Rapid colorimetric polymyxin B microelution directly from positive blood bottles: because patients with serious infections should not have to wait for results of culture-based methodologies.

PURPOSE: To evaluate the performance of the rapid colorimetric polymyxin B microelution (RCPEm) in determining polymyxin B resistance directly from Enterobacterales-positive blood cultures. METHODS: A set volume of positive blood culture bottles (diluted 1:10) was inoculated into a glucose-broth-phenol red solution (NP solution), where a polymyxin B disk was previously eluted (final concentration of 3 µg/mL). Test was read each 1 h for up to 4 h. Color change from red/orange to yellow indicated resistant isolates. Results were compared to the reference method, broth microdilution (BMD), performed from colonies grown on solid media from the same blood culture bottle. RESULTS: One hundred fifty-two Enterobacterales-positive blood cultures were evaluated, 22.4% (34/152) of them resistant to polymyxin B (including 6.6% with borderline MICs). When performing directly from positive blood cultures (RCPEm-BC), specificity and sensitivity were 99.1% and 94.1%, respectively. Of note, 79.4% (27/34) of truly resistant isolates required 3 h of incubation, compared to the 18 ± 2 h incubation that microtiter plates of BMD demand before reading can be performed. CONCLUSIONS: RCPEm directly from blood cultures has great potential to be part of the routine of clinical microbiology laboratories to establish polymyxin B susceptibility, impacting outcome of patients with bloodstream infections caused by carbapenem-resistant Enterobacterales.

Detection of KPC enzyme by MALDI-TOF MS from bacteria impregnated in filter paper.

The main mechanism that causes resistance to carbapenem, one of the most potent antibiotic available, in Enterobacterales bacterial isolates, is due to Klebsiella pneumoniae carbapenemase (KPC) production by the bacterium. KPC is spread worldwide, requiring laboratories to be capable of identifying this enzyme, however some methods can be expensive for small laboratories, especially in developing countries. Therefore, the development of methods with low cost of reagents for the detection of KPC enzyme is necessary. The objective of this study was to evaluate the detection of KPC enzyme by MALDI-TOF MS from inactivated bacteria impregnated in filter paper. A total of 129 Enterobacterales isolates were impregnated in filter paper, and after 7 days at room temperature, they were subjected to a protein extraction protocol and spectra acquisition, in triplicates, by MALDI-TOF MS. The spectra were evaluated and KPC was identified according to the presence of a peak of 28,712.62 ± 27.80 m/z. Considering the presence of the KPC peak in at least one spectrum of the triplicates, this method presented 60.8% sensitivity and 96.4% specificity. However, considering the presence of KPC peak in at least two spectra of the triplicate, a specificity of 100% was achieved. The detection of KPC enzyme from inactivated bacteria impregnated in filter paper can be used as a method to confirm the presence of KPC, which could be very significant for small laboratories with limited resources.

Hospital wastewater as source of human pathogenic bacteria: A phenotypic and genomic analysis of international high-risk clone VIM-2-producing Pseudomonas aeruginosa ST235/O11.

Pseudomonas aeruginosa belong to the special pathogen group capable of causing serious infections, with high mortality rates. The aim of this study was to describe the antibiotic resistance and genomic characteristics of Pseudomonas aeruginosa belonging to international high-risk clone ST235 (GPAE0131 isolate), obtained from hospital wastewater. P. aeruginosa GPAE0131 was isolated from ward tertiary hospital in Brazil and the antibiotic resistance profile was determined by the disc-diffusion method. Genomic characteristics related to antibiotic resistance and virulence factors were evaluated by genomic DNA sequencing on the Illumina MiSeq platform and bioinformatic analysis. GPAE0131 isolate showed resistance to piperacillin-tazobactam, cefepime, ceftazidime, imipenem, meropenem, ciprofloxacin, levofloxacin and tobramycin. Resistome comprehend of resistance genes to ß-lactams (blaVIM-2, blaOXA-4, blaOXA-488, blaPDC-35), aminoglycosides (aph(3')-IIb, aac(6')-IIc, aac(6')-Ib9, aadA1), fosfomycin (fosA), chloramphenicol (catB7) and sulfonamides (sul1). Genome comparisons evidence insertion of blaVIM-2 and blaOXA-4 genes. GPAE0131 isolate was predicted to be pathogenic to humans and several virulence factors were found, including encoding gene for ExoU and exotoxin A. All of these features into a pathogenic international high-risk clone (ST235), classified as critical priority, stands out as public health concern due to the widespread dispersal of human pathogens through wastewater. It is suggested that mitigating measures be implemented, such as the treatment of hospital sewage and the addition of tertiary treatment, to prevent the escape of pathogens at this level into the environment.

