Chronic rhinosinusitis with nasal polyps does not harbor KRAS, BRAF, and EGFR mutations.

BACKGROUND: Chronic rhinosinusitis is a chronic inflammation of the nasal mucosa and nasal polyps are present in ~25%-30% of cases (chronic rhinosinusitis with nasal polyps [CRSwNP]). CRSwNP is associated with significant morbidity and decreased quality of life, making it clinically important. Inflammation leads to DNA damage and DNA mutations occur in some inflammatory diseases. Notably, mutations in KRAS, BRAF, and EGFR have been reported in different human benign and malignant neoplastic lesions. In addition, KRAS mutations have also been reported in non-neoplastic tissues under chronic inflammatory conditions. Importantly, KRAS mutations have been reported in oncocytic sinonasal papillomas and sinonasal squamous cell carcinoma associated with oncocytic sinonasal papilloma and EGFR mutations have been reported in sinonasal adenocarcinoma, inverted sinonasal papilloma, and sinonasal squamous cell carcinoma associated with inverted sinonasal papilloma. The molecular pathogenesis of nasal polyps remains unclear. Therefore, the present study aimed to investigate the presence of KRAS, BRAF, and EGFR pathogenic mutations in CRSwNP. METHODS: Fourteen chronic rhinosinusitis-associated nasal polyp samples were direct sequenced, targeting KRAS exons 2, 3, and 4 (encompassing important hotspot mutations, including codons 12, 13, 61 and 146), BRAF exons 11 and 15, and EGFR exons 19 and 20. RESULTS: No pathogenic mutations were detected in the sequenced regions of KRAS, BRAF, and EGFR genes. CONCLUSION: This finding suggests that mutations in these genes are not a frequent event in CRSwNP, and, if they occur, they might represent marginal events at best.

Photobiomodulation Reduces the Cytokine Storm Syndrome Associated with COVID-19 in the Zebrafish Model.

Although the exact mechanism of the pathogenesis of coronavirus SARS-CoV-2 (COVID-19) is not fully understood, oxidative stress and the release of pro-inflammatory cytokines have been highlighted as playing a vital role in the pathogenesis of the disease. In this sense, alternative treatments are needed to reduce the level of inflammation caused by COVID-19. Therefore, this study aimed to investigate the potential effect of red photobiomodulation (PBM) as an attractive therapy to downregulate the cytokine storm caused by COVID-19 in a zebrafish model. RT-qPCR analyses and protein-protein interaction prediction among SARS-CoV-2 and Danio rerio proteins showed that recombinant Spike protein (rSpike) was responsible for generating systemic inflammatory processes with significantly increased levels of pro-inflammatory (il1b, il6, tnfa, and nfkbiab), oxidative stress (romo1) and energy metabolism (slc2a1a and coa1) mRNA markers, with a pattern similar to those observed in COVID-19 cases in humans. On the other hand, PBM treatment was able to decrease the mRNA levels of these pro-inflammatory and oxidative stress markers compared with rSpike in various tissues, promoting an anti-inflammatory response. Conversely, PBM promotes cellular and tissue repair of injured tissues and significantly increases the survival rate of rSpike-inoculated individuals. Additionally, metabolomics analysis showed that the most-impacted metabolic pathways between PBM and the rSpike treated groups were related to steroid metabolism, immune system, and lipid metabolism. Together, our findings suggest that the inflammatory process is an incisive feature of COVID-19 and red PBM can be used as a novel therapeutic agent for COVID-19 by regulating the inflammatory response. Nevertheless, the need for more clinical trials remains, and there is a significant gap to overcome before clinical trials can commence.

Adenoid ameloblastoma harbors beta-catenin mutations.

