Periodontal pathogen Aggregatibacter actinomycetemcomitans JP2 correlates with colonic leukocytes decrease and gut microbiome imbalance in mice.

AIM: Evidence suggests that translocation of oral pathogens through the oral-gut axis may induce intestinal dysbiosis. This study aimed to evaluate the impact of a highly leukotoxic Aggregatibacter actinomycetemcomitans (Aa) strain on the gut microbiota, intestinal mucosal integrity and immune system in healthy mice. METHODS: Eight-week-old male C57BL6 mice were divided into control (n = 16) and JP2 groups (n = 19), which received intragastric gavage with PBS and with a suspension of Aa JP2 (HK921), respectively, twice a week for 4 weeks. Colonic lamina propria, fecal material, serum, gingival tissues, and mandibles were obtained for analyses of leukocyte populations, inflammatory mediators, mucosal integrity, alveolar bone loss, and gut microbiota. Differences between groups for these parameters were examined by non-parametric tests. RESULTS: The gut microbial richness and the number of colonic macrophages, neutrophils, and monocytes were significantly lower in Aa JP2-infected mice than in controls (p < .05). In contrast, infected animals showed higher abundance of Clostridiaceae, Lactobacillus taiwanensis, Helicobacter rodentium, higher levels of IL-6 expression in colonic tissues, and higher splenic MPO activity than controls (p < .05). No differences in tight junction expression, serum endotoxin levels, and colonic inflammatory cytokines were observed between groups. Infected animals presented also slightly more alveolar bone loss and gingival IL-6 levels than controls (p < .05). CONCLUSION: Based on this model, intragastric administration of Aa JP2 is associated with changes in the gut ecosystem of healthy hosts, characterized by less live/recruited myeloid cells, enrichment of the gut microbiota with pathobionts and decrease in commensals. Negligible levels of colonic pro-inflammatory cytokines, and no signs of mucosal barrier disruption were related to these changes.

Synergism Between Essential Oils and Evaluation of Their Activities with a Focus on Malassezia furfur Control.

Seborrheic dermatitis is a chronic inflammatory disease caused by Malassezia yeast species that affects the regions of the body where the sebaceous glands are present. The combined use of different essential oils (EOs) can increase their spectrum of action. Thus, the present study aimed to evaluate the action of EOs alone and in combination with each other on M. furfur, in planktonic and biofilm form, and their anti-inflammatory and mutagenic potential, in addition to the effects on the viability of cells lines. Of the 40 evaluated EOs, 22 showed activity against M. furfur at 0.5 - 2.0 mg/mL concentrations. Among the most active species, a blend of essential oils (BEOs) composed of Cymbopogon martini (Roxb.) Will. Watson (MIC = 0.5 mg/mL) and Mentha × piperita L. (MIC = 1.0 mg/mL) was selected, which showed a synergistic effect against yeast when evaluated through the checkerboard assay. The fungicidal activity was maintained by the addition of anti-inflammatory oil from Varronia curassavica Jacq. to BEOs. The BEOs also showed activity in the inhibition of biofilm formation and in the eradication of the biofilm formed by M. furfur, being superior to the action of fluconazole. Furthermore, it did not show mutagenic potential and did not interfere with the cell viability of both evaluated cell lines (HaCaT and BMDMs). TNF-α levels were reduced only by C. martini; however, this property was maintained when evaluating BEOs. BEOs had no effect on IL-8 levels. Thus, the BEOs may be indicated for alternative treatments against seborrheic dermatitis.

Prevalence of COVID-19 Complications during a Program of Homeopathic Camphora Officinalis Distribution to City Populations of Santa Catarina, Brazil: An Ecological Study.

