Wastewater surveillance for antibiotic resistance genes during the late 2020 SARS-CoV-2 peak in two different populations.

The United States Centers for Disease Control and Prevention reported a rise in resistant infections after the coronavirus disease 2019 (COVID-19) pandemic started. How and if the pandemic contributed to antibiotic resistance in the larger population is not well understood. Wastewater treatment plants are good locations for environmental surveillance because they can sample entire populations. This study aimed to validate methods used for COVID-19 wastewater surveillance for bacterial targets and to understand how rising COVID-19 cases from October 2020 to February 2021 in Portugal (PT) and King County, Washington contributed to antibiotic resistance genes in wastewater. Primary influent wastewater was collected from two treatment plants in King County and five treatment plants in PT, and hospital effluent was collected from three hospitals in PT. Genomic extracts were tested with the quantitative polymerase chain reaction for antibiotic resistance genes conferring resistance against antibiotics under threat. Random-effect models were fit for log-transformed gene abundances to assess temporal trends. All samples collected tested positive for multiple resistance genes. During the sampling period, mecA statistically significantly increased in King County and PT. No statistical evidence exists of correlation between samples collected in the same Portuguese metro area.

Bladder training compared to bladder training associated with pelvic floor muscle training for overactive bladder symptoms in women: A randomized clinical trial.

AIMS: To compare the effects of bladder training (BT) versus BT with pelvic floor muscle training (PFMT) in women with overactive bladder (OAB) symptoms. METHODS: Randomized controlled clinical trial including women with OAB symptoms, randomized into two groups: BT versus BT + PFMT. For 12 consecutive weeks, the women received home BT. The BT + PFMT performed supervised PFMT, once/week, associated at home PFMT protocol. Primary outcomes were urinary urgency, daytime voiding frequency, nocturia and urgency urinary incontinence assisted by both 3-day bladder diary and International Consultation on Incontinence OAB (ICIQ-OAB) questionnaire. Secondary outcomes were 24-h pad test and Patient Global Impression of Improvement. T-test, analysis of variance, Mann-Whitney (SPSS 20.0) and power/effect size (G-power) were applied in data analyses. RESULTS: Sixty-three women were included (B = 31; BT + PFMT = 32). There was no significant statistical difference between groups in terms of urinary symptoms: daytime frequency (BT: pre: 11.59 [±5.80], post: 9.10 [±4.05]; BT + PFMT: pre: 10.67 [±3.73], post: 8.08 [±3.38]) p = 0.75; nocturia: (BT: pre: 1.46 [±0.91], post: 0.82 [±0.82]; BT + PFMT: pre: 1.80 [±2.26], post: 0.82 [±1.15]) p = 0.70; urinary urgency (BT: pre: 3.22 [±4.70], post: 4.49 [±4.32]; BT + PFMT: pre: 6.87 [±5.60], post: 6.15 [±4.52]) p = 0.10; ICIQ-OAB total score: (BT: pre: 9.16 [±2.55], post: 6.32 [±3.77]; (BT + PFMT: pre: 9.75 [±2.06], post: 5.06 [±3.44] p = 0.30. CONCLUSIONS: Supervised PFMT added to BT did not provide further improvements than isolated BT in women with OAB symptoms.

Impact of water reuse on agricultural practices and human health.

Climate change is altering the habits of the population. Extensive drought periods and overuse of potable water led to significant water shortages in many different places. Therefore, new water sources are necessary for usage in applications where the microbiological and chemical water quality demands are less stringent, as for agriculture. In this study, we planted, germinated, and grew vegetables/fruits (cherry tomato, lettuce, and carrot) using three types of potential waters for irrigation: secondary-treated wastewater, chlorine-treated wastewater, and green wall-treated greywater, to observe potential health risks of foodstuff consumption. In this study the waters and crops were analyzed for three taxonomic groups: bacteria, enteric viruses, and protozoa. Enteric viruses, human Norovirus I (hNoVGI) and Enterovirus (EntV), were detected in tomato and carrots irrigated with secondary-treated and chlorine-treated wastewater, in concentrations as high as 2.63 log genome units (GU)/g. On the other hand, Aichi viruses were detected in lettuce. Bacteria and protozoa remained undetected in all fresh produce although being detected in both types of wastewaters. Fresh produce irrigated with green wall-treated greywater were free from the chosen pathogens. This suggests that green wall-treated greywater may be a valuable option for crop irrigation, directly impacting the cities of the future vision, and the circular and green economy concepts. On the other hand, this work demonstrates that further advancement is still necessary to improve reclaimed water to the point where it no longer constitutes risk of foodborne diseases and to human health.