Detection of KPC directly from positive blood cultures by MALDI-TOF: From research to the clinical microbiology laboratory routine.

Bloodstream infections (BSI) caused by carbapenem-resistant Gram-negative bacilli (CR-GNB) are a subject of major clinical concern, mainly those associated with carbapenemase-producing isolates. Matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) has been proposed to detect specific ß-lactamases, including KPC. We aimed to detect KPC enzyme directly from positive blood cultures using MALDI-TOF MS. Overall, 146 clinical Gram-negative bacilli (46 CR-GNB) recovered from consecutive blood cultures were evaluated. Proteins were extracted using formic acid, isopropyl alcohol, and water and spotted onto a steel target plate using the double-layer sinapinic acid method. The relative ions intensity ≥120 arbitrary units (a.u.) of a peak close to 28,700 m/z indicated the presence of KPC. The results were compared to HRM-qPCR methodology. This specific peak was observed in 11/14 blood bottles with blaKPC positive isolates (78.6% sensitivity), with 3 false-positive results (97.7% specificity). Analysis from colonies reached identical sensitivity (78.6%), but higher specificity (100%). The detection of KPC peaks directly from positive blood cultures using MALDI-TOF MS is feasible and rapid. It's excellent specificity indicates that positive results are consistently associated with the presence of a KPC producer in positive blood culture.

Determination of aztreonam/ceftazidime-avibactam synergism and proposal of a new methodology for the evaluation of susceptibility in vitro.

We proposed a new methodology, the microelution ATM/CZA (mATM/CZA), based on the antibiotic disc elution and the use of resazurin, for rapid (<4h) determination of in vitro susceptibility to aztreonam combined with ceftazidime-avibactam among Enterobacterales. The mATM/CZA presented excellent accuracy with 1.9 %, 98.1 % and 100 % of major error, specificity and sensitivity, respectively. Furthermore, we assessed synergism between aztreonam and ceftazidime-avibactam in Enterobacterales and Pseudomonas aeruginosa, which was observed in 37/55 Enterobacterales and 31/56 P. aeruginosa. As reference methodologies (checkerboard, time-kill curve) are not compatible with the routine of the clinical microbiology laboratories, mATM/CZA is an important alternative to evaluate susceptibility of the combination in a scenario where its clinical use is increasingly important.

Detection of Carbapenem Resistance in Enterobacterales Directly From Positive Blood Cultures Using Matrix-Assisted Laser Desorption Ionization Time-of-Flight Mass Spectrometry.

