associated with extensive intratumoral fibrosis, advanced clinical stages, and shorter overall survival. GLUT1 was noticeable in nodules with extensive intratumoral steatosis, higher intratumoral fibrosis, and advanced clinical stages. Immunohistochemical expression of the metabolic biomarkers MCT4 and GLUT1 was higher in patients with Barcelona-clinic liver cancer B or C. GLUT1 correlated with higher degree of steatosis, marked ballooning, intratumoral fibrosis, and higher parenchymal necroinflammatory activity. CONCLUSION: Our data indicate that the expression of the glycolytic phenotype of metabolic markers, especially GLUT1 and MCT4, correlates with a more severe course of HCC occurring in NASH patients.

Low-Level Laser Therapy for Management of Hypersensitivity in Molar-Incisor Hypomineralization and Oral Health-Related Quality of Life: Case Report.

The aim was to report the use low-level laser therapy (LLLT) in the management of hypersensitivity in an adolescent with molar incisor hypomineralization (MIH) and the impact of LLLT on his oral health-related quality of life (OHRQoL). Clinical examination revealed severe MIH with hypersensitivity, in all first permanent molars and incisors. The treatment proposed was desensitization with fluoride and esthetic rehabilitation of the affected teeth. Then, LLLT was applied perpendicularly in a continuous mode (wavelength of 808 nm, power of 100 mW, dose of 1 J, and fluence of 35 J/cm2). The visual analogue scale was applied each session of LLLT. The child perceptions questionnaire (CPQ11-14) was administered at the beginning and the end of the treatment. It was concluded that LLLT can be indicated in the management of hypersensitivity in an adolescent with severe MIH to control pain and to improve his OHRQoL.

The Relationship Between COVID-19 Confinement, Psychological Adjustment, and Sexual Functioning, in a Sample of Portuguese Men and Women.

BACKGROUND: Past research has revealed the detrimental effects of social isolation and physical distancing measures in health related outcomes. However, only recently COVID-19 confinement measures provided a context to test whether such detrimental effects exist in the human sexuality domain. AIM: This study was aimed at testing the relationship between COVID-19 confinement levels and sexual functioning domains in men and women, while accounting for the mediating role of psychological adjustment during lockdown. METHODS: Two hundred and forty five men and 417 women completed a web survey on the effects of COVID-19 in sexual health. The reference period includes the first confinement in Portugal (March 19-June 1, 2020). Data were handled under Hays' procedures for simple mediation analysis. OUTCOMES: Measurement outcomes included self-reported levels of confinement, IIEF and FSFI scores, and psychological adjustment during lockdown. RESULTS: Psychological adjustment during lockdown mediated the relationship between confinement levels and most sexual functioning domains in men but not in women. Also, while confinement levels were not directly related with most sexual functioning domains, psychological adjustment during lockdown did predict lower sexual functioning in both genders. CLINICAL TRANSLATION: Findings support that human sexual functioning should be framed within a contextual perspective, emphasizing how psychological adjustment and situational life stressors, such as lockdown situations, shape individuals' sexuality. Such perspective should be considered in treatment protocols implemented during current pandemic. STRENGTHS & LIMITATIONS: Data add to the literature on the effects of lockdown in the sexuality domain. However, the study precludes inference of causality and targets only a little proportion of all the dynamics involved in sexuality during the current historical moment. CONCLUSION: Psychological adjustment during COVID-19 seems to have a role on human sexual functioning, over the single effects of confinement. Carvalho J, Campos P, Carrito M, et al. The Relationship Between COVID-19 Confinement, Psychological Adjustment, and Sexual Functioning, in a Sample of Portuguese Men and Women. J Sex Med 2021;18:1191-1197.

Brucella abortus Cyclic Dinucleotides Trigger STING-Dependent Unfolded Protein Response That Favors Bacterial Replication.

