Genomic Comparative of Pseudomonas aeruginosa Small Colony Variant, Mucoid and Non-mucoid Phenotypes Obtained from a Patient with Cystic Fibrosis During Respiratory Exacerbations.

Pseudomonas aeruginosa, the most prevalent opportunistic pathogen in chronic obstructive pulmonary disease, associated with high morbidity and mortality in patients with cystic fibrosis (CF), is practically impossible to be eradicated from the airways in chronicity. Its extraordinary genomic plasticity is possibly associated with high antimicrobial resistance, virulence factors, and its phenotypic diversity. The occurrence of P. aeruginosa isolates promoting airway infection, showing mucoid, non-mucoid, and small colony variant (SCV) phenotypes, was observed simultaneously, in the present study, in sputum cultures obtained from a male CF young patient with chronic pulmonary infection for over a decade. The isolates belonged to a new ST (2744) were obtained in two moments of exacerbation of the respiratory disease, in which he was hospitalized. Genetic background and phenotypic analysis indicated that the isolates exhibited multi- and pan-antimicrobial resistant profiles, as well as non-susceptible to polymyxin and predominantly hypermutable (HPM) phenotypes. Whole genome sequencing showed variations in genome sizes, coding sequences and their determinants of resistance and virulence. The annotated genomes were compared for antimicrobial resistance, hypermutability, and SCV characteristics. We highlight the lack of reported genetic determinants of SCV emergence and HPM phenotypes, which can be explained in part due to the very short time between collections of isolates. To the best of our knowledge, this is the first report of genome sequencing of P. aeruginosa SCV from a CF patient in Brazil.

Genetic determinants of antimicrobial resistance in polymyxin B resistant Pseudomonas aeruginosa isolated from airways of patients with cystic fibrosis.

Pseudomonas aeruginosa is the main pathogen associated with pulmonary exacerbation in patients with cystic fibrosis (CF). CF is a multisystemic genetic disease caused by mutations in the cystic fibrosis transmembrane conductance regulator gene, which mainly affects pulmonary function. P. aeruginosa isolated from individuals with CF in Brazil is not commonly associated with multidrug resistance (MDR), especially when compared to global occurrence, where the presence of epidemic clones, capable of expressing resistance to several drugs, is often reported. Due to the recent observations of MDR isolates of P. aeruginosa in our centers, combined with these characteristics, whole-genome sequencing was employed for analyses related to antimicrobial resistance, plasmid identification, search for phages, and characterization of CF clones. All isolates in this study were polymyxin B resistant, exhibiting diverse mutations and reduced susceptibility to carbapenems. Alterations in mexZ can result in the overexpression of the MexXY efflux pump. Mutations in oprD, pmrB, parS, gyrA and parC may confer reduced susceptibility to antimicrobials by affecting permeability, as observed in phenotypic tests. The phage findings led to the assumption of horizontal genetic transfer, implicating dissemination between P. aeruginosa isolates. New sequence types were described, and none of the isolates showed an association with epidemic CF clones. Analysis of the genetic context of P. aeruginosa resistance to polymyxin B allowed us to understand the different mechanisms of resistance to antimicrobials, in addition to subsidizing the understanding of possible relationships with epidemic strains that circulate among individuals with CF observed in other countries.

Susceptibility of solid organ transplant recipients to viral pathogens with zoonotic potential: A mini-review.

A substantial number of zoonotic diseases are caused by viral pathogens, representing a significant menace to public health, particularly to susceptible populations, such as pregnant women, the elderly, and immunocompromised individuals. Individuals who have undergone solid organ transplantation frequently experience immunosuppression, to prevent organ rejection, and, thus are more prone to opportunistic infections. Furthermore, the reactivation of dormant viruses can threaten transplant recipients and organ viability. This mini-review examines the up-to-date literature covering potential zoonotic and organ rejection-relevant viruses in solid organ transplant recipients. A comprehensive list of viruses with zoonotic potential is highlighted and the most important clinical outcomes in patients undergoing transplantation are described. Moreover, this mini-review calls attention to complex multifactorial events predisposing viral coinfections and the need for continuous health surveillance and research to understand better viral pathogens' transmission and pathophysiology dynamics in transplanted individuals.