CONTEXT.­: Carbapenem-resistant Enterobacterales are disseminated worldwide and associated with infections with high rates of morbidity and mortality. Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) is a useful tool for identification of pathogens directly from blood cultures in clinical microbiology laboratories. Furthermore, it has been applied for the detection of carbapenemase production, by evaluating carbapenem hydrolysis. OBJECTIVE.­: To determine meropenem hydrolysis to detect carbapenemase production directly from positive blood cultures, using logRQ to establish a quantitative measure of hydrolysis. DESIGN.­: We evaluated 100 Enterobacterales from positive blood cultures, with 81 carrying a carbapenemase gene (blaKPC, blaGES, blaNDM-1, blaIMP, blaVIM, and blaOXA-48-like), as determined by real-time multiplex polymerase chain reaction with high-resolution melting (HRM-qPCR). Bacterial proteins extracted from positive blood culture bottles were incubated in a meropenem solution (2-4 hours) followed by centrifugation for MALDI-TOF MS analysis. The intensity of peaks of the hydrolyzed and nonhydrolyzed forms were used to calculate the logRQ value. RESULTS.­: Overall, sensitivity was 86.8% and specificity, 89.5%. Of note, sensitivity varied depending on enzyme type. For blaKPC-positive isolates, sensitivity was 97.9%, while it reduced significantly for blaNDM-1 and blaOXA-48-like isolates: 62.5% (10 of 16) and 66.7% (6 of 9), respectively. Indeed, logRQ was higher in blaKPC-positive isolates (0.37-1.97) than in blaNDM-1 (-1.37 to 0.83) and blaOXA-48-like isolates (-1.08 to 1.79). CONCLUSIONS.­: This is an inexpensive and rapid test to identify carbapenemase activity directly from blood culture bottles, which contributes to early adequate antimicrobial therapy and implementation of infection control measures.

Evaluation of the ceftazidime/avibactam and meropenem susceptibility by MALDI-TOF MS directly from positive blood cultures.

The purpose of this study was to evaluate the MBT-ASTRA to determine susceptibility to ceftazidime/avibactam (CZA) and meropenem (MEM) of Enterobacterales directly from positive blood cultures (BC). Bacterial suspension was incubated with antibiotic and analyzed by MALDI-TOF MS. The relative growth was calculated and cutoff values were determined to categorize isolates as "S," "I," and "R." Klebsiella spp. with CZA 20/8 mg/L and 1.5-h incubation presented 1 (5.9%) major discrepancy and 96.3% category agreement; other species required 2.5 h for 100% category agreement. For MEM, 4 mg/L and 1.5h were necessary, demonstrating 2 (6.67%) minor discrepancies and 93.3% categorical agreement.

The role of ankle and knee muscle characteristics in spatiotemporal gait parameters at different walking speeds: A cross-sectional study.

BACKGROUND: Understanding the intricate interplay between ankle and knee muscle characteristics and their impact on gait parameters is crucial for enhancing our comprehension of human locomotion, particularly in the context of varying walking speeds among healthy young adults. RESEARCH QUESTION: The study aimed to identify the relative importance of ankle and knee flexor and extensor muscle characteristics (e.g., strength estimated by peak torque [PT] and rate of torque development [RTD]) in the spatiotemporal gait parameters and variability in self-selected (SSWS) and fast walking speeds (FWS) in healthy young adults. METHODS: One hundred and thirty-nine adults (75 men - 54% and 64 women - 46%; 29.04 ± 9.55 years) were assessed about their muscle characteristics (PT and RTD by an isokinetic dynamometer) and spatiotemporal gait parameters at different walking speeds (SSWS and FWS by an instrumented walkway). RESULTS: Data analysis indicated a weak relationship between the PT and RTD of the ankle and knee and spatiotemporal gait parameters and variability in both walking conditions (SSWS: R2 0.14-0.05; FWS: R2 0.40-0.05). The strength of the knee muscles was more relevant when walking at a self-selected speed, while the strength of the ankle muscles played a more prominent role when walking at a fast pace. SIGNIFICANCE: The findings underscore the critical role of ankle muscles (plantar and dorsiflexors) at fast walking speeds. Therefore, targeted interventions for strength and optimization of these muscles are paramount.

Combined ultrasound and electric field stimulations improve pain and functional capacity in immediate cesarean delivery: a randomized clinical trial, double-blind, and placebo-controlled / Terapia combinada de estimulação elétrica por meio do ultrassom melhora a dor e a capacidade funcional no pós-parto imediato de cesariana: ensaio clínico randomizado, duplo-cego e placebo

