Paving the way forward: Escherichia coli bacteriophages in a One Health approach.

Escherichia coli is one of the most notorious pathogens for its ability to adapt, colonize, and proliferate in different habitats through a multitude of acquired virulence factors. Its presence affects the food-processing industry and causes food poisoning, being also a major economic burden for the food, agriculture, and health sectors. Bacteriophages are emerging as an appealing strategy to mitigate bacterial pathogens, including specific E. coli pathovars, without exerting a deleterious effect on humans and animals. This review globally analyzes the applied research on E. coli phages for veterinary, food, and human use. It starts by describing the pathogenic E. coli pathotypes and their relevance in human and animal context. The idea that phages can be used as a One Health approach to control and interrupt the transmission routes of pathogenic E. coli is sustained through an exhaustive revision of the recent literature. The emerging phage formulations, genetic engineering and encapsulation technologies are also discussed as a means of improving phage-based control strategies, with a particular focus on E. coli pathogens.

Levodopa-induced dyskinesia is preceded by increased levels of anxiety and motor impairment in Parkinson's disease patients.

Introduction: Dopamine replacement therapy with levodopa is the gold standard treatment of Parkinson's disease (PD); however long-term levodopa use is associated with abnormal involuntary movements known as levodopa-induced dyskinesia (LID) in most patients. LID is not preventable and represents the major limitation of PD treatment.Objective: This study was aimed to find clinical and behavioral features that could be used to identify, years in advance, PD patients that are at high risk of developing LID in the future. Method: Data from PD patients enrolled in The Parkinson's progression markers initiative (PPMI, Michael J. Fox Foundation) that developed dyskinesia during their participation in the study were compared with those who did not, and with healthy controls.Result: LID was preceded byhigher levels of trait anxiety and increased motor impairment in PD patients. Additionally, younger age at PD diagnosis, earlier need for dopaminergic therapy and higher initial levodopa dose, were associated with future development of dyskinesia.Conclusion: These findings suggest that easily detectable clinical and behavioral alterations may help to identify PD patients that are more susceptible to develop LID.

Effect of brewery effluent inhibitors on Rhodotorula toruloides NCYC 921 cells grown in pure and mixed cultures at pH 4 and 6.

The presence of inhibitor compounds in the culture medium can cause severe effects on the microorganisms cells. Brewery wastewaters present organic acids (acetic, propionic and butyric acids) which can severely affect yeast cells metabolism, when grown in pure cultures, although in mixed cultures they are able to develop. To understand the physiological changes on Rhodotorula toruloides (formerly Rhodosporidium toruloides) cells when fermenting in the presence of the organic acids present in brewery wastewater, pure and mixed cultures with the microalga Tetradesmus obliquus were performed in a synthetic medium containing the same organic acids concentrations that are present in brewery wastewater at pH 4 and 6. It was concluded that, at pH 4, the organic acids effects in the yeast cells were much more toxic than at pH 6. Moreover, mixed cultures can be an advantage over heterotrophic pure cultures as the microalga is able to contribute for the consumption of potential inhibitors for the yeast.

Rhodosporidium toruloides and Tetradesmus obliquus Populations Dynamics in Symbiotic Cultures, Developed in Brewery Wastewater, for Lipid Production.