Adenoid ameloblastoma is a very rare benign epithelial odontogenic tumor characterized microscopically by epithelium resembling conventional ameloblastoma, with additional duct-like structures, epithelial whorls, and cribriform architecture. Dentinoid deposits, clusters of clear cells, and ghost-cell keratinization may also be present. These tumors do not harbor BRAF or KRAS mutations and their molecular basis appears distinct from conventional ameloblastoma but remains unknown. We assessed CTNNB1 (beta-catenin) exon 3 mutations in a cohort of 11 samples of adenoid ameloblastomas from 9 patients. Two of the 9 patients were female and 7 male and in 7/9 patients the tumors occurred in the maxilla. Tumors of 4 of these 9 patients harbored CTNNB1 mutations, specifically p.Ser33Cys, p.Gly34Arg, and p.Ser37Phe. Notably, for one patient 3 samples were analyzed including the primary tumour and two consecutive recurrences, and results were positive for the mutation in all three tumors. Therefore, 6/11 samples tested positive for the mutation. In the 6 mutation-positive samples, ghost cells were present in only 2/6, indicating beta-catenin mutations are not always revealed by ghost cell formation. Dentinoid matrix deposition was observed in 5/6 mutation-positive samples and clear cells in all 6 cases. None of the cases harbored either BRAF or KRAS mutations. Beta-catenin immunoexpression was assessed in the samples of 8 patients. Except for one wild-type case, all cases showed focal nuclear expression irrespective of the mutational status. Together with the absence of BRAF mutation, the detection of beta-catenin mutation in adenoid ameloblastomas supports its classification as a separate entity, and not as a subtype of ameloblastoma. The presence of this mutation may help in the diagnosis of challenging cases.

Analysis of Cardiovascular Hemodynamic and Autonomic Variables in Individuals with Systemic Arterial Hypertension, Type 2 Diabetes Mellitus, and Parkinson's Disease: A Comparative Study.

BACKGROUND: Systemic arterial hypertension (SAH), type 2 diabetes mellitus (T2DM), and Parkinson's disease (PD) are highly prevalent chronic diseases that can significantly impact the cardiovascular system. AIM: The aim of this study was to compare hemodynamic and autonomic variables at rest in individuals with SAH, T2DM, or PD. METHODS: Fifty sedentary or insufficiently active individuals (22 men) with SAH (age = 66 ± 5.0 yr), T2DM (age = 52 ± 10 yr) or PD (age = 68 ± 8.0 yr) had their resting blood pressure (BP), arterial stiffness, endothelial function, and heart rate variability (HRV) assessed and compared. RESULTS: Systolic and diastolic BP were higher in SAH (130 ± 10 / 80 ± 10 mmHg) than T2DM (110 ± 14 / 75 ± 11 mmHg) and PD, and (123 ± 20 / 70 ± 11 mmHg) respectively. T2DM individuals showed lower arterial stiffness (8.4 ± 1.1 m/s), when compared to SAH (10.3 ± 2.3 m/s) and PD (10.6 ± 3.0 m/s). T2DM had greater resting tachycardia showed by the mean RR (759 ± 79 ms), than SAH (962 ± 169 ms) and PD (976 ± 134 ms), which was accompanied by higher sympathetic modulation (low frequency [LF]: 62 ± 19 nu) and lower parasympathetic modulation (high frequency [HF]: 32 ± 16 nu) when compared to SAH (LF: 40 ± 16 nu; HF: 61 ± 33 nu). No differences among groups were found on non-linear HRV markers and endothelial reactivity indexes. CONCLUSIONS: Individuals with T2DM showed impaired levels of cardiac autonomic markers when compared to individuals with SAH and PD, despite of having lower levels of BP and arterial stiffness.

Acute high-intensity interval exercise versus moderate-intensity continuous exercise in heated water-based on hemodynamic, cardiac autonomic, and vascular responses in older individuals with hypertension.

OBJECTIVES: This crossover study design aimed to assess hemodynamic, cardiac autonomic, and vascular responses to high-intensity interval (HIIE) vs moderate-intensity continuous exercise (MICE) in older individuals with hypertension. METHODS: Twenty (67 ± 7 y) older individuals with hypertension were randomly assigned to perform HIIE, MICE, or control (CON) sessions in the heated swimming pool (30-32°C). Blood pressure (BP), arterial stiffness, endothelial reactivity, and heart rate variability (HRV) were measured pre, post, and 45 min (recovery) after each intervention followed by 24-h ambulatory BP and HRV. RESULTS: One single aerobic exercise session was not effective to provoke post-exercise hypotension and vascular improvements. HIIE was superior to MICE and CON to increasing parasympathetic modulation at post and recovery. Exercise sessions showed to disturb the autonomic system at nighttime compared to CON. CONCLUSIONS: These results may have important implications in water-based therapy and the elderly with hypertension.

