Your browser doesn't support javascript.
loading
Mostrar: 20 | 50 | 100
Resultados 1 - 20 de 9.214
Filtrar
1.
Crit Care Med ; 49(1): e31-e40, 2021 01 01.
Artigo em Inglês | MEDLINE | ID: mdl-33122577

RESUMO

OBJECTIVES: We aimed to assess the frequency of ICU-acquired bloodstream infections in coronavirus disease 2019 patients. DESIGN: Retrospective observational study. SETTING: The emergency expansion of an ICU from eight general beds to 30 coronavirus disease 2019 beds. PARTICIPANTS: Patients with coronavirus disease 2019 admitted to the ICU of Luigi Sacco Hospital (Milan, Italy) for greater than or equal to 48 hours between February 21, 2020, and April 30, 2020. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: The frequency of bloodstream infections per 1,000 days of ICU stay was calculated in 89 coronavirus disease 2019 patients, and the cumulative probability of bloodstream infection was estimated using death and ICU discharge as competing events. Sixty patients (67.4%) experienced at least one of the 93 recorded episodes of bloodstream infection, a frequency of 87 per 1,000 days of ICU stay (95% CI, 67-112).The patients who experienced a bloodstream infection had a higher Sequential Organ Failure Assessment score upon ICU admission (9.5; interquartile range, 8-12 vs 8, interquartile range, 5-10; p = 0.042), a longer median ICU stay (15 d; interquartile range, 11-23 vs 8, interquartile range, 5-12; p < 0.001), and more frequently required invasive mechanical ventilation (98.3% vs 82.8%; p = 0.013) than those who did not. The median time from ICU admission to the first bloodstream infection episode was 10 days. Gram-positive bacteria accounted for 74 episodes (79.6%), with Enterococcus species being the most prevalent (53 episodes, 55.8%). Thirty-two isolates (27.3%) showed multidrug resistance. CONCLUSIONS: Coronavirus disease 2019 seemed to increase the frequency of bloodstream infections (particularly Enterococcus-related bloodstream infection) after ICU admission. This may have been due to enteric involvement in patients with severe coronavirus disease 2019 and/or limitations in controlling the patient-to-patient transmission of infectious agents in extremely challenging circumstances.


Assuntos
/microbiologia , Enterococcus/isolamento & purificação , Infecções por Bactérias Gram-Positivas/microbiologia , Tempo de Internação/estatística & dados numéricos , Sepse/microbiologia , Adulto , Idoso , Estado Terminal , Feminino , Infecções por Bactérias Gram-Positivas/epidemiologia , Humanos , Unidades de Terapia Intensiva , Itália , Masculino , Pessoa de Meia-Idade , Estudos Retrospectivos , Sepse/epidemiologia , Resultado do Tratamento
2.
Int J Dermatol ; 60(1): 44-52, 2021 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-32686136

RESUMO

BACKGROUND: Toxic epidermal necrolysis (TEN) is a life-threatening severe cutaneous adverse reaction. Data on pediatric TEN is limited. METHODS: Case records of 44 children, 1 month-12 years with a diagnosis of TEN (>30% body surface area [%BSA] detachment) admitted to a tertiary pediatric intensive care unit (PICU) between 2009 and 2018 were analyzed retrospectively. The primary outcome was mortality, and secondary outcomes were organ dysfunction, length of stay (LOS), and long-term sequelae. RESULTS: Median (IQR) age was 6.5 (3.6, 8.0) years, and 25 (57%) were boys. Median (IQR) %BSA involved, SCORTEN score, and PRISM-III were 65% (45, 80); 2 (2, 3) and 13 (10, 16), respectively. Antiepileptics (n = 24, 54.6%) and antimicrobials (n = 8, 18.2%) were the most common offending agents. Twenty-four (54.5%) children had culture positive sepsis. Immunomodulatory therapy was provided in 35 (79.5%) and conservative management in nine (20.5%) children. Intravenous immunoglobulin (IVIG) was given in 22 (50%), steroids in three (6.8%), and both IVIG and steroids in 10 (22.7%) children. Respiratory failure (n = 14, 31.8%) was the commonest organ failure. Mortality was 15.9% (n = 7), and median (IQR) PICU-LOS in survivors was 8 (4, 11.75) days. There was no association between IVIG, steroids, or conservative management with mortality or LOS. Ocular sequelae (n = 20, 54.1%) were the most common long-term complication followed by skin (18, 40.1%). CONCLUSION: Immunomodulation with IVIG or steroids was not associated with any mortality benefit as compared to conservative management alone. Further research is required to determine the most effective treatment in pediatric TEN.


Assuntos
Cuidados Críticos , Síndrome de Stevens-Johnson/terapia , Anti-Inflamatórios/uso terapêutico , Criança , Pré-Escolar , Tratamento Conservador , Dexametasona/uso terapêutico , Oftalmopatias/etiologia , Feminino , Humanos , Imunoglobulinas Intravenosas/uso terapêutico , Fatores Imunológicos/uso terapêutico , Imunomodulação , Lactente , Unidades de Terapia Intensiva Pediátrica , Tempo de Internação , Masculino , Metilprednisolona/uso terapêutico , Insuficiência Respiratória/etiologia , Estudos Retrospectivos , Sepse/microbiologia , Índice de Gravidade de Doença , Esteroides/uso terapêutico , Síndrome de Stevens-Johnson/complicações , Síndrome de Stevens-Johnson/mortalidade , Taxa de Sobrevida , Fatores de Tempo , Resultado do Tratamento
3.
J Vis Exp ; (165)2020 11 06.
Artigo em Inglês | MEDLINE | ID: mdl-33226026

