RESUMO
The immune response to SARS-CoV-2 has been extensively studied following the pandemic outbreak in 2020; however, the presence of specific T cells against SARS-CoV-2 before vaccination has not been evaluated in Mexico. In this study, we estimated the frequency of T CD4+ and T CD8+ cells that exhibit a specific response to S (spike) and N (nucleocapsid) proteins in a Mexican population. We collected 78 peripheral blood samples from unvaccinated subjects, and the presence of antibodies against spike (RBD) and N protein was determined. Peripheral blood mononuclear cells were isolated and stimulated with a pool of S or N protein peptides (Wuhan-Hu-1 strain). IL-1ß, IL-4, IL-6, IL-10, IL-2, IL-8, TNF-α, IFN-γ, and GM-CSF levels were quantified in the supernatant of the activated cells, and the cells were stained to assess the activation and memory phenotypes. Differential activation frequency dependent on serological status was observed in CD4+ cells but not in CD8+ cells. The predominantly activated population was the central memory T CD4+ cells. Only 10% of the population exhibited the same phenotype with respect to the response to nucleocapsid peptides. The cytokine profile differed between the S and N responses. S peptides induced a more proinflammatory response compared with the N peptides. In conclusion, in a Mexican cohort before vaccination, there was a significant response to the S and N SARS-CoV-2 proteins resulting from previous infections with seasonal coronaviruses or previous undetected exposure to SARS-CoV-2.
Assuntos
Linfócitos T CD4-Positivos , Linfócitos T CD8-Positivos , COVID-19 , SARS-CoV-2 , Glicoproteína da Espícula de Coronavírus , Vacinação , Humanos , México/epidemiologia , SARS-CoV-2/imunologia , COVID-19/imunologia , COVID-19/epidemiologia , COVID-19/prevenção & controle , Feminino , Masculino , Adulto , Linfócitos T CD8-Positivos/imunologia , Pessoa de Meia-Idade , Linfócitos T CD4-Positivos/imunologia , Glicoproteína da Espícula de Coronavírus/imunologia , Citocinas/metabolismo , Vacinas contra COVID-19/imunologia , Proteínas do Nucleocapsídeo de Coronavírus/imunologia , Anticorpos Antivirais/sangue , Anticorpos Antivirais/imunologia , Adulto Jovem , Fosfoproteínas/imunologia , Idoso , Ativação Linfocitária/imunologiaRESUMO
In this study, lignin was chemically modified to promote hydrogel degradation as a source of carbon and nitrogen for a bacterial consortium consisting of P. putida F1, B. cereus and, B. paramycoides. A hydrogel was synthesized using acrylic acid (AA), acrylamide (AM), and 2-acrylamido-2-methyl-1-propanesulfonic acid (AMPS) and cross-linked with the modified lignin. The structural changes and mass loss in the hydrogel, as well as its final composition, were evaluated as functions of the growth of the selected strains in a culture broth with the powdered hydrogel. The average loss was 18.4% wt. The hydrogel was characterized using FTIR spectroscopy, scanning electronic microscopy (SEM), elemental analysis (EA), and thermogravimetric analysis (TGA) before and after bacterial treatment. FTIR showed that the carboxylic groups present in both the lignin and the acrylic acid of the hydrogel decreased during bacterial growth. The bacteria showed a preference for the biomaterial components of the hydrogel. SEM demonstrated superficial morphological changes in the hydrogel. The results reveal that the hydrogel was assimilated by the bacterial consortium while preserving the water retention capacity of the material and that the microorganisms carried out a partial biodegradation of the hydrogel. The results of the EA and TGA confirm that the bacterial consortium not only degraded the biopolymer (lignin), but also used the synthetic hydrogel as a carbon source to degrade its polymeric chains and modified original properties. This modification with lignin as a crosslinker (which is a waste product of the paper industry) is therefore proposed to promote hydrogel degradation.
RESUMO
Background: The axolotl, Ambystoma mexicanum is a unique biological model for complete tissue regeneration. Is a neotenic endangered species and is highly susceptible to environmental stress, including infectious disease. In contrast to other amphibians, the axolotl is particularly vulnerable to certain viral infections. Like other salamanders, the axolotl genome is one of the largest (32 Gb) and the impact of genome size on Ig loci architecture is unknown. To better understand the immune response in axolotl, we aimed to characterize the immunoglobulin loci of A. mexicanum and compare it with other model vertebrates. Methods: The most recently published genome sequence of A. mexicanum (V6) was used for alignment-based annotation and manual curation using previously described axolotl Ig sequences or reference sequences from other vertebrates. Gene models were further curated using A. mexicanum spleen RNA-seq data. Human, Xenopus tropicalis, Danio rerio (zebrafish), and eight tetrapod reference genomes were used for comparison. Results: Canonical A. mexicanum heavy chain (IGH), lambda (IGL), sigma (IGS), and the putative surrogate light chain (SLC) loci were identified. No kappa locus was found. More than half of the IGHV genes and the IGHF gene are pseudogenes and there is no clan I IGHV genes. Although the IGH locus size is proportional to genome size, we found local size restriction in the IGHM gene and the V gene intergenic distances. In addition, there were V genes with abnormally large V-intron sizes, which correlated with loss of gene functionality. Conclusion: The A. mexicanum immunoglobulin loci share the same general genome architecture as most studied tetrapods. Consistent with its large genome, Ig loci are larger; however, local size restrictions indicate evolutionary constraints likely to be imposed by high transcriptional demand of certain Ig genes, as well as the V(D)J recombination over very long genomic distance ranges. The A. mexicanum has undergone an extensive process of Ig gene loss which partially explains a reduced potential repertoire diversity that may contribute to its impaired antibody response.