Brucella abortus is a facultative intracellular bacterium that causes brucellosis, a prevalent zoonosis that leads to abortion and infertility in cattle, and undulant fever, debilitating arthritis, endocarditis, and meningitis in humans. Signaling pathways triggered by B. abortus involves stimulator of IFN genes (STING), which leads to production of type I IFNs. In this study, we evaluated the pathway linking the unfolded protein response (UPR) and the endoplasmic reticulum-resident transmembrane molecule STING, during B. abortus infection. We demonstrated that B. abortus infection induces the expression of the UPR target gene BiP and XBP1 in murine macrophages through a STING-dependent pathway. Additionally, we also observed that STING activation was dependent on the bacterial second messenger cyclic dimeric GMP. Furthermore, the Brucella-induced UPR is crucial for induction of multiple molecules linked to type I IFN signaling pathway, such as IFN-ß, IFN regulatory factor 1, and guanylate-binding proteins. Furthermore, IFN-ß is also important for the UPR induction during B. abortus infection. Indeed, IFN-ß shows a synergistic effect in inducing the IRE1 axis of the UPR. In addition, priming cells with IFN-ß favors B. abortus survival in macrophages. Moreover, Brucella-induced UPR facilitates bacterial replication in vitro and in vivo. Finally, these results suggest that B. abortus-induced UPR is triggered by bacterial cyclic dimeric GMP, in a STING-dependent manner, and that this response supports bacterial replication. In summary, association of STING and IFN-ß signaling pathways with Brucella-induced UPR unravels a novel link between innate immunity and endoplasmic reticulum stress that is crucial for bacterial infection outcome.

Dissonance-based eating disorder prevention delivered in-person after an online training: A randomized controlled trial for Brazilian men with body dissatisfaction.

OBJECTIVE: Few prevention programs have been developed and empirically evaluated to address eating disorder (ED) and muscle dysmorphia (MD) symptoms in men. Furthermore, new strategies for the broad implementation of available programs are needed. We investigated the acceptability and efficacy of a dissonance-based (DB) intervention for Brazilian undergraduate men with body dissatisfaction to target risk and protective factors for ED and MD symptoms (the Body Project: More Than Muscles) after an online training for facilitators. METHOD: Participants were randomized to a two-session DB-intervention (n = 89) or assessment-only control (AOC) (n = 91), and completed validated measures assessing ED and MD risk and protective factors pre-intervention, post-intervention, and at 4- and 24-week follow-up post-intervention. RESULTS: Acceptability ratings were highly favorable. Regarding efficacy, the DB condition demonstrated significantly greater decreases in ED and MD risk factors compared to AOC from pre-intervention to 4-week (p-values <.05, between-condition Cohen's d = 0.35-1.10) and 24-week follow-up (p-values <.05, between-condition Cohen's d = 0.33-0.78). Results at post-intervention were not significant, with the exception that body appreciation showed significantly greater improvements in the DB condition (post-intervention: p < .01, between-condition Cohen's d = 0.40; 4-week: p < .001, between-condition Cohen's d = 0.80; and 24-week follow-up: p < .001, between-condition Cohen's d = 0.58). DISCUSSION: Results support the acceptability and efficacy of a DB-intervention delivered in-person after an online training for facilitators up to 24-week follow-up in Brazilian men.

Brucella abortus nitric oxide metabolite regulates inflammasome activation and IL-1ß secretion in murine macrophages.

NLRP3 inflammasome is a protein complex crucial to caspase-1 activation and IL-1ß and IL-18 maturation. This receptor participates in innate immune responses to different pathogens, including the bacteria of genus Brucella. Our group recently demonstrated that Brucella abortus-induced IL-1ß secretion involves NLRP3 inflammasome and it is partially dependent on mitochondrial ROS production. However, other factors could be involved, such as P2X7-dependent potassium efflux, membrane destabilization, and cathepsin release. Moreover, there is increasing evidence that nitric oxide acts as a modulator of NLRP3 inflammasome. The aim of this study was to unravel the mechanism of NLRP3 inflammasome activation induced by B. abortus, as well as the involvement of bacterial nitric oxide (NO) as a modulator of this inflammasome pathway. We demonstrated that NO produced by B. abortus can be used by the bacteria to modulate IL-1ß secretion in infected murine macrophages. Additionally, our results suggest that B. abortus-induced IL-1ß secretion depends on a P2X7-independent potassium efflux, lysosomal acidification, cathepsin release, mechanisms clearly associated to NLRP3 inflammasome. In summary, our results help to elucidate the molecular mechanisms of NLRP3 activation and regulation during an intracellular bacterial infection.

Performance of light-emitting diode device in detecting occlusal caries in the primary molars.

