RESUMO
Inflammatory bowel disease, inclusive of Crohn's disease and ulcerative colitis, consists of immunologically mediated disorders involving the microbiota in the gastrointestinal tract. Lavender oil is a traditional medicine used to relieve many gastrointestinal disorders. The goal of this study was to examine the therapeutic effects of the essential oil obtained from a novel lavender cultivar, Lavandula×intermedia cultivar Okanagan lavender (OLEO), in a mouse model of acute colitis caused by Citrobacter rodentium. In colitic mice, oral gavage with OLEO resulted in less severe disease, including decreased morbidity and mortality, reduced intestinal tissue damage, and decreased infiltration of neutrophils and macrophages, with reduced levels of TNF-α, IFN-γ, IL-22, macrophage inflammatory protein-2α, and inducible nitric oxide synthase expression. This was associated with increased levels of regulatory T cell populations compared with untreated colitic mice. Recently, we demonstrated that the composition of the enteric microbiota affects susceptibility to C. rodentium-induced colitis. Here, we found that oral administration of OLEO induced microbiota enriched with members of the phylum Firmicutes, including segmented filamentous bacteria, which are known to protect against the damaging effects of C. rodentium. Additionally, during infection, OLEO treatment promoted the maintenance of microbiota loads, with specific increases in Firmicutes bacteria and decreases in γ-Proteobacteria. We observed that Firmicutes bacteria were intimately associated with the apical region of the intestinal epithelial cells during infection, suggesting that their protective effect was through contact with the gut wall. Finally, we show that OLEO inhibited C. rodentium growth and adherence to Caco-2 cells, primarily through the activities of 1,8-cineole and borneol. These results indicate that while OLEO promoted Firmicutes populations, it also controlled pathogen load through antimicrobial activity. Overall, our results reveal that OLEO can protect against colitis through the microbial-immunity nexus and that a pharmacological agent, in this case OLEO, alters the normal enteric microbiota.
Assuntos
Carga Bacteriana , Citrobacter rodentium , Colite , Lavandula , Óleos de Plantas/administração & dosagem , Administração Oral , Animais , Anti-Infecciosos/administração & dosagem , Carga Bacteriana/efeitos dos fármacos , Carga Bacteriana/fisiologia , Quimiocina CXCL2/metabolismo , Citrobacter rodentium/efeitos dos fármacos , Citrobacter rodentium/fisiologia , Colite/tratamento farmacológico , Colite/imunologia , Colite/metabolismo , Colite/microbiologia , Colo/imunologia , Colo/metabolismo , Colo/microbiologia , Modelos Animais de Doenças , Avaliação Pré-Clínica de Medicamentos , Interferon gama/metabolismo , Interleucinas/metabolismo , Macrófagos/metabolismo , Metagenoma/efeitos dos fármacos , Metagenoma/fisiologia , Camundongos , Camundongos Endogâmicos C57BL , Monoterpenos/administração & dosagem , Óxido Nítrico Sintase Tipo II/metabolismo , Fator de Necrose Tumoral alfa/metabolismo , Interleucina 22RESUMO
More than 30 years ago, auranofin was developed for the treatment of rheumatoid arthritis as a substitution for the injectable gold compounds aurothiomalate and aurothioglucose. Both the ease of oral administration over intramuscular injections and more potent anti-inflammatory effects in vitro made auranofin seem like an excellent substitute for the traditional injectable gold compounds. Despite efficacy in the treatment of both rheumatoid arthritis and psoriasis, currently, auranofin is seldom used as a treatment for patients with rheumatoid arthritis as more novel anti-rheumatic medications have become available. Despite the decline in its clinical applications, research on auranofin has continued as it shows promise in the treatment of several different diseases. In recent years, advances in technology have allowed researchers to use molecular techniques to identify novel mechanisms of action of auranofin. Additionally, researchers are discovering potential new applications of auranofin. Dual inhibition of inflammatory pathways and thiol redox enzymes by auranofin makes it a new candidate for cancer therapy and treating microbial infections. This review will summarize recently obtained data on the mechanisms of action of auranofin, and potential new applications of auranofin in the treatment of various diseases, including several types of leukaemia, carcinomas, and parasitic, bacterial, and viral infections.
