RESUMEN
Spinal muscular atrophy (SMA) is a motor neuron disease associated with progressive muscle weakness, ventilatory failure, and reduced survival. Onasemnogene abeparvovec is the first gene replacement therapy (GT) approved to treat this condition. An observational retrospective study was conducted to assess adverse events and efficacy of GT in SMA patients. Forty-one patients with SMA (58.5% females and 80.1% SMA type 1) were included. The mean age at GT dosing was 18 (±6.4) months. Thirty-six patients (87.8%) were under previous treatment with nusinersen, and 10 (24.4%) continued nusinersen after GT. Mean CHOP-INTEND increased 13 points after 6 months and this finding did not differ between groups according to nusinersen maintenance after GT (p = 0.949). Among SMA type 1 patients, 14 (46.6%) reached the ability to sit alone. Liver transaminases elevation at least two times higher than the upper limit of normal value occurred in 29 (70.7%) patients. Thrombocytopenia occurred in 13 (31.7%) patients, and one presented thrombotic microangiopathy. Older age (>2 years) was associated with more prolonged use of corticosteroids (p = 0.021). GT is effective in SMA patients, combined nusinersen after GT did not appear to add gain in motor function and older age is associated with prolonged corticosteroid use.
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Terapia Genética , Oligonucleótidos , Humanos , Femenino , Masculino , Terapia Genética/métodos , Estudios Retrospectivos , Oligonucleótidos/uso terapéutico , Lactante , Brasil , Atrofia Muscular Espinal/terapia , Atrofia Muscular Espinal/genética , Resultado del Tratamiento , Preescolar , Atrofias Musculares Espinales de la Infancia/genética , Atrofias Musculares Espinales de la Infancia/terapia , Productos Biológicos/uso terapéutico , Proteínas Recombinantes de FusiónRESUMEN
Primary coenzyme Q10 (CoQ10) deficiency is a group of inborn errors of metabolism caused by defects in CoQ10 biosynthesis. Biallelic pathogenic variants in COQ7, encoding mitochondrial 5-demethoxyubiquinone hydroxylase, have been reported in nine patients from seven families. We identified five new patients with COQ7-related primary CoQ10 deficiency, performed clinical assessment of the patients, and studied the functional effects of current and previously reported COQ7 variants and potential treatment options. The main clinical features included a neonatal-onset presentation with severe neuromuscular, cardiorespiratory and renal involvement and a late-onset disease presenting with progressive neuropathy, lower extremity weakness, abnormal gait, and variable developmental delay. Baker's yeast orthologue of COQ7, CAT5, is required for growth on oxidative carbon sources and cat5Δ strain demonstrates oxidative growth defect. Expression of wild-type CAT5 could completely rescue the defect; however, yeast CAT5 harboring equivalent human pathogenic variants could not. Interestingly, cat5Δ yeast harboring p.Arg57Gln (equivalent to human p.Arg54Gln), p.Arg112Trp (equivalent to p.Arg107Trp), p.Ile69Asn (equivalent to p.Ile66Asn) and combination of p.Lys108Met and p.Leu116Pro (equivalent to the complex allele p.[Thr103Met;Leu111Pro]) partially rescued the growth defects, indicating these variants are hypomorphic alleles. Supplementation with 2,4 dihydroxybenzoic acid (2,4-diHB) rescued the growth defect of both the leaky and severe mutants. Overexpression of COQ8 and 2,4-diHB supplementation synergistically restored oxidative growth and respiratory defect. Overall, we define two distinct disease presentations of COQ7-related disorder with emerging genotype-phenotype correlation and validate the use of the yeast model for functional studies of COQ7 variants.
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Enfermedades Mitocondriales , Ubiquinona , Humanos , Recién Nacido , Mitocondrias/metabolismo , Enfermedades Mitocondriales/metabolismo , Ubiquinona/metabolismoRESUMEN
Aim: This study aimed to evaluate the effect of physiological testosterone replacement on male aged rats with orchiectomy-induced osteoporosis in advanced stage.Methods: Thirty male rats (Rattus norvegicus albinus, Holtzman lineage) were randomly distributed into 3 groups (n = 10): 1-sham, 2-orchiectomy (OCX), 3-OCX + testosterone replacement (OCX + T). On day 0, a sham or orchiectomy surgery was performed according to the groups. Thirty and sixty days after surgeries, the animals from OCX + T group received testosterone intramuscularly, and the rats in all groups were euthanized on day 77. The femurs were removed for micro-CT scanning and biomechanical test.Results: Orchiectomy resulted in a marked trabecular bone damage (p < 0.05), which was not reversed with testosterone treatment (OCX + T group). The femoral strength was lower in orchiectomized animals (p < 0.05), while the bone strength in OCX + T group was similar to that observed in the sham animals (p > 0.05) and correlated to this parameter the deformation of rupture was smaller in OCX + T group.Conclusion: In conclusion, testosterone depletion induced by orchiectomy established an osteoporotic environment, mainly affecting the trabecular bone. Moreover, even though testosterone treatment did not enhance these variables, the hormonal replacement improved the femoral fracture strength and promoted beneficial effects on the biomechanical parameters compromised by castration in femoral bone.