RESUMO

In rodent models, tail vein injections are important methods for intravenous administration of experimental agents. Tail vein injections typically involve warming of the animal to promote vasodilation, which aids in both the identification of the blood vessels and positioning of the needle into the vessel lumen while securely restraining the animal. Although tail vein injections are common procedures in many protocols and are not considered highly technical if performed correctly, accurate and consistent injections are crucial to obtain reproducible results and minimize variability. Conventional methods for inducing vasodilation prior to tail vein injections generally depend on the use of a heat source such as a heat lamp, electrical/rechargeable heat pads, or pre-heated water at 37 °C. Despite being readily accessible in a standard laboratory setting, these tools evidently suffer from poor/limited thermo-regulatory capacity. Similarly, although various forms of restraining devices are commercially available, they must be used carefully to avoid trauma to the animals. These limitations of the current methods create unnecessary variables in experiments or result in varying outcomes between experiments and/or laboratories. In this article, we demonstrate an improved protocol using an innovative device that combines an independent, thermally regulated, warming device with an adjustable restraining unit into one system for efficient streamlined tail vein injection. The example we use is an intravenous model of fungal bloodstream infection that results in sepsis. The warming apparatus consists of a heat-reflective acrylic box installed with an adjustable automatic thermostat to maintain the internal temperature at a pre-set threshold. Likewise, the width and height of the cone restraining apparatus can be adjusted to safely accommodate various rodent sizes. With the advanced and versatile features of the device, the technique shown here could become a useful tool across a range of research areas involving rodent models that employ tail vein injections.


Assuntos
Temperatura Alta , Injeções/instrumentação , Sepse/microbiologia , Cauda/irrigação sanguínea , Veias/patologia , Animais , Candida/imunologia , Modelos Animais de Doenças , Vacinas Fúngicas/imunologia , Injeções Intravenosas , Camundongos Endogâmicos C57BL , Agulhas , Ratos , Sepse/complicações , Vacinação
4.
J Med Microbiol ; 69(12): 1398-1404, 2020 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-33156750

RESUMO

Introduction. Rapid identification of the causative agent of sepsis is crucial for patient outcomes.Aim. The Sepsityper sample preparation method enables direct microbial identification of positive blood culture samples via matrix-assisted laser desorption ionization/time-of-flight mass spectrometry (MALDI-TOF MS).Hypothesis/Gap statement. The implementation of the Sepsityper method in the routine practice could represent a fundamental tool to achieve a prompt identification of the causative agent of bloodstream infections, and therefore accelerate the adoption of the proper antibiotic treatment.Methodology. In this study, the novel rapid workflow of the MALDI Biotypr Sepsityper kit (Bruker Daltonik GmbH, Germany) was evaluated using routine samples from a 2-year period (n=6918), and dedicated optimized protocols for the microbial groups that were more difficult to identify were developed. Moreover, the use of the residual bacterial pellet to perform susceptibility testing using different methods (commercial broth microdilution, disc diffusion, gradient diffusion) was investigated.Results. The rapid Sepsityper protocol allowed the identification of 5470/6338 (86.3 %) monomicrobial samples at species level, with very good performance for all of the clinically most significant pathogens (2510/2592 enterobacteria, 631/669 Staphylococcus aureus and 223/246 enterococci were identified). Streptococcus pneumoniae, Bacteroides fragilis and yeasts were the most troublesome to identify, but the application of specific optimized protocols significantly improved their rate of identification (from 14.7-71.5 %, 47.8-89.7 % and 37.1-89.5 %, respectively). Specificity was 100 % (no identification was made for the false-positive samples). Further, the residual pellet proved to be suitable to investigate susceptibility to antimicrobials, enabling us to simplify the workflow and shorten the time to report.Conclusion. The Rapid Sepsityper workflow proved to be a reliable sample preparation method for identification and susceptibility testing directly from positive blood cultures, providing novel approaches for accelerated diagnostics of bloodstream infections.


Assuntos
Bactérias/classificação , Técnicas Bacteriológicas/métodos , Sepse/microbiologia , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz/métodos , Bactérias/isolamento & purificação , Humanos , Sepse/diagnóstico , Manejo de Espécimes
5.
BMC Med Genet ; 21(1): 229, 2020 11 19.
Artigo em Inglês | MEDLINE | ID: mdl-33213396

RESUMO

BACKGROUND: Peroxisome biogenesis disorders (PBDs) are a group of metabolic diseases caused by dysfunction of peroxisomes. Different forms of PBDs are described; the most severe one is the Zellweger syndrome (ZS). We report on an unusual presentation of Zellweger syndrome manifesting in a newborn with severe and fulminant sepsis, causing death during the neonatal period. CASE PRESENTATION: A term male Caucasian neonate presented at birth with hypotonia and poor feeding associated with dysmorphic craniofacial features and skeletal abnormalities. Blood tests showed progressive leukopenia; ultrasounds revealed cerebral and renal abnormalities. He died on the fourth day of life because of an irreversible Gram-negative sepsis. Post-mortem tests on blood and urine samples showed biochemical alterations suggestive of ZS confirmed by genetic test. CONCLUSIONS: ZS is an early and severe forms of PBDs. Peroxisomes are known to be involved in lipid metabolism, but recent studies suggest their fundamental role in modulating immune response and inflammation. In case of clinical suspicion of ZS it is important to focus the attention on the prevention and management of infections that can rapidly progress to death.


Assuntos
ATPases Associadas a Diversas Atividades Celulares/genética , Infecções por Bactérias Gram-Negativas/genética , Mutação , Peroxissomos/imunologia , Sepse/genética , Síndrome de Zellweger/genética , ATPases Associadas a Diversas Atividades Celulares/deficiência , ATPases Associadas a Diversas Atividades Celulares/imunologia , Evolução Fatal , Expressão Gênica , Infecções por Bactérias Gram-Negativas/imunologia , Infecções por Bactérias Gram-Negativas/microbiologia , Infecções por Bactérias Gram-Negativas/patologia , Humanos , Imunidade Inata , Recém-Nascido , Masculino , Peroxissomos/microbiologia , Peroxissomos/patologia , Sepse/imunologia , Sepse/microbiologia , Sepse/patologia , Síndrome de Zellweger/imunologia , Síndrome de Zellweger/microbiologia , Síndrome de Zellweger/patologia
6.
Zhonghua Fu Chan Ke Za Zhi ; 55(11): 770-777, 2020 Nov 25.
Artigo em Chinês | MEDLINE | ID: mdl-33228348