Application of optical coherence tomography angiography for microvascular changes in patients treated with hydroxychloroquine: a systematic review and meta-analysis.

BACKGROUND: Retinal toxicity with long-term hydroxychloroquine (HCQ) treatment is a major concern. This systematic review aims to assess the application of optical coherence tomography angiography (OCTA) to detect microvascular alterations in patients under HCQ. METHODS: PubMed, Scopus, Web of Science, and Cochrane Library databases were systematically searched until January 14, 2023. Studies using OCTA as a primary diagnostic method to evaluate the macular microvasculature of HCQ users were included. Primary outcomes were macular vessel density (VD) and foveal avascular zone (FAZ) at the superficial (SCP) and deep (DCP) capillary plexus. Meta-analysis was performed using a random-effects model. RESULTS: Of 211 screened abstracts, 13 were found eligible, enrolling 989 eyes from 778 patients. High-risk patients due to longer duration of treatment presented lower VD in the retinal microvasculature than those with low-risk in SCP (P = 0.02 in fovea; P = 0.004 in parafovea) and in DCP (P = 0.007 in fovea; P = 0.01 in parafovea). When compared with healthy controls, HCQ users had lower VD in both plexus-no quantitative synthesis was presented. CONCLUSIONS: Microvascular changes were found in autoimmune patients under HCQ treatment without any documented retinopathy. However, the evidence produced so far does not allow to draw conclusion concerning the effect of drug as studies were not controlled for disease duration.

Trypanosoma cruzi: Does the intake of nanoencapsulated benznidazole control acute infections?

Chagas Disease (CD) affects around eight million people worldwide. It is considered a neglected disease that presents few treatment options with efficacy only in the acute phase. Nanoparticles have many positive qualities for treating parasite infections and may be effectively and widely employed in clinical medicine. This research aimed to evaluate the nanoencapsulated benznidazole treatment in animals experimentally infected with Trypanosoma cruzi. To analyze the treatment efficacy, we evaluated survival during thirty days, parasitemia, genotoxicity, and heart and liver histopathology. Thirty-five female Swiss mice were organized into seven groups characterizing a dose curve: A - Negative control (uninfected animals), B - Positive control (infected animals), C - Benznidazole (BNZ) 100 mg/kg (infected animals), D - 5 mg/kg Benznidazole nanocapsules (NBNZ) (infected animals), E - 10 mg/kg Benznidazole nanocapsules (infected animals), F - 15 mg/kg Benznidazole nanocapsules (infected animals), G - 20 mg/kg Benznidazole nanocapsules (infected animals). The animals were infected with the Y strain of T. cruzi intraperitoneally. The treatment was administered for eight days by oral gavage. It was possible to observe that the treatment with the highest NBNZ dose presented efficacy similar to the standard benznidazole drug. The 20 mg/kg NBNZ dose was able to reduce parasitemia, increase survival, and drastically reduce heart and liver tissue damage compared to the 100 mg/kg BNZ dose. Moreover, it showed a lower DNA damage index than the BNZ treatment. In conclusion, the nanoencapsulation of BNZ promotes an improvement in parasite proliferation control with a five times smaller dose relative to the standard dose of free BNZ, thus demonstrating to be a potential innovative therapy for CD.

Safety and efficacy of Lucilia cuprina maggots on treating an induced infected wound in Wistar rats.