Asunto(s)
Orquiectomía , Osteoporosis , Animales , Fémur , Masculino , Osteoporosis/tratamiento farmacológico , Osteoporosis/etiología , Proyectos Piloto , Ratas , TestosteronaRESUMEN
OBJECTIVES: NDM carbapenemases have spread worldwide. However, little information exists about the impact of NDM-producing Enterobacteriaceae in Spain. By WGS, we sought to elucidate the population structure of NDM-like-producing Klebsiella pneumoniae and Escherichia coli in Spain and to determine the plasmids harbouring blaNDM-like genes. METHODS: High-resolution SNP typing, core-genome MLST and plasmid reconstruction (PlasmidID) were performed on 59 NDM-like-producing K. pneumoniae and 8 NDM-like-producing E. coli isolated over an 8 year period in Spain. RESULTS: Five major epidemic clones of NDM-producing K. pneumoniae caused five important nationwide outbreaks: ST437/NDM-7, ST437/NDM-1, ST147/NDM-1, ST11/NDM-1 and ST101/NDM-1; in contrast, the spread of NDM-producing E. coli was polyclonal. Three blaNDM types were identified: blaNDM-1, 61.2%; blaNDM-7, 32.8%; and blaNDM-5, 6%. Five K. pneumoniae isolates co-produced other carbapenemases (three blaOXA-48 and two blaVIM-1). The average number of acquired resistance genes was higher in K. pneumoniae than in E. coli. The plasmids encoding blaNDM-like genes belonged to IncFII, IncFIB, IncX3, IncR, IncN and IncC types, of which IncF, IncR and IncC were associated with MDR. The genetic surroundings of blaNDM-like genes showed a highly variable region upstream of ISAba125. CONCLUSIONS: In recent years NDM-producing K. pneumoniae and E. coli have emerged in Spain; the spread of a few high-risk K. pneumoniae clones such as ST437/NDM-7, ST437/NDM-1, ST147/NDM-1, ST11/NDM-1 and ST101/NDM-1 have caused several interregional outbreaks. In contrast, the spread of NDM-producing E. coli has been polyclonal. Plasmid types IncFII, IncFIB, IncX3, IncR, IncN and IncC carried blaNDM, and the same IncX3 plasmid was detected in K. pneumoniae and E. coli.
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Escherichia coli/genética , Klebsiella pneumoniae/genética , Filogenia , Plásmidos/genética , beta-Lactamasas/genética , Antibacterianos/farmacología , Técnicas de Tipificación Bacteriana , Farmacorresistencia Bacteriana Múltiple , Infecciones por Enterobacteriaceae/microbiología , Escherichia coli/clasificación , Escherichia coli/enzimología , Genoma Bacteriano , Humanos , Klebsiella pneumoniae/clasificación , Klebsiella pneumoniae/enzimología , Pruebas de Sensibilidad Microbiana , Tipificación de Secuencias Multilocus , Polimorfismo de Nucleótido Simple , España , Virulencia/genética , Secuenciación Completa del GenomaRESUMEN
Objective: to investigate the association between caries incidence in children and the number of clinical preventive visits and follow-up period (intervals). Study Design: a 30-year cohort composed of the dental records of 600 children who were 12-23 months old at their first dental appointment and who were followed up to 12 years of age (200 followed since 1981, Group 1; 200 followed since 1991, Group 2; and 200 followed since 2001, Group 3) was analytically and quantitatively evaluated. Random sample calculation was performed with 95% confidence, a maximum error of 2.95% and a ratio of 50%. Results: association was found between caries incidence and the number of dental visits and consultation intervals. Children who attended a smaller number of preventive visits had a higher incidence of the disease. Children who had an interval between returns greater than 12 months had an 18.7-times greater caries risk compared to children who had intervals no longer than 8 months between return visits. Conclusions: preventive consultations with an average interval of up to 8 months seemed to increase family adherence to preventive procedures and acted as a protective factor against caries incidence in children.
Asunto(s)
Caries Dental , Clínicas Odontológicas , Niño , Preescolar , Estudios de Cohortes , Humanos , Incidencia , LactanteRESUMEN
Forkhead box-O (FOXO) transcription factors have a fundamental role in the development and differentiation of immune cells. FOXO1 and FOXO3 are FOXO members that are structurally similar and bind to the same conserved consensus DNA sequences to induce transcription. FOXO1 has been studied in detail in the activation of dendritic cells (DCs), where it plays an important role through the regulation of target genes such as ICAM-1, CCR7, and the integrin αvß3. FOXO1 is activated by bacteria challenge in DCs and promotes DC bacterial phagocytosis, migration, homing to lymph nodes, DC stimulation of CD4+ T cells and resting B cells, and antibody production. Deletion of FOXO1 in DCs enhances susceptibility to bacteria-induced periodontal disease. FOXO1 and FOXO3 maintain naive T cell quiescence and survival. FOXO1 and FOXO3 enhance the formation of regulatory T cells and inhibit the formation of T-helper 1 (Th1) and Th17 cells. FOXO1 promotes differentiation, proliferation, survival, immunoglobulin gene rearrangement, and class switching in B cells, but FOXO3 has little effect. Both FOXO1 and FOXO3 are important in the maintenance of hematopoietic stem cells by protecting them from oxidative stress. This review examines FOXO1/FOXO3 in the adaptive immune response, key target genes, and FOXO inhibition by the phosphoinositide 3-kinase/AKT pathway.