In this work, primary brewery wastewater (PBWW) and secondary brewery wastewater (SBWW) separately, or mixed at the ratios of 1:1 (PBWW:SBWW) and 1:7 (PBWW:SBWW), with or without supplementation with sugarcane molasses (SCM), were used as culture media for lipid production by a mixed culture of the oleaginous yeast Rhodosporidium toruloides NCYC 921 and the microalgae Tetradesmus obliquus (ACOI 204/07). Flow cytometry was used to understand the dynamics of the two micro-organisms during the mixed cultures evolution, as well as to evaluate the physiological states of each micro-organism, in order to assess the impact of the different brewery effluent media composition on the microbial consortium performance. Both brewery wastewaters (primary and secondary) without supplementation did not allow R. toruloides heterotrophic growth. Nevertheless, all brewery wastewater media, with and without SCM supplementation, allowed the microalgae growth, although the yeast was the dominant population. The maximum total biomass concentration of 2.17 g L-1 was achieved in the PBWW mixed cultivation with 10 g L-1 of SCM. The maximum lipid content (14.86% (w/w DCW)) was obtained for the mixed culture developed on SBWW supplemented with 10 g L-1 of SCM. This work demonstrated the potential of using brewery wastewater supplemented with SCM as a low-cost culture medium to grow R. toruloides and T. obliquus in a mixed culture for brewery wastewater treatment with concomitant lipid production.

Effect of glutamate on Pyricularia oryzae infection of rice monitored by changes in photosynthetic parameters and antioxidant metabolism.

Considering the importance of blast caused by Pyricularia oryzae in the decrease of rice yield worldwide, this study aimed to assess the photosynthetic performance [leaf gas exchange and chlorophyll (Chl) a fluorescence parameters as well as the photosynthetic pigments concentration], the activities of antioxidant enzymes [ascorbate peroxidase, catalase (CAT), peroxidase (POX), superoxide dismutase (SOD), glutathione peroxidase (GPX), glutathione reductase (GR) and glutathione-S-transferase] and concentrations of hydrogen peroxide (H2 O2 ) and malondialdehyde (MDA) in the leaves of rice plants non-supplied (-Glu) or supplied (+Glu) with glutamate (Glu) and non-infected or infected by P. oryzae. Blast severity was reduced in the leaves of +Glu plants. On the infected leaves of +Glu plants, the values for internal CO2 concentration were lower while the values for net carbon assimilation rate, stomatal conductance as well as for the concentrations of Chl a, Chl b and carotenoids were higher in comparison to infected leaves of -Glu plants. The functionality of the photosynthetic apparatus was preserved in the infected leaves of +Glu plants. The activities of CAT, GPX, GR, POX and SOD increased in the infected leaves of both -Glu and +Glu plants compared to their non-inoculated counterparts, but their activities were lower for +Glu plants. The lower activity of these antioxidative enzymes was triggered by the reduced hydrogen peroxide concentration in the infected leaves of +Glu plants resulting in lower MDA concentration. It can be concluded that photosynthesis was less impaired in infected plants supplied with glutamate due to the lower biochemical constraints for CO2 fixation. Moreover, there was a need for lower activity of reactive oxygen species scavenging enzymes in infected leaves of plants supplied with glutamate due to the lower oxidative stress imposed by P. oryzae infection.

Virulence, attachment and invasion of Caco-2 cells by multidrug-resistant bacteria isolated from wild animals.

Wild animals may be considered important reservoirs for bacterial pathogens and, consequently, possible sources of infection for humans. In this study, selected multidrug-resistant bacteria (Acinetobacter spp., Aeromonas salmonicida, Klebsiella pneumoniae, Pseudomonas fluorescens and Shewanella putrefaciens) isolated from wild animals were characterized on their ability to attach and invade/internalize human colonic carcinoma (Caco-2) cells. In addition, the viability of these bacteria to survive under simulated human gastrointestinal tract conditions as well as the production of virulence factors (homoserine lactones signal molecules, gelatinases, proteases, siderophores and biofilm formation) were studied. The results suggests that all the bacteria presented the capacity to attach and internalize into Caco-2 cells. A. salmonicida and P. fluorescens exhibited the highest ability to internalize. These bacteria were also found to be the highest proteases producers. A. salmonicida and K. pneumoniae survived under simulated human gastrointestinal conditions. These were the bacteria with the highest capacity to produce biofilms. K. pneumoniae was the only bacterium producing siderophores. Taken together, the present results reinforce the need for the "One Health" initiative, underscoring the environment and the animals as important reservoirs of infectious determinants.