RESUMO

Objective: To investigate the clinical features, etiology, and prognosis of sepsis during pregnancy and the postpartum period. Methods: Sixty-eight pregnant women with maternal sepsis treated in Peking Union Medical College Hospital from January 1997 to December 2019 were collected, and divided into obstetric infection group (30 cases) and non-obstetric infection group (38 cases) according to different infection sources. Clinical manifestations, types of infection sources, microbiological characteristics, treatment and outcomes were studied and analyzed. Results: (1) General conditions and clinical features: sepsis occurrence rate was 57% (39/68) and 43% (29/68) in prenatal and postpartum period, repectively. Statistical analysis showed that incidence of respiratory, renal, liver and coagulation dysfunction in non-obstetric infection group were significantly higher than those in obstetric infection group, and multiple organ dysfunction, cardiac arrest and blood lactate≥4 mmol/L were more common (all P<0.05). Sequential organ failure score in non-obstetric infection group was also significantly higher than that in obstetric infection group (P<0.05). (2) Types of infection sources and microbiological characteristics: the most common maternal sepsis was genital tract sepsis (37%, 25/68). Chorioamnionitis was the most common cause in obstetric sepsis (40%, 12/30), while intra-abdominal infection was the most common cause in non-obstetric sepsis (34%, 13/38). Thirty-seven patients (54%, 37/68) were diagnosed as bloodstream infection (BSI). Gram-negative bacteremia accounted for 70% (26/37), the most common pathogen of which was Escherichia coli. BSI was most commonly secondary to a genital tract infection (65%, 17/26). (3) Treatment: the ICU hospitalization rates and the utilization rate of mechanical ventilation and vasoactive agents in non-obstetric group were higher than those in obstetric group with significant differences (all P<0.05). Thirty-two patients (47%, 32/68) underwent surgery to remove the infection sources, including 5 cases of hysterectomy. (4) Prognosis: the case fatality rate of maternal sepsis was 19% (13/68), which was significantly higher in the non-obstetric infection group (29%,11/38) compared with the obstetric infection group (7%,2/30; P=0.020). The time from diagnosis of sepsis to termination of pregnancy was (5.5±8.6) days in prenatal women, and time in obstetric infection group [(1.9±2.2) days] was significantly less than that of non-obstetric infection group [(7.7±10.3) days, P=0.029]. Adverse pregnancy outcomes were higher in the first and second trimester (72%, 18/25) than in the third trimester (21%, 3/14), and the difference was statistically significant (P=0.002). Conclusions: Sepsis during pregnancy and the postpartum period is a potentially life-threatening disease. Pregnant women with non-obstetric sepsis have more complications, more serious condition and worse prognosis than those with obstetric infection. Timely detection of risk factors, early identification and active treatment are helpful to improve maternal and fetal prognosis.


Assuntos
Infecções por Bactérias Gram-Negativas/epidemiologia , Infecções por Bactérias Gram-Positivas/epidemiologia , Unidades de Terapia Intensiva/estatística & dados numéricos , Complicações Infecciosas na Gravidez/microbiologia , Sepse/microbiologia , Adulto , Estudos de Casos e Controles , China/epidemiologia , Corioamnionite/epidemiologia , Feminino , Bactérias Gram-Negativas , Bactérias Gram-Positivas , Hospitalização , Humanos , Período Pós-Parto , Gravidez , Complicações Infecciosas na Gravidez/diagnóstico , Complicações Infecciosas na Gravidez/epidemiologia , Resultado da Gravidez , Infecções do Sistema Genital/microbiologia , Infecções do Sistema Genital/mortalidade , Estudos Retrospectivos , Sepse/diagnóstico , Sepse/epidemiologia
7.
PLoS One ; 15(11): e0242533, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-33226995

RESUMO

PURPOSE: In the management of COVID-19, knowledge is lacking on the frequency of secondary bacterial infections and on how empirical antibiotic therapy should be used. In the present study, we aimed to compare blood culture (BC) results of a COVID-19 patient cohort with two cohorts of patients without detected COVID-19. METHODS: Using a retrospective cohort study design of patients subjected to BC in six tertiary care hospitals, SARS-CoV-2 positive patients from March 1 to April 30 in 2020 (COVID-19 group) were compared to patients without confirmed SARS-CoV-2 during the same period (control group-2020) and with patients sampled March 1 to April 30 in 2019 (control group-2019). The outcomes studied were proportion of BC positivity, clinically relevant growth, and contaminant growth. RESULTS: In total 15,103 patients and 17,865 BC episodes were studied. Clinically relevant growth was detected in 197/3,027 (6.5%) BC episodes in the COVID-19 group compared to 717/6,663 (10.8%) in control group-2020 (p<0.0001) and 850/8,175 (10.4%) in control group-2019 (p<0.0001). Contamination was present in 255/3,027 (8.4%) BC episodes in the COVID-19 group compared to 330/6,663 (5.0%) in control group-2020 (p<0.0001) and 354/8,175 (4.3%) in control group-2019 (p<0.0001). CONCLUSION: In COVID-19 patients, the prevalence of bloodstream bacterial infection is low and the contamination rate of BC is high. This knowledge should influence guidelines regarding blood culture sampling and empirical antibiotic therapy in COVID-19 patients.


Assuntos
Hemocultura/métodos , Coinfecção/epidemiologia , Sepse/epidemiologia , Adulto , Idoso , Idoso de 80 Anos ou mais , /virologia , Coinfecção/diagnóstico , Coinfecção/virologia , Reações Falso-Positivas , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Prevalência , Estudos Retrospectivos , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Sepse/diagnóstico , Sepse/microbiologia , Suécia/epidemiologia
8.
PLoS Genet ; 16(10): e1009065, 2020 10.
Artigo em Inglês | MEDLINE | ID: mdl-33112851