BACKGROUND: Coronavirus disease 2019 (COVID-19), a disease caused by the SARS-CoV-2 coronavirus, was declared a pandemic in March 2020, posing significant challenges globally. Homeopathy has historical relevance in epidemic management. In response, the government of the state of Santa Catarina, Brazil, distributed Camphora 1M as a potential prophylactic intervention for COVID-19. OBJECTIVE: This study aimed to investigate the possible effects of Camphora 1M as an adjunctive prophylactic measure in managing COVID-19, focusing on mortality and hospitalization rates, during the period April 28 to July 31, 2020, within designated COVID-19 in-patient units in Santa Catarina. METHODS: An ecological study design was applied to this epidemiological research. Five case municipalities (Itajaí, Atalanta, Entre Rios, Rio do Campo, Trombudo Central) were compared with five control municipalities (São José, Galvão, Pedras Grandes, Grão-Pará, Ascurra). RESULTS: No statistically significant differences were observed in predictor variables between municipalities that received Camphora 1M and the respective controls. Similarly, no statistically significant differences were observed in outcomes: deaths (p = 0.879), hospitalized cases (p = 0.537), daily ward admissions (p = 0.730) and ICU admissions (p = 0.072). CONCLUSION: For the first wave of the pandemic in the state of Santa Catarina, Brazil, city-wide distribution of Camphora 1M was not associated with reduced numbers, severity or mortality among the population hospitalized in designated public hospitals for COVID-19.

Psychosocial changes during COVID-19 lockdown on nursing home residents, their relatives and clinical staff: a prospective observational study.

BACKGROUND: Previous works have observed an increase of depression and other psychological disorders on nursing home residents as a consequence of coronavirus disease 2019 (COVID-19) lockdown; however, there are few studies that have performed a comprehensive evaluation of all people involved in nursing homes environment. The objective of the work was to analyse the impact of lockdown on psychosocial factors of nursing home residents, relatives and clinical staff and how these variables have influenced residents' survival. METHODS: A prospective study was designed. Evaluations were performed at three different times: a) at the beginning of Spanish confinement, in March 2020; b) just before the second wave of the pandemic, with relaxation of security measures but in lockdown, and c) in January-February 2021, at the end of the second wave, when visits were already allowed. The study was conducted on three different nursing homes. Three hundred and one residents, 119 clinical staff and 51 relatives took part in the study. Anxiety and depression were evaluated in all participants. A scale on the meaning of suffering was also performed. In addition, burnout status was also determined in the clinical staff. RESULTS: All participants showed lower depression during lockdown, while at the beginning and at the end of the confinement, these values were significantly increased. In residents, these changes were dependent of cognitive status (p = 0.012). Anxiety was significantly higher in residents. The evolution of anxiety was similar than with depression, with lower values during confinement, although clinical staff showed higher anxiety levels at the beginning. The feeling of suffering was significantly lower in the clinical staff than in resident and relative groups. Residents' survival was dependent of cognitive status (p = 0.018) and voluntary confinement (p < 0.001). CONCLUSIONS: During the first COVID-19 lockdown, psychological wellbeing of residents cared in nursing homes, their relatives and staff did not seem to be seriously affected. Previous mental health in relatives and staff together with a resilient approach to the adversity might partly be protecting factors. The lack of consequences on residents' anxiety, depression and perception of social support may reflect the special attention and care they received. Finally, as in the current study only data of the first two COVID-19 waves were analysed, its findings might be partly generalized to all the pandemic.

What Are the Ready-to-Use Endoscope Channels Hiding?: Unraveling the Risks of Safe Reuse.

Contamination due to failures or omissions in the reprocessing steps of gastrointestinal endoscopes is common in clinical practice. Ensuring the proper execution of each step is a challenge for reprocessing personnel. This cross-sectional study was conducted in an endoscopy setting between March and May 2021. We performed interviews about reprocessing practices, analyzed the life history of the equipment, and performed inspections through a borescope video of gastrointestinal endoscope channels that were stored and ready for use. A borescope is a complementary tool used to validate endoscope reprocessing, evaluate the internal visualization of channels, and identify changes that can compromise the safety of its use, which are often not detected in the leak test. Thirteen biopsy channels from stored gastrointestinal endoscopes were inspected. We found that 85% had stains and grooves, 69% contained moisture, and 46% had debris. There was at least one noncompliance issue in all of the channels inspected.