Infection is one of the main complications that hinder wound healing. Currently, antibiotic-resistant bacteria, such as Methicilin-resistant Staphylococcus aureus (MRSA), are a concern worldwide for both humans and animals. Maggot therapy is re-emerging as an alternative to intractable wounds and may be an option to the traditional antibiotic treatment. Although the species of choice is Lucilia sericata, reports of clinical use have led us to evaluate the efficacy and safety of using Lucilia cuprina larvae on induced infected wounds in Wistar rats. In short, 32 male Wistar rats were divided into 4 groups: Group I - saline solution treated; Group II - antibiotic-treated; Group III - treated with larval debridement, and Group IV - without wound and treatment. Skin wounds were induced in groups I, II and III. All treatments were performed once and held for 48 h. Clinical, microbiological, histopathological, hematological, and biochemical analyses were done. Significant wound area contraction was found (>95%) in group III on day 9 compared to day 15 in group II. Complete elimination (0.0 ± 0.0 CFU/mL) of bioburden was achieved after the second treatment (day 6) in both the II and III groups, compared to an increase in Group I (6.51 ± 0.37 CFU/mL). A cleaner wound was also observed in the histopathological evaluation of group III, with adequate collagen formation and re-epithelialization on day 15. Furthermore, larvae increased blood platelet levels after the first treatment. L. cuprina larvae have proven safe and effective in accelerating wound treatment and eliminating MRSA.

Eucalyptus oil nanoemulsions against eggs and larvae of Haemonchus contortus.

Haemonchus contortus is a highly pathogenic and prevalent helminth that causes many deaths in sheep herds. Anthelmintics are usually employed to overcome this issue; however, they do not guarantee immediate and lasting efficacy because of the occurrence of drug-resistant parasites. Among substances that are used in scientific studies for parasitic control, essential oils are known to have different pharmacological properties. However, they demonstrate instability owing to several factors, and therefore, nanoemulsification is considered an alternative to control the instability and degradability of these compounds. The objective of this study was to evaluate the cytotoxicity of nanoemulsions containing essential oil of Eucalyptus globulus against the blood of healthy sheep and to verify their activity against the parasite H. contortus in sheep. The results presented adequate nanotechnological characteristics (diameter 72 nm, PDI 0.2, zeta -11 mV, and acidic pH) and adequate morphology. Further, the corona effect and cytotoxic profiles of the free oil and nanoemulsion against blood cells from healthy sheep were evaluated. The tests results did not present a toxicity profile. For evaluating efficacy, we observed an important anthelmintic action of the nanoemulsion containing oil in comparison to the free oil; the results demonstrate a potential role of the nanoemulsion in the inhibition of egg hatchability and the development of larvae L1 to L3 (infective stage). Based on these results, we developed an important and potential anthelmintic alternative for the control of the parasite H. contortus.

Recovery of SARS-CoV-2 from large volumes of raw wastewater is enhanced with the inuvai R180 system.

Wastewater-based epidemiology (WBE) for severe acute respiratory syndrome Coronavirus 2 (SARS-CoV-2) is a powerful tool to complement syndromic surveillance. Although detection of SARS-CoV-2 in raw wastewater may be prompted with good recoveries during periods of high community prevalence, in the early stages of population outbreaks concentration procedures are required to overcome low viral concentrations. Several methods have become available for the recovery of SARS-CoV-2 from raw wastewater, generally involving filtration. However, these methods are limited to small sample volumes, possibly missing the early stages of virus circulation, and restrained applicability across different water matrices. The aim of this study was thus to evaluate the performance of three methods enabling the concentration of SARS-CoV-2 from large volumes of wastewater: i) hollow fiber filtration using the inuvai R180, with an enhanced elution protocol and polyethylene glycol (PEG) precipitation; ii) PEG precipitation; and iii) skimmed milk flocculation. The performance of the three approaches was evaluated in wastewater from multiple wastewater treatment plants (WWTP) with distinct singularities, according to: i) effective volume; ii) percentage of recovery; iii) extraction efficiency; iv) inhibitory effect; and v) the limits of detection and quantification. The inuvai R180 system had the best performance, with detection of spiked control across all samples, with average recovery percentages of 68% for porcine epidemic diarrhea virus (PEDV), with low variability. Mean recoveries for PEG precipitation and skimmed milk flocculation were 9% and 14%, respectively. The inuvai R180 enables the scalability of volumes without negative impact on the costs, time for analysis, and recovery/inhibition. Moreover, hollow fiber ultrafilters favor the concentration of different microbial taxonomic groups. Such combined features make this technology attractive for usage in environmental waters monitoring.

