RESUMEN
In 2020, the world had to adapt to a pandemic caused by a then novel coronavirus. In addition to its direct impact on morbidity and mortality, the COVID-19 pandemic brought unprecedented control measures and challenges to both individuals and society. Sweden has been seen by many as an outlier in the management of the pandemic. It is therefore of special interest to document the actual management of the pandemic in Sweden during its first 2 years and how public health was affected. In the authors opinion, within the Swedish context, it has been possible to achieve a similar level of effect on mortality and morbidity through recommendations as was achieved through stringent legal measures in comparable countries. This is supported by comparisons of excess mortality that have been published. Furthermore, we see in the available data that the consequences on mental health and living habits were very limited for the majority of the population. Trust in public institutions is high in Sweden, which has been important and is part of the context that made it possible to manage a pandemic with relatively 'soft' measures. We acknowledge challenges in protecting certain vulnerable groups, particularly during the first and second wave.
Asunto(s)
COVID-19 , Humanos , COVID-19/epidemiología , Suecia/epidemiología , Salud Pública , Pandemias/prevención & control , SARS-CoV-2RESUMEN
In November 2010, ≈27,000 (≈45%) inhabitants of Östersund, Sweden, were affected by a waterborne outbreak of cryptosporidiosis. The outbreak was characterized by a rapid onset and high attack rate, especially among young and middle-aged persons. Young age, number of infected family members, amount of water consumed daily, and gluten intolerance were identified as risk factors for acquiring cryptosporidiosis. Also, chronic intestinal disease and young age were significantly associated with prolonged diarrhea. Identification of Cryptosporidium hominis subtype IbA10G2 in human and environmental samples and consistently low numbers of oocysts in drinking water confirmed insufficient reduction of parasites by the municipal water treatment plant. The current outbreak shows that use of inadequate microbial barriers at water treatment plants can have serious consequences for public health. This risk can be minimized by optimizing control of raw water quality and employing multiple barriers that remove or inactivate all groups of pathogens.
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Criptosporidiosis/epidemiología , Cryptosporidium/aislamiento & purificación , Adolescente , Adulto , Anciano , Anciano de 80 o más Años , Niño , Preescolar , Criptosporidiosis/microbiología , Brotes de Enfermedades , Femenino , Humanos , Lactante , Recién Nacido , Masculino , Persona de Mediana Edad , Suecia/epidemiología , Agua , Microbiología del Agua , Purificación del Agua/métodos , Abastecimiento de Agua , Adulto JovenRESUMEN
Microbial colonization of mammals is an evolution-driven process that modulate host physiology, many of which are associated with immunity and nutrient intake. Here, we report that colonization by gut microbiota impacts mammalian brain development and subsequent adult behavior. Using measures of motor activity and anxiety-like behavior, we demonstrate that germ free (GF) mice display increased motor activity and reduced anxiety, compared with specific pathogen free (SPF) mice with a normal gut microbiota. This behavioral phenotype is associated with altered expression of genes known to be involved in second messenger pathways and synaptic long-term potentiation in brain regions implicated in motor control and anxiety-like behavior. GF mice exposed to gut microbiota early in life display similar characteristics as SPF mice, including reduced expression of PSD-95 and synaptophysin in the striatum. Hence, our results suggest that the microbial colonization process initiates signaling mechanisms that affect neuronal circuits involved in motor control and anxiety behavior.