Assuntos
Antirreumáticos/uso terapêutico , Auranofina/uso terapêutico , Animais , Infecções Bacterianas/tratamento farmacológico , Humanos , Inflamação/tratamento farmacológico , Leucemia/tratamento farmacológico , Neoplasias/tratamento farmacológico , Doenças Parasitárias/tratamento farmacológico , Viroses/tratamento farmacológicoRESUMO
Individuals vary in their resistance to enteric infections. The role of the intestinal microbiota in altering susceptibility to enteric infection is relatively unknown. Previous studies have identified that C3H/HeOuJ mice suffer 100% mortality during Citrobacter rodentium-induced colitis, whereas C57BL/6 mice recover from infection. The basis for their differences in susceptibility is unclear and has been mainly attributed to differences in host genetics. This study investigated the role of the intestinal microbiota in altering susceptibility to C. rodentium-induced colitis. When the feces of C57BL/6 mice were gavaged into antibiotic treated C3H/HeOuJ mice, the C57BL/6 microflora led to a complete reversal in mortality patterns where 100% of the C3H/HeOuJ mice survived infection. This protection corresponded with reduced colonic pathology and less systemic pathogen load and was associated with increased inflammatory and redox responses with reduced epithelial cell death. C3H/HeOuJ mice are normally susceptible to infection-induced dehydration due to defective expression of colonic ion transporters such as Dra, CA IV, and CA I; expression of these genes was normalized when C3H/HeOuJ mice were colonized with the C57BL/6 microflora. Together, these data reveal that the colonic microbiota play a critical role in protecting against intestinal infection by inducing proinflammatory and prooxidant responses that control pathogen load as well as ion transporter gene expression previously shown to prevent fatal dehydration. Protection of mice from lethal colitis was associated with higher levels of bacteria from Bacteroidetes. This study reveals that the microbiota is sufficient to overcome inherent genetic susceptibility patterns in C3H/HeOuJ mice that cause mortality during C. rodentium infection.
Assuntos
Citrobacter rodentium , Colite/microbiologia , Colo/microbiologia , Infecções por Enterobacteriaceae/microbiologia , Metagenoma , Animais , Antiporters/genética , Bacteroidetes/isolamento & purificação , Anidrases Carbônicas/genética , Colite/patologia , Colo/patologia , Suscetibilidade a Doenças , Infecções por Enterobacteriaceae/patologia , Fezes/microbiologia , Feminino , Expressão Gênica , Transporte de Íons/genética , Camundongos , Camundongos Endogâmicos C3H , Camundongos Endogâmicos C57BL , Transportadores de SulfatoRESUMO
AIMS/HYPOTHESIS: Increased exposure to enteric microbes as a result of intestinal barrier disruption is thought to contribute to the development of several intestinal inflammatory diseases; however, it less clear whether such exposure modulates the development of extra-intestinal inflammatory and autoimmune diseases. The goal of this study was to examine the potential role of pathogenic enteric microbes and intestinal barrier dysfunction in the pathogenesis of type 1 diabetes. METHODS: Using NOD mice, we assessed: (1) intrinsic barrier function in mice at different ages by measuring serum levels of FITC-labelled dextran; and (2) the impact on insulitis development of infection by strains of an enteric bacterial pathogen (Citrobacter rodentium) either capable (wild-type) or incapable (lacking Escherichia coli secreted protein F virulence factor owing to deletion of the gene [DeltaespF]) of causing intestinal epithelial barrier disruption. RESULTS: Here we demonstrate that prediabetic (12-week-old) NOD mice display increased intestinal permeability compared with non-obese diabetes-resistant and C57BL/6 mice. We also found that young (4-week-old) NOD mice infected with wild-type C. rodentium exhibited accelerated development of insulitis in concert with infection-induced barrier disruption. In contrast, insulitis development was not altered in NOD mice infected with the non-barrier-disrupting DeltaespF strain. Moreover, C. rodentium-infected NOD mice demonstrated increased activation and proliferation of pancreatic-draining lymph node T cells, including diabetogenic CD8(+) T cells, compared with uninfected NOD mice. CONCLUSIONS/INTERPRETATION: This is the first demonstration that a loss of intestinal barrier integrity caused by an enteric bacterial pathogen results in the activation of diabetogenic CD8(+) T cells and modulates insulitis.