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Linfocitos B/inmunología , Células Dendríticas/inmunología , Proteína Forkhead Box O1/inmunología , Células Madre Hematopoyéticas/inmunología , Linfocitos T/inmunología , Animales , Humanos , Transducción de Señal/inmunologíaRESUMEN
OBJECTIVE: The aim of this study is to assess oral manifestations in patients with mucopolysaccharidosis IV (MPS IVA) and mucopolysaccharidosis VI (MPS VI). MATERIALS AND METHODS: Seventeen patients were assessed, nine with MPS IVA and eight with MPS VI, treated at the Medical Genetics Outpatient Clinic of Hospital Universitário Alcides Carneiro (HUAC) in Campina Grande, Paraíba State, Brazil. Assessments included clinical and intraoral examinations, analysis of occlusal function, and panoramic X-rays. RESULTS: Among all, 64.7% of the patients had unerupted teeth and 52.9% of them had enamel hypoplasia. Regarding the temporomandibular joint, 88.2% of the patients showed flattening of the mandibular condyle, 52.9% showed condylar hypoplasia, and all of them showed decreased mobility. Enamel hypoplasia was observed only in patients with MPS IVA (p < 0.05). On the other hand, only MPS VI patients presented with anterior open bite, restricted mouth opening (p < 0.05), and a higher rate of unerupted teeth, hyperplastic tooth follicle, and condylar defects (p < 0.05). CONCLUSIONS: Enamel hypoplasia was observed only in patients with MPS IVA, whereas anterior open bite was observed only in patients with MPS VI. Abnormal findings in the maxillomandibular complex were more frequent in patients with MPS VI. CLINICAL RELEVANCE: The relevant frequency of MPS VI and IVA in the sample allows us to compare the changes occurring in both groups of patients, therefore enabling us to further comprehend the oral manifestations in specific types of MPS.
Asunto(s)
Enfermedades de la Boca/etiología , Mucopolisacaridosis IV/complicaciones , Mucopolisacaridosis VI/complicaciones , Adolescente , Adulto , Niño , Estudios Transversales , Femenino , Humanos , Masculino , Enfermedades de la Boca/diagnóstico , Radiografía PanorámicaRESUMEN
INTRODUCTION: In this study, we aimed to analyze craniofacial morphology by assessing the skeletal cephalometric profiles of HIV-positive patients receiving antiretroviral therapy. METHODS: For this study, 21 HIV-positive patients aged between 6 and 17 years (study group) were selected and compared with 21 normoreactive patients (control group), paired by sex and age. The patients were also divided into 3 age ranges (6-8, 9-12, and 13-17 years) considering the pubertal growth spurt as the central event. Eighteen (linear and angular) measurements were traced on teleradiographs by using 2 methodologies. The mean values of each measurement were compared between the study and control groups by age range. RESULTS: The majority of the measurements checked in the HIV-positive children and adolescents for the 13-to-17 year age range were diminished, but not enough to generate a statistically significant difference in craniofacial growth. Statistically significant differences (P <0.05) were found only in the inclination of the palatal plane (6-8 years) and the position of the maxilla in the anteroposterior direction (13-17 years). CONCLUSIONS: These results led us to conclude that some cephalometric measurements of HIV-positive children and adolescents may be similar to those of normoreactive subjects.
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Terapia Antirretroviral Altamente Activa , Cefalometría , Cara/anatomía & histología , Seropositividad para VIH/tratamiento farmacológico , Cráneo/anatomía & histología , Adolescente , Estudios de Casos y Controles , Niño , Femenino , Humanos , Masculino , Proyectos PilotoRESUMEN
We studied in parallel the population structure of 90 carbapenemase-producing and 88 carbapenemase-susceptible Klebsiella pneumoniae isolates collected in 20 Spanish hospitals, in the context of the EuSCAPE project. Fourteen and 50 multilocus sequence types (MLSTs) were detected among the carbapenemase-producing and carbapenem-susceptible isolates, respectively. ST11 and ST15 clones were more frequent in the carbapenemase-producing group than in the carbapenemase-susceptible group (P < 0.0001). Among the members of the carbapenem-suceptible group, the cefotaxime-resistant population showed population parameters that differed between the populations of the wild-type strains and the carbapenemase producers.
