Farfantepenaeus gene-encoded antimicrobial peptides: Identification, molecular characterization and gene expression in response to fungal infections.

The aim of this study was to identify and characterize, at the molecular and transcriptional levels, sequences encoding the different members of the four families of shrimp antimicrobial peptides (AMPs) in species of the genus Farfantepenaeus. The identification of the AMP sequences was performed by in silico analysis as well as by molecular cloning and nucleotide sequencing. We identified all seven shrimp ALFs (ALF-A to ALF-G), both Type IIa and Type IIb crustins as well as two stylicins (STY1 and STY2) in Farfantepenaeus. Only two genes (PEN1/2 and PEN4) of the four-member penaeidin family (PEN1/2 to PEN5) were found and this is the first report of stylicins as well as of several additional members of ALFs, crustins and penaeidins in species of the genus Farfantepenaeus. All AMP genes have shown to be constitutively transcribed in the shrimp immune cells (hemocytes), except for ALF-G. Finally, the transcriptional profile of the different AMPs was assessed in the hemocytes of F. paulensis (pink shrimp) following an experimental infection with the opportunistic filamentous fungus Fusarium solani. We found that while the expression of ALF-B was induced at 24 h, the STY2 gene was down-regulated at 48 h post-challenge. These results provide evidence of the molecular diversity of AMPs from shrimp of the genus Farfantepenaeus in terms of sequences, biochemical properties and expression profiles in response to infectious diseases.

Infections and antimicrobial resistance in an adult intensive care unit in a Brazilian hospital and the influence of drug resistance on the thirty-day mortality among patients with bloodstream infections.

INTRODUCTION: The present study aimed to determine the incidence of health care-associated infections (HCAIs) and identify the main resistant microorganisms in intensive care unit (ICU) patients in a Brazilian university hospital. METHODS: A retrospective cohort study was conducted in a Brazilian teaching hospital between 2012 and 2014. RESULTS: Overall, 81.2% of the infections were acquired in the ICU. The most common resistant pathogenic phenotypes in all-site and bloodstream infections were oxacillin-resistant coagulase-negative staphylococci and carbapenem-resistant Acinetobacter spp. (89.9% and 87.4%; 80.6% and 70.0%), respectively. CONCLUSIONS: There is an urgent need to focus on HCAIs in ICUs in Brazil.

Comparação de diferentes comprimentos de onda do laser de baixa potência no sóleo de ratos Wistar após lesão nervosa / Comparación entre diferentes longitudes de onda de láser de baja potencia en el sóleo de ratas Wistar tras lesión nerviosa / Comparison of different low-level laser therapy wavelengths in the soleus of Wistar rats after nerve injury

RESUMO Os músculos esqueléticos podem ser afetados por lesões do sistema nervoso periférico, levando a fraqueza e atrofia muscular. Na tentativa de recuperar a funcionalidade dos músculos, existem vários recursos terapêuticos utilizados, dentre os quais o laser de baixa potência (LBP). Este estudo comparou o efeito do LBP em dois comprimentos de onda (660 nm e 830 nm), em características morfológicas do tecido muscular após axonotmese de nervos isquiáticos de ratos Wistar. Para tanto, foram utilizados 32 ratos Wistar, divididos em quatro grupos, sendo G1 (controle), G2 (lesão), G3 (lesão e tratamento com LBP de 660 nm) e G4 (lesão e tratamento com LBP de 830 nm). Os animais de G2, G3 e G4 foram submetidos à lesão do nervo isquiático e, três dias após a lesão, G3 e G4 realizaram tratamento com LBP de 660 nm e 830 nm, respectivamente. Após o tratamento, todos os animais foram eutanasiados e os músculos sóleos coletados para confecção das lâminas histológicas, visando a realização de análises morfológicas do tecido. Constatou-se que os animais submetidos à lesão sofreram alterações morfológicas na fibra, resultando em sua atrofia. Foi percebido também que o LBP com comprimento de onda de 830 nm apresentou ligeiros sinais de recuperação das características morfométricas analisadas.