New Perspectives on the Use of Phytochemicals as an Emergent Strategy to Control Bacterial Infections Including Biofilms.

The majority of current infectious diseases are almost untreatable by conventional antibiotic therapy given the advent of multidrug-resistant bacteria. The degree of severity and the persistence of infections are worsened when microorganisms form biofilms. Therefore, efforts are being applied to develop new drugs not as vulnerable as the current ones to bacterial resistance mechanisms, and also able to target bacteria in biofilms. Natural products, especially those obtained from plants, have proven to be outstanding compounds with unique properties, making them perfect candidates for these much-needed therapeutics. This review presents the current knowledge on the potentialities of plant products as antibiotic adjuvants to restore the therapeutic activity of drugs. Further, the difficulties associated with the use of the existing antibiotics in the treatment of biofilm-related infections are described. To counteract the biofilm resistance problems, innovative strategies are suggested based on literature data. Among the proposed strategies, the use of phytochemicals to inhibit or eradicate biofilms is highlighted. An overview on the use of phytochemicals to interfere with bacterial quorum sensing (QS) signaling pathways and underlying phenotypes is provided. The use of phytochemicals as chelating agents and efflux pump inhibitors is also reviewed.

Antimicrobial activity of isothiocyanates from cruciferous plants against methicillin-resistant Staphylococcus aureus (MRSA).

Purified isothiocyanates from cruciferous plants (Brassicacea, Syn. Cruciferae) plants were evaluated against 15 isolates of methicillin-resistant S. aureus isolated from diabetic foot-ulcer patients aiming the study of the potential usage of allyl-isothiocyanate, benzyl-isothiocyanate and 2-phenylethyl-isothiocyanate against this important bacteria. Disc diffusion and minimum inhibitory concentration methods were used to access the antimicrobial activity. The index (Ia) and rate (Ra) of the antibacterial activity for each compound were calculated. The results showed a highly dose-dependent compound and chemical structure antibacterial effectiveness. The results showed a strong relation between the chemical structure of isothiocyanates and its antibacterial effectiveness. The benzyl-isothiocyanate was the most effective with a minimum inhibitory concentration varying between 2.9 and 110 µg·mL(-1) with an antibacterial activity rate up to 87%. Moreover, their antibacterial activity was mainly bactericidal. This study provides scientific evidence that isothiocyanates have an interesting biological value and must be considered as an important tool to be used against MRSA.

Serotonin modulates infraslow oscillation in the dentate gyrus during Non-REM sleep.

Synchronous neuronal activity is organized into neuronal oscillations with various frequency and time domains across different brain areas and brain states. For example, hippocampal theta, gamma and sharp wave oscillations are critical for memory formation and communication between hippocampal subareas and the cortex. In this study, we investigated the neuronal activity of the dentate gyrus (DG) with electrophysiological and optical imaging tools during sleep-wake cycles. We found that the activity of major glutamatergic cell populations in the DG is organized into in-fraslow oscillations (0.01 - 0.03 Hz) during NREM sleep. Although the DG is considered a sparsely active network during wakefulness, we found that 50% of granule cells and about 25% of mossy cells exhibit increased activity during NREM sleep. Further experiments revealed that the infraslow oscillation in the DG is modulated by rhythmic serotonin release during sleep, which oscillates at the same frequency but in an opposite phase. Genetic manipulation of 5-HT receptors revealed that this neuromodulatory regulation is mediated by 5-HT1a receptors and the knockdown of these receptors leads to memory impairment. Together, our results provide novel mechanistic insights into how the 5-HT system can influence hippocampal activity patterns during sleep.

Intrauterine insemination: prognostic factors.