Criteria for the Use, Processing, and Disposal of Flexible Endoscope Cleaning Brushes.

Objectives: To ensure effectiveness in the endoscope channel cleaning process, using functional brushes that are in good condition is necessary. This study sought to identify the criteria for acquiring, using, and disposing of cleaning brushes at endoscopy facilities in Brazil. We further sought to evaluate the conditions of the cleaning brushes in use in the facilities. Methods: This cross-sectional study was conducted between July 2021 and January 2022. Personnel responsible for processing endoscopes were interviewed regarding the use, processing, and disposal of flexible endoscope cleaning brushes. In addition, the brushes used to clean the equipment were inspected. Results: All participants interviewed reported the practice of brushing endoscope channels. Of them, 60% noted the use of disposable brushes, with 40% using reusable brushes. None of the facilities interviewed reported discarding disposable brushes after use. The protocols for disposal of brushes included disposing due to bristle wear (70%), disposal at the end of the day (20%), and an absence of disposal protocols (10%). In addition, 30% of facilities did not clean the bristles before reintroducing them into the channel/lumen, and no facility had an established routine for cleaning brushes between uses. Inspection of brushes revealed that only 20% of facilities had new brushes with no signs of wear or damage. Conclusion: The use of inappropriate brushes/sponges for cleaning endoscope channels and the lack of criteria for the reuse and disposal of brushes increases the risk of cross-contamination, internal damage to channels, and biofilm formation.

Clinical isolates of E. faecalis and E. faecium harboring virulence genes show the concomitant presence of CRISPR loci and antibiotic resistance determinants.

In this study, we evaluated the antimicrobial susceptibility, the presence of gene-encoding virulence factors and CRISPR systems, as well as the ability to produce lytic enzymes among clinical E. faecalis and E. faecium isolates (n = 44). All enterococci isolates showed phenotypes of multidrug resistance. E. faecalis and E. faecium isolates exhibited high-level aminoglycoside resistance phenotype, several of them harboring the aac(6')Ie-aph(2″)Ia and aph(3')-IIIa genes. The gene vanA was the most frequent among vancomycin-resistant E. faecium. High prevalence of the virulence genes esp and efaA were observed; hyl gene was more associated with E. faecium, while ace and efaA genes were more frequently detected in E. faecalis. Caseinase activity was frequently detected among the isolates. Gelatinase and DNAse activities predominated among E. faecalis, while hemolytic capability was frequent among E. faecium isolates. Twenty-nine isolates showed at least one CRISPR system investigated. Several enterococci isolates harbored the aac(6')-Ie-aph(2″)-Ia or aph(3')-IIIa genes and a CRISPR loci. CRISPR loci were positively correlated to efaA and gelE genes, and gelatinase and DNAse activities, while CRISPR loci absence was related to hyl gene presence. These results show that clinical isolates of E. faecalis and E. faecium harboring virulence genes show the concomitant presence of CRISPR loci and antibiotic resistance determinants.

Oral-gut bacterial profiles discriminate between periodontal health and diseases.