Assuntos
Infecções Bacterianas/complicações , Animais , Infecções Bacterianas/microbiologia , Linfócitos T CD8-Positivos/imunologia , Citrobacter rodentium/imunologia , Citrobacter rodentium/patogenicidade , Enterobacteriaceae/imunologia , Enterobacteriaceae/patogenicidade , Infecções por Enterobacteriaceae/imunologia , Infecções por Enterobacteriaceae/patologia , Citometria de Fluxo , Rearranjo Gênico , Hiperinsulinismo/microbiologia , Inflamação/imunologia , Intestinos/microbiologia , Intestinos/fisiologia , Intestinos/fisiopatologia , Ativação Linfocitária , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Endogâmicos NOD , Estado Pré-Diabético/microbiologia , Estado Pré-Diabético/fisiopatologia , Receptores de Antígenos de Linfócitos T/genética , Especificidade da EspécieRESUMO
Myeloid differentiation factor (MyD)88, an adaptor protein shared by the Toll-interleukin 1 receptor superfamily, plays a critical role in host defence during many systemic bacterial infections by inducing protective inflammatory responses that limit bacterial growth. However, the role of innate responses during gastrointestinal (GI) infections is less clear, in part because the GI tract is tolerant to commensal antigens. The current study investigated the role of MyD88 following infection by the murine bacterial pathogen, Citrobacter rodentium. MyD88-deficient mice suffered a lethal colitis coincident with colonic mucosal ulcerations and bleeding. Their susceptibility was associated with an overwhelming bacterial burden and selectively impaired immune responses in colonic tissues, which included delayed inflammatory cell recruitment, reduced iNOS and abrogated production of TNF-alpha and IL-6 from MyD88-deficient macrophages and colons cultured ex vivo. Immunostaining for Ki67 and BrDU revealed that MyD88 signalling mediated epithelial hyper-proliferation in response to C. rodentium infection. Thus, MyD88-deficient mice could not promote epithelial cell turnover and repair, leading to deep bacterial invasion of colonic crypts, intestinal barrier dysfunction and, ultimately, widespread mucosal ulcerations. In conclusion, MyD88 signalling within the GI tract plays a critical role in mediating host defence against an enteric bacterial pathogen, by controlling bacterial numbers and promoting intestinal epithelial homeostasis.