This in vitro study aimed to compare the performance of a light-emitting diode (LED) device (Midwest Caries I.D.: MID), International Caries Detection and Assessment System (ICDAS) visual criteria, and fluorescence-based devices (DIAGNOdent: LF; DIAGNOdent pen: LFpen; and Quantitative Light-induced Fluorescence: QLF) in detecting occlusal caries in the primary molars. Eighty-eight primary molars with sound occlusal surfaces or carious lesions at different stages were assessed twice, with a 1-week interval in between, by one examiner using all three methods. Subsequently, the teeth were sectioned and lesion depth was verified using stereomicroscopy as a gold standard. Sensitivity, specificity, and accuracy were calculated at D1 (all carious lesions-enamel and dentin) and D3 (dentin lesions) thresholds. Correlation with histological analysis was evaluated using Spearman's rank correlation coefficients (rho). Weighted Kappa and intraclass-correlation (ICC) coefficients were calculated to assess intra-examiner reproducibility. At D1 threshold, ICDAS and LFpen showed higher sensitivity than the other methods, whereas ICDAS, LF, and QLF showed higher specificity (p < 0.05), and MID showed lower accuracy. At D3 threshold, ICDAS, LFpen, and QLF showed higher sensitivity than MID, whereas ICDAS, LF, and MID showed higher specificity (p < 0.05). All methods, except MID, showed statistically similar accuracy values (p < 0.05). Correlations with histopathological analysis varied from 0.15 (MID) to 0.57 (ICDAS). Intra-examiner reproducibility varied from 0.30 (MID) to 0.92 (ICDAS, LF, and QLF). The MID device exhibited a poor performance in detecting occlusal carious lesions in the primary molars, and ICDAS visual criteria exhibited greater accuracy than LF, LFpen, and QLF devices.

Immunoproteasome Subunits Are Required for CD8+ T Cell Function and Host Resistance to Brucella abortus Infection in Mice.

The immunoproteasome is a specific proteasome isoform composed of three subunits, termed ß1i, ß2i, and ß5i. Its proteolytic activity enhances the quantity and quality of peptides to be presented by major histocompatibility complex class I (MHC-I) molecules to CD8+ T cells. However, the role of the combined deficiency of the three immunoproteasome subunits in protective immunity against bacterial pathogens has not been investigated. In this study, we addressed the role of the immunoproteasome during infection by Brucella abortus, an intracellular bacterium that requires CD8+ T cell responses for the control of infection. Here, we demonstrate that immunoproteasome triple-knockout (TKO) mice were more susceptible to Brucella infection. This observed susceptibility was accompanied by reduced interferon gamma (IFN-γ) production by mouse CD4+ and CD8+ T lymphocytes. Moreover, the absence of the immunoproteasome had an impact on MHC-I surface expression and antigen presentation by dendritic cells. CD8+ T cell function, which plays a pivotal role in B. abortus immunity, also presented a partial impairment of granzyme B expression and, consequently, reduced cytotoxic activity. In conclusion, these results strongly suggest that immunoproteasome subunits are important components in host resistance to B. abortus infection by impacting both the magnitude and quality of CD8+ T cell responses.

NLRP12 negatively regulates proinflammatory cytokine production and host defense against Brucella abortus.

Brucella abortus is the causative agent of brucellosis, which causes abortion in domestic animals and undulant fever in humans. This bacterium infects and proliferates mainly in macrophages and dendritic cells, where it is recognized by pattern recognition receptors (PRRs) including Nod-like receptors (NLRs). Our group recently demonstrated the role of AIM2 and NLRP3 in Brucella recognition. Here, we investigated the participation of NLRP12 in innate immune response to B. abortus. We show that NLRP12 inhibits the early production of IL-12 by bone marrow-derived macrophages upon B. abortus infection. We also observed that NLRP12 suppresses in vitro NF-κB and MAPK signaling in response to Brucella. Moreover, we show that NLRP12 modulates caspase-1 activation and IL-1ß secretion in B. abortus infected-macrophages. Furthermore, we show that mice lacking NLRP12 are more resistant in the early stages of B. abortus infection: NLRP12-/- infected-mice have reduced bacterial burdens in the spleens and increased production of IFN-γ and IL-1ß compared with wild-type controls. In addition, NLRP12 deficiency leads to reduction in granuloma number and size in mouse livers. Altogether, our findings suggest that NLRP12 plays an important role in negatively regulating the early inflammatory responses against B. abortus.