Susceptibility of solid organ transplant recipients to viral pathogens with zoonotic potential: A mini-review

ABSTRACT A substantial number of zoonotic diseases are caused by viral pathogens, representing a significant menace to public health, particularly to susceptible populations, such as pregnant women, the elderly, and immunocompromised individuals. Individuals who have undergone solid organ transplantation frequently experience immunosuppression, to prevent organ rejection, and, thus are more prone to opportunistic infections. Furthermore, the reactivation of dormant viruses can threaten transplant recipients and organ viability. This mini-review examines the up-to-date literature covering potential zoonotic and organ rejection-relevant viruses in solid organ transplant recipients. A comprehensive list of viruses with zoonotic potential is highlighted and the most important clinical outcomes in patients undergoing transplantation are described. Moreover, this mini-review calls attention to complex multifactorial events predisposing viral coinfections and the need for continuous health surveillance and research to understand better viral pathogens' transmission and pathophysiology dynamics in transplanted individuals.

Methanolic Extracts of Chiococca alba in Aedes aegypti Biorational Management: Larvicidal and Repellent Potential, and Selectivity against Non-Target Organisms.

The use of formulations containing botanical products for controlling insects that vector human and animal diseases has increased in recent years. Plant extracts seem to offer fewer risks to the environment and to human health without reducing the application strategy's efficacy when compared to synthetic and conventional insecticides and repellents. Here, we evaluated the potential of extracts obtained from caninana, Chiococca alba (L.) Hitchc. (Rubiaceae), plants as a tool to be integrated into the management of Aedes aegypti, one of the principal vectors for the transmission of arborviruses in humans. We assessed the larvicidal and repellence performance against adult mosquitoes and evaluated the potential undesired effects of the extracts on non-target organisms. We assessed the susceptibility and predatory abilities of the nymphs of Belostoma anurum, a naturally occurring mosquito larva predator, and evaluated the C. alba extract's cytotoxic effects in mammalian cell lines. Our chromatographic analysis revealed 18 compounds, including rutin, naringin, myricetin, morin, and quercetin. The methanolic extracts of C. alba showed larvicidal (LC50 = 82 (72-94) mg/mL) activity without killing or affecting the abilities of B. anurum to prey upon mosquito larvae. Our in silico predictions revealed the molecular interactions between rutin and the AeagOBP1 receptor to be one possible mechanism for the repellent potential recorded for formulations containing C. alba extracts. Low cytotoxicity against mammalian cell lines reinforces the selectivity of C. alba extracts. Collectively, our findings highlight the potential of C. alba and one of its constituents (rutin) as alternative tools to be integrated into the management of A. aegypti mosquitoes.

Recognizing the enemy: do predator cues influence hatching in Neotropical annual killifish?

Annual fish species have evolved complex adaptations to survive in temporary wetlands. The main adaptation of these fish is the ability to produce embryos that survive dry periods. Embryo development of this fish can show variation at multiple levels influenced by many environmental factors, such as photoperiod and temperature. Predator cues are another factor that can influence the embryonic stage. One way in which annual fish could adapt to predators is by using risk-spreading strategies (through bet-hedging). Nonetheless, this strategy depends on the coevolutionary history between predators and preys and on the degree of environmental unpredictability, resulting in different responses across different species. This study investigated the influence of predator cues on the embryonic development and hatching of two Austrolebias species that inhabit ponds that present differences in hydroperiod and the risk of predator presence. The results confirmed a differentiated response between the two annual fish species tested, corroborating the modulation of hatching against the risk of predation by native predatory fish. The authors further showed that development times varied between the two annual fish species, regardless of the presence of predators. They highlight that the variation in embryonic development is strongly affected by different levels of hydroperiod unpredictability faced by the two species. To unravel finer-scale local adaptations in the annual fish embryo development, future studies should focus on a region with greater spatial gradient.