Asunto(s)
Encéfalo/metabolismo , Conducta Exploratoria/fisiología , Tracto Gastrointestinal/microbiología , Aprendizaje por Laberinto/fisiología , Actividad Motora/fisiología , Análisis de Varianza , Animales , Cromatografía Líquida de Alta Presión , Homólogo 4 de la Proteína Discs Large , Electroforesis en Gel de Poliacrilamida , Vida Libre de Gérmenes , Guanilato-Quinasas , Hibridación in Situ , Péptidos y Proteínas de Señalización Intracelular/metabolismo , Masculino , Proteínas de la Membrana/metabolismo , Ratones , Análisis por Micromatrices , Reacción en Cadena de la Polimerasa , Análisis de Secuencia de ARN , Organismos Libres de Patógenos Específicos , Sinaptofisina/metabolismoRESUMEN
BACKGROUND: A remarkable variety of restriction-modification (R-M) systems is found in Helicobacter pylori. Since they encompass a large portion of the strain-specific H. pylori genes and therefore contribute to genetic variability, they are suggested to have an impact on disease outcome. Type I R-M systems comprise three different subunits and are the most complex of the three types of R-M systems. AIMS: We investigated the genetic diversity and distribution of type I R-M systems in clinical isolates of H. pylori. MATERIAL AND METHODS: Sixty-one H. pylori isolates from a Swedish hospital based case-control study and 6 H. pylori isolates of a Swedish population-based study were analyzed using polymerase chain reaction for the presence of the three R-M systems' subunits. Representative gene variants were sequenced. RESULTS: Although the hsdM and hsdR genes appeared conserved in our clinical H. pylori isolates, the sequences of the hsdS loci were highly variable. Despite their sequence diversity, the genes per se were present at high frequencies. We identified a number of novel allelic hsdS variants, which are distinct from corresponding hsdS loci in the sequenced H. pylori strains 26695, J99 and HPAG1. In analyses of paired H. pylori isolates, obtained from the same individuals with a 4-year interval, we observed genetic modifications of hsdS genes in patients with atrophic gastric mucosa. DISCUSSION: We propose that the genetic variability of hsdS genes in a bacterial population will give rise to new specificities of these enzymes, which might lead to adaptation to an ever-changing gastric environment.
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Proteínas Bacterianas/genética , Técnicas de Tipificación Bacteriana , Enzimas de Restricción-Modificación del ADN/genética , Infecciones por Helicobacter/microbiología , Helicobacter pylori/clasificación , Helicobacter pylori/genética , Polimorfismo Genético , Secuencia de Aminoácidos , Análisis por Conglomerados , ADN Bacteriano/química , ADN Bacteriano/genética , Genotipo , Helicobacter pylori/aislamiento & purificación , Humanos , Datos de Secuencia Molecular , Filogenia , Reacción en Cadena de la Polimerasa/métodos , Alineación de Secuencia , Análisis de Secuencia de ADN , SueciaRESUMEN
BACKGROUND: The liver is the central organ for xenobiotic metabolism (XM) and is regulated by nuclear receptors such as CAR and PXR, which control the metabolism of drugs. Here we report that gut microbiota influences liver gene expression and alters xenobiotic metabolism in animals exposed to barbiturates. PRINCIPAL FINDINGS: By comparing hepatic gene expression on microarrays from germfree (GF) and conventionally-raised mice (SPF), we identified a cluster of 112 differentially expressed target genes predominantly connected to xenobiotic metabolism and pathways inhibiting RXR function. These findings were functionally validated by exposing GF and SPF mice to pentobarbital which confirmed that xenobiotic metabolism in GF mice is significantly more efficient (shorter time of anesthesia) when compared to the SPF group. CONCLUSION: Our data demonstrate that gut microbiota modulates hepatic gene expression and function by altering its xenobiotic response to drugs without direct contact with the liver.