Assuntos
Citrobacter rodentium/imunologia , Colite/imunologia , Células Epiteliais/microbiologia , Fator 88 de Diferenciação Mieloide/deficiência , Fator 88 de Diferenciação Mieloide/fisiologia , Animais , Medula Óssea/microbiologia , Colo/química , Colo/microbiologia , Colo/patologia , Contagem de Colônia Microbiana , Ensaio de Imunoadsorção Enzimática , Interleucina-6/análise , Antígeno Ki-67/análise , Macrófagos/imunologia , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Óxido Nítrico Sintase Tipo II/análise , Técnicas de Cultura de Órgãos , Análise de Sobrevida , Fator de Necrose Tumoral alfa/análiseRESUMO
The Salmonella rdar (red, dry, and rough) morphotype is an aggregative and resistant physiology that has been linked to survival in nutrient-limited environments. Growth of Salmonella enterica serovar Typhimurium was analyzed in a variety of nutrient-limiting conditions to determine whether aggregation would occur at low cell densities and whether the rdar morphotype was involved in this process. The resulting cultures consisted of two populations of cells, aggregated and nonaggregated, with the aggregated cells preferentially displaying rdar morphotype gene expression. The two groups of cells could be separated based on the principle that aggregated cells were producing greater amounts of thin aggregative fimbriae (Tafi or curli). In addition, the aggregated cells retained some physiological characteristics of the rdar morphotype, such as increased resistance to sodium hypochlorite. Competitive infection experiments in mice showed that nonaggregative DeltaagfA cells outcompeted rdar-positive wild-type cells in all tissues analyzed, indicating that aggregation via the rdar morphotype was not a virulence adaptation in Salmonella enterica serovar Typhimurium. Furthermore, in vivo imaging experiments showed that Tafi genes were not expressed during infection but were expressed once Salmonella was passed out of the mice into the feces. We hypothesize that the primary role of the rdar morphotype is to enhance Salmonella survival outside the host, thereby aiding in transmission.
Assuntos
Aderência Bacteriana/fisiologia , Salmonella typhimurium/fisiologia , Estruturas Animais/microbiologia , Animais , Antibacterianos/farmacologia , Contagem de Colônia Microbiana , Feminino , Proteínas de Fímbrias/genética , Fímbrias Bacterianas/fisiologia , Deleção de Genes , Genes Reporter , Luciferases/genética , Luciferases/metabolismo , Luminescência , Camundongos , Camundongos Endogâmicos C57BL , Salmonelose Animal/microbiologia , Salmonella typhimurium/efeitos dos fármacos , Salmonella typhimurium/patogenicidade , Hipoclorito de Sódio/farmacologia , VirulênciaRESUMO
While probiotics are a multi-billion dollar industry, there is little evidence to show that supplementing infants provides any health benefits. We conducted an observational study where 35 of 86 participating mothers self-administered probiotics during breastfeeding, as well as directly to their infants. The primary objective was to determine if probiotic exposure influenced the infants' fecal microbiome while the secondary objective assessed associated changes to the mothers' breast milk immunity and infant health. Analysis of infant fecal microbiome throughout the first 6 months of life revealed that probiotics were associated with higher abundances of Bifidobacterium at week 1 only. Short-chain fatty acid production and predicted metagenomic functions of the microbial communities were not altered. While probiotics did not alter breast milk immune markers, fecal sIgA responses were higher among probiotic supplemented infants. Surprisingly, this was not associated with better health outcomes, as the probiotic cohort had higher incidences of mucosal-associated illnesses as toddlers. This retrospective clinical comparison suggests that probiotic exposure during infancy has limited effects on gut microbial composition yet is associated with increased infection later in life. These correlative findings caution against probiotic supplementation during infancy until rigorous controlled follow-up studies determining their safety and efficacy have occurred.
Assuntos
Microbioma Gastrointestinal/efeitos dos fármacos , Probióticos/efeitos adversos , Probióticos/metabolismo , Adulto , Bifidobacterium , Aleitamento Materno , Suplementos Nutricionais , Ácidos Graxos Voláteis , Fezes/microbiologia , Feminino , Microbioma Gastrointestinal/fisiologia , Humanos , Imunoglobulina A Secretora/análise , Lactente , Recém-Nascido , Masculino , Microbiota , Leite Humano , Mães , Estudos RetrospectivosRESUMO
Intestinal goblet cells are potentially key players in controlling susceptibility to ulcerative colitis (UC). Although impaired mucin (Muc2) production by goblet cells increases microbial stimulation of the colonic mucosa, goblet cells secrete other mediators that may influence or promote UC development. Correspondingly, Muc2-deficient ((-/-)) mice develop spontaneous colitis, concurrent with the dramatic upregulation of the goblet cell mediator, resistin-like molecule-beta (RELM-ß). Testing RELM-ß's role, we generated Muc2(-/-)/Retnlb(-/-) mice, finding that RELM-ß deficiency significantly attenuated colitis development and symptoms compared with Muc2(-/-) mice. RELM-ß expression in Muc2(-/-) mice strongly induced the production/secretion of the antimicrobial lectin RegIIIß, that exerted its microbicidal effect predominantly on Gram-positive Lactobacillus species. Compared with Muc2(-/-)/Retnlb(-/-) mice, this worsened intestinal microbial dysbiosis with a selective loss of colonic Lactobacilli spp. in Muc2(-/-) mice. Orally replenishing Muc2(-/-) mice with murine Lactobacillus spp., but not with a probiotic formulation containing several human Lactobacillus spp. (VSL#3), ameliorated their spontaneous colitis in concert with increased production of short-chain fatty acids. These studies demonstrate that the goblet cell mediator RELM-ß drives colitis in Muc2(-/-) mice by depleting protective commensal microbes. The ability of selective commensal microbial replacement to ameliorate colitis suggests that personalized bacterial therapy may prove beneficial for treatment of UC.