Anabolic androgenic steroid-induced hypogonadism, a reversible condition in male individuals? A systematic review.

The anabolic-androgenic steroids (AAS) are clinically used as an androgen replacement, in hypogonadism treatment, to induce puberty, and also in the treatment of chronic degenerative diseases. The AAS use out of clinical context is becoming massively, being used merely for aesthetic reasons. AAS abuse may cause severe disarrangement on the HPG axis and generate a significant decrease in testosterone synthesis and secretion by the testes. This review aims to evaluate whether the hypogonadism induced by AAS abuse is reversible and under what circumstances the reversibility is possible. For this, PRISMA guidelines and several databases are used between July and September 2020. Altogether, this systematic review identified and analysed 179 cases of AAS users. Of these, 168 cases had the hypogonadism clearly diagnosed and proven to be linked exclusively to AAS abuse. However, between these 168 cases, only 38 cases presented fully known outcomes and among these, merely in 4, the hypogonadism was completely reversible (2 based on drug therapy) with HPG axis recovery. In conclusion, this review presents evidences that AAS-induced hypogonadism is a seriously underestimated problem, and in the majority of cases, full recovery is very difficult to succeed.

Carbapenem-Resistant Pseudomonas aeruginosa in Chronic Lung Infection: Current Resistance Profile and Hypermutability in Patients with Cystic Fibrosis.

Pseudomonas aeruginosa is associated with chronic and progressive lung disease and is closely related to increased morbidity and mortality in cystic fibrosis (CF) patients. Hypermutable (HPM) P. aeruginosa isolates have been described in these patients and are usually associated with antibiotic resistance. This study aimed to investigate the occurrence of carbapenem resistance and hypermutable phenotype in 179 P. aeruginosa isolates from 8 chronically CF patients assisted at two reference centers in Rio de Janeiro, Brazil. Using disk diffusion test, non-susceptible (NS) rates higher than 40% were observed for imipenem, amikacin, and gentamicin. A total of 79 isolates (44.1%), 71 (39.6%), and 8 (4.4%) were classified as carbapenem-resistant (CR resistance to at least one carbapenem), multidrug-resistant (MDR), and extensively drug-resistant (XDR), respectively. Minimal inhibitory concentration was determined for 79 CR P. aeruginosa and showed the following variations: 4 and 128 µg/mL to imipenem, 4 and 64 µg/mL to meropenem, and 4 and ≥ 32 µg/mL to doripenem. We have found only four (2.23%) HPM isolates from 4 patients. Analyzing the genetic relationship among the HPM isolates, 3 pulsed-field gel electrophoresis/pulsotypes (D, M, and J) were observed. Only M pulsotype was recovered from two patients in different years. Polymerase chain reaction screening for blaGES, blaIMP, blaKPC, blaNDM, blaOXA-48, blaSPM, and blaVIM genes was performed for all CR isolates and none of them were positive. Our results demonstrate a high occurrence of CR and MDR P. aeruginosa of CF patients follow-up in both centers studied, while the presence of HPM is still unusual.

Effect of high-fluoride dentifrice and bracket bonding composite material on enamel demineralization in situ.

OBJECTIVE: This in situ study evaluated the effect of high-fluoride dentifrice (5000 µg F-/g) and fluoride-containing bonding composite resin on enamel demineralization adjacent to orthodontic brackets. METHODS: Ten volunteers wore palatal appliances containing bovine enamel blocks with metallic brackets bonded with fluoride-free or fluoride-containing composite resin. During three phases of 14 days each, three dentifrices with different fluoride concentrations (0, 1100, and 5000 µg F-/g) were tested. The cariogenic challenge consisted of 20% sucrose solution dripped 8x/day onto the dental blocks. At the end of each phase, biofilm formed was collected for fluoride analysis. Cross section hardness was performed in enamel blocks, and the lesion area was calculated. Data were analyzed by two-way ANOVA followed by Tukey post hoc test (α = 5%). RESULTS: The only signicant factor for all the variables under study was the dentifrice. Smaller lesion area and higher fluoride concentration on biofilm were found in 5000 µg F-/g group, irrespective of bonding composite resin (p < 0.001). Neither bracket-bonding composite resin nor the interaction between the factors was statistically significant (p > 0.05) for all the variables. CONCLUSION: High-fluoride dentifrice is effective in reducing demineralization on enamel adjacent to orthodontic brackets, while the fluoride-containing bonding composite resin does not influence it. CLINICAL SIGNIFICANCE: Since high-fluoride dentifrice was able to reduce demineralization adjacent to brackets, it can be an option to caries management in orthodontics patients.