RESUMEN Las lesiones en el sistema nervioso periférico pueden afectar los músculos esqueléticos y provocar debilidad y atrofia muscular. Para recuperar la funcionalidad de los músculos, se utilizan varios recursos terapéuticos, entre los cuales el láser de baja potencia (LBP). Este estudio comparó el efecto del LBP en dos longitudes de onda (660 nm y 830 nm) sobre las características morfológicas del tejido muscular después de la axonotmesis de los nervios ciáticos en ratas Wistar. Para ello, se utilizaron 32 ratas Wistar, divididas en cuatro grupos: G1 (control), G2 (lesión), G3 (lesión y tratamiento con LBP de 660 nm) y G4 (lesión y tratamiento con LBP de 830 nm). Los animales de G2, G3 y G4 se sometieron a lesión del nervio ciático y, tres días después de la lesión, el G3 y G4 se sometieron al tratamiento con LBP de 660 nm y 830 nm, respectivamente. Después del tratamiento, todos los animales fueron sacrificados y se recogieron los músculos sóleos para la preparación de placas histológicas, con el fin de realizar análisis morfológicos del tejido. Se encontró que los animales sometidos a lesión sufrieron cambios morfológicos en la fibra, lo que resultó en atrofia. También se observó que el LBP con la longitud de onda de 830 nm presentó leves signos de recuperación de las características morfométricas analizadas.

ABSTRACT Skeletal muscles may be affected by peripheral nervous system injuries, leading to muscle weakness and atrophy. Several therapeutic resources may be used in the attempt to recover the functionality of muscles, such as low-level laser therapy (LLLT). This study compared the effect of LLLT of two wavelengths (660 nm and 830 nm) on morphological characteristics of muscle tissue after axonotmesis of ischiatic nerves of Wistar rats. A total of 32 Wistar rats were divided into four groups: G1 (control), G2 (injury), G3 (injury and treatment with 660 nm LLLT) and G4 (injury and treatment with 830 nm LLLT). G2, G3, and G4 animals were submitted to sciatic nerve damage and, three days after the injury, G3 and G4 were treated with LLLT of 660 nm and 830 nm, respectively. After the treatment, all animals were euthanized, and the soles muscles were collected to perform morphological analyzes of the tissue using histological slides. We verified that animals submitted to the lesion underwent morphological changes in the fiber, resulting in their atrophy. We also noticed that LLLT with a wavelength of 830 nm presented slight signs of recovery of the morphometric characteristics analyzed.

Novel small IncX3 plasmid carrying the blaKPC-2 gene in high-risk Klebsiella pneumoniae ST11/CG258.

This study used whole-genome sequencing (WGS) and PFGE to analysis KPC-2-producing Klebsiella pneumoniae strains from clinical specimens collected in Brazilian hospitals. The study identifies the emergence of a novel small IncX3 plasmid (pKPB11), 12,757-bp in length, in a high-risk K. pneumoniae ST11/CG258 lineage, a successful clonal group in Brazil, carrying the blaKPC-2 gene on a non-Tn4401 genetic element (NTEKPC-Ic). Comparative analysis of the pKPB11 showed that this plasmid reduced its size, losing part of its conjugation apparatus. The pKPB11 was also compared to another strain sequenced in this study (KPC89) that had the hybrid IncX3-IncU plasmid (pKP89), of approximately 45 kb in length, similarly carrying the blaKPC-2 gene on NTEKPC-Ic. To the best of our knowledge, pKPB11 is the first example of small IncX3 plasmid found in a high-risk KPC-2-producing K. pneumoniae ST11/CG258.

Infections and antimicrobial resistance in an adult intensive care unit in a Brazilian hospital and the influence of drug resistance on the thirty-day mortality among patients with bloodstream infections

Abstract INTRODUCTION: The present study aimed to determine the incidence of health care-associated infections (HCAIs) and identify the main resistant microorganisms in intensive care unit (ICU) patients in a Brazilian university hospital. METHODS: A retrospective cohort study was conducted in a Brazilian teaching hospital between 2012 and 2014. RESULTS: Overall, 81.2% of the infections were acquired in the ICU. The most common resistant pathogenic phenotypes in all-site and bloodstream infections were oxacillin-resistant coagulase-negative staphylococci and carbapenem-resistant Acinetobacter spp. (89.9% and 87.4%; 80.6% and 70.0%), respectively. CONCLUSIONS: There is an urgent need to focus on HCAIs in ICUs in Brazil.

The action of sericin protein on initial nerve repair, associated or not with swimming in wistar rats / Ação da proteína sericina no reparo nervoso inicial, associada ou não à natação, em ratos wistar / Acción de la proteína sericina en la reparación nerviosa inicial, asociada o no a la natación, en ratones wistar