Assuntos
Ambystoma mexicanum , Imunoglobulinas , Animais , Ambystoma mexicanum/genética , Genoma , Genômica , Imunoglobulinas/genéticaRESUMO
Mexican Lake Chapala is used as water supply for human consumption. Consequently, water quality of this lake is of paramount importance for the lake's wellbeing. The contribution presented in this paper investigates monitoring and assessment of lake water quality using water quality index (WQI), metal chemical speciation, and multivariate statistical techniques. Descriptive statistics shows total metal concentrations undetected conferring the lake a healthy status. Dissolved Cd and Pb exceed criterion continuous concentration limit, whereas Zn is below this limit indicating that water quality is satisfactory for aquatic life. However, WQI indicates poor water quality attributed to failure of conductivity, total solids, nitrogen, and phosphates, due to industrial and agro-industrial effluents. Metal speciations indicate that the presence of low concentrations of dissolved metals reflect interactions with gills of fish through metal-biotic ligand complexes affecting water quality. Positive correlations are obtained between conductivity and nitrates, indicating that agricultural activities and fertilizer runoffs increase the conductivity and that the environmental state of lake is being altered by human activities. Factors F1 (31%), F2 (19%), and F3 (11%) represent 61% of variability; F1 and F2 corroborate the pressure exerted by pollutants related with fertilizers and agrochemicals; F3 contains Zn and Pb with positive loads attributed to influx of tourist visitors. Sites S4, S5, S6, and S9 are identified as the most environmentally affected by COD, Alk*, pH, Cl-, nitrites, phosphates, and TS. Multivariate techniques permit to conclude that environmental stress of Lake Chapala is caused by variables pertaining to agrochemical, fertilizers and municipal wastes.
Assuntos
Metais Pesados , Poluentes Químicos da Água , Animais , Monitoramento Ambiental , Humanos , Lagos , Metais Pesados/análise , México , Poluentes Químicos da Água/análise , Qualidade da ÁguaRESUMO
Hypermucoviscosity (hmv) is a capsule-associated phenotype usually linked with hypervirulent Klebsiella pneumoniae strains. The key components of this phenotype are the RmpADC proteins contained in non-transmissible plasmids identified and studied in K. pneumoniae. Klebsiella variicola is closely related to K. pneumoniae and recently has been identified as an emergent human pathogen. K. variicola normally contains plasmids, some of them carrying antibiotic resistance and virulence genes. Previously, we described a K. variicola clinical isolate showing an hmv-like phenotype that harbors a 343-kb pKV8917 plasmid. Here, we investigated whether pKV8917 plasmid carried by K. variicola 8917 is linked with the hmv-like phenotype and its contribution to virulence. We found that curing the 343-kb pKV8917 plasmid caused the loss of hmv, a reduction in capsular polysaccharide (P < 0.001) and virulence. In addition, pKV8917 was successfully transferred to Escherichia coli and K. variicola strains via conjugation. Notably, when pKV8917 was transferred to K. variicola, the transconjugants displayed an hmv-like phenotype, and capsule production and virulence increased; these phenotypes were not observed in the E. coli transconjugants. These data suggest that the pKV8917 plasmid carries novel hmv and capsule determinants. Whole-plasmid sequencing and analysis revealed that pKV8917 does not contain rmpADC/rmpA2 genes; thus, an alternative mechanism was searched. The 343-kb plasmid contains an IncFIB backbone and shares a region of â¼150 kb with a 99% identity and 49% coverage with a virulence plasmid from hypervirulent K. variicola and multidrug-resistant K. pneumoniae. The pKV8917-unique region harbors a cellulose biosynthesis cluster (bcs), fructose- and sucrose-specific (fru/scr) phosphotransferase systems, and the transcriptional regulators araC and iclR, respectively, involved in membrane permeability. The hmv-like phenotype has been identified more frequently, and recent evidence supports the existence of rmpADC/rmpA2-independent hmv-like pathways in this bacterial genus.
RESUMO
BACKGROUND: Colorectal cancer is one of the most prevalent pathologies. Its prognosis is linked to the early detection and treatment. Currently diagnosis is performed by histological analysis from polyp biopsies, followed by morphological classification. Kudo's pit pattern classification is frequently used for the differentiation of neoplastic colorectal lesions using hematoxylin-eosin stained samples. Few articles have reported this classification with image software processing, using exogenous markers over the samples. The processing of autofluorescence images is an alternative that could allow the characterization of the pits from the crypts of Lieberkühn, bypassing staining techniques. OBJECTIVE: Processing and analysis of widefield autofluorescence microscopy images obtained by fresh colon tissue samples from a murine model of colorectal cancer in order to quantify and characterize the pits morphology by measuring morphology parameters and shape descriptors. METHODS: Adult male BALB/cCmedc strain mice (n=27), ranging from 20 to 30 g, were randomly assigned to four and five groups of treated and control animals. Colon samples were collected at day zero and at fourth, eighth, sixteenth and twentieth weeks after treatmentwith azoxymethane. Two-dimensional (2D) segmentation, quantification and morphological characterization of pits by image processing applied using macro programming from FIJI. RESULTS: Type I is the pit morphology prevailing between 53 and 81% in control group weeks. III-L and III-S types were detected in reduced percentages. Between the 33 and 56% of type I was stated as the prevailing morphology for the 4th, 8th and 20th weeks of treated groups, followed by III-L type. For the 16th week, the 39% of the pits was characterized as III-L type, followed by type I. Further, pattern types as IV, III-S and II were also found mainly in that order for almost all of the treated weeks. CONCLUSION: These preliminaries outcomes could be considered an advance in two-dimensional pit characterization as the whole image processing, comparing to the conventional procedure, takes a few seconds to quantify and characterize non-pathological colon pits as well as to estimate early pathological stages of colorectal cancer.