OBJECTIVE: This investigation explored oral-gut microbial signatures with potential to distinguish among periodontal conditions. BACKGROUND DATA: The interplay between the oral and gut microbiomes may be a critical pathway linking periodontal diseases and systemic inflammatory disorders. The mechanisms by which oral microorganisms translocate to the gut and cause microbial dysbiosis, favoring an inflammatory state, are still unknown. As a first approach, characterization of oral-gut microbial profiles associated with periodontal health and diseases can provide insights on such mechanisms of etiology and pathogenesis. METHODS: Fecal and saliva samples from individuals with periodontal health (PH, 8), gingivitis (GG, 17), and periodontitis (PD, 24) were analyzed for their microbial composition by 16S rRNA gene sequencing. Microbial taxa were compared and correlated to periodontal parameters. Multivariate discriminant analysis (MDA) was carried out to identify profiles related to health and disease. RESULTS: Few significant differences in oral-gut taxa were detected among clinical groups, although increase in fecal Fusobacterium nucleatum ss vincentii and salivary Aggregatibacter actinomycetemcomitans, Parvimonas micra, and Fretibacterium sp. HMT358 were strongly correlated with deep pockets and inflammation (p < .01). Over 50% of the fecal microbiota comprised microorganisms shared between oral and gut sites, whereas oral taxa were detected in approximately 9%, particularly enriched in GG fecal samples (p = .04). Trends for lower fecal richness and higher salivary diversity in PD compared to PH were observed. MDA was able to classify correctly 82% of the patients into the clinical groups. Main classifiers of periodontitis were high BMI, older age, and enrichment of oral-fecal Leptotrichia sp. HMT4, Peptostreptococcus stomatis, Dialister invisus, and a novel Lautropia sp. HMTC89-like organism. CONCLUSION: Within the limitations of an exploratory investigation, specific profiles of oral-gut taxa, including known and potential novel organisms, combined with social-demographic features were able to discriminate individuals with periodontal diseases in this study population.

In Vitro Cytotoxic and Leishmanicidal Activity of Isolated and Semisynthetic ent-Pimaranes from Aldama arenaria.

Two pimaranes ent-pimara-8(14),15-dien-19-oic acid (1: ) and ent-8(14),15-pimaradien-3ß-ol (2: ), isolated from Aldama arenaria, and six semi-synthetic derivatives methyl ester of the ent-pimara-8(14),15-dien-19-oic acid (3: ), ent-pimara-8(14),15-dien-19-ol (4: ), acetate of ent-pimara-8(14),15-dien-19-ol (5: ), ent-pimara-8(14),15-dien-19-ol succinic acid (6: ), acetate of ent-8(14),15-pimaradien-3ß-ol (7: ), ent-8(14),15-pimaradien-3ß-ol succinic acid (8: ) were evaluated in vitro for their cytotoxic activities to childhood leukemia cell lines and leishmanicidal activity against the parasite Leishmania amazonensis. Among these compounds, 1: to 6: presented moderate cytotoxic activity, with compound 4: being the most active (GI50 of 2.6 µM for the HL60 line) and the derivatives 7: and 8: being inactive. Against the parasite Leishmania amazonensis, the most promising derivative was the acetate of ent-pimara-8(14),15-dien-19-ol (5: ), with EC50 of 20.1 µM, selectivity index of 14.5, and significant reduction in the parasite load. Pimarane analogues 1: , ent-pimara-8(14),15-dien-19-oic acid, and 2: , ent-8(14),15-pimaradien-3ß-ol, presented different activities, corroborating the application of such molecules as prototypes for the design of other derivatives that have greater cytotoxic or leishmanicidal potential.

Thermofluor-Based Optimization Strategy for the Stabilization of Recombinant Human Soluble Catechol-O-Methyltransferase.

Catechol-O-methyltransferase (COMT) has been involved in a number of medical conditions including catechol-estrogen-induced cancers and a great range of cardiovascular and neurodegenerative diseases such as Parkinson's disease. Currently, Parkinson's disease treatment relies on a triple prophylaxis, involving dopamine replacement by levodopa, the use of aromatic L-amino acid decarboxylase inhibitors, and the use of COMT inhibitors. Typically, COMT is highly thermolabile, and its soluble isoform (SCOMT) loses biological activity within a short time span preventing further structural and functional trials. Herein, we characterized the thermal stability profile of lysate cells from Komagataella pastoris containing human recombinant SCOMT (hSCOMT) and enzyme-purified fractions (by Immobilized Metal Affinity Chromatography-IMAC) upon interaction with several buffers and additives by Thermal Shift Assay (TSA) and a biological activity assessment. Based on the obtained results, potential conditions able to increase the thermal stability of hSCOMT have been found through the analysis of melting temperature (Tm) variations. Moreover, the use of the ionic liquid 1-butyl-3-methylimidazolium chloride [C4mim]Cl (along with cysteine, trehalose, and glycerol) ensures complete protein solubilization as well as an increment in the protein Tm of approximately 10 °C. Thus, the developed formulation enhances hSCOMT stability with an increment in the percentage of activity recovery of 200% and 70% when the protein was stored at 4 °C and -80 °C, respectively, for 12 h. The formation of metanephrine over time confirmed that the enzyme showed twice the productivity in the presence of the additive. These outstanding achievements might pave the way for the development of future hSCOMT structural and biophysical studies, which are fundamental for the design of novel therapeutic molecules.