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Antibacterianos/farmacología , Proteínas Bacterianas/metabolismo , Carbapenémicos/farmacología , Klebsiella pneumoniae/efectos de los fármacos , beta-Lactamasas/metabolismo , Proteínas Bacterianas/genética , Cefotaxima/farmacología , Farmacorresistencia Bacteriana/genética , Klebsiella pneumoniae/enzimología , Pruebas de Sensibilidad Microbiana , Tipificación de Secuencias Multilocus , beta-Lactamasas/genéticaRESUMEN
OBJECTIVES: There is little information about carbapenemase-producing (CP) Citrobacter spp. We studied the molecular epidemiology and microbiological features of CP Citrobacter spp. isolates collected in Spain (2013-15). METHODS: In total, 119 isolates suspected of being CP by the EUCAST screening cut-off values were analysed. Carbapenemases and ESBLs were characterized using PCR and sequencing. The genetic relationship among Citrobacter freundii isolates was studied by PFGE. RESULTS: Of the 119 isolates, 63 (52.9%) produced carbapenemases, of which 37 (58.7%) produced VIM-1, 20 (31.7%) produced OXA-48, 12 (19%) produced KPC-2, 2 (3.2%) produced NDM-1 and 1 (1.6%) produced VIM-2; 9 C. freundii isolates co-produced VIM-1 plus OXA-48. Fourteen isolates (22.2%) also carried ESBLs: 8 CTX-M-9 plus SHV-12, 2 CTX-M-9, 2 SHV-12 and 2 CTX-M-15. Fifty-seven isolates (90.5%) were C. freundii, 4 (6.3%) were Citrobacter koseri, 1 (1.6%) was Citrobacter amalonaticus and 1 (1.6%) was Citrobacter braakii. By EUCAST breakpoints, eight (12.7%) of the CP isolates were susceptible to the four carbapenems tested. In the 53 CP C. freundii analysed by PFGE, a total of 44 different band patterns were observed. Four PFGE clusters were identified: cluster 1 included eight isolates co-producing VIM-1 and OXA-48; blaVIM-1 was carried in a class 1 integron (intI-blaVIM-1-aacA4-dfrB1-aadA1-catB2-qacEΔ1/sul1) and blaOXA-48 was carried in a Tn1999.2 transposon. CONCLUSIONS: We observed the clonal and polyclonal spread of CP Citrobacter spp. across several Spanish geographical areas. Four species of Citrobacter spp. produced up to five carbapenemase types, including co-production of VIM-1 plus OXA-48. Some CP Citrobacter spp. isolates were susceptible to the four carbapenems tested, a finding with potential clinical implications.
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Proteínas Bacterianas/genética , Enterobacteriaceae Resistentes a los Carbapenémicos/enzimología , Enterobacteriaceae Resistentes a los Carbapenémicos/genética , Citrobacter/enzimología , Citrobacter/genética , Infecciones por Enterobacteriaceae/microbiología , Variación Genética , beta-Lactamasas/genética , Enterobacteriaceae Resistentes a los Carbapenémicos/clasificación , Enterobacteriaceae Resistentes a los Carbapenémicos/aislamiento & purificación , Citrobacter/clasificación , Citrobacter/aislamiento & purificación , Electroforesis en Gel de Campo Pulsado , Infecciones por Enterobacteriaceae/epidemiología , Genotipo , Humanos , Epidemiología Molecular , Tipificación Molecular , Reacción en Cadena de la Polimerasa , Análisis de Secuencia de ADN , España/epidemiologíaRESUMEN
A rapid and sensitive (100%) matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) assay was developed to detect OXA-48-type producers, using 161 previously characterized clinical isolates. Ertapenem was monitored to detect carbapenem resistance, and temocillin was included in the assay as a marker for OXA-48-producers. Structural analysis of temocillin is described. Data are obtained within 60 min.
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Técnicas de Tipificación Bacteriana , Enterobacteriaceae/clasificación , Enterobacteriaceae/enzimología , Espectrometría de Masa por Láser de Matriz Asistida de Ionización Desorción , beta-Lactamasas/biosíntesis , Antibacterianos/farmacología , Enterobacteriaceae/efectos de los fármacos , Ertapenem , Humanos , Sensibilidad y Especificidad , Espectrometría de Masa por Láser de Matriz Asistida de Ionización Desorción/métodos , beta-Lactamas/farmacologíaRESUMEN
OBJECTIVES: The objective of this study was to analyse the microbiological traits and the population structure of carbapenemase-producing (CP) Escherichia coli isolates collected in Spain between 2012 and 2014. METHODS: Two-hundred-and-thirty-nine E. coli isolates non-susceptible to carbapenems were studied. The carbapenemase genes and the phylogenetic groups were characterized using PCR. MLST was carried out using the typing schemes of the University of Warwick and the Institut Pasteur. The diversity of the population structure was estimated by calculating a simple diversity index (SDI). RESULTS: One-hundred-and-twenty-one isolates (50.6%) produced carbapenemases, of which 87 (71.9%) were OXA-48, 27 (22.3%) were VIM-1, 4 (3.3%) were KPC-2, 2 (1.7%) were NDM and 1 (0.8%) was IMP-22; 4 isolates were collected in 2012, 40 in 2013 and 77 in 2014. Ertapenem was more sensitive than imipenem or meropenem for screening for OXA-48-producing E. coli. Using the Warwick typing scheme, 59 different STs were identified, the most prevalent being ST131 (16.5%). The population diversity was higher among VIM-1-producing isolates (SDIâ=â81.5%) than among OXA-48-producing isolates (SDIâ=â44.8%). The Pasteur scheme had a higher discrimination capability (SDIâ=â55.4%) than the Warwick scheme (SDIâ=â48.8%). CONCLUSIONS: A progressive increase in the prevalence of CP E. coli was observed, mainly due to the dissemination of OXA-48 producers. The most sensitive method for detecting decreased susceptibility of CP E. coli to carbapenems was disc diffusion with ertapenem using the EUCAST screening cut-offs. The spread of CP E. coli was due to a polyclonal population. The Pasteur scheme showed the highest discrimination power. Surveillance is crucial for the early detection of CP E. coli.