ABSTRACT BACKGROUND AND OBJECTIVES: Wound complications and pharmacological pain relief methods used at the skin surgical site after cesarean delivery may result in women's physical and emotional burden. Thus, nonpharmacological treatments must be explored to avoid these complications and side effects on maternal health. The objective of this study was to investigate the effects of Combined Ultrasound and Electric Field Stimulation (CUSEFS) on cicatricial pain and functional capacity in immediate cesarean delivery. METHODS: This study has a randomized clinical trial, double-blind, and placebo-controlled design. Thirty women (25.7±5.0 years) in immediate postpartum were randomly assigned to three groups: Control (CG, n:9), CUSEFS (TG, n:11), and Placebo (PG, n:10). CUSEFS was performed once for 20 minutes. Cicatricial pain (McGill Pain Questionnaire) and functional capacity (Functional Capacity Check) was assessed at baseline, after the intervention, and after 30 minutes. Cohen's (d) and Mixed-design analysis of variance were used to compare groups. RESULTS: Immediately after the intervention, TG showed a decrease in cicatricial pain compared with CG in sensory (d:3.8 to 4.0), affective (d:4.0), and total categories (d:3.9). In functional capacity, TG had less difficulty than CG at walking (d:0.6) and lying down (d:1.1), and PG at rest (d: 0.9). CONCLUSION: CUSEFS might be a resource for managing cicatricial pain and functional capacity in immediate cesarean delivery. Further studies with longer duration and different CUSEFS doses/parameters are required.

RESUMO JUSTIFICATIVA E OBJETIVOS: As complicações na ferida e o uso de métodos farmacológicos de alívio da dor no local cirúrgico após a cesariana podem resultar em sobrecarga física e emocional para a mulher. Assim, tratamentos não farmacológicos devem ser explorados para evitar essas complicações e efeitos adversos à saúde materna. O objetivo deste estudo foi investigar os efeitos da terapia combinada de estimulação elétrica por meio do ultrassom (CUSEFS) na dor cicatricial e na capacidade funcional no pós-parto imediato de cesariana. MÉTODOS: Este estudo possui um desenho de ensaio clínico randomizado, duplo-cego e controlado por placebo. Trinta mulheres (25,7±5,0 anos) em pós-parto imediato de cesariana foram distribuídas aleatoriamente em três grupos: Controle (CG, n:9), CUSEFS (TG:11) e Placebo (PG, n:10). O CUSEFS foi realizado uma vez por 20 minutos. A dor cicatricial (Questionário de Dor McGill) e a capacidade funcional (Functional Capacity Check) foram avaliadas no início, após a intervenção e após 30 minutos. As análises de variância de design misto e Cohen (d) foram usadas para comparar os grupos. RESULTADOS: Imediatamente após a intervenção, o TG apresentou diminuição na dor cicatricial em relação ao CG nas categorias sensorial (d:3,8 a 4,0), afetiva (d:4,0) e total (d:3,9). Na capacidade funcional, o TG apresentou menor dificuldade que o CG na marcha (d:0,6) e deitado (d:1,1), e que o PG em repouso (d:0,9). CONCLUSÃO: O CUSEFS pode ser um recurso para o manejo da dor cicatricial e da capacidade funcional imediatamente após a cesariana. Além disso, são necessários mais estudos com maior duração e diferentes doses/parâmetros de CUSEFS.

Detection of polymyxins resistance among Enterobacterales: evaluation of available methods and proposal of a new rapid and feasible methodology.

BACKGROUND: Fast and accurate detection of polymyxins resistance is necessary as they remain the last resources to treat infections caused by Carbapenem-resistant Enterobacterales in many regions. We evaluated the rapid colorimetric polymyxin B elution (RCPE) and developed its miniaturized version, RCPE microelution (RCPEm), aiming to detect polymyxins resistance among Enterobacterales. METHODS: The methodologies consist of exposing the bacterial population in a solution (NP solution) where polymyxin B disks were previously eluted to obtain a concentration of 2 µg/mL for RCPE and 3 µg/mL for RCPEm. RESULTS: Two hundred sixty-seven Enterobacterales were evaluated, 90 (33.7%) resistant to polymyxin B by broth microdilution. It was observed 0.6% of major error (ME) by RCPE, with a specificity of 99.4%. The miniaturized version (RCPEm) presented the same ME and specificity values, but slightly higher sensitivity (97.8% vs. 95.6%) with 2.2% of very major error (VME). CONCLUSIONS: RCPE and RCPEm proved to be useful alternatives to determine polymyxin B susceptibility in clinical microbiology laboratories, presenting low cost, being easy to perform, and demanding short incubation time.