Physicochemical Properties of Zinc and Lactose in Solid Mixtures: Influence of Trituration Process.

BACKGROUND: Recent experimental results supporting the dynamization process show modification in the characteristics of solid mixtures. OBJECTIVE: The present work aims to evaluate the physicochemical properties of metallic zinc and lactose, evidencing the interactions between all chemical components presented in dynamized solid mixtures by analytical techniques. METHODS: Mixtures of zinc and lactose (1:9 w/w) were successively triturated at the same proportion according to the Brazilian Homeopathic Pharmacopoeia, receiving the designation of 10-1 - 10-6 (1dH - 6dH). All samples were submitted to the following characterization techniques: Atomic Absorption Spectrometry (AAS), Scanning Electron Microscopy (SEM), X-ray Diffraction (XRD), Differential Scanning Calorimetry (DSC), Thermogravimetry (TG), and Raman Spectroscopy (RS). RESULTS: AAS results detected 97.0% of zinc in the raw material, and the triturated zinc lactose system (ZnMet) presented mean values similar to those expected for the physical mixtures: i.e., 9.94%, 1.23%, and 0.11% in the three first proportions (10-1, 10-2, 10-3), respectively. SEM images showed particle size reduction due to the trituration process. The XRD assays of ZnMet 10-3 and 10-6 indicated peak changes at 12.3° and 43.26°, probably associated with modifications of inter-atomic crystalline spacing. The thermal analysis results of dynamized samples suggest modifications in the chemical interaction between zinc and lactose induced by the physical forces applied. RS experiments showed variation in vibration frequencies due to the dynamization procedure, in which marked ZnMet 10-6 spectral modifications were detected at 357, 477, 1086 and 1142 cm-1, and in the wavelength range 860-920 cm-1. CONCLUSION: These results highlight the importance of applying suitable characterization methods to improve our understanding of the properties of homeopathic solid mixtures, whereas the uses of sensitive tools evidence the influence of trituration on the crystalline properties and in the enthalpy variation of dynamized samples.

The distribution and chemical coding of enteroendocrine cells in Trypanosoma cruzi-infected individuals with chagasic megacolon.

Chagas disease is caused by the parasite, Trypanosoma cruzi that causes chronic cardiac and digestive dysfunction. Megacolon, an irreversible dilation of the left colon, is the main feature of the gastrointestinal form of Chagas disease. Patients have severe constipation, a consequence of enteric neuron degeneration associated with chronic inflammation. Dysmotility, infection, neuronal loss and a chronic exacerbated inflammation, all observed in Chagas disease, can affect enteroendocrine cells (EEC) expression, which in turn, could influence the inflammatory process. In this study, we investigated the distribution and chemical coding of EEC in the dilated and non-dilated portion of T. cruzi-induced megacolon and in non-infected individuals (control colon). Using immunohistochemistry, EECs were identified by applying antibodies to chromogranin A (CgA), glucagon-like peptide 1 (GLP-1), 5-hydroxytryptamine (5-HT), peptide YY (PYY) and somatostatin (SST). Greater numbers of EEC expressing GLP-1 and SST occurred in the dilated portion compared to the non-dilated portion of the same patients with Chagas disease and in control colon, but numbers of 5-HT and PYY EEC were not significantly different. However, it was noticeable that EEC in which 5-HT and PYY were co-expressed were common in control colon, but were rare in the non-dilated and absent in the dilated portion of chagasic megacolon. An increase in the number of CgA immunoreactive EEC in chagasic patients reflected the increases in EEC numbers summarised above. Our data suggests that the denervation and associated chronic inflammation are accompanied by changes in the number and coding of EEC that could contribute to disorders of motility and defence in the chagasic megacolon.

