Cytotoxicity of glucoevatromonoside alone and in combination with chemotherapy drugs and their effects on Na+,K+-ATPase and ion channels on lung cancer cells.

Cardiac glycosides (CGs) are useful drugs to treat cardiac illnesses and have potent cytotoxic and anticancer effects in cultured cells and animal models. Their receptor is the Na+,K+ ATPase, but other plasma membrane proteins might bind CGs as well. Herein, we evaluated the short- and long-lasting cytotoxic effects of the natural cardenolide glucoevatromonoside (GEV) on non-small-cell lung cancer H460 cells. We also tested GEV effects on Na+,K+ -ATPase activity and membrane currents, alone or in combination with selected chemotherapy drugs. GEV reduced viability, migration, and invasion of H460 cells spheroids. It also induced cell cycle arrest and death and reduced the clonogenic survival and cumulative population doubling. GEV inhibited Na+,K+-ATPase activity on A549 and H460 cells and purified pig kidney cells membrane. However, it showed no activity on the human red blood cell plasma membrane. Additionally, GEV triggered a Cl-mediated conductance on H460 cells without affecting the transient voltage-gated sodium current. The administration of GEV in combination with the chemotherapeutic drugs paclitaxel (PAC), cisplatin (CIS), irinotecan (IRI), and etoposide (ETO) showed synergistic antiproliferative effects, especially when combined with GEV + CIS and GEV + PAC. Taken together, our results demonstrate that GEV is a potential drug for cancer therapy because it reduces lung cancer H460 cell viability, migration, and invasion. Our results also reveal a link between the Na+,K+-ATPase and Cl- ion channels.

Dissemination of Multidrug-Resistant Proteus mirabilis Clones Carrying a Novel Integron-Borne blaIMP-1 in a Tertiary Hospital.

This study aimed to characterize multidrug-resistant Proteus mirabilis clones carrying a novel class 1 integron-borne blaIMP-1 In1359 was inserted into a large conjugative plasmid that also carried blaCTX-M-2 The production of carbapenemases in Enterobacteriaceae that are intrinsically resistant to polymyxins and tigecycline is very worrisome, representing a serious challenge to clinicians and infection control teams.

Cytotoxic effects of the cardenolide convallatoxin and its Na,K-ATPase regulation.

Cardenolides are cardiac glycosides, mostly obtained from natural sources. They are well known for their inhibitory action on the Na,K-ATPase, an effect that regulates cardiovascular alterations such as congestive heart failure and atrial arrhythmias. In recent years, they have also sparked new interest in their anticancer potential. In the present study, the cytotoxic effects of the natural cardenolide convallatoxin (CON) were evaluated on non-small cell lung cancer (A549 cells). It was found that CON induced cytostatic and cytotoxic effects in A549 cells, showing essentially apoptotic cell death, as detected by annexin V-propidium iodide double-staining, as well as changes in cell form. In addition, it prompted cell cycle arrest in G2/M and reduced cyclin B1 expression. This compound also increased the number of cells in subG1 in a concentration- and time-dependent manner. At a long term, the reduction of cumulative population doubling was shown along with an increase of ß-galactosidase positive cells and larger nucleus, indicative of senescence. Subsequently, CON inhibited the Na,K-ATPase in A549 cells at nM concentrations. Interestingly, at the same concentrations, CON was unable to directly inhibit the Na,K-ATPase, either in pig kidney or in red blood cells. Additionally, results of docking calculations showed that CON binds with high efficiency to the Na,K-ATPase. Taken together, our data highlight the potent anticancer effects of CON in A549 cells, and their possible link with non-classical inhibition of Na,K-ATPase.

Influence of Culture Media on Detection of Carbapenem Hydrolysis by Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry.