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Perfilación de la Expresión Génica , Regulación de la Expresión Génica , Intestinos/microbiología , Hígado/microbiología , Xenobióticos/metabolismo , Animales , Barbitúricos/farmacología , Núcleo Celular/metabolismo , Hígado/metabolismo , Masculino , Ratones , Modelos Biológicos , Familia de Multigenes , Análisis de Secuencia por Matrices de Oligonucleótidos , Pentobarbital/farmacología , Factores de TiempoRESUMEN
The human gastric pathogen Helicobacter pylori colonizes the stomachs of half of the human population, and causes development of peptic ulcer disease and gastric adenocarcinoma. H. pylori-associated chronic atrophic gastritis (ChAG) with loss of the acid-producing parietal cells, is correlated with an increased risk for development of gastric adenocarcinoma. The majority of H. pylori isolates produce lipopolysaccharides (LPS) decorated with human-related Lewis epitopes, which have been shown to phase-vary in response to different environmental conditions. We have characterized the adaptations of H. pylori LPS and Lewis antigen expression to varying gastric conditions; in H. pylori isolates from mice with low or high gastric pH, respectively; in 482 clinical isolates from healthy individuals and from individuals with ChAG obtained at two time points with a four-year interval between endoscopies; and finally in isolates grown at different pH in vitro. Here we show that the gastric environment can contribute to a switch in Lewis phenotype in the two experimental mouse models. The clinical isolates from different human individuals showed that intra-individual isolates varied in Lewis antigen expression although the LPS diversity was relatively stable within each individual over time. Moreover, the isolates demonstrated considerable diversity in the levels of glycosylation and in the sizes of fucosylated O-antigen chains both within and between individuals. Thus our data suggest that different LPS variants exist in the colonizing H. pylori population, which can adapt to changes in the gastric environment and provide a means to regulate the inflammatory response of the host during disease progression.
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Helicobacter pylori/metabolismo , Lipopolisacáridos/metabolismo , Anciano , Animales , Estudios de Casos y Controles , Cartilla de ADN/química , Progresión de la Enfermedad , Epítopos/química , Humanos , Concentración de Iones de Hidrógeno , Inflamación , Antígenos del Grupo Sanguíneo de Lewis/química , Ratones , Persona de Mediana Edad , FenotipoRESUMEN
BACKGROUND: Campylobacter jejuni is widespread in the environment and is the major cause of bacterial gastroenteritis in humans. In the present study we use microarray-based comparative genomic hybridizations (CGH), pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing (MLST) to analyze closely related C. jejuni isolates from chicken and human infection. RESULTS: With the exception of one isolate, the microarray data clusters the isolates according to the five groups determined by PFGE. In contrast, MLST defines only three genotypes among the isolates, indicating a lower resolution. All methods show that there is no inherit difference between isolates infecting humans and chicken, suggesting a common underlying population of C. jejuni. We further identify regions that frequently differ between isolates, including both previously described and novel regions. Finally, we show that genes that belong to certain functional groups differ between isolates more often than expected by chance. CONCLUSION: In this study we demonstrated the utility of 70-mer oligonucleotide microarrays for genotyping of Campylobacter jejuni isolates, with resolution outperforming MLST.
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Infecciones por Campylobacter/microbiología , Infecciones por Campylobacter/veterinaria , Campylobacter jejuni/clasificación , Campylobacter jejuni/genética , Análisis de Secuencia por Matrices de Oligonucleótidos/métodos , Enfermedades de las Aves de Corral/microbiología , Animales , Técnicas de Tipificación Bacteriana , Campylobacter jejuni/aislamiento & purificación , Pollos/microbiología , ADN Bacteriano/análisis , ADN Bacteriano/genética , Electroforesis en Gel de Campo Pulsado , Genotipo , Humanos , Hibridación de Ácido Nucleico , Análisis de Secuencia de ADNRESUMEN
Separating the large intestine from gut flora is a robust layer of epithelial cells. This barrier is armed with an array of recognizing receptors that collectively set the host innate response. Here, we use nuclear receptors (NRs) and Toll-like receptors (TLRs), suggested to act as second messengers in the communication between microorganisms and epithelial cells, as probes to assess the impact of gut flora on innate immunity in germ-free (GF) mice. Using quantitative real-time polymerase chain reaction analyses, we show that 37/49 NRs are expressed in colonic cells of GF mice. Of these, 5 can be modulated by resident flora: LXRalpha, RORgamma and CAR show reduced expression and Nur77 and GCNF display elevated expression in conventionally raised mice compared with GF. Moreover, increased expression levels of TLR-2 and TLR-5 are observed in specific pathogen-free (SPF) mice compared with GF mice, and CAR expression is connected to the TLR-2 signalling pathway. Infections of GF or SPF mice with Yersinia pseudotuberculosis, show that GF intestinal epithelial cells fail to respond, except for CAR, which is downregulated. In contrast, SPF epithelial cells show a downregulation of all the NRs except CAR, which appears to be unaffected. Our findings indicate that gut flora contributes to the development of an intact barrier function.