Functional, biochemical and molecular impact of sediment plumes from deep-sea mining on Mytilus galloprovincialis under hyperbaric conditions.

The deep-sea is the biggest ecosystem in the world and despite the extreme conditions that characterize it, is highly biodiverse and complex. Deep-sea mining has been foreseen as a potential and concerning new stressor, and among the deep-sea mining associated stressors, sediment plumes, likely to be released into the water column as a side effect of mining, can reach habitats within a radius of more than a hundred kilometers. The present study examined the effects of suspended sediments of different grain sizes (63-125 µm, 125-250 µm and 250-500 µm) in the model species Mytilus galloprovincialis, at 4 bar, as a proxy to address the potential effects of sediment plumes, in the water column, with different grain sizes under high pressure conditions. Functional (filtration rate - FR), biochemical (catalase - CAT, glutathione s-transferase - GST, lipid peroxidation - LPO) and molecular (gene expression of [actin (ACTN), glutathione S-transferase alpha (GSTA), superoxide dismutase 2 (SOD2), catalase (CAT), heat shock protein 60 (HSP60), cytochrome c oxidase (COI) and DNA mismatch repair protein (MSH6)]) endpoints were studied in juvenile organisms. The FR decreased significantly for all tested grain size ranges, with a more severe effect for the particles with a diameter between 63 and 125 µm. In addition to the FR, significant changes were also observed for all tested biomarkers. Gene expression was significantly downregulated for CAT and ACTN. Overall, this study demonstrated that the smaller sized particles are the ones leading to more severe effects. Given their high dispersion potential and longer suspension periods under mining operation scenarios, particular attention should be given to the release of sediment plumes that may affect deep-sea environments and the water column. It is, therefore, vital to create standards and guidelines for sustainable mining practices.

Swine wastewater treatment in high rate algal ponds: Effects of Cu and Zn on nutrient removal, productivity and biomass composition.

This study aimed to evaluate the simultaneous interferences of Cu and Zn found in swine wastewater (SW) in the development of microalgae considering real conditions of cultivation in high rate algal ponds (HRAPs). Ten HRAPs on a pilot scale were fed with SW with different mixtures of Cu (0.5-3.0 mg/L) and Zn (5.0-25.0 mg/L). The interferences of these metals in removing nutrients (N-NH4+ and soluble phosphorus (Ps)) from the SW were determined. In addition, this study evaluated the effects on biomass growth and biochemical composition. Chlorella sp. was dominant in all HRAPs and the condition that potentiated its growth occurred in medium containing 1.8 mg Cu/L + 15.0 mg Zn/L, while higher concentrations conferred inhibition. Only Cu compromised the removal rates of N-NH4+ while the effects of Zn were not significant. Contrary, Zn interfered with Ps removal rates, but the impact of Cu was not significant. The greatest Cu applications increased the protein levels by biomass (50.5-55.2 %). Carbohydrate accumulation was favored by conditions that inhibited the development of microalgae due to either limitation or excess of metals. Copper and Zn compromised the levels of lipids, and the control treatment had the highest content (24.5 %). The presence of Cu and Zn changed the dynamics of HRAPs regarding nutrient removal, productivity, and biochemical composition of the biomass.

Effects of Artemisia annua alcohol extract on physiological and innate immunity of Nile tilapia (Oreochromis niloticus) to improve health status.