In this study, we evaluated the influence of distinct bacterial growth media on detection of carbapenemase hydrolysis by matrix-assisted laser desorption ionization-time of flight mass spectrometry. False-negative results were observed for OXA-25-, OXA-26-, and OXA-72-producing Acinetobacter baumannii isolates grown on MacConkey agar medium. The other culture media showed 100% sensitivity and 100% specificity for detecting carbapenemase.

Voiced High-Frequency Oscillation or Lax Vox Technique? Immediate Effects in Dysphonic Individuals.

OBJECTIVES: To analyze the immediate effects of voiced high-frequency oscillation (VHFO) and Lax Vox technique on vocal quality and self-reported intensity of vocal and laryngeal symptoms in individuals with behavioral dysphonia. METHODS: This experimental, prospective, randomized cross-over study, investigated thirty adults (15 women and 15 men) with behavioral dysphonia (vocal complaints, altered voice on auditory-perceptual evaluation, vocal nodules or mucosal thickening, and incomplete glottic closure). The outcome variables analyzed were auditory-perceptual analysis, acoustic analysis (voice quality characteristics), and self-reported intensities of vocal and laryngeal symptoms. Each participant performed two exercises-VHFO and Lax Vox technique-in a random sequence for 3 minutes. A 7-day washout period was provided between the exercises. The data were analyzed using the paired t-test and Wilcoxon test (P < 0.05). RESULTS: After VHFO, no significant difference was observed on auditory-perceptual evaluation in all participants, whereas the Lax Vox technique worsened breathiness among women (P = 0.027). VHFO significantly increased the fundamental frequency (P = 0.014) and decreased the noise harmonic ratios for women (P = 0.026). Among men, there was a decrease in shimmer parameter (P = 0.035). Moreover, symptoms such as "lump in the throat" (P = 0.005), "voice loss" (P = 0.017), and "high-pitched voice" (P = 0.023) decreased in women after VHFO, whereas in men, "itchiness" and "hoarseness" (P < 0.001) decreased after VHFO. The Lax Vox technique decreased "hoarseness" (P = 0.003) in women, without any effect in men. CONCLUSION: The VHFO exercise provided more positive immediate effects results than the Lax Vox technique regarding vocal quality and self-reported symptom intensity in participants with behavioral dysphonia.

Ultrasound Protocols to Assess Skeletal and Diaphragmatic Muscle in People Who Are Critically Ill: A Systematic Review.

This study aims to review published studies that use protocols and ultrasound measurements to evaluate skeletal and diaphragmatic muscles in patients who are critically ill. We searched for references on databases through September 2020 and included in our systematic review studies that used muscular ultrasound to assess skeletal or diaphragm muscles in patients who are critically ill. Seventy-six studies were included, 32 (1720 patients) using skeletal-muscle ultrasound and 44 (2946 patients) using diaphragmatic-muscle ultrasound, with a total of 4666 patients. The population is predominantly adult men. As for designs, most studies (n = 62) were cohort studies. B-mode B was dominant in the evaluations. Medium-to-high frequency bands were used in the analysis of peripheral muscles and medium-to-low frequency bands for diaphragmatic muscles. Evaluation of the echogenicity, muscle thickness and pennation angle of the muscle was also reported. These variables are important in the composition of the diagnosis of muscle loss. Studies demonstrate great variability in their protocols, and sparse description of the important variables that can directly interfere with the quality and validity of these measures. Therefore, a document is needed that standardizes these parameters for ultrasound assessment in patients who are critically ill.

Phylogeographical structure of the neotropical forest tree Hymenaea courbaril (Leguminosae: Caesalpinioideae) and its relationship with the Vicariant Hymenaea stigonocarpa from Cerrado.