RESUMO

The genus Escherichia is composed of several species and cryptic clades, including E. coli, which behaves as a vertebrate gut commensal, but also as an opportunistic pathogen involved in both diarrheic and extra-intestinal diseases. To characterize the genetic determinants of extra-intestinal virulence within the genus, we carried out an unbiased genome-wide association study (GWAS) on 370 commensal, pathogenic and environmental strains representative of the Escherichia genus phylogenetic diversity and including E. albertii (n = 7), E. fergusonii (n = 5), Escherichia clades (n = 32) and E. coli (n = 326), tested in a mouse model of sepsis. We found that the presence of the high-pathogenicity island (HPI), a ~35 kbp gene island encoding the yersiniabactin siderophore, is highly associated with death in mice, surpassing other associated genetic factors also related to iron uptake, such as the aerobactin and the sitABCD operons. We confirmed the association in vivo by deleting key genes of the HPI in E. coli strains in two phylogenetic backgrounds. We then searched for correlations between virulence, iron capture systems and in vitro growth in a subset of E. coli strains (N = 186) previously phenotyped across growth conditions, including antibiotics and other chemical and physical stressors. We found that virulence and iron capture systems are positively correlated with growth in the presence of numerous antibiotics, probably due to co-selection of virulence and resistance. We also found negative correlations between virulence, iron uptake systems and growth in the presence of specific antibiotics (i.e. cefsulodin and tobramycin), which hints at potential "collateral sensitivities" associated with intrinsic virulence. This study points to the major role of iron capture systems in the extra-intestinal virulence of the genus Escherichia.


Assuntos
Infecções por Escherichia coli/genética , Escherichia coli/genética , Ferro/metabolismo , Sepse/genética , Sideróforos/genética , Animais , Modelos Animais de Doenças , Escherichia coli/classificação , Escherichia coli/patogenicidade , Infecções por Escherichia coli/microbiologia , Infecções por Escherichia coli/patologia , Variação Genética/genética , Estudo de Associação Genômica Ampla , Ilhas Genômicas/genética , Humanos , Camundongos , Fenóis/metabolismo , Filogenia , Sepse/microbiologia , Sepse/patologia , Sideróforos/metabolismo , Tiazóis/metabolismo , Virulência/genética
9.
Discov Med ; 29(157): 129-137, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-33002409

RESUMO

Sepsis is a life-threatening clinical condition demanding accurate and rapid diagnosis of the culprit pathogen, thereby to improve prognosis. Pathogen determination through blood culture is the gold standard for diagnosis but has limitations due to low sensitivity. Recently, circulating DNAs derived from pathogenic organisms were found in the plasma of patients with sepsis and were further proved to be more sensitive biomarkers for the diagnosis of the pathogen origin in sepsis. However, the fundamental molecular characteristics of circulating DNA in patients with sepsis remain unclear. Here, we used specific PCR and Sanger sequencing to verify the microbiology culture results via the corresponding plasma circulating DNA. We analyzed the composition and molecular characteristics of circulating DNA in septic patients using next-generation sequencing technology. We showed the presence of pathogen-derived circulating DNA in the plasma of patients with sepsis. The sizes of circulating DNA fragments derived from pathogenic bacteria showed a skewed unimodal distribution, while those derived from host cells showed a normal unimodal distribution. Lengths of fragments at peak concentration for both origins ranged from 150 bp to 200 bp, and reads mapping to pathogenic bacteria genome distributed uniformly on the reference. Our findings have improved our understanding of microbial circulating DNA in patients with sepsis as a potential methodology for the accurate diagnosis of sepsis, especially in light of an urgent need for such a diagnosis associated with the COVID-19 infection.


Assuntos
Infecções Bacterianas/microbiologia , Ácidos Nucleicos Livres/sangue , DNA Bacteriano/sangue , Sepse/microbiologia , Adulto , Idoso , Infecções Bacterianas/complicações , Infecções Bacterianas/diagnóstico , Betacoronavirus , Ácidos Nucleicos Livres/análise , Técnicas de Laboratório Clínico , Infecções por Coronavirus/diagnóstico , Técnicas de Cultura , DNA Bacteriano/análise , Feminino , Sequenciamento de Nucleotídeos em Larga Escala , Humanos , Masculino , Pessoa de Meia-Idade , Neoplasias/complicações , Pandemias , Pneumonia Viral , Reação em Cadeia da Polimerase , Sepse/complicações , Sepse/diagnóstico , Análise de Sequência de DNA
10.
Am J Med Sci ; 360(6): 650-655, 2020 12.
Artigo em Inglês | MEDLINE | ID: mdl-32868035

RESUMO

BACKGROUND: Inappropriate antibiotic therapy in sepsis is associated with poor outcomes, clinicians often provide routine coverage for multidrug resistant (MDR) bacteria. However, these regimens may contribute to problems related to antibiotic overuse. To understand the incidence and related factors of multidrug resistant bacterial infections in ED patients with sepsis, we examined how often patients with sepsis in our emergency department had MDR infections. We also explored risk factors for, and outcomes from, MDR bacterial infections. METHODS: We reviewed records of patients presenting to our emergency department (ED) meeting criteria for severe sepsis or septic shock from March 2012 to July 2013. Patient demographics, comorbidities, preadmission location, and APACHE II scores were analyzed, as were clinical outcomes. RESULTS: A total of 191 episodes were examined. 108 (57%) cases were culture-positive, and of these, 23 (12.0%) had an MDR pathogen recovered. Among patients with positive cultures, MDR patients used mechanical ventilation more often 29% vs. 52% (P = 0.03) and had longer mean ICU and hospital length of stays: 4.0 vs 9.3 (P < 0.08) and 10.6 vs 20.8 (P = 0.01), respectively. We did not identify statistically significant predictors of MDR infection. CONCLUSIONS: The overall number of infections due to MDR bacteria was low, and MDR gram-negative infections were uncommon. The use of multiple empiric antibiotics for resistant gram-negative infections in the ED may be beneficial in only a small number of cases. Additionally, empiric coverage for vancomycin-resistant enterococci may need to be considered more often. Larger studies may help further elucidate the rates of MDR infections in ED patients, and identify specific risk factors to rationally guide empiric antibiotic treatment.