Plants are a potential source of active molecules and are environmentally safer and cheaper than synthetic antibiotics. Bioactive compounds of Artemisia annua have shown pharmacological activities and are used globally as a supplement. The present study tested whether dietary supplementation with alcohol extract of the plant A. annua (ae-Aa; patent BR10201902707) improves the health status of juvenile Nile tilapia and increases resistance to diseases when fish are challenged with the bacteria Aeromonas hydrophila. The experimental design was completely randomized with four experimental groups (0.0, 0.1, 0.25, and 0.5% ae-Aa in the diets) with five repetitions (12 fish per repetition/experimental unit). We assessed serum glucose and cortisol levels in plasma, leukocyte respiratory activity, total plasma protein, serum lysozyme levels, as well as the number of circulating red blood cells and fish leukocytes at the end of the 30 days of feeding (phase I) and 24h after exposure to bacteria (phase II). The levels of lipid peroxidation, catalase activity and glutathione S-transferase of fish were also analyzed. The supplementation of 0.5% of ae-Aa was sufficient to increase the respiratory burst of leukocyte and lysozyme activity, total plasma protein, blood thrombocytes, neutrophils and monocytes after bacterial challenge (P < 0.05), and minimized stress response with decreases in plasmatic glucose and cortisol, and reduction in lipid peroxidation levels (P < 0.05). Results of the present study suggest that ae-Aa as a dietary supplement has benefits, including supplementation with 0.5% A. annua extract for 30 days to minimize the stress response and modulate innate immunity in Nile tilapia, providing fish with greater resistance and disease protection.

Beta-lactams susceptibility testing of penicillin-resistant, ampicillin-susceptible Enterococcus faecalis isolates: a comparative assessment of Etest and disk diffusion methods against broth dilution.

This study aimed to evaluate the accuracy of disk diffusion and Etest methods, compared to that of the broth dilution reference method for identifying beta-lactam susceptibilities of Penicillin-Resistant, Ampicillin-Susceptible Enterococcus faecalis (PRASEF) isolates. Fifty-nine PRASEF and 15 Penicillin-Susceptible, Ampicillin-Susceptible E. faecalis (PSASEF) clinical nonrepetitive isolates were evaluated. The effectiveness of five beta-lactams (ampicillin, amoxicillin, imipenem, penicillin, and piperacillin) was tested. All antimicrobial susceptibility tests were performed and interpreted according to the Clinical and Laboratory Standards Institute guidelines. Interpretative discrepancies, such as essential agreement, categorical agreement, and errors, were assessed. The acceptability was ≥ 90% for both categorical agreement and essential agreement. Etest proved to be an accurate method for testing beta-lactam susceptibilities of the emerging PRASEF isolates, disk diffusion presented poor performance, particularly for imipenem and piperacillin.

Prospecting and Identifying Phyllanthus amarus Lignans with Antileishmanial and Antitrypanosomal Activity.

Ten lignans (1:  - 10: ) were isolated from the hexane-ethyl acetate extract of Phyllanthus amarus leaves. Three of them, cubebin dimethyl ether (3: ), urinatetralin (4: ), and lintetralin (7: ) are described for the first time in this species, while phyllanthin (1: ), niranthin (2: ), 5-demethoxyniranthin (5: ), isolintetralin (6: ), hypophyllanthin (8: ), nirtetralin (9: ), and phyltetralin (10: ) have been already reported from P. amarus. Among the lignans tested against Trypanosoma cruzi intracellular amastigotes, 2: was the most active with an EC50 of 35.28 µM. Lignans 2, 5, 7: , and 9: showed inhibitory effects against Leishmania amazonensis promastigotes with EC50 of 56.34, 51.86, 23.57, and 43.27 µM, respectively. During in vitro infection assays, 5: reduced amastigotes by 91% at 103.68 µM concentration, whereas 7: and 9: reduced amastigotes by approximately 84% at 47.5 and 86.04 µM, respectively. Lignans 5, 7: , and 9: were more potent in intracellular amastigotes with EC50 of 2.76, 8.30, and 15.83 µM, respectively, than in promastigotes. CC50 for all samples was > 100 µg/mL, thus revealing low cytotoxicity against macrophages, and selectivity against the parasite. L. amazonensis promastigotes treated with compounds 2: and 9: showed decreased respiratory control of 38% and 25%, respectively, suggesting a change in mitochondrial membrane potential and lower ATP production.