Efficacy and safety of noninvasive focused ultrasound for treatment of subcutaneous adiposity in healthy women.

INTRODUCTION: In recent years, a new method focused ultrasound (FUS) has been used in the treatment of localized fat. The objectives of this work were to evaluate the efficacy and safety of the FUS in the treatment of abdominal subcutaneous fat. MATERIALS AND METHODS: Thirty-one healthy women were divided into two groups: G1 (N = 7), 6 sessions, 3 passes, once a week and G2 (N = 23), 10 sessions, 2 passes, twice a week. Outcome measures were reduction of circumference and fat thickness. Safety monitoring included laboratory testing (serum lipids profile and liver function tests) and adverse events were also assessed. Patient satisfaction and tolerance questionnaires were also applied. One patient underwent abdominoplasty and received a single session of FUS 24 h before surgery and a skin sample was collected for histological analysis. RESULTS: The results showed improvement in body contouring and reduction of the thickness of the fat layer observed by ultrasonography in both groups: G1 (P < 0.001) and G2 (P < 0.0001). The histology showed disrupted adipocytes and collapsed membranes 24 h after the FUS treatment. CONCLUSION: FUS represents an effective and safe treatment for reducing localized adipose tissue by adipocytolysis.

Mutant Brucella abortus membrane fusogenic protein induces protection against challenge infection in mice.

Brucella species can cause brucellosis, a zoonotic disease that causes serious livestock economic losses and represents a public health threat. The mechanism of virulence of Brucella spp. is not yet fully understood. Therefore, it is crucial to identify new molecules that serve as virulence factors to better understand this host-pathogen interplay. Here, we evaluated the role of the Brucella membrane fusogenic protein (Mfp) and outer membrane protein 19 (Omp19) in bacterial pathogenesis. In this study, we showed that B. abortus Δmfp::kan and Δomp19::kan deletion mutant strains have reduced persistence in vivo in C57BL/6 and interferon regulatory factor 1 (IRF-1) knockout (KO) mice. Additionally, 24 h after macrophage infection with a Δmfp::kan or Δomp19::kan strain expressing green fluorescent protein (GFP) approximately 80% or 65% of Brucella-containing vacuoles (BCVs) retained the late endosomal/lysosomal marker LAMP-1, respectively, whereas around 60% of BCVs containing wild-type S2308 were found in LAMP-1-negative compartments. B. abortus Δomp19::kan was attenuated in vivo but had a residual virulence in C57BL/6 and IRF-1 KO mice, whereas the Δmfp::kan strain had a lower virulence in these same mouse models. Furthermore, Δmfp::kan and Δomp19::kan strains were used as live vaccines. Challenge experiments revealed that in C57BL/6 and IRF-1 KO mice, the Δmfp::kan strain induced greater protection than the vaccine RB51 and protection similar that of vaccine S19. However, a Δomp19::kan strain induced protection similar to that of RB51. Thus, these results demonstrate that Brucella Mfp and Omp19 are critical for full bacterial virulence and that the Δmfp::kan mutant may serve as a potential vaccine candidate in future studies.

Critical role of ASC inflammasomes and bacterial type IV secretion system in caspase-1 activation and host innate resistance to Brucella abortus infection.

Pathogens are detected by innate immune receptors that, upon activation, orchestrate an appropriate immune response. Recent studies revealed the intracellular signaling cascades involved in the TLR-initiated immune response to Brucella abortus infection. However, no report has elucidated the role of inflammasome receptors in Brucella recognition. Therefore, we decided to investigate the function of NLRC4, NLRP3, and AIM2 in sensing Brucella. In this study, we showed that NLRC4 is not required to induce caspase-1 activation and further secretion of IL-1ß by B. abortus in macrophages. In contrast, we determined that AIM2, which senses Brucella DNA, and NLRP3 are partially required for caspase-1 activation and IL-1ß secretion. Additionally, mitochondrial reactive oxygen species induced by Brucella were implicated in IL-1ß production. Furthermore, AIM2, NLRP3, ASC, and caspase-1 knockout mice were more susceptible to B. abortus infection than were wild-type animals, suggesting that multiple ASC-dependent inflammasomes contribute to host protection against infection. This protective effect is due to the inflammatory response caused by IL-1ß and IL-18 rather than pyroptosis, because we observed augmented bacterial burden in IL-1R and IL-18 knockout mice. Finally, we determined that bacterial type IV secretion system VirB and live, but not heat-killed, Brucella are required for full inflammasome activation in macrophages during infection. Taken together, our results indicate that Brucella is sensed by ASC inflammasomes that collectively orchestrate a robust caspase-1 activation and proinflammatory response.