Assuntos
Colite Ulcerativa/imunologia , Células Caliciformes/imunologia , Hormônios Ectópicos/imunologia , Mucosa Intestinal/imunologia , Lactobacillus/imunologia , Mucina-2/imunologia , Animais , Colite Ulcerativa/genética , Colite Ulcerativa/microbiologia , Colite Ulcerativa/prevenção & controle , Colo/imunologia , Colo/microbiologia , Disbiose , Ácidos Graxos Voláteis/biossíntese , Regulação da Expressão Gênica , Células Caliciformes/microbiologia , Hormônios Ectópicos/genética , Peptídeos e Proteínas de Sinalização Intercelular , Mucosa Intestinal/microbiologia , Camundongos , Camundongos Knockout , Mucina-2/deficiência , Mucina-2/genética , Proteínas Associadas a Pancreatite , Probióticos/administração & dosagem , Proteínas/genética , Proteínas/imunologia , Índice de Gravidade de Doença , Transdução de Sinais , Simbiose/imunologiaRESUMO
The agfBAC operon of Salmonella enteritidis encodes thin aggregative fimbriae, fibrous, polymeric structures primarily composed of AgfA fimbrins. Although uncharacterized, AgfB shows a 51 % overall amino acid sequence similarity to AgfA. Using AgfB epitope-specific antiserum, AgfB was detected as a minor component of whole, purified fimbriae. Like AgfA, AgfB was released from purified fimbriae by >70 % formic acid, whereupon both AgfA-AgfA and AgfA-AgfB dimers as well as monomers were detected. This suggested that AgfB may form specific, highly stable, structural associations with AgfA in native fimbrial filaments, associations that were weakened in structurally unstable fibers derived from AgfA chimeric fimbrial mutants. Detailed sequence comparisons between AgfA and AgfB showed that AgfB harbored a similar fivefold repeated sequence pattern (x(6)QxGx(2)NxAx(3)Q), and contained structural motifs similar to the parallel beta helix model proposed for AgfA. Molecular modeling of AgfB revealed a 3D structure remarkably similar to that of AgfA, the structures differing principally in the surface disposition of non-conserved, basic, acidic and non-polar residues. Thus AgfB is a fimbrin-like structural homologue of AgfA and an integral, minor component of native thin aggregative fimbrial fibers. AgfB from an agfA deletion strain was detected as a non-fimbrial, SDS-insoluble form in the supernatant and was purified. AgfA from an agfB deletion strain was found in both SDS-soluble and insoluble, non-fimbrial forms. No AgfA-AgfA dimers were detected in the absence of AgfB. Fimbriae formation by intercellular complementation between agfB and agfA deletion strains could not be shown under a variety of conditions, indicating that AgfA and AgfB are not freely diffusible in S. enteritidis. This has important implications on the current assembly hypothesis for thin aggregative fimbriae.