OBJECTIVE: To evaluate the impact of possible maternal and paternal prognostic factors and ovarian stimulation protocols on clinical pregnancy and live birth rates in intrauterine insemination (IUI) cycles. METHODS: Retrospective observational study of 341 IUI cycles performed from January 2016 to November 2020 at the Assisted Reproduction Service of the Clinics Hospital of the Ribeirão Preto Medical School, University of São Paulo. Clinical pregnancy and live birth rates and their potential prognostic factors were evaluated. Wilcoxon's non-parametric test was used to compare quantitative variables, and the chi-square test to compare qualitative variables, adopting a significance level of p<0.05. A logistic regression model was performed to verify which exploratory variables are predictive factors for pregnancy outcome. RESULTS: The ovulation induction protocol using gonadotropins plus letrozole (p=0.0097; OR 4.3286, CI 1.3040 - 14.3684) and post-capacitation progressive sperm ≥ 5million/mL (p=0.0253) showed a statistically significant correlation with the live birth rate. Female and male age, etiology of infertility, obesity, multifollicular growth, endometrial thickness ≥ 7 mm, and time between human chorionic gonadotropin administration and IUI performance were not associated with the primary outcomes. In the group of patients with ideal characteristics (women aged< 40 years, BMI < 30 kg/m2, antral follicle count ≥ 5, partner aged< 45 years, and post-capacitation semen with progressive spermatozoa ≥ 5 million/mL), the rate of clinical pregnancy was 14.8%, while that of live birth, 9.9%. CONCLUSIONS: In this study, the ovulation induction protocol with gonadotropins plus letrozole and post-capacitation progressive sperm ≥ 5 million/mL were the only variables that significantly correlated with intrauterine insemination success.

Oncology nursing specialty across Latin America: struggle and achievements past and present.

BACKGROUND: Latin America (LATAM) is a large region from Mexico to southern Patagonia in Chile and includes most islands in the Caribbean where Spanish is an official language. Efforts to address nursing specialization in the care of patients with cancer throughout the cancer continuum are described in narratives of oncology nursing struggles and achievements from eight Latin American countries. METHODS: Contributions by authors from Mexico, Costa Rica, Ecuador, Brazil and Chile are complemented by interview data (in Spanish by a bilingual medical anthropologist) to share the history and present status in the Dominican Republic, Peru and Argentina. RESULTS: We present stories of the pioneers of oncology nursing and a brief description of health systems in several of the included countries to show the disparate approaches towards cancer control and the context in which the oncology nurses work. We include key demographics and cancer data from the highlighted countries. We follow with descriptions of the formation and importance of oncology nursing associations/societies and share the history of oncology nursing education efforts, scopes of practice and recommendations for improvement in oncology nursing. CONCLUSIONS: Despite the challenges, oncology nurse leaders across LATAM are determined to raise the standards of care with structured specialization education/training, and to create advanced practice oncology nursing roles. It is critical that the achievements of oncology nurses are nationally acknowledged and appropriately compensated by governments and national and local authorities. The future is now for oncology nursing to be recognized as a critical specialized healthcare workforce to effectively address the growing threat of cancer and to ensure optimal control of this public health crisis across the region.

Ameloblastomas: clinical-histopathological evaluation of 85 cases with emphasis on squamous metaplasia and keratinization aspects.

OBJECTIVES: Ameloblastoma is a benign odontogenic neoplasm with an origin reputed to reactivation of odontogenic structures. Histological classification is based on microscopic features and architectural distribution of neoplastic cells. The importance of squamous metaplasia and keratinization has been disputed in ameloblastomas. Clinical and histopathological aspects were evaluated of 85 ameloblastomas, with attention to keratinization and squamous metaplasia features. STUDY DESIGN: Clinical-demographical information of 85 ameloblastomas were gleaned from the medical records. Microscopic analysis of all cases was carried out with emphasis on keratinization aspects of each tumor. RESULTS: Most ameloblastomas (54.12%) were diagnosed in males with a mean age of 37 years. Fifty-six patients were Caucasians (65.88%) and the mandible was affected in 68 (89.4%) cases. Most cases analyzed presented areas of squamous metaplasia/keratinization. Recurrence was detected in 16 cases; this was not related to keratinization aspects of the tumor. CONCLUSIONS: Keratinization is a common feature in ameloblastomas with no impact in tumor behavior.