Assuntos
Infecções Bacterianas/epidemiologia , Farmacorresistência Bacteriana Múltipla , Serviço Hospitalar de Emergência/estatística & dados numéricos , Sepse/epidemiologia , Adulto , Idoso , Idoso de 80 Anos ou mais , Antibacterianos/uso terapêutico , Bactérias/efeitos dos fármacos , Infecções Bacterianas/microbiologia , Feminino , Humanos , Incidência , Masculino , Pessoa de Meia-Idade , Philadelphia/epidemiologia , Sepse/microbiologia , Choque Séptico/epidemiologia , Choque Séptico/microbiologia
11.
PLoS One ; 15(9): e0239147, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32960928

RESUMO

Ever decreasing efficiency of antibiotic treatment due to growing antibiotic resistance of pathogenic bacteria is a critical issue in clinical practice. The two generally accepted major approaches to this problem are the search for new antibiotics and the development of antibiotic adjuvants to enhance the antimicrobial activity of known compounds. It was therefore the aim of the present study to test whether alkylresorcinols, a class of phenolic lipids, can be used as adjuvants to potentiate the effect of various classes of antibiotics. Alkylresorcinols were combined with 12 clinically used antibiotics. Growth-inhibiting activity against a broad range of pro- and eukaryotic microorganisms was determined. Test organisms did comprise 10 bacterial and 2 fungal collection strains, including E. coli and S. aureus, and clinical isolates of K. pneumoniae. The highest adjuvant activity was observed in the case of 4-hexylresorcinol (4-HR), a natural compound found in plants with antimicrobial activity. 50% of the minimal inhibitory concentration (MIC) of 4-HR caused an up to 50-fold decrease in the MIC of antibiotics of various classes. Application of 4-HR as an adjuvant revealed its efficiency against germination of bacterial dormant forms (spores) and prevented formation of antibiotic-tolerant persister cells. Using an in vivo mouse model of K. pneumoniae-induced sepsis, we could demonstrate that the combination of 4-HR and polymyxin was highly effective. 75% of animals were free of infection after treatment as compared to none of the animals receiving the antibiotic alone. We conclude that alkylresorcinols such as 4-HR can be used as an adjuvant to increase the efficiency of several known antibiotics. We suggest that by this approach the risk for development of genetically determined antibiotic resistance can be minimized due to the multimodal mode of action of 4-HR.


Assuntos
Adjuvantes Farmacêuticos/farmacologia , Antibacterianos/farmacologia , Hexilresorcinol/farmacologia , Infecções por Klebsiella/tratamento farmacológico , Sepse/tratamento farmacológico , Adjuvantes Farmacêuticos/uso terapêutico , Animais , Antibacterianos/uso terapêutico , Modelos Animais de Doenças , Farmacorresistência Bacteriana Múltipla , Sinergismo Farmacológico , Quimioterapia Combinada/métodos , Escherichia coli/efeitos dos fármacos , Feminino , Hexilresorcinol/uso terapêutico , Humanos , Infecções por Klebsiella/microbiologia , Klebsiella pneumoniae/efeitos dos fármacos , Klebsiella pneumoniae/isolamento & purificação , Camundongos , Testes de Sensibilidade Microbiana , Polimixinas/farmacologia , Polimixinas/uso terapêutico , Sepse/microbiologia , Staphylococcus aureus/efeitos dos fármacos
12.
Nat Commun ; 11(1): 4774, 2020 09 22.
Artigo em Inglês | MEDLINE | ID: mdl-32963224

RESUMO

Detection of microbial nucleic acids in body fluids has become the preferred method for rapid diagnosis of many infectious diseases. However, culture-based diagnostics that are time-consuming remain the gold standard approach in certain cases, such as sepsis. New culture-free methods are urgently needed. Here, we describe Single MOLecule Tethering or SMOLT, an amplification-free and purification-free molecular assay that can detect microorganisms in body fluids with high sensitivity without the need of culturing. The signal of SMOLT is generated by the displacement of micron-size beads tethered by DNA probes that are between 1 and 7 microns long. The molecular extension of thousands of DNA probes is determined with sub-micron precision using a robust and rapid optical approach. We demonstrate that SMOLT can detect nucleic acids directly in blood, urine and sputum at sub-femtomolar concentrations, and microorganisms in blood at 1 CFU mL-1 (colony forming unit per milliliter) threefold faster, with higher multiplexing capacity and with a more straight-forward protocol than amplified methodologies. SMOLT's clinical utility is further demonstrated by developing a multiplex assay for simultaneous detection of sepsis-causing Candida species directly in whole blood.


Assuntos
Líquidos Corporais/química , Técnicas de Diagnóstico Molecular/métodos , Ácidos Nucleicos/isolamento & purificação , Sepse/diagnóstico , Candida/genética , Candida/isolamento & purificação , Candidíase/diagnóstico , Contagem de Colônia Microbiana , Doenças Transmissíveis/diagnóstico , DNA/isolamento & purificação , Humanos , Ácidos Nucleicos/sangue , Ácidos Nucleicos/urina , Reação em Cadeia da Polimerase/métodos , RNA/isolamento & purificação , Sensibilidade e Especificidade , Sepse/microbiologia , Infecções Estafilocócicas/diagnóstico , Staphylococcus aureus/genética , Staphylococcus aureus/isolamento & purificação , Urina
13.
PLoS One ; 15(9): e0239069, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32915919

RESUMO

The diagnosis of leptospirosis remains a challenge due to its non-specific symptoms and the biphasic nature of the illness. A comprehensive diagnosis that includes both molecular (polymerase chain reaction (PCR)) and serology is vital for early detection of leptospirosis and to avoid misdiagnosis. However, not all samples could be subjected to both tests (serology and molecular) due to budget limitation, infrastructure, and technical expertise at least in resource-limited countries. We evaluated the usefulness of testing the clinically suspected leptospirosis cases with both techniques on all samples collected from the patients on the day of admission. Among the 165 patient's blood/serum samples tested (from three hospitals in Central Malaysia), 43 (26%) showed positivity by microscopic agglutination test (MAT), 63 (38%) by PCR, while 14 (8%) were positive by both MAT and PCR. For PCR, we tested two molecular targets (lipL32 by qPCR and 16S rDNA or rrs by nested PCR) and detected lipL32 in 47 (29%) and rrs gene in 63 (38%) patients. The use of more than one target gene for PCR increased the detection rates. Hence, a highly sensitive multiplex PCR targeting more than one diagnostic marker is recommended for the early detection of Leptospira in suspected patients. When the frequencies for positivity detected either by MAT or PCR combined, leptospirosis was diagnosed in a total of 92 (56%) patients, a higher frequency compared to when samples were only tested by a single method (MAT or PCR). The results from this study suggest the inclusion of both serology and molecular methods for every first sample irrespective of the days post-onset of symptoms (DPO) collected from patients for early diagnosis of leptospirosis.