The phylogeography of Hymenaea courbaril var. stilbocarpa from Atlantic Forest and riverine forests of the Cerrado biome in central and southeastern Brazil was investigated. The data were compared with those of its congeneric Hymenaea stigonocarpa, a typical tree from savanna. In the Cerrado, H. courbaril var. stilbocarpa is found in sites contiguous with those of H. stigonocarpa, and they share common life-history attributes. The psbC/trnS3 region of the chloroplast DNA was sequenced in 149 individuals of H. courbaril var. stilbocarpa. High genetic variation was found in this species, with the identification of 18 haplotypes, similarly to what was found in H. stigonocarpa with 23 haplotypes in the same geographic region. Populations of H. courbaril var. stilbocarpa could be structured in 3 phylogeographic groups. Spatial analysis of molecular variation indicated that 46.4% of the genetic variation was due to differences among these groups. Three haplotypes were shared by H. courbaril var. stilbocarpa and H. stigonocarpa, and only 10.5% of the total genetic variation could be attributed to between-species difference. We surmise that during the glacial times, H. courbaril var. stilbocarpa populations must have gone extinct in most parts of the southern of its present-day occurrence area. After climate amelioration, these areas were probably recolonized from northern and eastern. The relatively similar phylogeographic structure of vicariant Hymenaea species suggests that they were subjected to the same impacts during the Quaternary climatic fluctuations. The sharing of haplotypes and the genetic similarity between the 2 Hymenaea species suggest the existence of ancestral polymorphism and/or hybridization.

High Frequency of Extended-Spectrum Beta-Lactamase-Producing Klebsiella pneumoniae Nosocomial Strains Isolated from a Teaching Hospital in Brazil.

The aim of this study was to investigate the frequency, antimicrobial sensitivity profile, and genetic characteristics of nosocomial strains of extended-spectrum ß-lactamase (ESBL)-producing Klebsiella pneumoniae isolated from inpatients at a teaching hospital in Brazil. The bacterial identification, phenotypic detection of ESBL, and antimicrobial susceptibility profile were performed by the VITEK 2 automated system. Matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF MS) mass spectrometry was used to confirm the identity of the species and genotyping of ESBL-producing K. pneumoniae was performed by pulsed-field gel electrophoresis (PFGE). Thirty-six ESBL-producing K. pneumoniae nosocomial strains isolated from November 2013 to August 2014 were analyzed. High resistance rates were observed for ceftriaxone, ceftazidime, cefepime, gentamicin, and ciprofloxacin. However, all isolates were susceptible to amikacin and meropenem. All strains harbored blaCTX-M-like and blaSHV-like genes. Molecular typing by PFGE showed a diversity of genotypes distributed among 25 clusters, but two isolates collected in different wards had the same genotypic profile and carried the same bla genes, so they were considered clones. The data showed that there was a high frequency of ESBL-producing K. pneumoniae multidrug-resistant among patients in the studied hospital. Furthermore, the detection of blaCTX-M-like genes in all isolates suggests that these enzymes are the major ESBL responsible for the beta-lactam resistance phenotypes among the analyzed strains.

Effectiveness of Voice Therapy Associated With Electromyographic Biofeedback in Women With Behavioral Dysphonia: Randomized Placebo-Controlled Double-Blind Clinical Trial.

OBJECTIVE: To analyze the effectiveness of vocal therapy associated with electromyographic biofeedback in women with behavioral dysphonia. MATERIALS AND METHODS: This is a randomized placebo-controlled double-blind clinical trial. Twenty-two adult women with behavioral dysphonia were randomly divided into two groups: Experimental Group-11 women participated in vocal therapy associated with the application of electromyographic biofeedback; Placebo Group-11 women participated in vocal therapy associated with the application of placebo electromyographic biofeedback. Both groups performed eight therapy sessions, twice a week, lasting 30 minutes. The vocal therapy of both groups was composed of semioccluded vocal tract exercises (trill, humming, and fricative). The evaluations were performed at four time points-before, after, one, and three months after the vocal therapy-and will consist of the following assessments: auditory-perceptual evaluation of voice, acoustic evaluation of voice, and surface electromyographic. The data were analyzed statistically comparing the groups and the time of evaluation (P < 0.05). RESULTS: The proposed vocal therapy promoted positive results in vocal quality and muscular electrical activity during rest in women with behavioral dysphonia for both groups. Electromyographic biofeedback promoted additional positive results in muscle electrical activity during phonatory tasks in women with behavioral dysphonia. CONCLUSION: In this study, the vocal therapy associated with electromyographic biofeedback had equivalent efficacy to traditional therapy in the voice. The biofeedback was more effective than traditional therapy on muscular electrical activity and had effects that remained for a longer time in women with behavioral dysphonia.