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Inmunidad Innata , Intestino Grueso/inmunología , Intestino Grueso/microbiología , Receptores Citoplasmáticos y Nucleares/inmunología , Receptores Toll-Like/inmunología , Animales , Colitis/inmunología , Células Epiteliales , Vida Libre de Gérmenes , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Transducción de Señal , Organismos Libres de Patógenos Específicos , Receptor Toll-Like 2/inmunología , Receptor Toll-Like 5/inmunología , Yersinia pseudotuberculosis/fisiologíaRESUMEN
A large number of genes encoding restriction-modification (R-M) systems are found in the genome of the human pathogen Helicobacter pylori. R-M genes comprise approximately 10% of the strain-specific genes, but the relevance of having such an abundance of these genes is not clear. The type II methyltransferase (MTase) M.HpyAIV, which recognizes GANTC sites, was present in 60% of the H. pylori strains analyzed, whereof 69% were resistant to restriction enzyme digestion, which indicated the presence of an active MTase. H. pylori strains with an inactive M.HpyAIV phenotype contained deletions in regions of homopolymers within the gene, which resulted in premature translational stops, suggesting that M.HpyAIV may be subjected to phase variation by a slipped-strand mechanism. An M.HpyAIV gene mutant was constructed by insertional mutagenesis, and this mutant showed the same viability and ability to induce interleukin-8 in epithelial cells as the wild type in vitro but had, as expected, lost the ability to protect its self-DNA from digestion by a cognate restriction enzyme. The M.HpyAIV from H. pylori strain 26695 was overexpressed in Escherichia coli, and the protein was purified and was able to bind to DNA and protect GANTC sites from digestion in vitro. A bioinformatic analysis of the number of GANTC sites located in predicted regulatory regions of H. pylori strains 26695 and J99 resulted in a number of candidate genes. katA, a selected candidate gene, was further analyzed by quantitative real-time reverse transcription-PCR and shown to be significantly down-regulated in the M.HpyAIV gene mutant compared to the wild-type strain. This demonstrates the influence of M.HpyAIV methylation in gene expression.
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Metilasas de Modificación del ADN/genética , Metilasas de Modificación del ADN/metabolismo , Helicobacter pylori/enzimología , Helicobacter pylori/genética , Línea Celular , Clonación Molecular , Enzimas de Restricción del ADN/metabolismo , ADN Bacteriano/metabolismo , Células Epiteliales/microbiología , Escherichia coli/genética , Eliminación de Gen , Expresión Génica , Regulación Bacteriana de la Expresión Génica , Humanos , Interleucina-8/biosíntesis , Viabilidad Microbiana , Mutagénesis Insercional , Unión Proteica , Proteínas Recombinantes/genética , Proteínas Recombinantes/aislamiento & purificación , Proteínas Recombinantes/metabolismoRESUMEN
The genetic variability of Helicobacter pylori is known to be high compared to that of many other bacterial species. H. pylori is adapted to the human stomach, where it persists for decades, and adaptation to each host results in every individual harboring a distinctive bacterial population. Although clonal variants may exist within such a population, all isolates are generally genetically related and thus derived from a common ancestor. We sought to determine the rate of genetic change of H. pylori over 9 years in two asymptomatic adult patients. Arbitrary primed PCR confirmed the relatedness of individual subclones within a patient. Furthermore, sequencing of 10 loci ( approximately 6,000 bp) in three subclones per time and patient revealed only two base pair changes among the subclones from patient I. All sequences were identical among the patient II subclones. However, PCR amplification of the highly divergent gene amiA revealed great variation in the size of the gene between the subclones within each patient. Thus, both patients harbored a single strain with clonal variants at both times. We also studied genetic changes in culture- and mouse-passaged strains, and under both conditions no genetic divergence was found. These results suggest that previous estimates of the rate of genetic change in H. pylori within an individual might be overestimates.