Degradation of antibiotic ciprofloxacin by different AOP systems using electrochemically generated hydrogen peroxide.

The present work reports the degradation of the antibiotic ciprofloxacin (CIP) by different advanced oxidative process systems (UV; Anodic Oxidation; H2O2; H2O2/UV; H2O2/Fe2+ and H2O2/UV/Fe2+) in an electrochemical cell using gas diffusion electrode (GDE) for the synthesis of hydrogen peroxide. CIP degradation and mineralization were evaluated by high efficiency liquid chromatography (HPLC) and total organic carbon (TOC) techniques. Of all the systems investigated, the photoelectro-Fenton system presented the best degradation efficiency; this system promoted highly significant mineralization percentages of 54.8% and 84.6% in 90 and 360 min, and relatively lower energy consumption rates of 4110.0 and 9808.2 kWh kg-1 TOC, respectively. In 6 h period of experiment, the main degradation products of ciprofloxacin were identified, and the aliphatic acids obtained helped confirm the rupture of the aromatic ring. The application of the photoelectro-Fenton process with in situ eletroctrogeneration of H2O2 using GDE has proved to be suitably promising for the treatment of organic pollutants.

Oxidative stress resistance in a short-lived Neotropical annual killifish.

Oxidative stress plays an important role in the evolution of aging and life history. High investments in life-history traits and environmental conditions can be associated with increased oxidative stress and aging process. However, to date, most studies that investigated variations in oxidative status were performed with long-lived vertebrates. Studies with short-lived vertebrates in wild are nonexistent. Annual killifishes have the shortest lifespans among vertebrates and inhabit temporary ponds subject to large variations in environmental conditions. In this sense, we investigated whether the high investment in growth and reproduction in a short-lived vertebrate and the large variations in environment has any cost in susceptibility to oxidative stress. We assessed the seasonal variation and the environmental correlates of four different oxidative status markers (lipid peroxidation and activity of the antioxidant enzymes Superoxide Dismutase, Catalase and Glutathione S-Transferase) along the life cycle of wild individuals of the Neotropical annual fish Austrolebias minuano. Males showed reduction in all biomarkers (except proteins) along their life cycle, while females showed increased oxidative stress only in the growth period. In addition, we showed that water physicochemical parameters, habitat structure and presence of co-occurring killifish species influenced the seasonal variation of the biomarkers. A. minuano showed an efficient antioxidant system for most part of their life cycle (mainly in males), suggesting a well-developed oxidative stress regulation system. We also show that annual fish mortality (mainly in males) apparently is not related to oxidative stress. Thus, environmental factors should drive annual fish aging and mortality.

Skin temperature as a clinical parameter for nursing care: a descriptive correlational study.

OBJECTIVE: To identify the skin temperature in different body areas of hospitalised individuals in the surgical unit, without risk of developing a pressure ulcer (PU). METHODS: A descriptive, correlational and cross-sectional study, carried out May-October 2017, in a surgical unit of a university hospital in southern Brazil. Temperature was measured at the bony prominences including scapula, elbow, trochanters and heels, on both sides of the body, as well as occipital and sacral regions. RESULTS: A total of 230 patients took part in the study. All regions of the body measured presented differences in temperatures. The sacral region presented the highest mean temperature (34.2±0.1°C). Patients (aged 18-59 years) had higher skin temperatures in the sacral region than older patients (aged 60-88 years). There was a symmetry in temperatures on both sides of the body. There was a low degree of correlation between age, room temperature, room humidity and skin temperature in some body regions. CONCLUSION: The study established mean values for skin temperature in specific body regions in patients without risk of developing a PU, hospitalised in a surgical unit. It also demonstrates how skin temperature can be used as a clinical parameter in practice to support the prevention of PUs.