Rapid detection of blaKPC directly from surveillance rectal swabs by EasyQ KPC.

The performance of EasyQ KPC assay was evaluated for the first time for blaKPC detection directly from surveillance rectal swabs without broth enrichment. Using conventional polymerase chain reaction as gold standard method, EasyQ KPC and culture-based molecular tests demonstrated a sensitivity/specificity of 100%/87.3% and 83.3%/98.2%, respectively.

Musculoskeletal Pain and Voice-related Quality of Life in Dysphonic and Non-dysphonic Subjects.

PURPOSE: This study aimed to compare and correlate musculoskeletal pain and voice-related quality of life of dysphonic and non-dysphonic individuals. METHOD: This is a retrospective case-control study. A total of 74 adults were divided into two groups: the experimental group (EG) comprising 37 individuals with vocal complaints and hyperfunctional dysphonia, and the control group (CG) comprising 37 individuals without vocal complaints and with healthy voices. Both groups presented similar gender and age (28 females and 9 males for each group; average age = 31.5). All the participants answered the protocols: Voice-Related Quality of Life and Musculoskeletal Pain Questionnaire. Statistical data were analyzed by Mann-Whitney U test (P ≤ 0.05) and Spearman correlation test (P ≤ 0.05). RESULTS: It was observed that the EG presented significantly lower scores of voice-related quality of life in the social-emotional (P < 0.001), physical (P < 0.001), and total (P < 0.001) fields. Concerning musculoskeletal pain, it was observed in the EG that there was a higher intensity in pain in the region of the larynx (P < 0.001), and a higher frequency of pain in the submandibular (P = 0.013), larynx (P < 0.001), and front of the neck (P = 0.002) regions, when compared with the CG. CONCLUSION: In the group of individuals studied, worst indexes of voice-related quality of life and higher frequency and intensity of pain in the larynx were observed, in addition to higher frequency of pain in regions near the larynx in dysphonic subjects. There was correlation between voice-related quality of life and the frequency and intensity of musculoskeletal pain.

A high mortality rate associated with multidrug-resistant Acinetobacter baumannii ST79 and ST25 carrying OXA-23 in a Brazilian intensive care unit.

The global spread of carbapenem-resistant Acinetobacter baumannii (A. baumannii) strains has restricted the therapeutic options available to treat infections due to this pathogen. Understanding the prevalence of such infections and the underlying genetic mechanisms of resistance may help in the implementation of adequate measures to control and prevent acquisition of nosocomial infections, especially in an intensive care unit setting. This study describes the molecular characteristics and risk factors associated with OXA-23-producing A. baumannii infections. A case-control study was undertaken from September/2013 to April/2015. Acquisition of OXA-23-producing A. baumannii was found to be associated with the use of nasogastric tubes, haemodialysis, and the use of cephalosporins. These isolates were only susceptible to amikacin, gentamicin, tigecycline, and colistin, and contained the ISAba1 insertion sequence upstream ofblaOXA-23 and blaOXA-51 genes. Twenty-six OXA-23-producing A. baumannii strains belonged to the ST79 (CC79) clonal group,and patients infected or colonised by these isolates had a higher mortality rate (34.6%). In conclusion, this study showed a dissemination of OXA-23-producing A. baumannii strains that was associated with several healthcare-related risk factors and high mortality rates among intensive care unit patients.