Rederivation of a mutant line (prop 1) of zebrafish Danio rerio infected with Pseudoloma neurophilia using in vitro fertilization with eggs from pathogen-free wild-type (AB) females and sperm from prop 1 males.

Along with the growing number of laboratories that work with zebrafish (Danio rerio), it is necessary to have animals with good sanitary quality. Specific pathogens can interfere with the experimental results and in the life quality of the animals. Pseudoloma neurophilia is a parasite with high potential for interference in behavioural, morphology, toxicological and genetic research, and is very common in zebrafish facilities. With that, we implemented a protocol for the pathogen elimination in a genetically modified lineage (prop 1) using eggs from specific pathogen-free (SPF) wild-type fish (AB line) for in vitro fertilization, along with water recirculation equipment disinfection, appropriate PCR screening and back crossing protocols. This resulted in SPF prop 1 heterozygotes, which allowed us to move forward with subsequent crossings to develop homozygote prop 1 mutants for our research. Hence, this demonstrates a useful strategy for an individual research laboratory to rederive a specific mutant free line that is not available from other SPF laboratories.

Protective effect of kavain in meristematic cells of Allium cepa L.

Kavain is one of the main kavalactones of Piper methysticum (Piperaceae) with anxiolytic, analgesic, and antioxidant activities. Therefore, the aim of the study was to evaluate the cytotoxic, mutagenic, and antimutagenic potential of kavain in Allium cepa cells. Roots of A. cepa were transferred to the negative (2% acetone) and positive (10 µg/mL of Methylmethanesulfonate, MMS) controls and to the concentrations of kavain (32, 64 and 128 µg/mL) for 48 h. A total of 5,000 meristematic cells were analyzed under an optical microscope to determine the mitotic index, mean number of chromosomal alterations and percentage of damage reduction. Data were analyzed by Kruskal-Wallis test (p <0.05). All concentrations of kavain were not cytotoxic and did not show significant chromosomal changes when compared to 2% acetone. Kavain showed a cytoprotective effect in the pre (128 µg/mL) and in the post-treatment (32 and 64 µg/mL) and reduced damage against the mutagenic action of MMS in all concentrations of the pre and simultaneous and at the highest of post (128 µg/mL). Kavain promoted a significant reduction in micronuclei, nuclear buds and chromosomal losses in relation to MMS. The observed data indicate the importance of kavain for the inhibition of damage and chemoprevention.

Prescribing and Self-Regulating Heated Water-Based Exercise by Rating of Perceived Exertion in Older Individuals With Hypertension.

We aimed to analyze the usefulness of the 6-20 rating of perceived exertion (RPE) scale for prescribing and self-regulating high-intensity interval (HIIE) and moderate-intensity continuous (MICE) aerobic exercise performed in a heated swimming pool (32 °C). Fifteen older individuals (65 ± 3 years) treated for hypertension underwent a symptom-limited maximal graded exercise test to determine their heart rate at anaerobic threshold, and respiratory compensation point. On different days, participants were randomized to HIIE (walking/jogging between 11 and 17 of RPE; 25 min) and MICE (walking at 11-13 of RPE; 30 min). Heart rate during the low-intensity intervals of HIIE and MICE remained below the graded exercise test's heart rate at anaerobic threshold (-7 ± 18 bpm/-16 ± 15 bpm) and respiratory compensation point (-18 ± 18 bpm/-30 ± 16 bpm), respectively, and maintained in the aerobic training zone during the high-intensity intervals of HIIE (+8 ± 18 bpm/-4 ± 19 bpm). The RPE scale at 15-17 is a useful tool for prescribing and self-regulating heated water-based HIIE and may have important implications for water-based exercise in older individuals with hypertension.

Prospection of the use of encapsulation in food packaging.

Food safety and extended shelf life linked to convenience were the major reasons for the development of the packaging field. However, advances in material science and the widespread encapsulation technologies are allowing the establishment of new concepts for packages, such as intelligent and active packages. Particulate systems have been developed in recent years for the most diverse area with several purposes that can be employed to improve packaging performance mainly focusing on the modification of barrier properties. This review analyzes the recent developments using encapsulation in food packaging and the main concepts about mass transfer evolved in the functionality of these packages, as well as discusses the research challenges faced by the food packaging sector.