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Evolución Biológica , Variación Genética , Helicobacter pylori/genética , Adulto , Antiinfecciosos/farmacología , Proteínas Bacterianas/genética , Proteínas Portadoras/genética , Farmacorresistencia Bacteriana , Mucosa Gástrica/microbiología , Helicobacter pylori/efectos de los fármacos , Humanos , Lipoproteínas/genética , Metronidazol/farmacología , Datos de Secuencia Molecular , Mutación , Reacción en Cadena de la Polimerasa , Secuencias Repetitivas de Ácidos Nucleicos , Análisis de Secuencia de ADNRESUMEN
Helicobacter pylori is acquired during childhood, but its mode of transmission remains unclear. A genotyped H. pylori isolate (Hp1) that expresses two classes of adhesins was introduced into the stomachs of three types of germ-free FVB/N mice to model factors that may affect spread of H. pylori in humans. Normal mice represented human hosts with normal gastric acid production. Transgenic animals expressing human alpha-1,3/4-fucosyltransferase in their gastric pit cells represented humans with normal acid production and the commonly encountered Lewis(b) histo-blood group receptor for the bacterium's BabA adhesin. tox176 transgenic mice have a genetically engineered ablation of their acid-producing parietal cells and increased proliferation of gastric epithelial lineage progenitors that express sialylated glycan receptors for the bacterium's SabA adhesin. These mice mimic features encountered in humans with H. pylori-associated chronic atrophic gastritis (CAG). Different combinations and numbers of 6-week-old germ-free normal and transgenic mice were housed together. At least one donor mouse per cage was infected with a single gavage of 10(7) colony-forming units of Hp1. All cagemates were sacrificed 8 weeks later. Cultures of gastric and cecal contents, plus quantitative PCR assays of cecal contents harvested from donors and potential recipients, revealed that transmission only occurred between tox176 donors and tox176 recipients, and that the distribution of Hp1 along the gastrointestinal tract was significantly broader in mice without parietal cells (P < 0.001). Transmission between tox176 mice was not attributable to any significant difference in the density of Hp1 colonization of the stomachs of tox176 versus normal donors. Our findings lead to the testable hypothesis that the relative hypochlorhydria of young children, and conditions that promote reduced acid production in infected adults (e.g. CAG), represent risk factors for spread of H. pylori.
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Infecciones por Helicobacter/patología , Helicobacter pylori/patogenicidad , Células Parietales Gástricas/microbiología , Células Parietales Gástricas/patología , Animales , Vida Libre de Gérmenes , Infecciones por Helicobacter/etiología , Infecciones por Helicobacter/microbiología , Infecciones por Helicobacter/transmisión , Helicobacter pylori/aislamiento & purificación , Ratones , Ratones TransgénicosRESUMEN
Research on Helicobacter pylori has been driven by the field of genomics since the release of the first of two complete genome sequences in 1997. In this review we highlight progress made in the last year. New bioinformatics tools and methods promise better functional and strain comparative analyses of individual genes. Sequence-based methods of strain comparison documented the coevolution of H. pylori with human populations. Several comprehensive analyses of the bacterial transcriptome were undertaken as well as two sophisticated studies of the transcriptional response of specific host tissues in response to H. pylori infection using different mouse models of H. pylori diseases. Some progress was made in developing genetic tools for mutational analysis of the genes required for infection. Finally, proteomic approaches were refined to delineate surface exposed and secreted proteins that represent potential antigens. In summary, while we do not have the full story of H. pylori, significant progress in deciphering the genome into functional biology has been made.