Assuntos
Proteínas de Bactérias/química , Proteínas de Fímbrias , Fímbrias Bacterianas/química , Salmonella enteritidis/química , Motivos de Aminoácidos , Sequência de Aminoácidos , Especificidade de Anticorpos , Antígenos de Bactérias/química , Antígenos de Bactérias/genética , Antígenos de Bactérias/imunologia , Proteínas de Bactérias/genética , Proteínas de Bactérias/imunologia , Proteínas de Bactérias/isolamento & purificação , Western Blotting , Dimerização , Mapeamento de Epitopos , Epitopos de Linfócito B/química , Epitopos de Linfócito B/genética , Epitopos de Linfócito B/imunologia , Fímbrias Bacterianas/genética , Modelos Moleculares , Estrutura Terciária de Proteína , Sequências Repetitivas de Aminoácidos , Salmonella enteritidis/genética , Análise de Sequência de Proteína , Deleção de Sequência/genética , Eletricidade EstáticaRESUMO
Our previous studies revealed that offspring from rat dams fed fish oil (at 8% and 18% energy), developed impaired intestinal barriers sensitizing the colon to exacerbated injury later in life. To discern the mechanism, we hypothesized that in utero exposure to fish oil, rich in n-3 polyunsaturated fatty acid (PUFA), caused abnormal intestinal reparative responses to mucosal injury through differences in intestinal microbiota and the presence of naïve immune cells. To identify such mechanisms, gut microbes and naïve immune cells were compared between rat pups born to dams fed either n-6 PUFA, n-3 PUFA or breeder chow. Maternal exposure to either of the PUFA rich diets altered the development of the intestinal microbiota with an overall reduction in microbial density. Using qPCR, we found that each type of PUFA differentially altered the major gut phyla; fish oil increased Bacteroidetes and safflower oil increased Firmicutes. Both PUFA diets reduced microbes known to dominate the infant gut like Enterobacteriaceae and Bifidobacteria spp. when compared to the chow group. Uniquely, maternal fish oil diets resulted in offspring showing blooms of opportunistic pathogens like Bilophila wadsworthia, Enterococcus faecium and Bacteroides fragilis in their gut microbiota. As well, fish oil groups showed a reduction in colonic CD8+ T cells, CD4+ Foxp3+ T cells and arginase+ M2 macrophages. In conclusion, fish oil supplementation in pharmacological excess, at 18% by energy as shown in this study, provides an example where excess dosing in utero can prime offspring to harbor intestinal pathobionts and alter immune cell homeostasis.
Assuntos
Óleos de Peixe/administração & dosagem , Microbioma Gastrointestinal/efeitos dos fármacos , Trato Gastrointestinal/imunologia , Trato Gastrointestinal/microbiologia , Exposição Materna , Animais , Citosol/química , Ácidos Graxos/análise , Feminino , Óleos de Peixe/efeitos adversos , Macrófagos/imunologia , Ratos Sprague-Dawley , Óleo de Cártamo/administração & dosagem , Óleo de Cártamo/efeitos adversos , Subpopulações de Linfócitos T/imunologiaRESUMO
A recent population-based study in the Canadian province of British Columbia showed that, since the mid-1960s, there has been a significant increase in the incidence of retinopathy of prematurity-induced blindness in infants weighing 750 to 999 g at birth. To determine the impact of changing birth weight-specific survival on this new epidemic, all infants born in the province in the period 1952 through 1986 and known to the British Columbia Health Surveillance Registry as having retinopathy of prematurity-induced blindness were identified. In addition, the birth registration records for the 1,299 740 infants born in British Columbia in the same period and the death records of the 22,940 British Columbia-born infants who died in the province before the end of their first year of life were linked using a combination of probabilistic and manual record linkage techniques. These linked records and the records from the Health Surveillance Registry were used to calculate birth weight-specific incidence rates of retinopathy of prematurity-induced blindness in liveborn infants and first-year-of-life survivors. The rates, in 5-year intervals, showed that, in both liveborn infants and first-year survivors, the highest birth weight-specific rates occurred during the first epidemic of retinopathy of prematurity, which ended in British Columbia in 1954. Since the mid-to late-1960s, the incidence of retinopathy of prematurity-induced blindness in liveborn infants weighing less than 1000 g increased steadily whereas in infants weighting 1000 to 1499 g, incidence decreased slightly since the original epidemic ended. However, the experience of first-year-of-life survivors is substantially different.(ABSTRACT TRUNCATED AT 250 WORDS)