Assuntos
Testes de Aglutinação , Leptospira/isolamento & purificação , Leptospirose/diagnóstico , Reação em Cadeia da Polimerase , Sepse/diagnóstico , DNA Bacteriano/isolamento & purificação , Erros de Diagnóstico/prevenção & controle , Diagnóstico Precoce , Estudos de Viabilidade , Humanos , Leptospira/genética , Leptospira/imunologia , Leptospirose/sangue , Leptospirose/imunologia , Leptospirose/microbiologia , Malásia , RNA Ribossômico 16S/genética , Sensibilidade e Especificidade , Sepse/sangue , Sepse/imunologia , Sepse/microbiologia , Fatores de Tempo
14.
BMC Infect Dis ; 20(1): 651, 2020 Sep 04.
Artigo em Inglês | MEDLINE | ID: mdl-32887563

RESUMO

BACKGROUND: Risk factors related to mortality due to invasive pneumococcal disease (IPD) have been unveiled previously, but early clinical manifestations of IPD based on prognosis remain uncovered. METHODS: The demographic characteristics, clinical features, serotype, antibiotic susceptibility, and outcomes of 97 hospitalized children with laboratory-confirmed IPD from Suzhou, China, were collected and analyzed retrospectively. RESULTS: The median age was 0.69 (0.49-1.55) years in the non-survivor group compared with 2.39 (0.90-3.81) years in the survivor group. The mortality of 97 children with laboratory-confirmed IPD was 17.5% (17/97), and 53.6% of them were aged less than 2 years. Pathogens were mainly from the blood and cerebrospinal fluid, and sepsis was the most frequent type. Statistically significant differences were found in hyperpyrexia, vomiting, anorexia, lethargy, poor perfusion of extremities, Hb level, and Plt count between the nonsurvival and survival groups. Further, the multivariate regression analysis showed that early signs, including hyperpyrexia, vomiting, anorexia, lethargy, and poor perfusion of extremities, were independent risk factors for the in-hospital mortality of children with laboratory-confirmed IPD. The mortality was also associated with antimicrobial sensitivity in pneumococcal isolates. The microbes in 1/17 (5.9%) children who were prescribed an antibiotic showed antimicrobial sensitivity in the nonsurvival group, compared with 21/80 (26.3%) children who survived. The most common serotypes identified were 6B (35.3%, 6/17), 14 (23.5%, 4/17), 19F (23.5%, 4/17), 19A (5.9%, 1/17), 23F (5.9%, 1/17), and 20 (5.9%, 1/17) in the nonsurvival group. The coverage of IPD serotypes of the 7-valent pneumococcal conjugate vaccine (PCV7) was 88.2% (15/17), while that of the 13-valent S. pneumoniae vaccine (PCV13) was 94.1% (16/17) of the coverage in the nonsurvival group. CONCLUSIONS: Recurrent hyperpyrexia, vomiting, anorexia, lethargy, and poor perfusion of extremities in the early stage were independent predictors for the in-hospital mortality of children with laboratory-confirmed IPD. Appropriate use of antibiotics and PCV immunization were the keys to improve the outcome of IPD.


Assuntos
Infecções Pneumocócicas/microbiologia , Infecções Pneumocócicas/mortalidade , Antibacterianos/farmacologia , Antibacterianos/uso terapêutico , Pré-Escolar , Feminino , Vacina Pneumocócica Conjugada Heptavalente/uso terapêutico , Mortalidade Hospitalar , Humanos , Lactente , Masculino , Testes de Sensibilidade Microbiana , Infecções Pneumocócicas/tratamento farmacológico , Infecções Pneumocócicas/prevenção & controle , Vacinas Pneumocócicas/uso terapêutico , Prognóstico , Estudos Retrospectivos , Fatores de Risco , Sepse/microbiologia , Streptococcus pneumoniae/isolamento & purificação , Cobertura Vacinal
15.
Mayo Clin Proc ; 95(11): 2509-2524, 2020 11.
Artigo em Inglês | MEDLINE | ID: mdl-32829901

RESUMO

Bloodstream infections are a leading cause of morbidity and mortality. Molecular rapid diagnostic tests (mRDTs) are transforming care for patients with bloodstream infection by providing the opportunity to dramatically shorten times to effective therapy and speeding de-escalation of overly broad empiric therapy. However, because of the novelty of these tests which provide information regarding microbial identification and whether specific antibiotic-resistance mutations were detected, many front-line providers still delay final decisions until complete phenotypic susceptibility results are available several days later. Thus the benefits of mRDTs have been largely limited to circumstances where antimicrobial stewardship programs closely monitor these tests and intervene as soon as the results are available. We searched PubMed and Google Scholar for articles published from 1980 to 2019 using the terms antibiotic, antifungal, bacteremia, bloodstream infection, candidemia, candidiasis, children, coagulase negative staphylococcus, consultation, contamination, costs, echocardiogram, endocarditis, enterobacteriaceae, enterococcus, Gram-negative, guidelines, IDSA, immunocompromised, infectious disease or ID, lumbar puncture, meningitis, mortality, MRSA, MSSA, neonatal, outcomes, pediatric, pneumococcal, polymicrobial, Pseudomonas, rapid diagnostic testing, resistance, risk factors, sepsis, Staphylococcus aureus, stewardship, streptococcus, and treatment. With the data from this search, we aim to provide guidance to front-line providers regarding the interpretation and immediate actions to be taken in response to the identification of common bloodstream pathogens by mRDTs. In addition to antimicrobial therapy, additional diagnostic or therapeutic interventions are recommended for particular organisms and clinical settings to either determine the extent of infection or control its source. Pediatric perspectives are offered for those bloodstream pathogens for which management differs from that in adults.