ABSTRACT Objective: To analyze the effects of sericin treatment, associated or not with swimming with load exercise, on initial sciatic nerve repair after compression in Wistar rats. Methods: Forty animals were divided into five groups: control, injury, injury-sericin, injury-swimming and injury-sericin-swimming. During the axonotmesis procedure, the sericin was applied to the injury-sericin and injury-sericin-swimming groups. The injury-swimming and injury-sericin-swimming groups performed the swimming with load exercise for five days, beginning on the third postoperative day (PO), and were evaluated for function, nociception and allodynia. Euthanasia was performed on the 8th PO day and fragments of the nerve were collected and prepared for quantitative and descriptive analysis in relation to the total amount of viable nerve fibers and non-viable nerve fibers, nerve fiber diameter, axon diameter and myelin sheath thickness. Results: There was no significant improvement in the sciatic functional index up to the eighth day. The Von Frey test of the surgical scar and plantar fascia indicated a reduction in pain and allodynia for the injury-swimming and injury-sericin-swimming groups. The morphological analysis presented similar characteristics in the injury-sericin, injury-swimming and injury-sericin-swimming groups, but there was a significant difference in the number of smaller non-viable nerve fibers in the injury-swimming and injury-sericin-swimming groups as compared to the others. Conclusions: Isolated sericin protein presented proinflammatory characteristics. There was improvement of allodynia and a decrease in the pain at the site of the surgical incision, possibly linked to an aquatic effect. There was no acceleration of nerve repair on the eighth day after the injury. Level of Evidence I; High quality randomized clinical trial with or without statistically significant difference, but with narrow confidence intervals.

RESUMO Objetivo: Analisar os efeitos do tratamento da sericina, associada ou não ao exercício de natação com sobrecarga, sobre o reparo inicial do nervo isquiático após compressão em ratos Wistar. Métodos: Foram separados 40 animais em cinco grupos, sendo eles: controle; lesão; lesão-sericina; lesão-natação e lesão-sericina-natação. Durante o procedimento de axonotmese, a sericina foi aplicada sobre a lesão nos grupos lesão-sericina e lesão-sericina-natação. Os grupos lesão-natação e lesão-sericina-natação realizaram o exercício de natação com sobrecarga durante cinco dias, iniciando no terceiro dia pós-operatório (PO), sendo avaliados quanto à função, nocicepção e alodinia. A eutanásia foi realizada no oitavo dia PO, sendo que dois fragmentos do nervo foram coletados e preparados para análise quantitativa e descritiva em relação a quantidade total de fibras nervosas viáveis, não viáveis, diâmetro da fibra nervosa, do axônio e espessura da bainha de mielina. Resultados: No índice funcional isquiático não houve melhora significativa até o oitavo dia. O teste de Von Frey na cicatriz cirúrgica e fáscia plantar indicaram redução do quadro álgico e alodinia para os grupos lesão-natação e lesão-sericina-natação. A análise morfológica apresentou características semelhantes nos grupos lesão-sericina, lesão-natação e lesão-sericina-natação, porém houve diferença significativa das fibras nervosas não viáveis menores nos grupos lesão-natação e lesão-sericina-natação em relação aos demais. Conclusões: A proteína sericina isolada apresentou características pró-inflamatórias. Houve melhora da alodinia e diminuição do quadro álgico no local da incisão cirúrgica relacionadas a possível efeito aquático. Não houve aceleração do reparo nervoso no oitavo dia após a lesão. Nível de Evidência I; Estudo clínico randomizado de alta qualidade com ou sem diferença estatisticamente significante, mas com intervalos de confiança estreitos.

RESUMEN Objetivo: Analizar los efectos del tratamiento de la sericina, asociada o no al ejercicio de natación con sobrecarga, sobre la reparación inicial del nervio isquiático después de compresión, en ratones Wistar. Métodos: Se separaron 40 animales en cinco grupos, siendo: control; lesión; lesión-sericina; lesión-natación y lesión-sericina-natación. Durante el procedimiento de axonotmesis, la sericina fue aplicada sobre la lesión en los grupos lesión-sericina y lesión-sericina-natación. Los grupos lesión-natación y lesión-sericina-natación realizaron el ejercicio de natación con sobrecarga durante cinco días, iniciándose en el tercer día postoperatorio (PO), siendo evaluados cuanto a la función, nocicepción y alodinia. La eutanasia fue realizada en el octavo día PO, siendo que dos fragmentos del nervio fueron recolectados y preparados para análisis cuantitativo y descriptivo, con relación a la cantidad total de fibras nerviosas viables, no viables, diámetro de la fibra nerviosa, del axón y espesor de la vaina de mielina. Resultados: En el índice funcional isquiático no hubo mejoría significativa hasta el octavo día. La prueba de "Von Frey" en la cicatriz quirúrgica y la fascia plantar indicaron reducción del cuadro álgico y alodinia, para los grupos lesión-natación y lesión-sericina-natación. El análisis morfológico presentó características similares en los grupos lesión-sericina, lesión-natación y lesión-sericina-natación, pero hubo diferencia significativa de las fibras nerviosas no viables menores en los grupos lesión-natación y lesión-sericina-natación con relación a los demás. Conclusiones: La proteína sericina aislada presentó características proinflamatorias. Hubo mejora de la alodinia y disminución del cuadro álgico en el lugar de la incisión quirúrgica, relacionadas al posible efecto acuático. No hubo aceleración de la reparación nerviosa en el octavo día después de la lesión. Nivel de Evidencia I; Ensayo clínico aleatorizado de alta calidad con o sin diferencia estadísticamente significativa, pero con intervalos de confianza estrechos.