Fungal rock phosphate solubilization using sugarcane bagasse.

The effects of different doses of rock phosphate (RP), sucrose, and (NH(4))(2)SO(4) on the solubilization of RP from Araxá and Catalão (Brazil) by Aspergillus niger, Penicillium canescens, Eupenicillium ludwigii, and Penicillium islandicum were evaluated in a solid-state fermentation (SSF) system with sugarcane bagasse. The factors evaluated were combined following a 2(3) + 1 factorial design to determine their optimum concentrations. The fitted response surfaces showed that higher doses of RP promoted higher phosphorus (P) solubilization. The addition of sucrose did not have effects on P solubilization in most treatments due to the presence of soluble sugars in the bagasse. Except for A. niger, all the fungi required high (NH(4))(2)SO(4) doses to achieve the highest level of P solubilization. Inversely, addition of (NH(4))(2)SO(4) was inhibitory to P solubilization by A. niger. Among the fungi tested, A. niger stood out, showing the highest solubilization capacity and for not requiring sucrose or (NH(4))(2)SO(4) supplementation. An additional experiment with A. niger showed that the content of soluble P can be increased by adding higher RP doses in the medium. However, P yield decreases with increasing RP doses. In this experiment, the maximal P yield (approximately 60 %) was achieved with the lower RP dose (3 g L(-1)). Our results show that SSF can be used to obtain a low cost biofertilizer rich in P combining RP, sugarcane bagasse, and A. niger. Moreover, sugarcane bagasse is a suitable substrate for SSF aiming at RP solubilization, since this residue can supply the C and N necessary for the metabolism of A. niger within a range that favors RP solubilization.

[Clinical Nurse Specialist: a model of advanced nursing practice in pediatric oncology in Brazil]. / Enfermeiro Clínico Especialista: um modelo de prática avançada de enfermagem em oncologia pediátrica no Brasil.

The objective of this paper is to report the experience of implementing a model of advanced nursing practice that integrates a clinical nurse specialist into the nursing staff. The authors describe the concept of this model and the process of its implementation in a pediatric oncology reference institution in the city of São Paulo, Brazil. The main result achieved in evaluating the clinical nurse specialist was strengthening of the role of the nurse as a reference professional for the articulation of the various demands of care in assisting the child, adolescent or young adult with cancer and his/her family. The satisfaction of these users due to the closer relationship between the patient, family and clinical nurse in all stages of the caring process also stands out.

In Vitro Effect of Eucalyptus Essential Oils and Antiseptics (Chlorhexidine Gluconate and Povidone-Iodine) against Bacterial Isolates from Equine Wounds.

Considering the increasing antibiotics resistance, there has been a propensity to replace them with antiseptics when it comes to wound management and treatment. Nevertheless, in recent years, there have been reports regarding resistance to antiseptics by some bacterial strains. There is also concern about the environmental impact of these substances. The aim of this study was to compare the antimicrobial efficacy of antiseptics and eucalyptus essential oils on bacterial strains from horse's wounds. We used twelve Escherichia coli, eight Staphylococcus aureus, two Staphylococcus pseudintermedius, one Staphylococcus vitulinus and one Staphylococcus saprophyticus strains from equine wounds. The effect of Eucalyptus radiata essential oil, Eucalyptus globulus essential oil, povidone-iodine and chlorhexidine gluconate against the isolated strains was evaluated applying the Kirby-Baüer method. Regarding the Escherichia coli strains, E. radiata and the mixture of E. radiata and E. globulus had a better inhibitory effect than antiseptics. E. globulus had a better effect against most Staphylococcus spp. compared to E. radiata. For both Gram-negative and Gram-positive strains tested, chlorhexidine gluconate had a better inhibitory effect than povidone-iodine. The antibacterial efficacy of essential oils highlights their potential to substitute or complement the use of antiseptics and so reduce resistance to antiseptics.