Copper-Catalyzed Synthesis of Pyrrolo[1,2-c]quinazolines and Pyrrolo[2,1-a]isoquinolines and Antiplasmodial Evaluation.

Reactions involving C(sp3)-H bonds of azaarenes have been widely studied in recent years as they allow direct functionalization of these N-heterocycles without the use of harsh reaction conditions. In this work, we describe the C(sp3)-H functionalization of 4-methylquinazolines and 1-benzylisoquinolines, employing α-substituted ß-nitrostyrenes catalyzed by inexpensive copper acetate. Under the optimized condition, 21 pyrrolo[1,2-c]quinazolines, as well as an imidazo[1,2-c]quinazoline and 4 pyrrolo[2,1-a]isoquinolines, were obtained in moderate to good yields. Furthermore, the biological activity of the pyrrolo[1,2-c]quinazolines was evaluated against Plasmodium falciparum, and promising results were obtained.

Multidrug-resistant Enterobacter spp. in wastewater and surface water: Molecular characterization of ß-lactam resistance and metal tolerance genes.

Among the ESKAPE group pathogens, Enterobacter spp. is an opportunistic Gram-negative bacillus, widely dispersed in the environment, that causes infections. In the present study, samples of hospital wastewater, raw and treated urban wastewater, as well as surface receiving water, were collected to assess the occurrence of multidrug-resistant (MDR) Enterobacter spp. A molecular characterization of ß-lactam antibiotic resistance and metal tolerance genes was performed. According to identification by MALDI-TOF MS, 14 isolates were obtained: 7 E. bugandensis, 5 E. kobei, and 2 E. cloacae. The isolates showed resistance mainly to ß-lactam antibiotics, including those used to treat infections caused by MDR bacteria. Multiple antibiotic resistance index was calculated for all isolates. It allowed verify whether sampling points showed a high risk due to antibiotic resistant Enterobacter spp., as well as to determine if the isolates have been in environments with a frequent antibiotic use. Twelve isolates showed ß-lactam antibiotic resistance gene, being the blaKPC widely detected. Regarding metal tolerance, 13 isolates showed at least two genes that encode metal tolerance mechanisms. Overall, metal tolerance mechanisms to silver, copper, mercury, arsenic and tellurium were found. New data on metal tolerance mechanisms dispersion and antibiotic-resistance characterization of the E. bugandensis and E. kobei species were here provided. The occurrence of MDR Enterobacter spp. in analyzed samples draws attention to an urgent need to put control measures into practice. It also evidences waterborne spread of clinically important antibiotic-resistant bacteria recognized as critical priority pathogens.

The Brazilian Portuguese version of the Pregnancy Mobility Index: Cross-cultural adaptation and psychometric evaluation - a validation study.

BACKGROUND: The Pregnancy Mobility Index (PMI) was developed to assess mobility in pregnant women in the Netherlands. At present, no similar questionnaire is available in Brazil. OBJECTIVE: The present study aimed to translate, cross-culturally adapt, and evaluate the psychometric properties of a Brazilian PMI. DESIGN AND SETTING: The present study was a validation study conducted at the Universidade Federal do Paraná and a public maternity ward in Curitiba, Brazil. METHODS: Text translation and cross-cultural adaptation followed international guidelines. Construct validity, internal consistency, and inter- and intra-rater reliability tests included 97 women. The Pelvic Girdle Questionnaire, Multidimensional Pain Evaluation Scale, Schober's test, and lumbar spine range of motion assessment were administered on the first day. Intra-rater reliability (n = 19) was measured after 15 days. Exploratory factor analysis was performed, and the correlation matrix was analyzed using Pearson's coefficient. RESULTS: Pregnant women (88%) understood the cultural adaptation process. The internal consistency was high (Cronbach's alpha > 0.90), construct validity was moderate, with significant correlation between lumbar spine range of motion (r = 0.283-0.369) and Schober's test (r = -0.314), and high correlation between the Multidimensional Pain Evaluation Scale (r = -0.650 and -0.499) and Pelvic Girdle Questionnaire (r = -0.737). Intra- and inter-rater reliabilities were excellent (intraclass correlation coefficient = 0.932 and 0.990, respectively). CONCLUSION: The Brazilian version of the PMI was successfully translated with excellent reliability and moderate-to-high construct validity. It is an important tool for assessing mobility in pregnant women. CLINICAL TRIAL: RBR-789tps (Validation study), https://ensaiosclinicos.gov.br/rg/RBR-789tps.