Resveratrol impacts in oxidative stress in liver during Trypanosoma cruzi infection.

Trypanosoma cruzi is the causative agent of Chagas disease, infecting the heart, intestines and liver tissues. There is growing evidence that oxidative stress, defined as a persistent imbalance between highly oxidative compounds and antioxidant defenses, is a marker of tissue inflammation; it is related to immune responses such as damage, as well as to strand breaks in DNA contributing to disease progression. Antioxidant agents help mitigate the damage caused by inflammation, preventing or slowing damage to cells caused by free radicals. In this sense, resveratrol (RSV) is an important polyphenol that demonstrates antioxidant effects. It reverses damage caused by several infectious diseases. The aim of the present study was to determine whether treatment with RSV would prevent or minimize oxidative damage caused by T. cruzi. The animals were divided into four groups (n = 5): A) control; B) control + RSV; C) infected and D) infected + RSV. The infected groups received 1 x 104 Y strain trypomastigotes via intraperitoneal injection; after confirmation of infection, the mice received RSV 100 mg/kg for seven days orally. On the 8th day post-infection, we collected liver tissue for analysis of oxidant/antioxidant status: superoxide dismutase (SOD), catalase (CAT), and glutathione s-transferase (GST) activities, as well as reactive oxygen species (ROS), non-protein thiols (NPSH), thiols, carbonyl protein, thiobarbituric acid reactive substance (TBARS), and finally, the nitrite/nitrate ratio (NOx) levels were determined. The administration of RSV did not exert direct effect on parasitemia. The infection produced high levels of TBARS, NOx, and ROS levels in liver tissue, suggesting cellular injury with production of free radicals in animals infected by T. cruzi. RSV positively modulated SOD and aumenting GST activities enzymes in infected animals. Protein thiols levels in infected animals were lower than those of control. Taken together, the data suggest T. cruzi causes hepatic oxidative stress, and RSV 100 mg/kg for seven days it's dosen't seem minimized these negative effects in the acute phase of disease.

Involvement of ectonucleotidases and purinergic receptor expression during acute Chagas disease in the cortex of mice treated with resveratrol and benznidazole.

Chagas disease (CD) is caused by the parasite Trypanosoma cruzi. CD affects people worldwide, primarily in tropical areas. The central nervous system (CNS) is an essential site for T. cruzi persistence during infection. The protozoan may pass through the blood-brain barrier and may cause motor and cognitive neuronal damage. Once in the CNS, T. cruzi triggers immune responses that the purinergic system can regulate. Treatment for CD is based on benznidazole (BNZ); however, this agent has negative side-effects and is toxic to the host. For this reason, we investigated whether resveratrol (RSV), a potent antioxidant and neuroprotective molecule, would modulate purinergic signaling and RSV alone or in combination with BNZ would prevent changes in purinergic signaling and oxidative damage caused by T. cruzi. We infected mice with T. cruzi and treated them with RSV or BNZ for 8 days. Increases in ATP and ADP hydrolysis by NTPDase in the total cortex of infected animals were observed. The treatment with RSV in infected group diminished ATP, ADP, and AMP hydrolysis compared to infected group. The combination of RSV + BNZ decreased AMP hydrolysis in infected animals compared to the INF group, exerting an anti-inflammatory effect. RSV acted as a neuroprotector, decreasing adenosine levels. Infected animals presented an increase of P2X7 and A2A density of purine receptors. RSV reduced P2X7 and A2A and increased A1 density receptors in infected animals. In addition, infected animals showed higher TBARS and reactive oxygen species (ROS) levels than control. RSV diminished ROS levels in infected mice, possibly due to antioxidant properties. In short, we conclude that resveratrol could act as a neuroprotective molecule, probably preventing inflammatory changes caused by infection by T. cruzi, even though the mice experienced high levels of parasitemia.