An Open-Source Virtual Reality System for the Measurement of Spatial Learning in Head-Restrained Mice.

Head-restrained behavioral experiments in mice allow neuroscientists to observe neural circuit activity with high-resolution electrophysiological and optical imaging tools while delivering precise sensory stimuli to a behaving animal. Recently, human and rodent studies using virtual reality (VR) environments have shown VR to be an important tool for uncovering the neural mechanisms underlying spatial learning in the hippocampus and cortex, due to the extremely precise control over parameters such as spatial and contextual cues. Setting up virtual environments for rodent spatial behaviors can, however, be costly and require an extensive background in engineering and computer programming. Here, we present a simple yet powerful system based upon inexpensive, modular, open-source hardware and software that enables researchers to study spatial learning in head-restrained mice using a VR environment. This system uses coupled microcontrollers to measure locomotion and deliver behavioral stimuli while head-restrained mice run on a wheel in concert with a virtual linear track environment rendered by a graphical software package running on a single-board computer. The emphasis on distributed processing allows researchers to design flexible, modular systems to elicit and measure complex spatial behaviors in mice in order to determine the connection between neural circuit activity and spatial learning in the mammalian brain.

Prediction of dyskinesia in Parkinson's disease patients using machine learning algorithms.

Dyskinesias are non preventable abnormal involuntary movements that represent the main challenge of the long term treatment of Parkinson's disease (PD) with the gold standard dopamine precursor levodopa. Applying machine learning techniques on the data extracted from the Parkinson's Progression Marker Initiative (PPMI, Michael J. Fox Foundation), this study was aimed to identify PD patients who are at high risk of developing dyskinesias. Data regarding clinical, behavioral and neurological features from 697 PD patients were included in our study. Our results show that the Random Forest was the classifier with the best and most consistent performance, reaching an area under the receiver operating characteristic (ROC) curve of up to 91.8% with only seven features. Information regarding the severity of the symptoms, the semantic verbal fluency, and the levodopa treatment were the most important for the prediction, and were further used to create a Decision Tree, whose rules may guide the pharmacological management of PD symptoms. Our results contribute to the identification of PD patients who are prone to develop dyskinesia, and may be considered in future clinical trials aiming at developing new therapeutic approaches for PD.

Biofilm formation in vitro by Leptospira interrogans strains isolated from naturally infected dogs and their role in antimicrobial resistance.

Leptospira interrogans is a biofilm-forming pathogen, however, there are few data involving Brazilian strains isolated from dogs and their antimicrobial sensitivity in planktonic and biofilm forms. The potential for biofilm formation and antimicrobial resistance in naturally infected dogs is a fundamental approach towards disease epidemiology and the establishment of consistent prophylaxis and control measures. The objective of this study was to evaluate in vitro biofilm formation of a reference strain (L. interrogans, sv. Copenhageni L1 130 - L20) and of L. interrogans isolated from dogs (C20, C29, C51, C82), with subsequent evaluation of antimicrobial susceptibility in planktonic and biofilm forms. The semi quantification of biofilm production revealed a dynamic process of development over time, with mature biofilm formation early on the seventh day of incubation. All strains were efficient for in vitro biofilm formation and, in this form, they were considerably more resistant compared to their planktonic form, with MIC90 of 1600 µg/mL for amoxicillin, 800 µg/mL for ampicillin, and >1600 µg/mL for doxycycline and ciprofloxacin. The strains studies were isolated on naturally infected dogs that might act as reservoirs and sentinels for human infections. The potential to antimicrobial resistance together with the close relation between dogs and humans indicates the need for greater actions on disease control and surveillance. Moreover, biofilm formation may contribute to the persistence of Leptospira interrogans in the host and these animals can act as chronic carriers, disseminating the agent in the environment.