Assuntos
Pólipos do Colo/diagnóstico por imagem , Neoplasias Colorretais/diagnóstico por imagem , Microscopia de Fluorescência , Animais , Pólipos do Colo/patologia , Neoplasias Colorretais/patologia , Modelos Animais de Doenças , Masculino , Camundongos , Camundongos Endogâmicos BALB CRESUMO
ABSTRACT BACKGROUND: Colorectal cancer is one of the most prevalent pathologies. Its prognosis is linked to the early detection and treatment. Currently diagnosis is performed by histological analysis from polyp biopsies, followed by morphological classification. Kudo's pit pattern classification is frequently used for the differentiation of neoplastic colorectal lesions using hematoxylin-eosin stained samples. Few articles have reported this classification with image software processing, using exogenous markers over the samples. The processing of autofluorescence images is an alternative that could allow the characterization of the pits from the crypts of Lieberkühn, bypassing staining techniques. OBJECTIVE: Processing and analysis of widefield autofluorescence microscopy images obtained by fresh colon tissue samples from a murine model of colorectal cancer in order to quantify and characterize the pits morphology by measuring morphology parameters and shape descriptors. METHODS: Adult male BALB/cCmedc strain mice (n=27), ranging from 20 to 30 g, were randomly assigned to four and five groups of treated and control animals. Colon samples were collected at day zero and at fourth, eighth, sixteenth and twentieth weeks after treatmentwith azoxymethane. Two-dimensional (2D) segmentation, quantification and morphological characterization of pits by image processing applied using macro programming from FIJI. RESULTS: Type I is the pit morphology prevailing between 53 and 81% in control group weeks. III-L and III-S types were detected in reduced percentages. Between the 33 and 56% of type I was stated as the prevailing morphology for the 4th, 8th and 20th weeks of treated groups, followed by III-L type. For the 16th week, the 39% of the pits was characterized as III-L type, followed by type I. Further, pattern types as IV, III-S and II were also found mainly in that order for almost all of the treated weeks. CONCLUSION: These preliminaries outcomes could be considered an advance in two-dimensional pit characterization as the whole image processing, comparing to the conventional procedure, takes a few seconds to quantify and characterize non-pathological colon pits as well as to estimate early pathological stages of colorectal cancer.
RESUMO CONTEXTO: O câncer colorretal é uma das patologias mais prevalentes. Seu prognóstico é ligado à detenção e ao tratamento precoces. Atualmente o diagnóstico é realizado por análise histológica de biópsias de pólipo, seguida de classificação morfológica. A classificação de padrões de Kudo é frequentemente utilizada para a diferenciação de lesões colorretais neoplásicas usando amostras coradas por hematoxilina-eosina. Poucos artigos relatam esta classificação com utilização de processamento por software de imagem, utilizando marcadores exógenos sobre as amostras. O processamento de imagens de autofluorescência é uma alternativa que pode permitir a caracterização do padrão das criptas de Lieberkühn, contornando técnicas de coloração. OBJETIVO: Analisar, quantificar e caracterizar a morfologia do padrão das criptas medindo os parâmetros morfológicos e descritores de forma, através do processamento e análise de imagens de microscopia de autofluorescência de campo de Widefield obtidas em amostras de tecido de cólon fresco a partir de um modelo murino de câncer colorretal. MÉTODOS: Camundongos machos adultos BALB/cCmedc (n=27), variando de 20 a 30 g, foram distribuídos aleatoriamente em quatro e cinco grupos de animais tratados e de controle. As amostras de cólon foram coletadas no dia zero e na 4ª, 8ª, 16ª e 20ª semanas após o tratamento com azoxometano. Segmentação bidimensional (2D), quantificação e caracterização morfológica do padrão das criptas por processamento de imagem aplicados utilizando programação macro de FIJI. RESULTADOS: O tipo I é a morfologia da cripta prevalente entre 53% e 81% semanas do grupo controle. Os tipos III-L e III-S foram detectados em porcentagens reduzidas. A morfologia do tipo I entre os 33% e 56% foi constatada como a predominante para as 4ª, 8ª e 20ª semanas de grupos tratados, seguidos pelo tipo III-L. Para a 16ª semana, os 39% dos padrões das criptas foram caracterizados como tipo III-L, seguidos pelo tipo I. Além disso, os tipos de padrão como IV, III-S e II também foram encontrados principalmente nessa ordem para quase todas as semanas tratadas. CONCLUSÃO: Estes resultados preliminares podem ser considerados um avanço na caracterização bidimensional da cripta como um processamento integral da imagem, comparando-se ao procedimento convencional; demora-se alguns segundos a mais para quantificar e caracterizar pontos não-patológicos, bem como para estimar estágios patológicos precoces do câncer colorretal.
Assuntos
Animais , Masculino , Neoplasias Colorretais/diagnóstico por imagem , Pólipos do Colo/diagnóstico por imagem , Microscopia de Fluorescência , Neoplasias Colorretais/patologia , Pólipos do Colo/patologia , Modelos Animais de Doenças , Camundongos Endogâmicos BALB CRESUMO
Carbapenem-resistant Gram-negative bacteria isolated in Venezuela have been poorly characterized. The present study characterized a total of 34 isolates obtained from 27 patients; five of these patients were multi-infected. The bacterial species identified were Klebsiella pneumoniae (17), Pseudomonas aeruginosa (9), and Acinetobacter baumannii (8). From these isolates, 85% were identified as carbapenemase-producing bacteria, and the identified carbapenemase genes were blaKPC-2 (10/29 [34.4%]), blaVIM-type (7/29 [24.1%]), blaOXA-23 (7/29 [24.1%]), blaNDM-1 (8/29 [27.5%]), and the coexistence of blaOXA-23/blaNDM-1 (2/29 [6.8%]). Patient 1 was multi-infected by K. pneumoniae ST11 and ST2413 isolates harbouring the blaNDM-1 and blaKPC-2 genes, respectively. The other patients were multi-infected by two or three different bacterial species such as ESBL-producing K. pneumoniae isolates, P. aeruginosa harbouring the blaVIM-type gene, K. pneumoniae ST147 harbouring the blaKPC-2 gene and by A. baumannii harbouring the blaOXA-23 gene. The blaNDM-1 gene in A. baumannii is flanked by an uncommon genetic structure, whereas blaNDM-1 gene in K. pneumoniae revealed a common structure described in different plasmids from Enterobacteriaceae isolates. This study provides new information about the epidemiology of carbapenemase-producing bacteria in clinical setting in Venezuela.