Choroidal neovascularization secondary to pathological myopia-macular Bruch membrane defects as prognostic factor to anti-VEGF treatment.

PURPOSE: To evaluate the prevalence and visual outcomes of macular Bruch membrane (BM) defects in patients treated with anti-vascular endothelial growth factors (VEGF) for choroidal neovascularization secondary to pathological myopia (mCNV). METHODS: Single-center retrospective observational case series of 68 eyes from 62 patients with mCNV treated with one anti-VEGF injection followed by a pro re nata (1 + PRN) regimen. A minimum follow-up of 6 months was defined. Chorioretinal atrophy was assessed by fundus examination, fluorescein angiography, and SD-OCT. RESULTS: Median follow-up was 28.5 (range 6-89) months with a median number of 5 anti-VEGF injections. At baseline, 27.9% of eyes had macular BM defects increasing to 36.8% during follow-up (p<0.001). Eyes without macular BM defects at the baseline had higher BCVA at the last observation than patients with BM defects (p=0.003). An increase of 5 or more ETDRS letters was more frequent in eyes without BM defects (p=0.001). At the end of follow-up, mCNV-related macular atrophy was present in 44.1%; out of which, 83.3% presented macular BM defects (p<0.001). Eyes with mCNV-related macular atrophy without BM defects had a significant increase of best-corrected visual acuity compared with eyes with mCNV-related macular atrophy and BM defect (p=0.002). CONCLUSIONS: Macular Bruch membrane defects are often seen in mCNV and have a significant impact in visual acuity and prognosis. Eyes with macular BM defects have a poorer response and worse visual outcomes after anti-VEGF therapy.

Left atrial functional assessment and mortality in patients with severe aortic stenosis with sinus rhythm.

BACKGROUND: Aortic valve stenosis (AS) is the most common primary valvular heart disease leading to surgical or percutaneous aortic valve replacement (AVR) in Europe and its prevalence keeps growing. While other risk factors in severe AS are well documented, little is known about the prognostic value of left atrial (LA) function in AS. Our aim is to clarify the relationship between LA function measured at severe AS diagnosis (evaluated by means of volumetric assessment) and all-cause mortality during follow-up. METHODS: We retrospectively evaluated patients diagnosed with severe AS for the first time at our echocardiography laboratory. We evaluated LA reservoir, conduit and pump function by measuring LA volumes at different timings of cardiac cycle. Treatment strategy was decided according to heart team consensus and patient decision. We divided patients into groups according to terciles of LA reservoir, conduit and pump function. Primary outcome was defined by the occurrence of all-cause mortality during follow-up. RESULTS: A total of 408 patients were included in the analysis, with a median follow-up time of 45 months (interquartile range 54 months). 57.9% of patients underwent AVR and 44.9% of patients registered the primary outcome during follow-up. Left atrial emptying fraction (LAEF) was the best LA functional parameter and the best overall parameter in discriminating primary outcome (AUC 0.845, 95%CI 0.81-0.88, P < 0.001). After adjustment for clinical, demographic and echocardiographic variables, cumulative survival of patients with LAEF < 37% and LAEF 37 to 53% relative to patients with LAEF ≥54% remained significantly lower (HR 13.91, 95%CI 6.20-31.19, P < 0.001 and HR 3.40, 95%CI 1.57-7.37, P = 0.002, respectively). After adjustment for AVR, excess risk of LAEF < 37% and LAEF 37 to 53% relative to LAEF ≥54% remained significant (HR 11.71, 95%CI 5.20-26.40, P < 0.001 and HR 3.59, 95%CI 1.65-7.78, P = 0.001, respectively). CONCLUSIONS: In patients with a first diagnosis of severe AS, LA function, evaluated by means of volumetric assessment, is an independent predictor of all-cause mortality and a more potent predictor of death compared to classical severity parameters. These data can be useful to identify high-risk patients who might benefit of AVR.