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Proteínas Bacterianas/metabolismo , Infecciones por Escherichia coli/epidemiología , Infecciones por Escherichia coli/microbiología , Proteínas de Escherichia coli/metabolismo , Escherichia coli/enzimología , Variación Genética , Filogenia , beta-Lactamasas/metabolismo , Anciano , Anciano de 80 o más Años , Antibacterianos/farmacología , Proteínas Bacterianas/genética , Pruebas Antimicrobianas de Difusión por Disco , Ertapenem , Escherichia coli/clasificación , Escherichia coli/genética , Escherichia coli/aislamiento & purificación , Proteínas de Escherichia coli/genética , Femenino , Humanos , Masculino , Persona de Mediana Edad , Epidemiología Molecular , Tipificación de Secuencias Multilocus , Reacción en Cadena de la Polimerasa , Prevalencia , España/epidemiología , beta-Lactamasas/genética , beta-Lactamas/farmacologíaRESUMEN
OBJECTIVES: We analysed the microbiological traits and population structure of KPC-producing Enterobacteriaceae isolates collected in Spain between 2012 and 2014. We also performed a comparative WGS analysis of the three major KPC-producing Klebsiella pneumoniae clones detected. METHODS: Carbapenemase and ESBL genes were sequenced. The Institut Pasteur MLST scheme was used. WGS data were used to construct phylogenetic trees, to identify the determinants of resistance and to de novo assemble the genome of one representative isolate of each of the three major K. pneumoniae clones. RESULTS: Of the 2443 carbapenemase-producing Enterobacteriaceae isolates identified during the study period, 111 (4.5%) produced KPC. Of these, 81 (73.0%) were K. pneumoniae and 13 (11.7%) were Enterobacter cloacae. Three major epidemic clones of K. pneumoniae were identified: ST11/KPC-2, ST101/KPC-2 and ST512/KPC-3. ST11/KPC-2 differed from ST101/KPC-2 and ST512/KPC-3 by 27â819 and 6924 SNPs, respectively. ST101/KPC-2 differed from ST512/KPC-3 by 28â345 SNPs. Nine acquired resistance genes were found in ST11/KPC-2, 11 in ST512/KPC-3 and 13 in ST101/KPC-2. ST101/KPC-2 had the highest number of virulence genes (20). An 11 bp deletion at the end of the mgrB sequence was the cause of colistin resistance in ST512/KPC-3. CONCLUSIONS: KPC-producing Enterobacteriaceae are increasing in Spain. Most KPC-producing K. pneumoniae isolates belonged to only five clones: ST11 and ST512 caused interregional spread, ST101 caused regional spread and ST1961 and ST678 produced independent hospital outbreaks. ST101/KPC-2 had the highest number of resistance and virulence genes. ST101/KPC-2 and ST512/KPC-3 were recently implicated in the spread of KPC in Italy.