Phytochemical and antioxidant characterization, cytogenotoxicity and antigenotoxicity of the fractions of the ethanolic extract of in Poincianella bracteosa (Tul.) L.P. Queiroz.

POINCIANELLA BRACTEOSA: has been widely used in folk medicine to treat catarrhal infections, diarrhea, and anemia; however, phytochemical and toxicogenetic data are still lacking. The objective of this study was to examine the phytochemical and antioxidant characteristics as well as assess cytogenotoxicity and antigenotoxicity in hexane (HF), ether (EF) and ethyl acetate (AF) fractions of P. bracteosa leaves using Allium cepa bioassay. Phytochemical analysis revealed the presence of saponins and phenolic groups. EF fraction contained a higher content of total phenolics (441.23 ± 1.82 mg GAE/g), while HF fraction showed a higher content of total flavonoids (84.77 ± 5.33 mg QE/g). Higher antioxidant activity was observed in EF (EC50 25.06 ± 0.07 µg/ml). Cytotoxic effect was verified for all fractions, but no chromosomal alterations were observed in the A. cepa assay. With respect to antigenotoxicity, the protective effect of EF and AF fractions was attributed to as evidenced by the modulation of mutagenic action of methyl methanesulfonate (MMS), mainly by inhibiting the development of micronuclei. Among the fractions, EF was considered the most promising, as it exhibited higher antioxidant activity, was not genotoxic, exerted protective activity against the damage induced by MMS and also presented cytotoxic activity, a desired quality in the search for natural anticarcinogenic compounds.

Hypotensive Effect of Heated Water-based Exercise in Older Individuals with Hypertension.

Blood pressure (BP) and hemodynamic response to heated water-based (HEx) vs. land-based exercise (LEx) were assessed in 15 (6 men) older hypertensives (age 66.4±4.9 yr) under pharmacological treatment. Participants were randomly assigned to perform 30 min of moderate-intensity HEx (walking inside the pool), LEx (walking on a treadmill) and non-exercise control (CON) intervention. Resting BP, arterial stiffness, endothelial reactivity and heart rate variability (HRV) were measured before, immediately after, and 45 min after interventions. 24-h ambulatory BP monitoring was performed after interventions. Resting systolic (but not diastolic) BP reduced 9.9±3.1 mmHg (P<0.01) 45 min after HEx only. 24-h systolic and diastolic, daytime diastolic and nightime systolic BP were lower (P<0.05) after HEx than both LEx and CON. Daytime systolic BP was also lower (P<0.05) after HEx than CON. Nighttime diastolic was not different between interventions. HEx-induced ambulatory BP reduction ranged 4.5±1.3 mmHg (24-h diastolic BP) to 9.5±3.0 mmHg (nighttime systolic BP), and persisted for 18/11 h in systolic/diastolic BP, when compared with CON. No significant changes in arterial stiffness, endothelial reactivity and HRV were found during any intervention. These results suggest that HEx may have important implications for managing BP in older hypertensive under pharmacological treatment.

Associations between a Brazilian suicide awareness campaign and suicide trends from 2000 to 2019: Joinpoint and regression discontinuity analysis.

BACKGROUND: Suicide is a global health concern whose rates are soaring in many low-and-middle-income countries. Public awareness campaigns have been implemented in Brazil; however, their impact is uncertain. METHODS: This was an ecological study using population data from DATA-SUS, a Brazilian official notification system, selecting only deaths that were voluntarily self-inflicted (CID-10 × 60-X84). We analyzed all available data from 2000 to 2019 to assess trends before and after the national Yellow September (YS) campaign started in 2014. Differences in trends were assessed by Joinpoint Analysis (JA) and Regression Discontinuity Analysis (RDD), unadjusted and adjusted for economic factors. RESULTS: Overall, there was a progressive increase in the rate of relative (per 100,000 inhabitants) number of suicides over time between 2000 and 2019 (57 % increase). The JA detected a change in the slope of the curve representing an acceleration in suicides starting in the year 2015. Adjusted RDD revealed the year that Yellow September started significantly change the slope of the association between time and rates of suicide (pinteraction < 0.01), and marginal analysis detected the coefficient increased from 0.07 (95%CI 0.04-0.10) to 0.27 (95%CI -0.07-0.60) suicides/year per 100,000 inhabitants. LIMITATIONS: The ecological nature of the manuscript compromises causational implications. CONCLUSIONS: We found an increase in suicidal trends in Brazil, against the global trend that coincides with the beginning of a large national awareness campaign. Although we cannot attribute causality, our results reinforce the need of further studies to better understand the role of awareness campaigns in suicide reduction interventions, including potential unintended effects.