Assuntos
Sepse/diagnóstico , Bacteriemia/diagnóstico , Bacteriemia/microbiologia , Candidíase Invasiva/diagnóstico , Candidíase Invasiva/microbiologia , Humanos , Infecções Meningocócicas/diagnóstico , Infecções Meningocócicas/microbiologia , Sepse/microbiologia , Infecções Estafilocócicas/diagnóstico , Infecções Estafilocócicas/microbiologia , Infecções Estreptocócicas/diagnóstico , Infecções Estreptocócicas/microbiologia , Fatores de Tempo
16.
Mil Med Res ; 7(1): 36, 2020 08 04.
Artigo em Inglês | MEDLINE | ID: mdl-32753048

RESUMO

Sepsis is a life-threatening condition that is characterized by multiple organ dysfunction due to abnormal host response to various pathogens, like bacteria, fungi and virus. The differences between viral and bacterial sepsis are indeed of great significance to deepen the understanding of the pathogenesis of sepsis, especially under pandemics of SARS-CoV-2 infection.


Assuntos
Infecções Bacterianas/complicações , Betacoronavirus , Infecções por Coronavirus/complicações , Pneumonia Viral/complicações , Sepse/microbiologia , Idoso , Técnicas de Laboratório Clínico/métodos , Infecções por Coronavirus/diagnóstico , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Pandemias
17.
PLoS One ; 15(8): e0237871, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32817720

RESUMO

Streptococcus pneumoniae is a common cause of infectious diseases such as pneumonia and sepsis. Its colonization is thought to be the first step in the development of invasive pneumococcal diseases. This study aimed to investigate pneumococcal colonization patterns in early childhood. A longitudinal birth cohort study was conducted for investigating nasopharyngeal colonized pneumococci at 1, 6, 12, 18, 24, and 36 months of age, particularly focusing on the serotype distribution and antimicrobial susceptibilities. Pneumococcal conjugate vaccine (PCV) effect on nasopharyngeal colonization was also assessed. During 2013-2017, 855 infants were enrolled and a total of 107 isolates were recovered from 95 infants during the first three years of life. In this period, the prevalence of pneumococcal colonization increased, with values ranging from 0.2% (2/834) at 1 month of age to 5.9% (19/323) at 36 months of age. The investigation of serotype revealed that 81.1% (73/90) belonged to the non-PCV13 serotypes-23A, 15A, 15C, and 15B. Moreover, PCV13 serotypes significantly decreased during 2014-2015, when routine PCV13 vaccination was initiated in Taiwan. PCV13 introduction may lead to the reduction in the rates of pneumococcal isolates resistant (R) to penicillin. Under conditional PCV13 vaccination, pneumococcal isolates primarily belonged to non-PCV13 serotypes. This non-PCV13 serotype replacement exhibited lower rates of penicillin R isolates, suggesting that PCV13 administration may reduce the antibiotic-nonsusceptible pneumococcal disease burden and antibiotic use.


Assuntos
Doenças Nasofaríngeas/tratamento farmacológico , Nasofaringe/efeitos dos fármacos , Infecções Pneumocócicas/tratamento farmacológico , Streptococcus pneumoniae/efeitos dos fármacos , Criança , Pré-Escolar , Estudos de Coortes , Feminino , Humanos , Lactente , Masculino , Doenças Nasofaríngeas/imunologia , Doenças Nasofaríngeas/microbiologia , Doenças Nasofaríngeas/patologia , Nasofaringe/microbiologia , Penicilinas/administração & dosagem , Infecções Pneumocócicas/imunologia , Infecções Pneumocócicas/microbiologia , Infecções Pneumocócicas/patologia , Vacinas Pneumocócicas/administração & dosagem , Pneumonia/microbiologia , Pneumonia/prevenção & controle , Sepse/microbiologia , Sepse/prevenção & controle , Sorogrupo , Streptococcus pneumoniae/genética , Streptococcus pneumoniae/patogenicidade , Taiwan , Vacinas Conjugadas/administração & dosagem
18.
Sci Rep ; 10(1): 12974, 2020 07 31.
Artigo em Inglês | MEDLINE | ID: mdl-32737397

RESUMO

Extended early antibiotic exposure in the neonatal intensive care unit is associated with an increased risk for the development of late-onset sepsis (LOS). However, few studies have examined the mechanisms involved. We sought to determine how the neonatal microbiome and intestinal immune response is altered by transient early empiric antibiotic exposure at birth. Neonatal mice were transiently exposed to broad-spectrum antibiotics from birth for either 3- (SE) or 7-days (LE) and were examined at 14-days-old. We found that mice exposed to either SE or LE showed persistent expansion of Proteobacteria (2 log difference, P < 0.01). Further, LE mice demonstrated baseline translocation of E. coli into the liver and spleen and were more susceptible K. pneumoniae-induced sepsis. LE mice had a significant and persistent decrease in type 3 innate lymphoid cells (ILC3) in the lamina propria. Reconstitution of the microbiome with mature microbiota by gavage in LE mice following antibiotic exposure resulted in an increase in ILC3 and partial rescue from LOS. We conclude that prolonged exposure to broad spectrum antibiotics in the neonatal period is associated with persistent alteration of the microbiome and innate immune response resulting in increased susceptibility to infection that may be partially rescued by microbiome reconstitution.


Assuntos
Antibacterianos/efeitos adversos , Escherichia coli/imunologia , Microbioma Gastrointestinal/imunologia , Imunidade Inata/efeitos dos fármacos , Klebsiella pneumoniae/imunologia , Linfócitos/imunologia , Sepse , Animais , Animais Recém-Nascidos , Antibacterianos/farmacologia , Translocação Bacteriana/efeitos dos fármacos , Translocação Bacteriana/imunologia , Suscetibilidade a Doenças , Infecções por Escherichia coli/induzido quimicamente , Infecções por Escherichia coli/imunologia , Infecções por Escherichia coli/microbiologia , Infecções por Escherichia coli/patologia , Feminino , Microbioma Gastrointestinal/efeitos dos fármacos , Infecções por Klebsiella/induzido quimicamente , Infecções por Klebsiella/imunologia , Infecções por Klebsiella/microbiologia , Infecções por Klebsiella/patologia , Linfócitos/patologia , Masculino , Camundongos , Sepse/induzido quimicamente , Sepse/imunologia , Sepse/microbiologia , Sepse/patologia
19.
Am J Physiol Heart Circ Physiol ; 319(3): H705-H721, 2020 09 01.
Artigo em Inglês | MEDLINE | ID: mdl-32762560