Incidence of infections caused by carbapenem-resistant Acinetobacter baumannii.

BACKGROUND: Carbapenem-resistant Acinetobacter baumannii (CR-Ab) has become a worrying health care problem, mainly in developing countries, such as Brazil. The objective was to investigate the prevalence and prognostic factors for CR-Ab infections at a Brazilian university hospital and examine the impact of inappropriate antimicrobial therapy on patient outcome. METHODS: A retrospective study on hospitalized patients with CR-Ab infections was carried out from January 2013 to December 2017. An epidemiologic analysis was carried out to determine the frequency of infections, the epidemiologic indicators by year, the risk factors for 30-day mortality, and the impact of inappropriate therapy. RESULTS: A total of 489 patients were included in the study. A rate of 0.7 per 1,000 patient-day CR-Ab infections was observed, mostly in the lungs (54.7%), and predominantly in the adult intensive care unit. The occurrence of infections by CR-Ab per 1,000 patient-days in November 2014 exceeded the established control limit, confirming an outbreak. CONCLUSIONS: The prevalence of CR-Ab increased in the investigated hospital, passing to an endemic pathogen with a direct impact on mortality and the control of these strains.

Molecular Detection of Class 1 Integron-Associated Gene Cassettes in KPC-2-Producing Klebsiella pneumoniae Clones by Whole-Genome Sequencing.

The dissemination of antimicrobial resistance genes and the bacterium that harbor them have increasingly become a public concern, especially in low- and middle-income countries. The present study used whole-genome sequencing to analyze 10 KPC-2-producing Klebsiella pneumoniae isolates obtained from clinical specimens originated from Brazilian hospitals. The study documents a relevant "snapshot" of the presence of class 1 integrons in 90% of the strains presenting different gene cassettes (dfrA30, dfrA15, dfrA12, dfrA14, aadA1, aadA2, and aac(6')Iq), associated or not with transposons. Two strains presented nonclassical integron (lacking the normal 3'conserved segment). In general, most strains showed a complex resistome, characterizing them as highly resistant. Integrons, a genetically stable and efficient system, confer to bacteria as highly adaptive and low cost evolution potential to bacteria, even more serious when associated with high-risk clones, indicating an urgent need for control and prevention strategies to avoid the spread of resistance determinants in Brazil. Despite this, although the class 1 integron identified in the KPC-2-producing K. pneumoniae clones is important, our findings suggest that other elements probably have a greater impact on the spread of antimicrobial resistance, since many of these important genes were not related to this cassette.

Molecular characterization and clonal dynamics of nosocomial blaOXA-23 producing XDR Acinetobacter baumannii.

The emergence of infections associated to new antimicrobial resistance in Acinetobacter baumannii (Ab) genotypes represents a major challenge. In this context, this study aimed to determine the diversity of resistance mechanisms and investigate clonal dissemination and predominant sequence types (STs) in multidrug-resistant Ab strains of clinical (tracheal aspirate, n = 17) and environmental (surface, n = 6) origins. Additionally, the major clones found in clinical (A) and environmental (H) strains had their complete genomes sequenced. All strains were submitted to polymerase chain reactions (PCR) for the detection of the ISAba1/blaOXA-51-like and ISAba1/blaOXA-23-like genes, while the expression of genes encoding the carO porin, AdeABC (adeB), AdeFGH (adeG), and AdeIJK (adeJ) efflux pumps was determined by real time PCR (qPCR). Most of the strains were characterized as extensively drug-resistant (XDR) with high minimal inhibitory concentrations (MICs) detected for tigecycline and carbapenems. Associations between ISAba1/OXA-51 and ISAba1/OXA-23 were observed in 91.3% and 52.2% of the strains, respectively. Only the adeB gene was considered hyper-expressed. Furthermore, most of the strains analyzed by the MuLtilocus Sequence-Typing (MLST) were found to belong to the clonal complex 113 (CC113). In addition, a new ST, ST1399, belonging to CC229, was also discovered herein. Strains analyzed by whole genome sequencing presented resistance genes linked to multidrug-resistance phenotypes and confirmed the presence of Tn2008, which provides high levels carbapenem-resistance.