Recurrence of adolescent suicide attempt and self-harm (RASS study): Effectiveness of single therapeutic project.

OBJECTIVES: This retrospective cohort study aims to understand the effectiveness of the Singular Therapeutic Project (STP) implemented in a Child Psychosocial Care Center in preventing recurrence of self-harm and suicide attempt (SA), by comparing the group of patients who adhered to the STP with the group that did not adhere completely. METHOD: Data were collected from the medical records of adolescents with self-harm or SA during the period from 2015 to 2019. The primary outcomes analyzed were recurrence of SA, self-harm, and hospitalization; and the secondary outcomes analyzed were demographics, diagnosis, number of appointments, and negative life events. RESULTS: A total of 228 adolescents were included. After multivariate analysis, social service consultations decreased risk in 94.6% (HR .054, 95% CI: .004-.681) of the cases, but risk was increased by 23 times if there was an episode of self-harm among family members and/or friends (HR 23.641, 95% CI: 1.394-400.8). Additionally, in terms of SA, adherence to family interventions reduced the risk by 66.2% (HR .338, 95% CI: .125-.913). Victims of prejudice, racism, homophobia, transphobia presented a 3.7-fold increased risk (HR 3.766, 95% CI: 1.058-13.401). CONCLUSION: The STP interventions were effective in reducing the recurrence of self-harm and SAs in adolescents.

Evaluation of clinical and genetic factors in obstructive sleep apnoea.

Purpose: To evaluate the correlation between several presumed candidate genes for obstructive sleep apnoea (OSA) and clinical OSA phenotypes and propose a predictive comprehensive model for diagnosis of OSA. Methods: This case-control study compared polysomnographic patterns, clinical data, morbidities, dental factors and genetic data for polymorphisms in PER3, BDNF, NRXN3, APOE, HCRTR2, MC4R between confirmed OSA cases and ethnically matched clinically unaffected controls. A logistic regression model was developed to predict OSA using the combined data. Results: The cohort consisted of 161 OSA cases and 81 controls. Mean age of cases was 53.5 ± 14.0 years, mostly males (57%) and mean body mass index (BMI) of 27.5 ± 4.3 kg/m2. None of the genotyped markers showed a statistically significant association with OSA after adjusting for age and BMI. A predictive algorithm included the variables gender, age, snoring, hypertension, mouth breathing and number of T alleles of PER3 (rs228729) presenting 76.5% specificity and 71.6% sensitivity. Conclusions: No genetic variant tested showed a statistically significant association with OSA phenotype. Logistic regression analysis resulted in a predictive model for diagnosing OSA that, if validated by larger prospective studies, could be applied clinically to allow risk stratification for OSA.

Whole-exome identifies germline variants in families with obstructive sleep apnea syndrome.

Background: Obstructive sleep apnea syndrome (OSAS) (OMIM #107650) is characterized by complete or partial obstruction of the upper airways, resulting in periods of sleep associated apnea. OSAS increases morbidity and mortality risk from cardiovascular and cerebrovascular diseases. While heritability of OSAS is estimated at ∼40%, the precise underlying genes remain elusive. Brazilian families with OSAS that follows as seemingly autosomal dominant inheritance pattern were recruited. Methods: The study included nine individuals from two Brazilian families displaying a seemingly autosomal dominant inheritance pattern of OSAS. Whole exome sequencing of germline DNA were analyzed using Mendel, MD software. Variants selected were analyzed using Varstation® with subsequent analyses that included validation by Sanger sequencing, pathogenic score assessment by ACMG criteria, co-segregation analyses (when possible) allele frequency, tissue expression patterns, pathway analyses, effect on protein folding modeling using Swiss-Model and RaptorX. Results: Two families (six affected patients and three unaffected controls) were analyzed. A comprehensive multistep analysis yielded variants in COX20 (rs946982087) (family A), PTPDC1 (rs61743388) and TMOD4 (rs141507115) (family B) that seemed to be strong candidate genes for being OSAS associated genes in these families. Conclusion: Sequence variants in COX20, PTPDC1 and TMOD4 seemingly are associated with OSAS phenotype in these families. Further studies in more, ethnically diverse families and non-familial OSAS cases are needed to better define the role of these variants as contributors to OSAS phenotype.