Anti-Leishmania amazonensis activity of the terpenoid fraction from Eugenia pruniformis leaves.

Leishmaniasis is caused by protozoan parasites belonging to the genus Leishmania and includes cutaneous, mucocutaneous and visceral clinical forms. Drugs currently available for leishmaniasis treatment present high toxicity, and development of parasite resistance. Plants constitute an important source of compounds with leishmanicidal potential. This study aimed to evaluate the anti-Leishmania amazonensis activity of the terpenoid fraction of Eugenia pruniformis leaves (TF-EpL). TF-EpL was active against the promastigote and intracellular amastigote forms of L. amazonensis with IC50(24h) value of 43.60µg/mL and 44.77µg/mL, respectively. TF-EpL altered the cell cycle of the parasite, increasing 2.32-fold the cells in the Sub-G0/G1 phase. TF-EpL also changed the ΔΨm and increased ROS and the number of annexin-V-PI positive promastigotes, which suggests incidental death. ß-sitosterol, ursolic acid, corosolic acid and asiatic acid were isolated from TF-EpL. The results showed the antileishmanial activity of E. pruniformis terpenoids and its potential for further studies as a source of new drugs for leishmaniasis.

Macrobrachium amazonicum (Crustacea, Decapoda) Used to Biomonitor Mercury Contamination in Rivers.

Open-air landfill's may be are considered as a potential source of human environmental exposure to chemical substances such as, polycyclic aromatic hydrocarbons and toxic metals. Due to possible availability of mercury in the environment caused by open landfill emissions, this study evaluates the spatiality and seasonality of macroinvertebrates, in particular shrimps (Macrobrachium amazonicum), exposure to mercury (Hg). Information regarding Hg accumulation in this crustacean may be important for the development of public policies aiming conservation and preservation of ecosystems surrounding landfills in Amazon, and around the world. Sampling occurred quarterly in the following months: November/2015; February/2016; May/2016 and; August/2016. In each of these months, three points were selected: P1, P2 and P3. The samples were processed via acid digestion and the quantification of metal was performed by inductively coupled plasma mass spectrometry. The average concentration of total mercury (T-Hg) was 24.565 ± 6.610 µg kg-1 wet weight, with minimum and maximum limits of 12.742 ± 11.367 (P3) and 35.509 ± 14.761 µg kg-1 wet weight (P1) in November/2015 and August/2016, respectively. The concentration of total mercury (T-Hg) in shrimps was different between points (p = 0.004) and months (p = 0.000). The T-Hg concentrations were significantly higher in May and August 2016, which corresponds to the dry season. The presence of landfills promotes large accumulation of T-Hg in the aquatic biota and represents a risk to human health. However, seasonal changes in T-Hg levels were observed. In the wettest period, bioconcentration factor levels decrease in aquatic organisms.

Impact of Surgical Checklist on Mortality, Reoperation, and Readmission Rates in Brazil, a Developing Country, and Canada, a Developed Country.

PURPOSE: To compare the mortality, reoperation, and readmission rates before and after the implementation of a surgical checklist in Brazil and Canada. DESIGN: An epidemiological, retrospective study was conducted. METHODS: Preimplementation and postimplementation data were collected via patient chart reviews to determine mortality, reoperation, and readmission rates. FINDINGS: In Brazil, a decrease in readmission rate from 2.9% to 1.7% (P = .518) was observed after the implementation of the checklist. In Canada, reoperation rate decreased from 5.6% to 4.8% (P = .649) and mortality from 1.7% to 0.9% (P = .407) after implementation. In the Brazilian institution, patients with incomplete checklists had increased rates of readmission, from 1.4% to 2.4% (P = .671), and reoperation, from 6.8% to 10.4% (P = .232). CONCLUSIONS: The use of surgical checklist did not translate into improvements in the outcomes studied after its implementation in any of the scenarios evaluated. This result is possibly justified by the socioeconomic structure of each of these settings.