Association of Colistin-Resistant KPC Clonal Strains with Subsequent Infections and Colonization and Biofilm Production.

Carbapenemase-producing organisms are pandemic and a significant threat to public health. We investigated the clonal relatedness of colistin-resistant Klebsiella pneumoniae strains producing KPC-type carbapenemase (KPC-KP) causing subsequent infections or colonization. Moreover, we aimed to gain insight into the ability of biofilm production in K. pneumoniae strains producing carbapenemase. Twenty-two consecutive KPC-KP and one KPC-negative strain was identified from an adult intensive care unit in Brazil. Seventy-five percent of isolates that harbored the blaKPC gene exhibited genetic relatedness by pulsed-field gel electrophoresis, and none presented the plasmid-mediated mcr-1 and blaNDM genes. This study showed that the majority of repeated KPC infections in adults were caused by a clone that caused the previous infections/colonizations even after a long period of time and illustrates the capacity of multiple clones producing biofilms to coexist in the same patient at the same time, becoming a reservoir of KPC-KP in the hospital environment.

Arthroscopic surgical treatment of pigmented villonodular synovitis of the elbow: case report.

This case concerns a male patient complaining of pain and discomfort in his right elbow, associated with decreased range of motion. Elbow radiography showed an osteolytic lesion in the metaphyseal region of the distal humerus and magnetic resonance imaging showed intra-articular tumor formation with swelling that suggested pigmented villonodular synovitis. Arthroscopic treatment was performed in order to obtain a synovial biopsy and then to carry out total synovectomy. Anatomopathological examination confirmed the diagnosis. The patient presented excellent progress through the physiotherapeutic rehabilitation proposed and continued to be asymptomatic 12 months after the operation.

O caso diz respeito a um paciente do sexo masculino com queixa de dor e desconforto no cotovelo direito associados a diminuição da amplitude de movimento. Apresentava radiografia do cotovelo com lesão osteolítica da região metafisária do úmero distal e ressonância magnética que mostrava tumoração intra-articular com aumento de volume que sugeria sinovite vilonodular pigmentada. Foi feito tratamento artroscópico para biópsia sinovial e sinovectomia total. O estudo anatomopatológico confirmou o diagnóstico. O paciente apresentou ótima evolução com reabilitação fisioterápica proposta, até 12 meses de pós-operatorio apresentava-se assintomático.

Arthroscopic surgical treatment of pigmented villonodular synovitis of the elbow: case report / Tratamento cirúrgico artroscópico de sinovite vilonodular pigmentada de cotovelo: relato de caso

ABSTRACT This case concerns a male patient complaining of pain and discomfort in his right elbow, associated with decreased range of motion. Elbow radiography showed an osteolytic lesion in the metaphyseal region of the distal humerus and magnetic resonance imaging showed intra-articular tumor formation with swelling that suggested pigmented villonodular synovitis. Arthroscopic treatment was performed in order to obtain a synovial biopsy and then to carry out total synovectomy. Anatomopathological examination confirmed the diagnosis. The patient presented excellent progress through the physiotherapeutic rehabilitation proposed and continued to be asymptomatic 12 months after the operation.

RESUMO O caso diz respeito a um paciente do sexo masculino com queixa de dor e desconforto no cotovelo direito associados a diminuição da amplitude de movimento. Apresentava radiografia do cotovelo com lesão osteolítica da região metafisária do úmero distal e ressonância magnética que mostrava tumoração intra-articular com aumento de volume que sugeria sinovite vilonodular pigmentada. Foi feito tratamento artroscópico para biópsia sinovial e sinovectomia total. O estudo anatomopatológico confirmou o diagnóstico. O paciente apresentou ótima evolução com reabilitação fisioterápica proposta, até 12 meses de pós-operatorio apresentava-se assintomático.

Design of a spring-assisted exoskeleton module for wrist and hand rehabilitation.

This paper reports on the development of a low-profile exoskeleton module to enable training of the fingers and thumb in grasp and release tasks. The design has been made as an add-on module for use with the ArmAssist arm rehabilitation system (Tecnalia, Spain). Variable-position springs and adjustable link lengths provide adaptability to fit a variety of users. Additive manufacturing has been utilized for the majority of components allowing easy modifications. A few structural components were machined from aluminum or steel to produce a functional prototype with sufficient strength for direct evaluation. The design includes independent and adjustable assistance in finger and thumb extension using various width elastic bands, and measurement of user grasp/release forces in finger flexion/extension, thumb flexion/extension, and thumb adduction/abduction using low-profile force sensitive resistors.