RESUMO

Myeloperoxidase (MPO)-derived hypochlorous (HOCl) reacts with membrane plasmalogens to yield α-chlorofatty aldehydes such as 2-chlorofatty aldehyde (2-ClFALD) and its metabolite 2-chlorofatty acid (2-ClFA). Recent studies showed that 2-ClFALD and 2-ClFA serve as mediators of the inflammatory responses to sepsis by as yet unknown mechanisms. Since no scavenger for chlorinated lipids is available and on the basis of the well-established role of the MPO/HOCl/chlorinated lipid axis in inflammatory responses, we hypothesized that treatment with MPO inhibitors (N-acetyl lysyltyrosylcysteine amide or 4-aminobenzoic acid hydrazide) would inhibit inflammation and proinflammatory mediator expression induced by cecal ligation and puncture (CLP). We used intravital microscopy to quantify in vivo inflammatory responses in Sham and CLP rats with or without MPO inhibition. Small intestines, mesenteries, and lungs were collected to assess changes in MPO-positive staining and lung injury, respectively, as well as free 2-ClFA and proinflammatory mediators levels. CLP caused neutrophil infiltration, 2-ClFA generation, acute lung injury, leukocyte-/platelet-endothelium interactions, mast cell activation (MCA), plasminogen activator inhibitor-1 (PAI-1) production, and the expression of several cytokines, chemokines, and vascular endothelial growth factor, changes that were reduced by MPO inhibition. Pretreatment with a PAI-1 inhibitor or MC stabilizer prevented CLP-induced leukocyte-endothelium interactions and MCA, and abrogated exogenous 2-ClFALD-induced inflammatory responses. Thus, we provide evidence that MPO instigates these inflammatory changes in CLP and that chlorinated lipids may serve as a mechanistic link between the enzymatic activity of MPO and PAI-1- and mast cell-dependent adhesive interactions, providing a rationale for new therapeutic interventions in sepsis.NEW & NOTEWORTHY Using two distinct myeloperoxidase (MPO) inhibitors, we show for the first time that MPO plays an important role in producing increases in free 2-chlorofatty aldehyde (2-ClFALD)-a powerful proinflammatory chlorinated lipid in plasma and intestine-a number of cytokines and other inflammatory mediators, leukocyte and platelet rolling and adhesion in postcapillary venules, and lung injury in a cecal ligation and puncture model of sepsis. In addition, the use of a plasminogen activator inhibitor-1 (PAI-1) inhibitor or a mast cell stabilizer prevented inflammatory responses in CLP-induced sepsis. PAI-1 inhibition also prevented the proinflammatory responses to exogenous 2-ClFALD superfusion. Thus, our study provides some of the first evidence that MPO-derived free 2-ClFA plays an important role in CLP-induced sepsis by a PAI-1- and mast cell-dependent mechanism.


Assuntos
Ceco/microbiologia , Ácidos Graxos/metabolismo , Ácido Hipocloroso/metabolismo , Mediadores da Inflamação/metabolismo , Inflamação/enzimologia , Peroxidase/metabolismo , Sepse/enzimologia , Aldeídos/metabolismo , Animais , Anti-Inflamatórios/farmacologia , Ceco/cirurgia , Citocinas/metabolismo , Modelos Animais de Doenças , Inibidores Enzimáticos/farmacologia , Inflamação/imunologia , Inflamação/microbiologia , Inflamação/prevenção & controle , Mediadores da Inflamação/antagonistas & inibidores , Intestino Delgado/enzimologia , Intestino Delgado/imunologia , Ligadura , Pulmão/enzimologia , Pulmão/imunologia , Mastócitos/enzimologia , Mastócitos/imunologia , Mesentério/enzimologia , Mesentério/imunologia , Peroxidase/antagonistas & inibidores , Inibidor 1 de Ativador de Plasminogênio/metabolismo , Punções , Ratos Sprague-Dawley , Sepse/imunologia , Sepse/microbiologia , Sepse/prevenção & controle , Transdução de Sinais
20.
PLoS Negl Trop Dis ; 14(8): e0008381, 2020 08.
Artigo em Inglês | MEDLINE | ID: mdl-32804954

RESUMO

The world's most consequential pathogens occur in regions with the fewest diagnostic resources, leaving the true burden of these diseases largely under-represented. During a prospective observational study of sepsis in Takeo Province Cambodia, we enrolled 200 patients over an 18-month period. By coupling traditional diagnostic methods such as culture, serology, and PCR to Next Generation Sequencing (NGS) and advanced statistical analyses, we successfully identified a pathogenic cause in 46.5% of our cohort. In all, we detected 25 infectious agents in 93 patients, including severe threat pathogens such as Burkholderia pseudomallei and viral pathogens such as Dengue virus. Approximately half of our cohort remained undiagnosed; however, an independent panel of clinical adjudicators determined that 81% of those patients had infectious causes of their hospitalization, further underscoring the difficulty of diagnosing severe infections in resource-limited settings. We garnered greater insight as to the clinical features of severe infection in Cambodia through analysis of a robust set of clinical data.


Assuntos
Sepse/epidemiologia , Sepse/etiologia , Sepse/microbiologia , Adulto , Idoso , Idoso de 80 Anos ou mais , Bactérias/classificação , Infecções Bacterianas/diagnóstico , Infecções Bacterianas/epidemiologia , Camboja/epidemiologia , Feminino , Sequenciamento de Nucleotídeos em Larga Escala , Humanos , Masculino , Pessoa de Meia-Idade , Reação em Cadeia da Polimerase , Estudos Prospectivos , Sepse/virologia , Análise de Sequência de RNA , Testes Sorológicos , Viroses/diagnóstico , Viroses/epidemiologia , Vírus/classificação
SELEÇÃO DE REFERÊNCIAS
DETALHE DA PESQUISA