Cobertura vacinal e o movimento antivacina: o impacto na saúde pública no Brasil / Vaccination coverage and anti-vaccine movement: the impact on public health in Brazil / Cobertura de vacunación y movimiento antivacuna: el impacto en la salud pública en Brasil

A vacina constitui um dos principais métodos de prevenção contra doenças. Em 1973, o Brasil criou o Programa Nacional de Imunizações a fim de promover a imunização gratuita para a população, o que mais tarde tornou o país em referência mundial em vacinação. No entanto, a recusa vacinal ainda é um grande problema de saúde pública, sendo o movimento antivacina um dos destaques dessa realidade. Dessa forma, o objetivo deste artigo é avaliar como o movimento antivacina impacta na saúde pública no Brasil através da diminuição da cobertura vacinal. Trata-se de uma pesquisa de metodologia mista, com uma primeira etapa qualitativa, composta de uma revisão integrativa nas plataformas PubMed, LILACS e SciELO, no período de 2010 a 2020, e uma pesquisa documental em portais de movimentos antivacina; e uma segunda etapa quantitativa, em que foi realizado um estudo epidemiológico do tipo ecológico, com consulta nas bases eletrônicas do Datasus e no Sistema de Informações do Programa Nacional de Imunizações (SI-PNI), no período de 2010 a 2022. No período investigado, apenas em 2015 o Brasil alcançou a meta preconizada de cobertura vacinal, diferentemente dos anos seguintes, que apresentaram oscilações preocupantes. As publicações apresentam argumentos utilizados pelos grupos antivacina, evidenciados entre 2015 e 2019, período em que os dados de cobertura vacinal oscilaram. Assim, conclui-se que a ascensão do movimento antivacina é um dos fatores que influenciaram na queda da vacinação no Brasil, a exemplo do sarampo e da febre amarela.

The vaccine is one of the main methods of preventing diseases. Since 1973, Brazil created the National Immunization Program to ensure free immunization to the population, which later made the country a world reference in vaccination. However, vaccine refusal is still a great public health issue, and the anti-vaccine movement stand out in this reality. Thus, the purpose of this article is to evaluate how the anti-vaccine movement affects public health in Brazil with vaccination coverage reduction. This is a mixed methodology study, with first a qualitative step, composed of an integrative review in the platforms PubMed, LILACS, and SciELO, in the period from 2010 to 2020,and a documental research in portals of anti-vaccination movements; and a second quantitative step, where an epidemiological study of the ecological type was carried out, with consultation in the electronic databases of DATASUS and in the Information System of the National Immunization Program (SI-PNI) in the period of 2010 to 2022. In the investigative period, only in 2015 Brazil managed to reach the recommended vaccination coverage goal, unlike in the following years, which showed worrying fluctuations. The publications summarize arguments used by the anti-vaccination groups, evidenced between 2015 and 2019, a period in which the vaccination coverage data fluctuated. Therefore, it is clear that the rise of the anti-vaccination movement is a factor that influenced the drop in vaccination numbers in Brazil, with yellow fever and measles as examples.

La vacuna es uno de los principales métodos de prevención de enfermedades. En 1973, Brasil creó el Programa Nacional de Inmunización con el fin de promover la inmunización gratuita para la población, lo que luego convirtió al país en un referente mundial en vacunación. Sin embargo, la negativa de la vacuna sigue siendo un problema importante en la salud pública, y el movimiento antivacunas es uno de los aspectos más destacados de esta realidad. Así, el objetivo de este artículo es evaluar cómo el movimiento antivacunas impacta en la salud pública en Brasil mediante la disminución de la cobertura de vacunación. Se trata de un estudio epidemiológico mixto, con una primera etapa cualitativa, consistente en una revisión integradora en las plataformas PubMed, Lilacs y SciELO, en el período de 2010 a 2020, y una investigación documental en portales de movimientos antivacunas; y una segunda etapa cuantitativa, en la que se realizó un estudio epidemiológico de tipo ecológico, con consulta en las bases de datos electrónicas de DATASUS y en el Sistema de Información del Programa Nacional de Inmunización (SI-PNI), en el período de 2010 a 2022. Entre eses años, solo el año 2015 logró alcanzar la meta recomendada, a diferencia de los años siguientes, que mostraron fluctuaciones preocupantes en la cobertura de vacunación. Las publicaciones mostraron los argumentos utilizados por los grupos antivacina, evidenciados entre 2015 y 2019, período en que los datos de cobertura de la vacuna fluctuaron. Así, se concluye que la asunción del movimiento antivacunación es uno de los factores que influye en la caída de la vacunación en Brasil, como en el sarampión y la fiebre amarilla.