Assuntos
Proteínas de Bactérias/biossíntese , Bactérias Gram-Negativas/genética , Resistência beta-Lactâmica/genética , beta-Lactamases/biossíntese , Acinetobacter baumannii , Adulto , Feminino , Genes Bacterianos/genética , Bactérias Gram-Negativas/enzimologia , Humanos , Klebsiella pneumoniae , Masculino , Pessoa de Meia-Idade , Pseudomonas aeruginosa , VenezuelaRESUMO
Antibody class switch recombination (CSR) to IgG, IgA, or IgE is a hallmark of adaptive immunity, allowing antibody function diversification beyond IgM. CSR involves a deletion of the IgM/IgD constant region genes placing a new acceptor Constant gene, downstream of the VDJH exon. CSR depends on non-coding (CSRnc) transcription of donor Iµ and acceptor IH exons, located 5' upstream of each CH coding gene. Although, our knowledge of the role of CSRnc transcription has advanced greatly, its extension and importance in healthy and diseased humans is scarce. We analyzed CSRnc transcription in 70,603 publicly available RNA-seq samples, including GTEx, TCGA, and the Sequence Read Archive using recount2, an online resource consisting of normalized RNA-seq gene and exon counts, as well as, coverage BigWig files that can be programmatically accessed through R. CSRnc transcription was validated with a qRT-PCR assay for Iµ, Iγ3, and Iγ1 in humans in response to vaccination. We mapped IH transcription for the human IGH locus, including the less understood IGHD gene. CSRnc transcription was restricted to B cells and is widely distributed in normal adult tissues, but predominant in blood, spleen, MALT-containing tissues, visceral adipose tissue and some so-called "immune privileged" tissues. However, significant Iγ4 expression was found even in non-lymphoid fetal tissues. CSRnc expression in cancer tissues mimicked the expression of their normal counterparts, with notable pattern changes in some common cancer subsets. CSRnc transcription in tumors appears to result from tumor infiltration by B cells, since CSRnc transcription was not detected in corresponding tumor-derived immortal cell lines. Additionally, significantly increased Iδ transcription in ileal mucosa in Crohn's disease with ulceration was found. In conclusion, CSRnc transcription occurs in multiple anatomical locations beyond classical secondary lymphoid organs, representing a potentially useful marker of effector B cell responses in normal and pathological immune responses. The pattern of IH exon expression may reveal clues of the local immune response (i.e., cytokine milieu) in health and disease. This is a great example of how the public recount2 data can be used to further our understanding of transcription, including regions outside the known transcriptome.
Assuntos
Linfócitos B/imunologia , Genes de Cadeia Pesada de Imunoglobulina/imunologia , Switching de Imunoglobulina/imunologia , Transcrição Gênica/imunologia , Éxons VDJ/imunologia , Adulto , Linfócitos B/patologia , Linhagem Celular Transformada , Bases de Dados de Ácidos Nucleicos , Feminino , Humanos , Masculino , Neoplasias/imunologiaRESUMO
Segmented polyurethanes were prepared with polycaprolactone diol as soft segment and various amounts of 4,4´-Methylenebis(cyclohexyl isocyanate) and atorvastatin, a statin used for lowering cholesterol, in order to obtain SPU with different content of rigid segments. Polyurethanes with 35% or 50% of rigid segment content were physicochemically characterized and their biocompatibility assessed with L929 fibroblasts. High concentrations of atorvastatin were incorporated by increasing the content of rigid segments as shown by FTIR, Raman, NMR, XPS and EDX. Thermal and mechanical characterization showed that polyurethanes containing atorvastatin and 35% of rigid segments were low modulus (13 MPa) semicrystalline polymers as they exhibited a glass transition temperature (Tg) at -38 °C, melting temperature (Tm) at 46 °C and crystallinity close to 35.9% as determined by DSC. In agreement with this, X-ray diffraction showed reflections at 21.3° and 23.6° for PCL without reflections for atorvastatin suggesting its presence in amorphous form with higher potential bioavailability. Low content of rigid segments led to highly degradable polymer in acidic, alkaline and oxidative media with an acceptable fibroblast cytotoxicity up to 7 days possibly due to low atorvastatin content.
Assuntos
Atorvastatina/química , Materiais Biocompatíveis/química , Cianatos/química , Poliésteres/química , Poliuretanos/química , Animais , Atorvastatina/toxicidade , Materiais Biocompatíveis/toxicidade , Linhagem Celular , Sobrevivência Celular/efeitos dos fármacos , Camundongos , Estrutura Molecular , Microscopia Óptica não Linear , Poliésteres/toxicidade , Poliuretanos/toxicidade , Espectrofotometria Infravermelho , TemperaturaRESUMO
Chitosan, sodium alginate and gel of Aloe vera (Aloe barbadensis Miller) were employed for the preparation of polyelectrolyte complexes at pH 4 and 6. FT-IR spectroscopy analysis showed evidence on complexes formation and incorporation of the Aloe vera gel. The ζ potential determination of the polyelectrolyte complexes revealed the presence of surface charges in the range of -20 to -24â¯mV, which results in stable systems. The dynamic moduli exhibited a high dependence on angular frequency, which is commonly found in solutions of macromolecules. The materials showed human fibroblast and lymphocyte viabilities up to 90% in agreement with null cytotoxicity. The polyelectrolyte complexes at pH 6 with Ca2+ were stable, showed high water absorption, satisfactory morphology, pore size and rigidity, characteristics that allowed significant human fibroblast migration in wound closure in vitro assays.
Assuntos
Movimento Celular/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Fibroblastos/citologia , Linfócitos/citologia , Polieletrólitos/química , Polieletrólitos/farmacologia , Alginatos/química , Aloe/química , Quitosana/química , Fibroblastos/efeitos dos fármacos , Ácido Glucurônico/química , Ácidos Hexurônicos/química , Humanos , Linfócitos/efeitos dos fármacos , ReologiaRESUMO
A nano-composite from biologically obtained chitin nanofillers homogenously dispersed in a poly(ε-caprolactone) matrix was successfully achieved by an ultrasonication-assisted non-toxic and non-aqueous methodology. For this purpose, biological chitin was obtained from lactic acid fermentation of shrimp wastes and converted into chitin whiskers by acidic hydrolysis in a novel process at low temperature (4°C) that enhanced the distribution and yield. Additionally, the polyester matrix was enzymatically produced in a non-toxic compressed fluid (1,1,1,2-tetrafluoroethane at 25bar and 65°C) medium. The homogeneous distribution of the nanofiller in the matrix was corroborated by confocal and atomic force microscopies. Films of the nanocomposite were physicochemically characterized to assess its adequate properties. Additionally, the qualitative viability of human fibroblasts and osteoblasts cells was studied on the produced nanocomposite films showing good biocompatibility.