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Infecciones por Enterobacteriaceae/epidemiología , Infecciones por Enterobacteriaceae/microbiología , Enterobacteriaceae/enzimología , Genoma Bacteriano , Genotipo , Análisis de Secuencia de ADN , beta-Lactamasas/genética , Infección Hospitalaria/epidemiología , Infección Hospitalaria/microbiología , Enterobacteriaceae/clasificación , Enterobacteriaceae/genética , Enterobacteriaceae/aislamiento & purificación , Humanos , Epidemiología Molecular , Tipificación de Secuencias Multilocus , Prevalencia , España/epidemiologíaRESUMEN
OBJECTIVES: The global emergence of OXA-48-producing Klebsiella pneumoniae clones is a significant threat to public health. We used WGS and phylogenetic analysis of Spanish isolates to investigate the population structure of blaOXA-48-like-expressing K. pneumoniae ST11 and ST405 and to determine the distribution of resistance genes and plasmids encoding blaOXA-48-like carbapenemases. METHODS: SNPs identified in whole-genome sequences were used to reconstruct phylogenetic trees, identify resistance determinants and de novo assemble the genomes of 105 blaOXA-48-like-expressing K. pneumoniae isolates. RESULTS: Genome variation was generally lower in outbreak-associated isolates compared with those associated with sporadic infections. The relatively limited variation observed within the outbreak-associated isolates was on average 7-10 SNPs per outbreak. Of 24 isolates from suspected sporadic infections, 7 were very closely related to isolates causing hospital outbreaks and 17 were more diverse and therefore probably true sporadic cases. On average, 14 resistance genes were identified per isolate. The 17 ST405 isolates from sporadic cases of infection had four distinct resistance gene profiles, while the resistance gene profile differed in all ST11 isolates from sporadic cases. Sequence analysis of 94 IncL/M plasmids carrying blaOXA-48-like genes revealed an average of two SNP differences, indicating a conserved plasmid clade. CONCLUSIONS: Whole-genome sequence analysis enabled the discrimination of outbreak and sporadic isolates. Significant inter-regional spread within Spain of highly related isolates was evident for both ST11 and ST405 K. pneumoniae. IncL/M plasmids carrying blaOXA-48-like carbapenemase genes were highly conserved geographically and across the outbreaks, sporadic cases and clones.
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Klebsiella pneumoniae/efectos de los fármacos , Klebsiella pneumoniae/genética , Filogenia , Proteínas Bacterianas/biosíntesis , Proteínas Bacterianas/genética , Infección Hospitalaria , ADN Bacteriano/genética , Brotes de Enfermedades , Variación Genética , Humanos , Infecciones por Klebsiella/epidemiología , Infecciones por Klebsiella/microbiología , Klebsiella pneumoniae/enzimología , Epidemiología Molecular , Plásmidos/genética , España/epidemiología , beta-Lactamasas/biosíntesis , beta-Lactamasas/genéticaRESUMEN
BACKGROUND: The objective was to assess histopathological changes and the expression of proliferating cell nuclear antigen (PCNA), Bcl-2, suppressor of cytokine signaling (SOCS) 1 and 3, Vimentin, TWIST1, and Cdh 1 and 2 in early stages of experimental oral carcinogenesis process using a shorter period of exposure to 4-nitroquinoline oxide (4-NQO) model. METHODS: In this study, 20 rats were divided into control group (n = 10), sacrificed on the first day of the experiment, and experimental group (n = 10) treated with 50 ppm of 4-NQO solution dissolved in drinking water for 8 and 12 weeks. The histological sections were stained with H&E or subjected to immunohistochemistry for detecting PCNA, Bcl-2, SOCS 1 and 3, and STAT 3. Some specimens were used for verification of Vimentin expression, Cdh 1, Cdh 2, and TWIST1 by RT-qPCR. RESULTS: At both 8 and 12 weeks, morphological changes occurred mainly in the posterior portion of the tongue and were limited to the epithelial tissue, including moderate to severe dysplasia at 8 weeks, and severe dysplasia with exacerbation of atypical cells at 12 weeks. Expression of SOCS 1 and 3 increased from 8 to 12 weeks (P < 0.05), whereas STAT 3 expression was reduced mainly at 12 weeks (P < 0.05) in comparison with the control group. The expression of all epithelial-mesenchymal transition markers (EMT) was increased after 12 weeks, reaching statistical significance (P < 0.05) for Cdh 1 and 2. CONCLUSIONS: Together, the results suggested that overexpression of Bcl-2, SOCS 1 and 3, and Cdh 1 and 2 is associated with the early neoplasic changes in modified 4-nitroquinoline 1-oxide-induced murine oral cancer model.