Cytotoxic and cytostatic effects of digitoxigenin monodigitoxoside (DGX) in human lung cancer cells and its link to Na,K-ATPase.

Cardiac glycosides (CGs) are natural compounds widely used to treat several cardiac conditions and more recently have been recognized as potential antitumor agents. They are known as Na,K-ATPases ligands, which is a promising drug target in cancer. In this study, the short and long-lasting cytotoxic effects of the natural cardenolide digitoxigenin monodigitoxoside (DGX) were evaluated against two non-small cell lung cancer lines (A549 and H460 cells). It was found that DGX induced cytotoxic effects in both cells and the apoptotic effects were more pronounced on H460 cells. In long-term analysis, using the clonogenic and the cumulative population doubling (CPD) assays, DGX showed a reduction of cell survival, after 15days without re-treatment. To better understand DGX effects in A549 cells, several assays were conducted. In cell cycle analysis, DGX caused an arrest in S and G2/M phases. This compound also increased the number of cells in subG1 phase in a concentration- and time-dependent manner. The presence of ß-galactosidase positive cells, large nucleus and flattened cells indicated senescence. Additionally, DGX inhibited Na,K-ATPase activity in A549 cells, as well as in purified pig kidney and in human red blood cell membrane preparations, at nanomolar range. Moreover, results of molecular docking showed that DGX binds with high efficiency (-11.4Kcal/mol) to the Na,K-ATPase (PDB:4HYT). Taken together, our results highlight the potent effects of DGX both in A549 and H460 cells, and disclose its link with Na,K-ATPase inhibition.

Cytotoxicity of AMANTADIG - a semisynthetic digitoxigenin derivative - alone and in combination with docetaxel in human hormone-refractory prostate cancer cells and its effect on Na+/K+-ATPase inhibition.

Cardiac glycosides (CGs) are natural compounds used to treat congestive heart failure. They have garnered attention as a potential cancer treatment option, especially because they bind to Na+/K+-ATPase as a target and activate intracellular signaling pathways leading to a variety of cellular responses. In this study we evaluated AMANTADIG, a semisynthetic cardenolide derivative, for its cytotoxic activity in two human androgen-insensitive prostate carcinoma cell lines, and the potential synergistic effects with docetaxel. AMANTADIG induced cytotoxic effects in both cell lines, and a combination with docetaxel showed a moderate and strong synergism in DU145 and PC-3 cells, respectively, at concentrations considerably lower than their IC50 values. Cell cycle analyses showed that AMANTADIG and its synergistic combination induced G2/M arrest of DU145 and PC-3 cells by modulating Cyclin B1, CDK1, p21 and, mainly, survivin expression, a promising target in cancer therapy. Furthermore, AMANTADIG presented reduced toxicity toward non-cancerous cell type (PBMC), and computational docking studies disclosed high-affinity binding to the Na+/K+-ATPase α subunit, a result that was experimentally confirmed by Na+/K+-ATPase inhibition assays. Hence, AMANTADIG inhibited Na+/K+-ATPase activity in PC-3 cells, as well as in purified pig kidney at nanomolar range. Altogether, these data highlight the potent effects of AMANTADIG in combination with docetaxel and offer important insights for the development of more effective and selective therapies against prostate cancer.