Direct Detection of KPC Peak from Positive Blood Cultures Using MALDI-TOF MS: Are We There Yet?

Detecting carbapenemase-associated carbapenem resistance is a subject of major clinical and epidemiological concern as it influences therapeutic choice. Matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) has been proposed as a means to assess bacterial resistance mechanisms. We aimed to detect the KPC enzyme directly from positive blood cultures using MALDI-TOF MS. To do so, 102 clinical Enterobacteria were evaluated, including 59 blaKPC positives. Proteins were extracted using formic acid, isopropyl alcohol, and water (17:33:50) and spotted onto a steel target plate using the double-layer sinapinic acid technique. Two parameters were considered: (i) the visual detection of a clear peak with the expected KPC m/z and (ii) the evaluation of the relative intensity of the ions in the peak. A peak was observed in 56/59 blaKPC-positive isolates (94.9% sensitivity), with no false-positive results (100% specificity). When considering intensity, with a cut-off ≥120 (a.u.), sensitivity was 94.9% and specificity was 95.3%. We proposed a "buffer" zone, with intermediate values of intensity (115 to 125) reaching 100% sensitivity and specificity. The detection of KPC peaks directly from positive blood cultures using MALDI-TOF MS is feasible and rapid, which may improve appropriate patient therapy and antimicrobial stewardship.

Comparison of the Clinical-Functional Vulnerability Index and the frailty phenotype for the identification of falls in older individuals: A cross-sectional study.

BACKGROUND: Frailty increases the risk of falls, disability and death in older adults. The Cardiovascular Health Study identified a frailty phenotype (the Fried Phenotype) that was primarily based on physical domains. Instruments that incorporate additional domains (e.g., cognitive, disability or mood) may more accurately identify falls. OBJECTIVES: The study aimed i) to evaluate the association between falls and the number of phenotypes identified by the Fried Phenotype and CFVI-20 scores and ii) to compare the strength of the association between falls and each frailty instrument. METHODS: This study used the CFVI-20 and the Fried Phenotype and reported falls during the last twelve months. Logistic regression models, odds ratios (ORs), and ROC curves were used to identify associations and perform comparisons (p<0.05). The reporting of the study followed the Strobe guidelines. RESULTS: This study included 1,826 individuals (mean 70.9 (SD 7.3) years old). Prevalence of pre-frailty and low vulnerability was high (72% and 69%) and comparable between frailty instruments. The number of Fried phenotypes increased the odds of having fallen in the past 12 months (OR: 1.5 to 29.5) and the CFVI-20 scores (11% increase/unit change). The CFVI-20 identified falls more accurately than the Fried Phenotype (AUC: 0.68 vs. 0.60, p < 0.001). CONCLUSIONS: The number of phenotypes and the CFVI-20 scores were associated with falls; continuous scores identified falls more accurately than categorical classifications. The CFVI-20 was more strongly associated with falls in community-dwelling older adults than the Fried Phenotype.

Are the spatiotemporal gait parameters at different walking speeds capable of predicting variations in cognitive status by the mini-mental state examination?