Evaluation of Legiolert™ for the Detection of Legionella pneumophila and Comparison with Spread-Plate Culture and qPCR Methods.

Legionella pneumophila, the organism responsible for Legionnaires' disease, a potentially lethal pneumonia, is an opportunistic bacterium spread via inhalation of contaminated, aerosolized water. The detection and control of L. pneumophila is crucial to reduce the risk it poses to human health. L. pneumophila is generally detected and quantified by the plating method, ISO 11731:2017 and by qPCR. ISO 11731 is based on the filtration of the water sample through a membrane, which is placed on selective agar medium, and after colony growth, presumptive Legionella are then confirmed by subculturing, serology, or PCR. Quantitative Polymerase Chain Reaction (qPCR) is based on the amplification of a DNA sequence specific to L. pneumophila, usually within the mip gene. The objective of this study was to compare these methods to a new, liquid culture method based on the Most Probable Number (MPN) technique, Legiolert™/Quanti-Tray® with data obtained with ISO 11731 and a viability quantitative qPCR (v-qPCR), for quantification of L. pneumophila in potable and non-potable waters. Data showed that the Legiolert method revealed concentrations of L. pneumophila greater than ISO 11731 and generally similar results to those of v-qPCR. The Legiolert method was highly specific and easy to use, representing a significant advancement in the quantification of L. pneumophila from potable and non-potable waters.

Comparing the efficacy of nutmeg essential oil and a chemical pesticide against Musca domestica and Chrysomya albiceps for selecting a new insecticide agent against synantropic vectors.

The insecticidal activity of Myristica fragrans (Houtt) essential seed oil, (Nutmeg) was evaluated against Musca domestica (Linnaeus) and Chrysomya albiceps (Wiedemann); both important infectious pathogenic disease vectors. The oil was extracted by distillation, and 21 components were identified during chemical analysis; principally ß-pinene (26%), α-pinene (10.5%), Sabinene (9.1%) and γ-terpinen (8.5%). Insecticidal properties were identified through larvicide and adulticide tests. Using the immersion method, the oil at 5% was found to be very effective (90 ± 1%) against M. domestica larvae. The results for adulticide activity varied by fly species, dosage, time, and method of exposure. Topical application (on the insect thorax) was more toxic to C. albiceps, where the lethal concentration at 50% (LC50) was 2.02 ± 0.56, and 8.57 ± 2.41 for the common flies. When the insects were exposed to oil impregnated paper, the results were similar for M. domestica and C. albiceps adults with respective LC50 values of 2.74 ± 0.24, and 3.65 ± 0.48. Thus, the results demonstrated that M. fragrans oil presents insecticidal activity and can be used for control of Musca domestica and Chrysomya albiceps.

In vitro and in vivo trypanocidal activity of a benzofuroxan derivative against Trypanosoma cruzi.

Chagas disease, caused by Trypanosoma cruzi, is a major public health problem and is described as one of the most neglected diseases worldwide. It affects about 6-7 million people. Currently, only two drugs are available for the treatment of this disease: nifurtimox and benznidazole. However, both drugs are highly toxic and have several side effects, which lead many patients to discontinue treatment. Moreover, these compounds show a significant curative efficacy only in the acute phase of the disease. Therefore, searching for new drugs is necessary. The objective of this study was to evaluate the in vitro and in vivo activity of a benzofuroxan derivative (EA2) against T. cruzi, and to evaluate the hematological and biochemical changes induced by its treatment in animals infected with T. cruzi. The results were then compared with those of healthy controls. In vitro testing was first performed with T. cruzi epimastigote forms. In this experiment, EA2 was diluted at three different concentrations (0.25, 0.50, and 1%). In vitro evaluation of the trypanocidal activity was performed 24, 48, and 72 h after incubation. In vivo assays were performed using three different doses (10, 5, and 2,5 mg/kg). Mice were divided into 10 groups (five animals/group), wherein four groups comprised non-infected animals (A, G, H, I) and six groups comprised infected animals (B, C, D E, F, J). Groups B and J represented the negative and positive controls, respectively. Groups G, H, and I were used to confirm that EA2 was not toxic to non-infected animals. Parasitemia was measured in infected animals and the hematological and biochemical profiles (urea, creatinine, albumin, aspartate aminotransferase, alanine aminotransferase, and alkaline phosphatase) were evaluated in all animals. EA2 demonstrated in vitro trypanocidal activity at all concentrations tested. Although it did not demonstrate a curative effect in vivo, EA2 was able to retard the onset of parasitemia, and significantly reduced the parasite count in groups D and E (treated with 5 and 2.5 mg/kg, respectively). EA2 did not induce changes in hematological and biochemical parameters in non-infected animals, demonstrating that it is not toxic. However, further assessments should aim to confirm the safety of EA2 since this was the first in vitro and in vivo study conducted with this molecule.