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4-Nitroquinolina-1-Óxido , Biomarcadores de Tumor/biosíntesis , Carcinógenos , Neoplasias de la Boca/inducido químicamente , Neoplasias de la Boca/metabolismo , Animales , Biomarcadores de Tumor/genética , Biomarcadores de Tumor/metabolismo , Cadherinas/biosíntesis , Cadherinas/genética , Modelos Animales de Enfermedad , Transición Epitelial-Mesenquimal , Masculino , Neoplasias de la Boca/genética , Neoplasias de la Boca/patología , Proteínas del Tejido Nervioso/biosíntesis , Proteínas del Tejido Nervioso/genética , Antígeno Nuclear de Célula en Proliferación/biosíntesis , Antígeno Nuclear de Célula en Proliferación/genética , Proteínas Proto-Oncogénicas c-bcl-2/biosíntesis , Proteínas Proto-Oncogénicas c-bcl-2/genética , Ratas , Proteína 1 Supresora de la Señalización de Citocinas/biosíntesis , Proteína 1 Supresora de la Señalización de Citocinas/genética , Proteína 3 Supresora de la Señalización de Citocinas/biosíntesis , Proteína 3 Supresora de la Señalización de Citocinas/genética , Proteína 1 Relacionada con Twist/biosíntesis , Proteína 1 Relacionada con Twist/genética , Vimentina/biosíntesis , Vimentina/genéticaRESUMEN
Curcumin has therapeutic potential in preventing several types of cancer, including colon, liver, prostate, and breast. The goal of this study was to evaluate the chemopreventive activity of systemically administered curcumin on oral carcinogenesis induced by 4-nitroquinolone-1-oxide (4-NQO). A total of 50 male albino rats, Rattus norvegicus, (Holtzman), were divided into five groups (n = 10 per group). Four of these groups were exposed to 50 ppm 4-NQO in their drinking water ad libitum for 8 or 12 weeks, two groups were treated with curcumin by oral gavage at 30 or 100 mg/kg per day, and one group was treated with corn oil (vehicle) only. The negative control group was euthanized at baseline. Tongues of all animals were removed after euthanasia and used in the subsequent analysis because the tongue is the primary site of carcinogenesis in this model. Descriptive histological analysis and immunohistochemistry for PCNA, Bcl-2, SOCS1 e-3, and STAT3 were performed to assess the oncogenic process. The gene expression of Vimentin, E-cadherin, N-cadherin, or TWIST1 was assessed using RT-qPCR as a representative of epithelial-mesenchymal transition (EMT) events. The administration of curcumin at 100 mg/kg during the 12 weeks markedly decreased the expression of PCNA, Bcl-2, SOCS1 e -3, and STAT3. Curcumin also minimized the cellular atypia under microscopic analysis and diminished the expression of the genes associated with EMT. These findings demonstrate that the systemic administration of curcumin has chemopreventive activity during oral carcinogenesis induced by 4-NQO.
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Antineoplásicos/uso terapéutico , Curcumina/uso terapéutico , Neoplasias de la Boca/prevención & control , 4-Nitroquinolina-1-Óxido/metabolismo , Animales , Antineoplásicos/farmacología , Biomarcadores de Tumor/genética , Carcinógenos/metabolismo , Aceite de Maíz/uso terapéutico , Curcumina/farmacología , Modelos Animales de Enfermedad , Células Epiteliales , Transición Epitelial-Mesenquimal/efectos de los fármacos , Expresión Génica/efectos de los fármacos , Masculino , Neoplasias de la Boca/inducido químicamente , Neoplasias de la Boca/tratamiento farmacológico , Quinolonas/metabolismo , Ratas , Lengua/patologíaRESUMEN
The aim of this study was to determine the impact of carbapenemase-producing Enterobacteriaceae (CPE) in Spain in 2013 by describing the prevalence, dissemination, and geographic distribution of CPE clones, and their population structure and antibiotic susceptibility. From February 2013 to May 2013, 83 hospitals (about 40,000 hospital beds) prospectively collected nonduplicate Enterobacteriaceae using the screening cutoff recommended by EUCAST. Carbapenemase characterization was performed by phenotypic methods and confirmed by PCR and sequencing. Multilocus sequencing types (MLST) were determined for Klebsiella pneumoniae and Escherichia coli. A total of 702 Enterobacteriaceae isolates met the inclusion criteria; 379 (54%) were CPE. OXA-48 (71.5%) and VIM-1 (25.3%) were the most frequent carbapenemases, and K. pneumoniae (74.4%), Enterobacter cloacae (10.3%), and E. coli (8.4%) were the species most affected. Susceptibility to colistin, amikacin, and meropenem was 95.5%, 81.3%, and 74.7%, respectively. The most prevalent sequence types (STs) were ST11 and ST405 for K. pneumoniae and ST131 for E. coli. Forty-five (54.1%) of the hospitals had at least one CPE case. For K. pneumoniae, ST11/OXA-48, ST15/OXA-48, ST405/OXA-48, and ST11/VIM-1 were detected in two or more Spanish provinces. ST11 isolates carried four carbapenemases (VIM-1, OXA-48, KPC-2, and OXA-245), but ST405 isolates carried OXA-48 only. A wide interregional spread of CPE in Spain was observed, mainly due to a few successful clones of OXA-48-producing K. pneumoniae (e.g., ST11 and ST405). The dissemination of OXA-48-producing E. coli is a new finding of public health concern. According to the susceptibilities determined in vitro, most of the CPE (94.5%) had three or more options for antibiotic treatment.