BACKGROUND: Several studies have reported the association between cognition and gait; however, most are limited to investigating gait in just one speed, usually the self-selected one, and commonly, only the general score of cognitive tests is used. AIMS: Investigate the relationship between the spatiotemporal gait parameters at different speeds (self-selected and fast walking speeds) and cognitive status in older adults. METHODS: Cross-sectional study. Two hundred and ninety-five older adults (70.9 ± 7.2 years; 82.3% women) were evaluated according to cognitive status (Mini-Mental State Examination, MMSE) and spatiotemporal gait parameters. RESULTS: Data analysis indicates that the spatiotemporal gait parameters (stride length, step time, and single support time) were able to predict cognitive status, assessed by the MMSE general score and its subdomain (temporal and spatial orientation) at both walking speeds. CONCLUSIONS: The spatiotemporal gait parameters were associated with cognitive status, assessed by the MMSE (general score and temporal and spatial orientation), regardless of the walking conditions. The model's variance was significant and with a large effect size. Stride length and step time were identified as the variables with the best explanatory factors. Therefore, it is concluded that greater cognitive performance is associated with longer stride lengths and faster step times.

Nursing Home Residence, Overweight and Cognitive Status are Related to Falls in Older Adults: A Cross-sectional Study.

BACKGROUND: Nursing home residents are likely to differ from community older adults when their gait parameters are compared, as nursing home residents present more falls. AIM: The study aim was to identify the main fall (occurrence predictors) (anthropometrics, functional and gait-related parameters) between older adults living in community and nursing homes during self-selected (SSWS) and fast walking speeds (FWS). METHODS: A hundred and sixty-five older adults were selected from the community (n = 92) and nursing home (n = 73) with and without fall history. They were assessed for fall history, functionality, cognitive status, and several gait parameters in SSWS and FWS conditions. RESULTS: Fallers differed from non-fallers in the SSWS, while such differences were not evidenced during the FWS. Cadence and stride width did not differ when living backgrounds were compared. Nursing home residents walked slower than their non-institutionalized peers, regardless of fall history or walking speed. Besides, binary logistic regression analysis showed that living in a nursing home, age, body mass index (BMI), mini-mental state examination (MMSE), and step width were related to falls in the SSWS. On the other hand, living in a nursing home, having a larger BMI and having low MMSE scores were fall predictors in the FWS. CONCLUSION: Fall occurrence can be identified by factors related to living in nursing homes, cognitive status, BMI, and gait parameters, at the SSWS. Cognitive status and BMI are related to falls in the FWS for those living in nursing homes.

A set of antibiotic-resistance mechanisms and virulence factors in GES-16-producing Klebsiella quasipneumoniae subsp. similipneumoniae from hospital wastewater revealed by whole-genome sequencing.

Klebsiella quasipneumoniae subsp. similipneumoniae has emerged as a human pathogen and sporadic isolates from non-clinical sources were reported. Here, we described the phenotypic- and genomic-characteristics of a multidrug-resistant (MDR) and potentially hypervirulent (MDR-hv) Klebsiella quasipneumoniae subsp. similipneumoniae (KqA1) isolated from hospital wastewater. The antibiotic susceptibility profile of KqA1 was investigated using disk-diffusion method, broth microdilution method, and agar dilution method, and the genetic characteristics of antimicrobial resistance, mobile genetics elements, and virulence were evaluated by genomic DNA sequencing on the Illumina® NovaSeq6000 platform as well as by bioinformatic analysis. Resistome analyses revealed the presence of genes related to resistance to ß-lactams, aminoglycosides, quinolones, tetracyclines, sulfonamides, trimethoprim, chloramphenicol, macrolides, and fosfomycin. New genetic contexts to blaGES-16 (carbapenemase gene) and to fosA (fosfomycin resistance gene) were described. A set of mechanisms that can contribute to antibiotic resistance, commonly detected in Klebsiella spp., was also found including chromosomal mutations, efflux systems, proteins, and regulators. Moreover, KqA1 presented genes related to tolerance to metals (arsenic, copper, nickel, cobalt, magnesium, cadmium, zinc, tellurium, selenium) and to biocides (quaternary-ammonium compounds). The isolate was classified as potentially hypervirulent due to a wide range of virulence factors found associated to regulation, motility, biofilm, effector delivery systems, immune modulation, nutritional/metabolic factors, adherence, invasion, and competitive advantage. The occurrence of MDR-hv KqA1 in hospital wastewater points out how this environment matrix plays a crucial role in the maintenance and selection of critical bacterial pathogens. Regarding One Health perspective, it is evident the need for multidisciplinary implementation of control measures for antibiotic-resistant bacteria, not only in hospital settings but also in a general environmental context to mitigate the dissemination of MDR and hv bacteria.