Homozygous CDH2 variant may be associated with hypopituitarism without neurological disorders.

Context: Congenital hypopituitarism is a genetically heterogeneous condition. Whole exome sequencing (WES) is a promising approach for molecular diagnosis of patients with this condition. Objectives: The aim of this study is to conduct WES in a patient with congenital hypopituitarism born to consanguineous parents, CDH2 screening in a cohort of patients with congenital hypopituitarism, and functional testing of a novel CDH2 variant. Design: Genomic DNA from a proband and her consanguineous parents was analyzed by WES. Copy number variants were evaluated. The genetic variants were filtered for population frequency (ExAC, 1000 genomes, gnomAD, and ABraOM), in silico prediction of pathogenicity, and gene expression in the pituitary and/or hypothalamus. Genomic DNA from 145 patients was screened for CDH2 by Sanger sequencing. Results: One female patient with deficiencies in growth hormone, thyroid-stimulating hormone, adrenocorticotropic hormone, luteinizing hormone, and follicle-stimulating hormone and ectopic posterior pituitary gland contained a rare homozygous c.865G>A (p.Val289Ile) variant in CDH2. To determine whether the p.Val289Ile variant in CDH2 affects cell adhesion properties, we stably transfected L1 fibroblast lines, labeled the cells with lipophilic dyes, and quantified aggregation. Large aggregates formed in cells expressing wildtype CDH2, but aggregation was impaired in cells transfected with variant CDH2 or non-transfected. Conclusion: A homozygous CDH2 allelic variant was found in one hypopituitarism patient, and the variant impaired cell aggregation function in vitro. No disease-causing variants were found in 145 other patients screened for CDH2 variants. Thus, CDH2 is a candidate gene for hypopituitarism that needs to be tested in different populations. Significance statement: A female patient with hypopituitarism was born from consanguineous parents and had a homozygous, likely pathogenic, CDH2 variant that impairs cell aggregation in vitro. No other likely pathogenic variants in CDH2 were identified in 145 hypopituitarism patients.

A novel insight on SARS-CoV-2 S-derived fragments in the control of the host immunity.

Despite all efforts to combat the pandemic of COVID-19, we are still living with high numbers of infected persons, an overburdened health care system, and the lack of an effective and definitive treatment. Understanding the pathophysiology of the disease is crucial for the development of new technologies and therapies for the best clinical management of patients. Since the manipulation of the whole virus requires a structure with an adequate level of biosafety, the development of alternative technologies, such as the synthesis of peptides from viral proteins, is a possible solution to circumvent this problem. In addition, the use and validation of animal models is of extreme importance to screen new drugs and to compress the organism's response to the disease. Peptides derived from recombinant S protein from SARS-CoV-2 were synthesized and validated by in silico, in vitro and in vivo methodologies. Macrophages and neutrophils were challenged with the peptides and the production of inflammatory mediators and activation profile were evaluated. These peptides were also inoculated into the swim bladder of transgenic zebrafish larvae at 6 days post fertilization (dpf) to mimic the inflammatory process triggered by the virus, which was evaluated by confocal microscopy. In addition, toxicity and oxidative stress assays were also developed. In silico and molecular dynamics assays revealed that the peptides bind to the ACE2 receptor stably and interact with receptors and adhesion molecules, such as MHC and TCR, from humans and zebrafish. Macrophages stimulated with one of the peptides showed increased production of NO, TNF-α and CXCL2. Inoculation of the peptides in zebrafish larvae triggered an inflammatory process marked by macrophage recruitment and increased mortality, as well as histopathological changes, similarly to what is observed in individuals with COVID-19. The use of peptides is a valuable alternative for the study of host immune response in the context of COVID-19. The use of zebrafish as an animal model also proved to be appropriate and effective in evaluating the inflammatory process, comparable to humans.