Author Correction: Effects of the serine protease inhibitor rBmTI-A in an experimental mouse model of chronic allergic pulmonary inflammation.

An amendment to this paper has been published and can be accessed via a link at the top of the paper.

Semi-automated counting model for arbuscular mycorrhizal fungi spores using the Circle Hough Transform and an artificial neural network.

Arbuscular Mycorrhizae (AM) are mutualistic associations between Arbuscular Mycorrhizal Fungi (AMF) and the roots of many plant species. AMF spores give rise to filaments that develop in the root system of plants and contribute to the absorption of water and some nutrients. This article introduces a semi-automated counting model of AMF spores in slide images based on Artificial Neural Network (ANN). The semi-automated counting of AMF spores facilitates and accelerates the tasks of researchers, who still do the AMF spore counting manually. We built a representative database of spore images, processing images through the Circle Hough Transform (CHT) method and training an ANN to classify patterns automatically. The classification analysis and the performances of the proposed method against the manual method are presented in this paper. The accuracy for the identification of spores by CHT in conjunction to ANN classification in the images was 90%. The results indicate that this method can accurately detect the presence of AMF spores in images as well as count them with a high level of confidence.

Effects of the serine protease inhibitor rBmTI-A in an experimental mouse model of chronic allergic pulmonary inflammation.

To evaluate whether a recombinant serine protease inhibitor (rBmTI-A) modulates inflammation in an experimental model of chronic allergic lung inflammation. Balb/c mice were divided into four groups: SAL (saline), OVA (sensitized with ovalbumin), SAL + rBmTI-A (control treated with rBmTI-A) and OVA + rBmTI-A (sensitized with ovalbumin and treated with rBmTI-A). The animals received an intraperitoneal injection of saline or ovalbumin, according to the group. The groups received inhalation with saline or ovalbumin and were treated with rBmTI-A or saline by nasal instillation. After 29 days, we evaluated the respiratory mechanics; bronchoalveolar lavage fluid (BALF); cytokines; MMP-9, TIMP-1; eosinophils; collagen and elastic fibre expression in the airways; and the trypsin-like, MMP-1, and MMP-9 lung tissue proteolytic activity. Treatment with rBmTI-A reduced the trypsin-like proteolytic activity, the elastance and resistance maximum response, the polymorphonuclear cells, IL-5, IL-10, IL-13 and IL-17A in the BALF, the expression of IL-5, IL-13, IL-17, CD4+, MMP-9, TIMP-1, eosinophils, collagen and elastic fibres in the airways of the OVA + rBmTI-A group compared to the OVA group (p < 0.05). rBmTI-A attenuated bronchial hyperresponsiveness, inflammation and remodelling in this experimental model of chronic allergic pulmonary inflammation. This inhibitor may serve as a potential therapeutic tool for asthma treatment.

Age-associated liver alterations in wild populations of Austrolebias minuano, a short-lived Neotropical annual killifish.

Aging processes have become an attractive field for researchers and annual fish have been used as biological models. However, the study on the changes in age-associated markers during the normal aging in wild populations of annual fish remains open. Austrolebias is a genus of Neotropical annual killifishes, distributed mainly in ephemeral pools across grassland floodplains of temperate South America and represent an emerging biological model for aging research, but studies investigating rapid aging and senescence in this genus of annual fish are almost non-existent. This study was undertaken to examine the changes in age-associated liver markers at the different developmental stages in wild populations of Austrolebias minuano. We demonstrate that A. minuano has a number of liver alterations of different severities throughout the life cycle, suggesting that these changes tend to increase with age. Our results revealed that > 70% of the analyzed livers presented alterations. Thus, our study should instigate new approaches on aging using Neotropical annual fish, and could be useful to improve the knowledge already provided by consecrated biological aging models as e.g. Nothobranchius killifishes.