Assuntos
Quitina/química , Nanocompostos/química , Nanopartículas/química , Poliésteres/química , Adulto , Animais , Candida/enzimologia , Criança , Quitina/isolamento & purificação , Fibroblastos , Química Verde , Humanos , Hidrocarbonetos Fluorados/química , Hidrólise , Lactobacillus plantarum/química , Lipase/química , Osteoblastos , Tamanho da Partícula , Penaeidae/químicaRESUMO
BACKGROUND: Influenza viruses display a high mutation rate and complex evolutionary patterns. Next-generation sequencing (NGS) has been widely used for qualitative and semi-quantitative assessment of genetic diversity in complex biological samples. The "deep sequencing" approach, enabled by the enormous throughput of current NGS platforms, allows the identification of rare genetic viral variants in targeted genetic regions, but is usually limited to a small number of samples. METHODOLOGY AND PRINCIPAL FINDINGS: We designed a proof-of-principle study to test whether redistributing sequencing throughput from a high depth-small sample number towards a low depth-large sample number approach is feasible and contributes to influenza epidemiological surveillance. Using 454-Roche sequencing, we sequenced at a rather low depth, a 307 bp amplicon of the neuraminidase gene of the Influenza A(H1N1) pandemic (A(H1N1)pdm) virus from cDNA amplicons pooled in 48 barcoded libraries obtained from nasal swab samples of infected patients (n â=â 299) taken from May to November, 2009 pandemic period in Mexico. This approach revealed that during the transition from the first (May-July) to second wave (September-November) of the pandemic, the initial genetic variants were replaced by the N248D mutation in the NA gene, and enabled the establishment of temporal and geographic associations with genetic diversity and the identification of mutations associated with oseltamivir resistance. CONCLUSIONS: NGS sequencing of a short amplicon from the NA gene at low sequencing depth allowed genetic screening of a large number of samples, providing insights to viral genetic diversity dynamics and the identification of genetic variants associated with oseltamivir resistance. Further research is needed to explain the observed replacement of the genetic variants seen during the second wave. As sequencing throughput rises and library multiplexing and automation improves, we foresee that the approach presented here can be scaled up for global genetic surveillance of influenza and other infectious diseases.
Assuntos
Antivirais/uso terapêutico , Farmacorresistência Viral/genética , Vírus da Influenza A Subtipo H1N1/genética , Influenza Humana/epidemiologia , Neuraminidase/genética , Oseltamivir/uso terapêutico , Pandemias , Proteínas Virais/genética , Adolescente , Adulto , Idoso , Criança , Pré-Escolar , Farmacorresistência Viral/efeitos dos fármacos , Monitoramento Epidemiológico , Feminino , Variação Genética , Sequenciamento de Nucleotídeos em Larga Escala , Humanos , Vírus da Influenza A Subtipo H1N1/isolamento & purificação , Influenza Humana/diagnóstico , Influenza Humana/virologia , Masculino , México/epidemiologia , Pessoa de Meia-Idade , Mutação , Projetos PilotoRESUMO
BACKGROUND: Distal embolization of thrombus/platelet aggregates decreases myocardial reperfusion during primary percutaneous coronary intervention (PCI), and is associated with worse immediate and long-term prognosis of patients with ST-elevation myocardial infarction (STEMI). OBJECTIVE: Assess the efficacy of a mesh covered stent (MGuard™ stent, MGS) in preventing distal embolization and microvascular reperfusion impairment during primary PCI, compared with a bare metal stent (BMS). METHODS: Forty patients with STEMI referred for primary PCI were randomized for stenting the culprit lesion with the MGS (n = 20) or a BMS (n = 20). Blinded experts performed off-line measurements of angiographic epicardial and microvascular reperfusion criteria: TIMI flow grade, myocardial blush, corrected TIMI frame count (cTFC). RESULTS: At baseline clinical, angiographic and procedural variables were not different between groups. Post PCI TIMI flow grade was similar in both groups. We observed better myocardial Blush grade in group MGS compared to BMS (median value 3.0 vs 2.5, 2p = 0.006) and cTFC (mean cTFC: MGS 19.65 ± 4.07 vs BMS 27.35 ± 7.15, 2p < 0.001, cTFC mean difference MGS-BMS: 7.7, CI 95%: 3.94 to 11.46). MGS stent group had a higher percentage of successful angioplasty (cTFC ≤ 23: MGS 85% vs BMS 30%, 2p < 0.001). We had two cases of acute stent thrombosis (one for each group) at 30days follow up, but no clinical events at 6 months follow up. CONCLUSIONS: In this exploratory study, MGS significantly improved microvascular reperfusion criteria compared with a BMS in primary PCI. However its safety and impact on clinical outcomes should be verified in larger randomized clinical trials.
Assuntos
Circulação Coronária , Metais , Microcirculação , Infarto do Miocárdio/terapia , Intervenção Coronária Percutânea/instrumentação , Stents , Telas Cirúrgicas , Adulto , Idoso , Distribuição de Qui-Quadrado , Chile , Angiografia Coronária , Embolia/etiologia , Embolia/prevenção & controle , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Infarto do Miocárdio/diagnóstico por imagem , Infarto do Miocárdio/fisiopatologia , Intervenção Coronária Percutânea/efeitos adversos , Desenho de Prótese , Trombose/etiologia , Trombose/prevenção & controle , Fatores de Tempo , Resultado do TratamentoRESUMO
Presentamos un caso de infarto agudo al miocardio complicado con shock cardiogénico y arritmias ventriculares refractarias. El paciente requirió oxigenación por membrana extracorpórea (ECMO) antes de su traslado a un hospital con asistencia ventricular izquierda (LVAD) y programa de trasplante de corazón. El paciente tuvo una buena recuperación después de 16 días de soporte con LVAD seguido de trasplante cardiaco, lo que muestra los beneficios de la terapia multimodal en casos complejos de shock cardiogénico refractario.