Hemodynamic response to exercise is impaired in individuals with Parkinson's disease.

BACKGROUND: The aim of this study was to assess hemodynamic and cardiac autonomic response to high-intensity interval exercise (HIIE) versus moderate-intensity continuous exercise (MICE) in individuals with Parkinson's disease (PD). METHODS: Twelve individuals (six men) with PD were randomly assigned to perform HIIE (4 min of warm-up followed by 21 min alternating 1 min at levels 15-17 with 2 min at levels 9-11 of rating of perceived exertion [RPE] in a cycle ergometer), MICE (4 min of warm-up followed by 26 min at levels 11-14 of RPE in a cycle ergometer) and control (CON; 30 min of sitting rest) interventions in separate days. Heart rate (HR), blood pressure (BP), endothelial reactivity and heart rate variability (HRV) were assessed before, immediately after and 45 min after each intervention. HR and exercise workload were measured during each intervention. RESULTS: Despite the within (high- vs. low-intensity intervals of HIIE) and between (HIIE vs. MICE) differences in workload during exercise sessions, HR was not different between high- (average HR=98±18 bpm) and low-intensity (average HR 97±19 bpm) intervals of HIIE, as well as between HIIE (average HR=97±18 bpm) and MICE (average HR=93±19 bpm) throughout the exercise. There were significant, but small, increases (P<0.01) in HR and systolic BP at post HIIE and MICE, which returned to levels similar to preintervention during recovery. There were no within- and between-intervention differences in diastolic BP, endothelial reactivity and HRV. CONCLUSIONS: The present results suggest that hemodynamic response to exercise is impaired in individuals with PD.

The blockade of µ­opioid receptors in the lateral hypothalamus enhances panic attack­like behaviour and diminishes defensive antinociception.

The lateral hypothalamus (LH) sends neural pathways to structures involved on predator­related defensive behaviours, escape and antinociception. The aim of this study was to investigate the role played by µ-opioid receptors located on LH neurons in defensive behaviour and unconditioned fear­induced antinociception elicited by electric stimulation of LH. To achieve the goals, the µ1-opioid receptor selective antagonist naloxonazine was administered at different concentrations in the LH, and the defensive behaviour and fear­induced antinociception elicited by electrical stimulation of LH were evaluated. The electrical stimulation of LH caused escape behaviour followed by defensive antinociception. Microinjections of naloxonazine in a concentration of 5.0 µg/0.2 µL in the LH decreased the aversive stimulus­induced escape behaviour thresholds, but diminished defensive antinociception. These findings suggest that µ-opioid receptors of LH can be critical to panic attack­related symptoms and facilitate the unconditioned fear­induced antinociception produced by LH neurons activation.

Virulomic Analysis of Multidrug-Resistant Klebsiella pneumoniae Isolates and Experimental Virulence Model Using Danio rerio (Zebrafish).

This study evaluates a possible correlation between multidrug-resistant Klebsiella pneumoniae strains and virulence markers in a Danio rerio (zebrafish) model. Whole-genome sequencing (WGS) was performed on 46 strains from three Brazilian hospitals. All of the isolates were colistin-resistant and harbored blaKPC-2. Ten different sequence types (STs) were found; 63% belonged to CC258, 22% to ST340, and 11% to ST16. The virulence factors most frequently found were type 3 fimbriae, siderophores, capsule regulators, and RND efflux-pumps. Six strains were selected for a time-kill experiment in zebrafish embryos: infection by ST16 was associated with a significantly higher mortality rate when compared to non-ST16 strains (52% vs. 29%, p = 0.002). Among the STs, the distribution of virulence factors did not differ significantly except for ST23, which harbored a greater variety of factors than other STs but was not related to a higher mortality rate in zebrafish. Although several virulence factors are described in K. pneumoniae, our study found ST16 to be the only significant predictor of a virulent phenotype in an animal model. Further research is needed to fully understand the correlation between virulence and sequence types.