Effects of dietary inclusion of high concentrations of crude glycerin on meat quality and fatty acid profile of feedlot fed Nellore bulls.

Crude glycerin, the main by-product of biodiesel production, can replace dietary energy sources, such as corn. The objective of this study was to evaluate the inclusion of up to 30% of crude glycerin in dry matter (DM) of the total diets, and its effects on meat quality parameters of feedlot Nellore bulls. Thirty animals (227.7 ± 23.8 kg body weight; 18 months old) were housed in individual pens and fed 5 experimental diets, containing 0, 7.5, 15, 22.5 or 30% crude glycerin (DM basis). After 103 d (21 d adaptation) animals were slaughtered and the Longissimus muscle was collected. The characteristics assessed were chemical composition, fatty acid profile, cholesterol, shear force, pH, color, water-holding capacity, cooking loss and sensory properties. The increasing inclusion of crude glycerin in the diets did not affect the chemical composition of the Longissimus muscle (P > 0.10). A quadratic effect was observed when levels of crude glycerin were increased, on the concentration of pentadecanoic, palmitoleic and eicosenoic fatty acids in meat (P < 0.05), and on the activity of the delta-9 desaturase 16 and delta-9 desaturase 18 enzymes (P < 0.05). The addition of crude glycerin increased the gamma linolenic fatty acid concentration (P < 0.01), and altered the monounsaturated fatty acids in Longissimus muscle of animals (Pquad. < 0.05). Crude glycerin decreased cholesterol content in meat (P < 0.05), and promoted higher flavor score and greasy intensity perception of the meat (P < 0.01). The inclusion of up to 30% crude glycerin in Nellore cattle bulls`diets (DM basis) improves meat cholesterol and sensory attributes, such as flavor, without affecting significantly the physical traits, the main fatty acid concentrations and the chemical composition.

TipMT: Identification of PCR-based taxon-specific markers.

BACKGROUND: Molecular genetic markers are one of the most informative and widely used genome features in clinical and environmental diagnostic studies. A polymerase chain reaction (PCR)-based molecular marker is very attractive because it is suitable to high throughput automation and confers high specificity. However, the design of taxon-specific primers may be difficult and time consuming due to the need to identify appropriate genomic regions for annealing primers and to evaluate primer specificity. RESULTS: Here, we report the development of a Tool for Identification of Primers for Multiple Taxa (TipMT), which is a web application to search and design primers for genotyping based on genomic data. The tool identifies and targets single sequence repeats (SSR) or orthologous/taxa-specific genes for genotyping using Multiplex PCR. This pipeline was applied to the genomes of four species of Leishmania (L. amazonensis, L. braziliensis, L. infantum and L. major) and validated by PCR using artificial genomic DNA mixtures of the Leishmania species as templates. This experimental validation demonstrates the reliability of TipMT because amplification profiles showed discrimination of genomic DNA samples from Leishmania species. CONCLUSIONS: The TipMT web tool allows for large-scale identification and design of taxon-specific primers and is freely available to the scientific community at http://200.131.37.155/tipMT/ .

Genomic information on multidrug-resistant livestock-associated methicillin-resistant Staphylococcus aureus ST398 isolated from a Brazilian patient with cystic fibrosis.

Alarmingly, the isolation of methicillin-resistant Staphylococcus aureus (MRSA) has been increasing among patients with cystic fibrosis (CF). During a previous molecular characterisation of MRSA isolates obtained from patients with CF from Rio de Janeiro, Brazil, one isolate was identified as the ST398 clone, a livestock-associated (LA) MRSA. In this study, we report the draft genome sequence of an LA-MRSA ST398 clone isolated from a patient with CF.