A case of acute myocardial infarction complicated with cardiogenic shock and refractory ventricular arrhythmias is described. The patient required extracorporeal membrane oxygenation (ECMO) prior to transfer to a hospital with a left ventricular assist device (LVAD) and heart transplantation program. He made a good recovery after 16 days of LVAD support followed by heart transplantation, showing the benefits of multimodal therapy incomplex cases of refractory cardiogenic shock Palabras clave: Infarto agudo del miocardio, shock cardiogénico,asistencia biventricular, ECMO.
Assuntos
Humanos , Masculino , Pessoa de Meia-Idade , Choque Cardiogênico/terapia , Oxigenação por Membrana Extracorpórea , Transplante de Coração , Coração Auxiliar , Infarto do Miocárdio/terapiaRESUMO
Con el objeto de conocer los agentes implicados en las infecciones nosocomiales y su susceptibilidad antimicrobiana en pacientes del Servicio de Neonatología, se realizó un estudio prospectivo en 101 aislamientos bacterianos, analizándose los casos de sepsis nosocomial. Se identificó la susceptibilidad de las cepas por métodos bacteriológicos convencionales, con medios de cultivo universales y selectivos. Se calculó la frecuencia de infecciones por 100 ingresos y se analizó la sensibilidad. La incidencia de sepsis neonatal nosocomial fue 36.4%. Predominaron bacterias gramnegativas (Klebsiella pneumoniae, Escherichia coli y no fermentadores) (54.4%), seguido por bacterias grampositivas (Staphylococcus aureus y Staphylococcus coagulasa negativo) (37.6%) y hongos (7.9%). La susceptibilidad a aminoglucósidos fue 28%. Para la Ciprofloxacina la sensibilidad para las Klebsiella fue 56.2%. Con respecto al cefepime y cefalosporinas de 3ra generación, la sensibilidad para los no fermentadores fue 50%. En cuanto al meropenem, Escherichia coli fue 54.4% sensible, Klebsiella 65.5%, y los no fermentadores 50%. Para piperacilina-tazobactam, Escherichia coli, Klebsiella y no fermentadores fueron sensibles en 45.4%, 40.6 % y 50%, respectivamente. La susceptibilidad al Colistin fue de 83.3% en los no fermentadores; Klebsiella 71.8% y Escherichia coli 63.6%. La sensibilidad del Staphylococcus aureus a la oxacilina fue de 18,7%, al linezolid, teicoplamina y vancomicina de 100%. El Staphylococcus coagulasa negativo fue 100% sensible a linezolid, teicoplanina y vancomicina y 46,6 % a clindamicina. Se pudo constatar la estabilidad en la circulación de los agentes causales, predominando las bacterias gramnegativas, y la baja susceptibilidad a la terapia antimicrobiana utilizada.
In order to determine the agents involved in nosocomial infections and their antimicrobial susceptibility in neonatology patients, a prospective study was carried out on 101 bacterial isolates, and cases of nosocomial sepsis were analyzed. We identified the susceptibility of strains by conventional microbiology with universal and selective culture media. Sensitivity was analyzed for estimation of rates for each 100 admissions. The incidence of nosocomial neonatal sepsis was 36.4%. Gram-negative bacteria (Klebsiella pn., Escherichia coli and non-fermenting) were predominant, 54.4%, followed by Gram-positive bacteria (Staphylococcus aureus and coagulase-negative Staphylococcus) 37.6%, and 7.9% fungi. Susceptibility to aminoglycosides was 28%. Sensitivity to ciprofloxacin for Klebsiella was 56.2%. Sensitivity of non-fermenters to cefepime and 3rd generation cephalosporins was 50%. Regarding meropenem, Escherichia coli was 54.4% sensitive, Klebsiella 65.5%, and non-fermenters 50%. Sensitivity of Escherichia coli, Klebsiella and nonfermenters to piperacillintazobactam was 45.4%, 40.6% and 50%, respectively. Sensitivity to colistin was 83.3% for non-fermenters, 71.8% for Klebsiella and 63.6% for Escherichia coli. Sensitivity of Staphylococcus aureus to oxacillin was 18.7%, and 100% to linezolid and vancomycin teicoplamina. Coagulase-negative Staphylococcus was 100% sensitive to linezolid, teicoplanin and vancomycin, and 46.6% to clindamycin. Results show stability of causal agents in the circulation, predominantly gram-negative bacteria and low susceptibility to antimicrobial therapy used.
RESUMO
OBJECTIVE: The aim of this registry was to evaluate a new device designed to facilitate antegrade guidewire re-entry into the true lumen of a chronic total coronary occlusion (CTO) from the adjacent subintimal space. BACKGROUND: Successful recanalization of CTOs results in clinical improvement in appropriately selected patients. CTO intervention is time- and resource-consuming, and a simplified approach enabling antegrade guidewire re-entry into the distal true lumen might improve success. METHODS: Patients with CTO and ischemia were entered into a prospective registry regardless of lesion characteristics. If wire manipulation resulted in subintimal wire entrapment, a new re-entry tool (a 2.5-mm flat subintimal balloon with two exit ports offset by 180°) was used as a platform to attempt guidewire penetration into the distal true lumen. The primary endpoint assessed was successful device-guided re-entry. Standard techniques were then utilized to open the CTO. RESULTS: In 40 consecutive CTO lesions attempted, 19 resulted in subintimal wire entrapment (mean occlusion length 44 mm). Sixteen of these 19 were successfully crossed with an antegrade guidewire into the distal true lumen using the new device (84%). One patient with unsuccessful re-entry was subsequently recanalized with a retrograde technique. All crossed lesions were stented (17/17), resulting in TIMI 3 flow without major complications. Two cases were unsuccessful. One patient had a grade I coronary perforation requiring no treatment. CONCLUSIONS: A new device to recanalize CTOs complicated by subintimal wire entrapment can be used successfully by experienced operators. Further study of this coronary re-entry device is ongoing.