The Brazilian Portuguese version of the ICIQ-OABqol: cross-cultural adaptation and reliability.

INTRODUCTION AND HYPOTHESIS: The International Consultation on Incontinence Questionnaire Overactive Bladder Symptoms Quality of Life (ICIQ-OABqol) Module evaluates the quality of life of individuals with overactive bladder (OAB) symptoms, and its use in scientific studies and clinical practice is recommended by the International Continence Society. The aim was to conduct the cross-cultural adaptation and validation of the Brazilian Portuguese version of the ICIQ-OABqol (ICIQ-OABqol_portuguese) in individuals with OAB symptoms. METHODS: An observational cross-sectional study was performed at the Clinical Physiotherapy of PUC MINAS in Belo Horizonte, MG, Brazil, between March 2017 and October 2018. The translation was previously carried out by the Mapi Research Institute. After receiving the translated questionnaire, the cross-cultural adaptation process was conducted as follows: (1) review by an expert committee (13 experts); (2) pre-test (n = 30); (3) cross-cultural adaptation; (4) validation of the ICIQ-OABqol_portuguese. We analyzed the intraexaminer reliability validation (n = 118) and internal consistency measurement (Cronbach's α coefficient), test-retest reliability (ICC) and correlation between the ICIQ-OAB and ICIQ-OABqol_portuguese questionnaires through Pearson's correlation coefficient and Bland-Altman scatter plot and concordance. Confirmatory factor analysis was used to confirm the domains of the instrument. RESULTS: The ICIQ-OABqol was cross-culturally adapted to Brazilian Portuguese and presented satisfactory internal consistency (α-Cronbach coefficient 0.88), adequate construct validity, strong reliability considering the test-retest with an interval of 19.68 (±6.98) days and moderate correlation with the ICIQ-OAB. CONCLUSIONS: The Brazilian Portuguese version of the ICIQ-OABqol shows satisfactory psychometric properties and can be used to evaluate the quality of life of individuals of both sexes with OAB symptoms.

Incidence of enterococci resistant to clinically relevant antibiotics in environmental waters and in reclaimed waters used for irrigation.

Treated wastewater discharged into the environment or reused in different activities can be a major vehicle for the transmission of antibiotic-resistant bacteria and antibiotic-resistance genes. In this study, environmental and wastewater samples, collected at different stages of treatment, were studied to identify the possibility of a positive selection of antibiotic-resistant organisms in wastewater treatment plants (WWTPs). Enterococci were isolated, characterized into the main human species, and subjected to the Kirby-Bauer test using seven antibiotics (five classes): ampicillin, chloramphenicol, ciprofloxacin, gentamicin, linezolid, tetracycline, and vancomycin. Furthermore, vancomycin-resistant enterococci (VRE), a major cause of nosocomial infection, was identified, and the genes vanA and vanB detected directly in the samples and in all confirmed VRE. Data showed that WWTPs were able to reduce the levels of antibiotic resistance, although 72% of the disinfected wastewaters still presented antibiotic-resistant enterococci. VRE were detected in 6% of the samples, including in reclaimed waters. UV disinfection was not effective at removing VRE and multiple antibiotic-resistant (MAR) enterococci, most commonly Enterococcus faecalis. The use of reclaimed water containing VRE and MAR enterococci in crop production, irrigation of urban gardens, and street cleaning increases immensely the potential risk to human health.