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Antibacterianos/farmacología , Proteínas Bacterianas/metabolismo , Colistina/farmacología , Enterobacteriaceae/efectos de los fármacos , Enterobacteriaceae/enzimología , Tienamicinas/farmacología , beta-Lactamasas/metabolismo , Anciano , Escherichia coli/efectos de los fármacos , Escherichia coli/enzimología , Humanos , Klebsiella pneumoniae/efectos de los fármacos , Klebsiella pneumoniae/enzimología , Meropenem , Pruebas de Sensibilidad Microbiana , Tipificación de Secuencias Multilocus , Estudios Prospectivos , EspañaRESUMEN
OBJECTIVES: There are different methodological recommendations for in vitro testing of the co-amoxiclav combination. Performance of co-amoxiclav MIC testing for Escherichia coli by the standard ISO microdilution method (ISO 20776-1) was compared using EUCAST (fixed 2 mg/L clavulanate concentration) and CLSI (2â:â1 ratio) interpretive criteria. METHODS: MICs were determined by broth microdilution using a 2â:â1 ratio and fixed clavulanate concentrations (2 and 4 mg/L) for 160 clinical E. coli isolates with characterized resistance mechanisms. Essential agreements, categorical agreements and relative errors were determined. RESULTS: For all isolates, essential agreement between microdilution using 2 mg/L clavulanate and a 2â:â1 ratio was 25.6%. For ESBL-producing isolates, considering EUCAST breakpoints, 55% of isolates tested with 2 mg/L clavulanate were classified as resistant; conversely, 95% of isolates tested with 4 mg/L clavulanate were susceptible. When using CLSI breakpoints and a 2â:â1 ratio, 90% of isolates were susceptible and 10% were intermediate. CONCLUSIONS: Variation in the clavulanate concentration gave different susceptibility testing results, particularly among ESBL-producing E. coli isolates. The in vitro concentration of clavulanate that better correlates with clinical outcome is still under debate and should be established.
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Combinación Amoxicilina-Clavulanato de Potasio/farmacología , Antibacterianos/farmacología , Escherichia coli/efectos de los fármacos , Escherichia coli/aislamiento & purificación , Infecciones por Escherichia coli/microbiología , Humanos , Pruebas de Sensibilidad Microbiana/métodos , FenotipoRESUMEN
In a previous prospective multicenter study in Spain, we found that OXA-1 and inhibitor-resistant TEM (IRT) ß-lactamases constitute the most common plasmid-borne mechanisms of genuine amoxicillin-clavulanate (AMC) resistance in Escherichia coli. In the present study, we investigated the population structure and virulence traits of clinical AMC-resistant E. coli strains expressing OXA-1 or IRT and compared these traits to those in a control group of clinical AMC-susceptible E. coli isolates. All OXA-1-producing (n = 67) and IRT-producing (n = 45) isolates were matched by geographical and temporal origin to the AMC-susceptible control set (n = 56). We performed multilocus sequence typing and phylogenetic group characterization for each isolate and then studied the isolates for the presence of 49 virulence factors (VFs) by PCR and sequencing. The most prevalent clone detected was distinct for each group: group C isolates of sequence type (ST) 88 (C/ST88) were the most common in OXA-1 producers, B2/ST131 isolates were the most common in IRT producers, and B2/ST73 isolates were the most common in AMC-susceptible isolates. The median numbers of isolates per ST were 3.72 in OXA-1 producers, 2.04 in IRT producers, and 1.69 in AMC-susceptible isolates; the proportions of STs represented by one unique isolate in each group were 19.4%, 31.1%, and 48.2%, respectively. The sum of all VFs detected, calculated as a virulence score, was significantly higher in AMC-susceptible isolates than OXA-1 and IRT producers (means, 12.5 versus 8.3 and 8.2, respectively). Our findings suggest that IRT- and OXA-1-producing E. coli isolates resistant to AMC have a different and less diverse population structure than AMC-susceptible clinical E. coli isolates. The AMC-susceptible population also contains more VFs than AMC-resistant isolates.
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Combinación Amoxicilina-Clavulanato de Potasio/farmacología , Antibacterianos/farmacología , Farmacorresistencia Bacteriana/efectos de los fármacos , Infecciones por Escherichia coli/microbiología , Escherichia coli/genética , Escherichia coli/patogenicidad , Inhibidores de beta-Lactamasas/farmacología , beta-Lactamasas/biosíntesis , beta-Lactamasas/efectos de los fármacos , Escherichia coli/efectos de los fármacos , Humanos , Epidemiología Molecular , Tipificación de Secuencias Multilocus , Filogenia , Virulencia/efectos de los fármacos , Virulencia/genética , Factores de Virulencia/genética , beta-Lactamasas/genéticaRESUMEN
BACKGROUND: Moebius syndrome (MS) is a rare congenital condition that is characterised by facial hypomimia and congenital strabismus caused by complete or partial impairment of the 6th and 7th cranial nerves. MS may be further associated with other nerves or malformations, mainly involving the extremities. The objective of this study was to quantify the decrease in oral motor performance in people with MS compared with normoreactive individuals using the Oral Motor Assessment Scale (OMAS). METHODS: The study group comprised 33 subjects between the ages of 2 and 20 years (average age: 10 ± 5 years) with MS along with 46 age- and gender-matched control subjects. RESULTS: The study group displayed a lower average functional score than the control group (P < 0.0001). A significant lack of lip closure (P = 0.03) and anterior lingual seal during swallowing (P = 0.03) occurred in the study group; in most cases, the individuals with MS were classified as 'subfunctional'. In addition, individuals with MS in the older age group displayed better functional scores than those in the younger group (P = 0.05). CONCLUSIONS: Functional damage to oral motor function in individuals with MS is evident, but differs among patients with respect to severity and the movements that are compromised. However, overall, improvements in the functional patterns of these individuals can be observed as they mature in age.