Assuntos
Angioplastia Coronária com Balão/instrumentação , Cateteres Cardíacos , Oclusão Coronária/terapia , Idoso , Angioplastia Coronária com Balão/efeitos adversos , Chile , Doença Crônica , Angiografia Coronária , Oclusão Coronária/diagnóstico por imagem , Vasos Coronários/lesões , Desenho de Equipamento , Feminino , Humanos , Japão , Masculino , Pessoa de Meia-Idade , Estudos Prospectivos , Sistema de Registros , Resultado do Tratamento , Lesões do Sistema Vascular/etiologiaRESUMO
Las úlceras por presión son un problema de gran importancia en el hospital Guillermo Almenara Irigoyen, representan aproximadamente el 5 por ciento de los egresos hospitalarios de los servicios de Medicina Interna y producen elevada morbimortalidad. En la actualidad en el protocolo de tratamiento de la ulcera por presión (UPP), estadio III, se emplea los apósitos de Hidrocoloide, pero estos no son muy eficaces, solo un 39 por ciento de pacientes curan. Objetivo: Determinar la eficacia clínica (cicatrización) y seguridad del Jengibre iZingiber ojjicinale) en comparación con el Hidrocoloide en el tratamiento tópico de la Ulcera por Presión, sacro, Estadio III. Material y métodos: Ensayo clínico, controlado, no ciego, fue realizado en el Servicio de Medicina Interna No 5, del Hospital Guillermo Almenara Irigoyen, en Lima-Perú, 2003, 34 pacientes fueron tratados con Hidrocoloide y 34 pacientes fueron tratados con pasta de Jengibre. Resultados: El 52,9 por ciento de los pacientes que fue tratado con Hidrocoloide cicatrizo su UPP, estadio III, y el 66,2 por ciento de los pacientes que fue tratado con Jengibre cicatrizo su UPP, estadio III. En el grupo tratado con Hidrocoloide un 14,7 por ciento presentó infección de la úlcera, un 8,82 por ciento maceración de la piel, un 2,9 por ciento presenta hipergranulación y un 14,7 por ciento aparición de mal olor relacionado a la desintegración del Hidrocoloide. En el grupo tratado con jengibre un 5,8 por ciento presento infección de la ulcera, un 20,5 por ciento irritación cutánea, un 11,7 por ciento maceración de la piel. Los factores relacionados a una no cicatrización de la úlcera por presión fueron la desnutrición, la mayor severidad de compromiso del sensorio, el no control de los esfínteres y el mayor tamaño de la ulcera por presión. Conclusiones: 1) El tratamiento tópico para las úlceras por presión, estadio III; con Jengibre es más eficaz que el tratamiento tópico con Hidrocoloide. 2) El tratamiento con Jengibre presenta más efectos adversos. 3) Los factores que influyen en un retardo en la cicatrización de las ulceras por presión son la desnutrición, el no control de esfínteres, el mayor compromiso neurológico y el tamaño mayor al inicio de la ulcera por presión.
Skin ulcers caused by pressure are a very serious problem in the Guillermo Almenara Hospital, approximately represent 5 per cent of the hospitable debits of the Services of Internal Medicine and produce high morbimortality. Now a day a protocol of treatment of the pressure ulcer, stage III, use the hydrocolloid dressing, but it is not very effective, only 39 per cent of patients complete healing. Objective: Determine the clinical effectiveness and security of the ginger in comparison with the hydrocolloid dressing in the topical treatment of the pressure ulcer, sacred, (Stage III). Material and methods: A clinical trial, controlled, not blind, was made in the Service of Internal Medicine No 5, of the Guillermo Almenara Irigoyen Hospital, in Lima-Peru, 2003. A group of 34 patients were treatment with hidrocolloid dressing and 34 patients were treated with ginger paste. Results: 52,9 per cent complete healing with hydrocolloid, and 66,2 per cent complete healing with ginger. In the group treatment with hydrocolloid 14,7 per cent present infection of the ulcer, 8,82 per cent maceration of the skin, 2,9 per cent present hypergranulation and 14,7 per cent present a smell bad related to the disintegration of the hydrocolloid. In the group treatment with ginger 5,8 per cent present infection of the ulcer, 20,5 per cent cutaneous irritation, 11,7 per cent maceration of the skin. The related factors with the not complete healing ofthe pressure ulcer were the malnutrition, the greater severity of commitment of sense, the not control of the sphincters and the greater size of the pressure ulcer. Conclusions: l) Topical treatment for the pressure ulcer, stage III with ginger paste is more effective than the topical treatment with hidrocolloid dressing. 2) Treatment with ginger presents more adverse effects. 3) Factors that influence in a retardation in the cure of the pressure ulcer are the malnutrition, the not control of sphincters, the greater severity of commitment of sense and the greater size of the pressure ulcer.
Assuntos
Humanos , Masculino , Feminino , Pessoa de Meia-Idade , Idoso de 80 Anos ou mais , Extratos Vegetais , Zingiber officinale , Curativos Hidrocoloides , Úlcera por Pressão/terapia , Ensaios Clínicos Controlados como AssuntoRESUMO
Introducción: La revascularización de una estenosis coronaria que induce isquemia mejora el estatus funcional del paciente y su pronóstico. Por el contrario, la angioplastía de una lesión que no provoca isquemia puede implicar complicaciones sin beneficios. La medición de flujo de reserva coronario (FFR) es un índice del significado fisiopatológico de las estenosis coronarias. Objetivos: Comparar los eventos cardiovasculares adversos mayores (MACE) en pacientes en quienes la FFR permitió excluir una o más lesiones de angioplastia (Grupo FFR >0,75) con aquellos en que ésta obligó a la revascularización (grupo FFR < 0,75). Métodos: Estudio observacional en 74 pacientes derivados para angioplastía entre agosto de 2006 y julio de 2010. Se realizó medición de FFR con ade-nosina a lesiones de severidad intermedia (60-70 por ciento). Una FFR translesional <0,75 se consideró significativa para isquemia y subsidiaria de angioplastía. Se hizo un seguimiento de los MACE definidos como: muerte, infarto, necesidad de revascularización y/o presencia de angor. Resultados: 35 pacientes (47,3 por ciento) constituyeron el Grupo FFR >0,75, y en ellos disminuyó el porcentaje de lesiones múltiples angiográficamente significativas (dos o más vasos), desde un 51,4 por ciento (previo a la realización de FFR), a un 25,7 por ciento. En este grupo se utilizó un promedio de 0.7 stents por paciente, mientras que en el grupo FFR <0,75 (39 pacientes), este fue de 1.5 stents. Se efectuó un seguimiento promedio de 21,5 meses. Diecinueve pacientes (25,6 por ciento) desarrollaron MACE (28 eventos). No hubo fallecidos ni diferencias significativas en el análisis multifactorial en cuanto a infartos o angor, pero sí en la necesidad de nueva revascularización, siendo ésta significativamente mayor en el grupo FFR <0,75 (12,8 por ciento versus 2,9 por ciento, p=0,047). Conclusiones: La medición del FFR permite, en caso de descartar isquemia, disminuir la necesidad de angioplastías...