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Genómica , Helicobacter pylori/genética , Variación Genética , ProteómicaRESUMEN
Helicobacter pylori infection of the human stomach is common and typically benign, although a subset of hosts develops severe pathology. Infection occurs in an organ with distinct microenvironments characterized by pronounced differences in the composition of acid-producing parietal cells. In this study, we examine determinants of bacterial tropism to various gastric niches by using germ-free normal and transgenic mice with an engineered parietal cell ablation. Mice were colonized for 8 weeks with a clinical isolate (Hp1) that expresses adhesins recognized by epithelial NeuAcalpha2,3Galbeta1,4 glycan receptors. In normal mice, Hp1 has tropism for a parietal cell-deficient niche where sialylated glycans are expressed by a narrow band of pit cells positioned at the boundary between the squamous epithelium (forestomach) and the proximal glandular epithelium. Lymphoid aggregates that develop in this niche, but not elsewhere in the stomach, were analyzed by GeneChip and quantitative RT-PCR studies of laser capture microdissected mucosa and yielded a series of biomarkers indicative of immune cell activation and maturation. Genetic ablation of parietal cells produced a new source of NeuAcalpha2,3Galbeta1,4 glycans in amplified gastric epithelial lineage progenitors, with accompanying expansion of Hp1 within the glandular epithelium. Lymphoid aggregates that develop in this formerly acid-protected epithelium have molecular features similar to those observed at the forestomach/glandular junction. These findings demonstrate the important roles played by parietal cells and glycan receptors in determining the positioning of H. pylori within the gastric ecosystem, and emphasize the need to consider the evolution of pathology within a given host in a niche-specific context.
Asunto(s)
Gastritis/microbiología , Gastritis/patología , Infecciones por Helicobacter/microbiología , Infecciones por Helicobacter/patología , Helicobacter pylori/patogenicidad , Células Parietales Gástricas/microbiología , Células Parietales Gástricas/patología , Animales , Toxina Diftérica/genética , Gastritis/etiología , Perfilación de la Expresión Génica , Vida Libre de Gérmenes , Infecciones por Helicobacter/etiología , Helicobacter pylori/aislamiento & purificación , Humanos , Tejido Linfoide/patología , Ratones , Ratones Transgénicos , Análisis de Secuencia por Matrices de Oligonucleótidos , Fragmentos de Péptidos/genéticaRESUMEN
Helicobacter pylori infects the stomachs of half of all humans. It has a relatively benign relationship with most hosts but produces severe pathology, including gastric cancer, in others. Identifying the relative contributions of host, microbial, and environmental factors to the outcome of infection has been challenging. Here we describe one approach for identifying microbial genes that affect the magnitude of host responses to infection. Single colony purified H. pylori isolates were obtained from 25 cases and 71 controls in a Swedish case-control study of gastric cancer. Strains were first phenotyped based on their ability to produce adhesins that recognize two classes of human gastric epithelial receptors. Thirteen binding strains and two non-binding controls were then subjected to whole genome genotyping using H. pylori DNA microarrays. A cohort of "variable" genes was identified based on a microarray-determined call of "absent" in at least one member of the strain panel. Each strain was subsequently introduced into two types of germ-free transgenic mice, each programmed to express a different host factor postulated to pose increased risk for development of severe pathology. Expression of biomarkers of host defense was quantitated 4 weeks after inoculation, and the magnitude of the response correlated with bacterial genotype. The proportion of genes encoding HsdS homologs (specificity subunit of hetero-oligomeric type I restriction-modification systems) was significantly higher in the pool of 18 variable genes whose presence directly correlated with a robust host response than their proportion in the remaining 352 members of the variable gene pool. This suggests that the functions of these HsdS homologs may include control of expression of microbial determinants that affect the extent of gastric responses to this potentially virulent pathogen.