The Brazilian Portuguese version of the Pregnancy Mobility Index: Cross-cultural adaptation and psychometric evaluation - a validation study

ABSTRACT BACKGROUND: The Pregnancy Mobility Index (PMI) was developed to assess mobility in pregnant women in the Netherlands. At present, no similar questionnaire is available in Brazil. OBJECTIVE: The present study aimed to translate, cross-culturally adapt, and evaluate the psychometric properties of a Brazilian PMI. DESIGN AND SETTING: The present study was a validation study conducted at the Universidade Federal do Paraná and a public maternity ward in Curitiba, Brazil. METHODS: Text translation and cross-cultural adaptation followed international guidelines. Construct validity, internal consistency, and inter- and intra-rater reliability tests included 97 women. The Pelvic Girdle Questionnaire, Multidimensional Pain Evaluation Scale, Schober's test, and lumbar spine range of motion assessment were administered on the first day. Intra-rater reliability (n = 19) was measured after 15 days. Exploratory factor analysis was performed, and the correlation matrix was analyzed using Pearson's coefficient. RESULTS: Pregnant women (88%) understood the cultural adaptation process. The internal consistency was high (Cronbach's alpha > 0.90), construct validity was moderate, with significant correlation between lumbar spine range of motion (r = 0.283-0.369) and Schober's test (r = -0.314), and high correlation between the Multidimensional Pain Evaluation Scale (r = -0.650 and -0.499) and Pelvic Girdle Questionnaire (r = -0.737). Intra- and inter-rater reliabilities were excellent (intraclass correlation coefficient = 0.932 and 0.990, respectively). CONCLUSION: The Brazilian version of the PMI was successfully translated with excellent reliability and moderate-to-high construct validity. It is an important tool for assessing mobility in pregnant women. CLINICAL TRIAL: RBR-789tps (Validation study), https://ensaiosclinicos.gov.br/rg/RBR-789tps.

Novel Dual AChE and ROCK2 Inhibitor Induces Neurogenesis via PTEN/AKT Pathway in Alzheimer's Disease Model.

Alzheimer's disease (AD) is a progressive and complex neurodegenerative disease. Acetylcholinesterase inhibitors (AChEIs) are a major class of drugs used in AD therapy. ROCK2, another promising target for AD, has been associated with the induction of neurogenesis via PTEN/AKT. This study aimed to characterize the therapeutic potential of a novel donepezil-tacrine hybrid compound (TA8Amino) to inhibit AChE and ROCK2 protein, leading to the induction of neurogenesis in SH-SY5Y cells. Experiments were carried out with undifferentiated and neuron-differentiated SH-SY5Y cells submitted to treatments with AChEIs (TA8Amino, donepezil, and tacrine) for 24 h or 7 days. TA8Amino was capable of inhibiting AChE at non-cytotoxic concentrations after 24 h. Following neuronal differentiation for 7 days, TA8Amino and donepezil increased the percentage of neurodifferentiated cells and the length of neurites, as confirmed by ß-III-tubulin and MAP2 protein expression. TA8Amino was found to participate in the activation of PTEN/AKT signaling. In silico analysis showed that TA8Amino can stably bind to the active site of ROCK2, and in vitro experiments in SH-SY5Y cells demonstrate that TA8Amino significantly reduced the expression of ROCK2 protein, contrasting with donepezil and tacrine. Therefore, these results provide important information on the mechanism underlying the action of TA8Amino with regard to multi-target activities.