Phylogeography of the tree Hymenaea stigonocarpa (Fabaceae: Caesalpinioideae) and the influence of quaternary climate changes in the Brazilian cerrado.

BACKGROUND AND AIMS: Hymenaea stigonocarpa (Fabaceae: Caesalpinioideae) is an endemic tree from the Brazilian cerrado (savanna vegetation), a biome classified as a hotspot for conservation priority. This study investigates the phylogeographic structure of H. stigonocarpa, in order to understand the processes that have led to its current spatial genetic pattern. METHODS: The polymorphism level and spatial distribution of variants of the plastid non-coding region between the genes psbC and trnS were investigated in 175 individuals from 17 populations, covering the greater part of the total distribution of the species. Molecular diversity indices were calculated and intra-specific relationships were inferred by the construction of haplotype networks using the median-joining method. Genetic differentiation among populations and main geographical groups was evaluated using spatial analysis of molecular variance (SAMOVA). KEY RESULTS: Twenty-three different haplotypes were identified. The level of differentiation among the populations analysed was relatively high (F(ST) = 0.692). Phylogeographic analyses showed a clear association between the haplotype network and geographic distribution of populations, revealing three main geographical groups: western, central and eastern. SAMOVA corroborated this finding, indicating that most of the variation can be attributed to differences among these three groups (58.8 %), with little difference among populations within groups (F(SC) = 0.252). CONCLUSIONS: The subdivision of the geographic distribution of H. stigonocarpa populations into three genetically differentiated groups can be associated with Quaternary climatic changes. The data suggest that during glacial times H. stigonocarpa populations became extinct in most parts of the southern present-day cerrado area. Milder climatic conditions in the north and eastern portions of the cerrado resulted in maintenance of populations in these regions. Thus it is inferred that the most southern part of the present-day cerrado was re-colonized by different lineages from northern parts of this biome, after postglacial climate amelioration.

Detection and analysis of different interactions between resistance mechanisms and carbapenems in clinical isolates of Klebsiella pneumoniae.

Carbapenems are considered last-line agents for the treatment of serious infections caused by Klebsiella pneumoniae, and this microorganism may exhibit resistance to ß-lactam antibiotics due to different mechanisms of resistance. We evaluated 27 isolates of K. pneumoniae resistant to carbapenems recovered from inpatients at the University Hospital of Santa Maria-RS from July 2013 to August 2014. We carried out antimicrobial susceptibility, carbapenemase detection, testing for the presence of efflux pump by broth microdilution and loss of porin by sodium dodecyl sulfate polyacrylamide gel electrophoresis. Genetic similarity was evaluated by ERIC-PCR. High levels of resistance were verified by the minimum inhibitory concentration for the antimicrobials tested. The blaKPC gene was present in 89% of the clinical isolates. Blue-Carba and combined disk with AFB tests showed 100% concordance, while the combined disk test with EDTA showed a high number of false-positives (48%) compared with the gold-standard genotypic test. Four isolates showed a phenotypic resistance profile consistent with the overexpression of the efflux pump, and all clinical isolates had lost one or both porins. The ERIC-PCR dendrogram demonstrated the presence of nine clusters. The main mechanism of resistance to carbapenems found in the assessed isolates was the presence of the blaKPC gene.

Identification of São Paulo metallo-beta-lactamase-1-producing Pseudomonas aeruginosa in the Central-West region of Brazil: a case study.

Metallo-beta-lactamase production is an important mechanism for carbapenem resistance of Pseudomonas aeruginosa , which represents an emerging public health challenge. We report the case of a patient admitted to an intensive care unit, with sepsis caused by multidrug-resistant São Paulo Metallo-beta-lactamase-1-producing P. aeruginosa . This is the first case of infection by this pathogenic strain in the State of Mato Grosso do Sul, Brazil. Thus, infection control measures are required for preventing future spread and outbreaks.