Detection of toxins involved in foodborne diseases caused by Gram-positive bacteria.

Bacterial toxins are food safety hazards causing about 10% of all reported foodborne outbreaks in Europe. Pertinent to Gram-positive pathogens, the most relevant toxins are emetic toxin and diarrheal enterotoxins of Bacillus cereus, neurotoxins of Clostridium botulinum, enterotoxin of Clostridium perfringens, and a family of enterotoxins produced by Staphylococcus aureus and some other staphylococci. These toxins are the most important virulence factors of respective foodborne pathogens and a primary cause of the related foodborne diseases. They are proteins or peptides that differ from each other in their size, structure, toxicity, toxicological end points, solubility, and stability, types of food matrix to which they are mostly related to. These differences influence the characteristics of required detection methods. Therefore, detection of these toxins in food samples, or detection of toxin production capacity in the bacterial isolate, remains one of the cornerstones of microbial food analysis and an essential tool in understanding the relevant properties of these toxins. Advanced research has led into new insights of the incidence of toxins, mechanisms of their production, their physicochemical properties, and their toxicological mode of action and dose-response profile. This review focuses on biological, immunological, mass spectrometry, and molecular assays as the most commonly used detection and quantification methods for toxins of B. cereus, C. botulinum, C. perfringens, and S. aureus. Gathered and analyzed information provides a comprehensive blueprint of the existing knowledge on the principles of these assays, their application in food safety, limits of detection and quantification, matrices in which they are applicable, and type of information they provide to the user.

Effects of resveratrol on the differentiation fate of neural progenitor cells of mouse embryos infected with Trypanosoma cruzi.

Chagas disease (CD) affecting about 7 million people is caused by the flagellate protozoan Trypanosoma cruzi. The central nervous system (CNS) is an important site for T. cruzi persistence in the host during the chronic phase of infection, because the protozoan may pass the blood-brain barrier and may cause motor and cognitive neuronal damage. Thinking about avoiding or minimizing these negative effects, it is hypothesized that resveratrol (RSV), a component with several medicinal properties has beneficial effects on the CNS. The objective of this study was to investigate, whether T. cruzi infection interferes with neurogenesis and gliogenesis of embryos of infected mice females, and whether RSV would be able to avoid or minimize these changes caused by CD. RSV is a polyphenol found in grapes and widely studied for its neuroprotective and antioxidant properties. In addition, we investigated the role caused by the parasite during congenital infection and CNS development. Embryos and their brains were PCR-positive for T. cruzi. For this study, NPCs obtained from telencephalon of infected and uninfected embryos and were cultured in presence of resveratrol for forming neurospheres. The results demonstrated that the congenital transmission of T. cruzi influences CNS formation and neural fate, decreasing the number of neuroespheres and causing an elongation in the phases of the cell cycle. In addition, the parasite promoted an increase in neugliogenesis. Resveratrol was neuroprotective and prevented negative effects of the infection. Thus, we suggest the use of resveratrol as a therapeutic target for the treatment of neuroinflammation or as neuroprotective agent during Chagas disease, as it improves gliogenesis and restores neural migration.

Trypanosoma evansi impacts on embryonic neural progenitor cell functions.

Trypanosoma evansi appears to have a significant tropism for brain tissue in its chronic and acute phases. The most common symptoms of this brain infection are motor incoordination, meningoencephalitis, demyelination, and anemia. There have only been few studies of the effects of T. evansi infection on neuronal differentiation and brain plasticity. Here, we investigated the impact of the congenital T. evansi infection on brain development in mice. We collected telencephalon-derived neural progenitor cells (NPCs) from T. evansi uninfected and infected mice, and cultivated them into neurospheres. We found that T. evansi significantly decreased the number of cells during development of neurospheres. Analysis of neurosphere differentiation revealed that T. evansi infection significantly increased neural migration. We also observed that T. evansi promoted expression of glial fibrillary acidic protein (GFAP) in infected cells. These data suggest that congenital T. evansi infection may affect embryonic brain development.