The treatment of coronary stenosis causing myocardial ischemia improves functional capacity and prognosis. Treatment of non-ischemia inducing coronary stenosis may lead to complications with no benefit to the patient Measurement of coronary flow reserve (CFR) may be used to assess the significance of coronary artery stenosis. Aim: To compare major adverse cardiovascular events (MACE) in patients with significant stenosis (CFR < 0.75), with those in which one or more stenosis was not significant (CFR > 0.75) and thus were not subjected to PTCA in the corresponding artery. Methods: 74 patients were included from August 2006 to July 2010. CFR was measured in lesions exhibiting 6070 percent stenosis, using adenosine. A value <0.75 was considered significant and led to PTCA. Patients were followed for death, myocardial infarction, revascularization and/or angina. Results: 35 patients (47 percent) constituted the >0.75 CFR group. After evaluation of CFR the number of significant coronary stenosis decreased from 51.4 percent to 25.7 percent. They received a mean of 0.7 stents per patient. On the other hand, in the group with CFR <0.75 (39 patients) the mean number of stents was 1.5. Patients were followed for a mean of 21.5 months. 28 MACE events were observed in 19 patients (25.6 percent). No deaths were observed. Multivariable analysis revealed no significant difference between groups regarding myocardial infarction or angina. The need for myocardial revascularization was greater in the CFR <0.75 group compared to the CFR >0.75 group (12.8 percent vs 2.9 percent, respectively, p=0.047). Conclusion: The exclusion of myocardial ischemia as inferred by a CFR >0.75 allowed a significantly lower number of PTCAs without increase in MACE at a medium term follow up. A greater number of revascularization procedures was required in patients with CFR <0.75, which is expected from the greater number of lesions subjected to PTCA.
Assuntos
Humanos , Masculino , Feminino , Angioplastia Coronária com Balão/métodos , Circulação Coronária , Estenose Coronária/fisiopatologia , Reserva Fracionada de Fluxo Miocárdico , Isquemia Miocárdica/diagnóstico , Evolução Clínica , Estenose Coronária/terapia , Seguimentos , Índice de Gravidade de DoençaRESUMO
Antecedentes: La Vasculopatía del injerto (VDI) es la principal causa de muerte tardía del trasplante cardiaco (TX). Un diagnóstico precoz de esta complicación tendría un impacto en la terapia y pronóstico de esta afección. El ultrasonido intracoronario (IVUS), permite un diagnóstico precoz y certero de VDI, pero en la mayoría de los centros aún se utiliza la coronariografía. Objetivo: Evaluar la validez de la coronariografía en el diagnostico de VDI en pacientes trasplantados cardiacos, comparado con el IVUS. Metodología: Estudio transversal, en el cual se analizó una muestra de 36 pacientes con un tiempo promedio de trasplante de 3,7 +/- 3,7 años, a quienes se realizó simultáneamente un examen de coronario-grafía y un IVUS. Se evaluó la reproducibilidad contrastando los resultados de la interpretación visual de la coronariografía versus la coronariografía con análisis cuantitativo. Se calculó la Sensibilidad, Especificidad, Valores predictivos (VPP, VPN), Likelihoods y el coeficiente de equivalencia de Spearman-Brown. Resultados: La coronariografía cualitativa mostró ser más exacta que la cuantitativa, con una sensibilidad 30,4 por ciento [95 por ciento IC= 11,6 - 49,2], una especificidad 92,3 por ciento [95 por ciento IC= 77,8 - 106,7], VPP 87,5 por ciento [95 por ciento IC= 64,5 - 110,4], VPN 42,8 por ciento [95 por ciento IC= 24,5 - 61,1], LR (+) 3,9 [95 por ciento IC 0,55 - 28,7] y un LR (-) 0,75 [95 por ciento IC= 0,55 - 1,03]. La coronariografía cualitativa y cuantitativa son moderadamente equivalentes con un coeficiente de equivalencia Spearman Brown de 0,65. Conclusión: La validez y la reproducibilidad de la coronariografía en el paciente con TX es moderada y debería ser complementada con IVUS para el diagnóstico de VDI.
Background: Graft vasculopathy (GV) is the main cause of late death following cardiac transplantation (TX). Early diagnosis of this condition may have an impact upon treatment and prognosis of this complication. Intravascular ultrasound (IVUS) allows an early and accurate diagnosis of GV. However, most centers continue to use coronary angiography for this purpose. Aim: to evaluate coronary angiography for the diagnosis of GV in relation to IVUS in post TX patients. Methods: In a cross-sectional study the results of coronary angiography and IVUS, used as gold standard, were compared in 36 patients with a mean post TX follow up of 3.7 +/- 3.7 years. Results were compared between visual and quantitative coronary angiography. Sensitivity, specificity, positive and negative predictive values (PPV, NPV), likelihood ratios and the equivalence Spearman-Brown coefficient were calculated. Results: Visual evaluation of coronary angiography was more accurate than quantitative coronary angiography. The sensitivity for GV was 30.4 percent (95 percent C.I. 11.6 - 49.2), specificity 92.3 percent (95 percent C.I. 77.8 - 106.7), PPV 87.5 percent (95 percent C.I. 64.5 - 110.4, NPP 42.8 percent (95 percent C.I. 24.5 - 61.1), likelihood ratio (+) 3.9 (95 percent C.I. 0.55 - 28.7), likelihood ratio (-) 0.75 (95 percent C.I. 0.55 -1.03). The Spearman Brown coefficient between visual and quantitative coronary angiography evaluation was 0.65. Conclusion: Accuracy and reproducibility of coronary angiography in the evaluation of GV is limited. IVUS should be used for better identification of GV.