An alveolus lung-on-a-chip model of Mycobacterium fortuitum lung infection.

Lung disease due to non-tuberculous mycobacteria (NTM) is rising in incidence. While both two dimensional cell culture and animal models exist for NTM infections, a major knowledge gap is the early responses of human alveolar and innate immune cells to NTM within the human alveolar microenvironment. Here we describe development of a humanized, three-dimensional, alveolus lung-on-a-chip (ALoC) model of Mycobacterium fortuitum lung infection that incorporates only primary human cells such as pulmonary vascular endothelial cells in a vascular channel, and type I and II alveolar cells and monocyte-derived macrophages in an alveolar channel along an air-liquid interface. M. fortuitum introduced into the alveolar channel primarily infected macrophages, with rare bacteria inside alveolar cells. Bulk-RNA sequencing of infected chips revealed marked upregulation of transcripts for cytokines, chemokines and secreted protease inhibitors (SERPINs). Our results demonstrate how a humanized ALoC system can identify critical early immune and epithelial responses to M. fortuitum infection. We envision potential application of the ALoC to other NTM and for studies of new antibiotics.

Measurement invariance of the Intuitive Eating Scale-2 across country, ethnicity, sex, and sexual orientation: A cross-cultural study between Brazil and the U.S.

Intuitive eating is defined as being connected to internal hunger, satiety, and appetitive cues and flexibly using these cues to determine when, what, and how much to eat. The Intuitive Eating Scale-2 (IES-2) is a widely used measure of facets of intuitive eating. However, the scale has shown unstable factor structure in several validation studies and there is a lack of studies investigating the measurement invariance of the IES-2 beyond sex. We aimed to evaluate the psychometric properties of the IES-2, testing several factor structures among Brazilian and U.S. samples of men and women; to test measurement invariance across country of origin, ethnicity, sex, and sexual orientation; and to evaluate its internal consistency. Three models of the latent structure of the IES-2 were tested using confirmatory factor analyses (CFA) in a total of 1072 young adults (452 Brazilians and 620 Americans), aged 18-35 years. Results demonstrated that only a 3-factor solution with 11 items of the IES-2 showed adequate fit to the data for both countries. This model demonstrated scalar invariance across sex and sexual orientation, but only configural invariance was found across country of origin and ethnicity. Good internal consistencies were found for both the Brazilian and American samples. The present study provides support for a 3-factor solution with 11 items of the IES-2, to Brazilian and American samples. The study also offers evidence of internal consistency, and invariance between sex (i.e., male and female) and sexual orientation (i.e., heterosexual participants and sexual minority participants).

Apnea behavior in early- and late-stage mouse models of Parkinson's disease: Cineradiographic analysis of spontaneous breathing, acute stress, and swallowing.

This study aimed to evaluate the timing and frequency of spontaneous apneas during breathing and swallowing by using cineradiography on mouse models of early/initial or late/advanced Parkinson's disease (PD). C57BL/6 J mice received either 6-OHDA or vehicle injections into their right striatum, followed by respiratory movement recordings during spontaneous breathing and swallowing, and a stress challenge, two weeks later. Experimental group animals showed a significantly lower respiratory rate (158.66 ± 32.88 breaths/minute in late PD, 173.16 ± 25.19 in early PD versus 185.27 ± 25.36 in controls; p<0.001) and a significantly higher frequency of apneas (median 1 apnea/minute in both groups versus 0 in controls; p<0.001). Other changes included reduced food intake and the absence of swallow apneas in experimental mice. 6-OHDA-induced nigrostriatal degeneration in mice disrupted respiratory control, swallowing, stress responsiveness, and feeding behaviors, potentially hindering airway protection and elevating the risk of aspiration.