Assuntos
Peso ao Nascer , Cegueira/mortalidade , Surtos de Doenças , Retinopatia da Prematuridade/mortalidade , Cegueira/epidemiologia , Cegueira/etiologia , Colúmbia Britânica/epidemiologia , Humanos , Incidência , Lactente , Mortalidade Infantil , Recém-Nascido , Recém-Nascido Prematuro , Sistema de Registros/estatística & dados numéricos , Retinopatia da Prematuridade/complicações , Retinopatia da Prematuridade/epidemiologiaRESUMO
This study provides the first empiric evidence for the existence of a new epidemic of retinopathy of prematurity-induced blindness. Data from a population-based register of handicapping conditions in the Canadian province of British Columbia, and a birth weight-specific census of live-born infants in British Columbia, were used to determine annual, population-level incidences of retinopathy of prematurity-induced blindness during 1952 to 1983. Changes in incidence since the end of the original epidemic (1954) were determined by subdividing the 29-year period (1955 to 1983) into two intervals (1955 to 1964 and 1965 to 1983). Standardized incidence ratio analyses revealed a marginally significant increase in the overall incidence of retinopathy of prematurity-induced blindness in the later as compared with the earlier period. Infants weighing 750 to 999 g at birth had a significantly increased standardized incidence ratio of 3.07 (95% confidence interval 1.26, 11.06). No increases in risk were observed in heavier or lighter weight infants. Because ascertainment and diagnostic changes do not explain the weight-specific increases in incidence, these results provide the first population-level evidence for a new epidemic.
Assuntos
Surtos de Doenças , Doenças do Prematuro/epidemiologia , Doenças Retinianas/epidemiologia , Cegueira/epidemiologia , Cegueira/etiologia , Colúmbia Britânica , Humanos , Recém-Nascido de Baixo Peso , Recém-Nascido , Doenças do Prematuro/complicações , Oxigênio/efeitos adversos , Sistema de Registros , Doenças Retinianas/complicaçõesRESUMO
The establishment of a pharmacist-managed out-patient anticoagulation clinic in a private community hospital is described. Discussions by pharmacy with office-based physicians at a 187-bed, private, nonprofit community medical center indicated that the traditional system of anticoagulation management was not ideal for the physicians or their patients. Development of a pharmacist-managed anticoagulation clinic began in fall 1993; operations began in spring 1994. Planning included analyzing existing practices, reviewing the relevant literature, obtaining physician input, visiting an established anticoagulation clinic, formulating a business plan, and developing clinical protocols. Collaborative relationships were established with the hospital laboratory, business office, and risk management, information services, and medical records departments. Two pharmacists were trained to work in the clinic and provide coverage 24 hours a day. Services include patient assessment, monitoring of anticoagulation, warfarin dosage adjustment, medication management, patient education, follow-up care, and providing feedback to referring and attending physicians. The clinic has met with physician and patient satisfaction, has reduced the number of admissions to treat warfarin-related bleeding, and has been able to cover its direct costs. A pharmacist-managed anti-coagulation clinic was successfully established in a private community hospital.
Assuntos
Anticoagulantes/administração & dosagem , Ambulatório Hospitalar/organização & administração , Anticoagulantes/economia , Hospitais com 100 a 299 Leitos , Hospitalização/economia , Hospitais Comunitários/organização & administração , Hospitais Privados/organização & administração , Humanos , Ambulatório Hospitalar/economia , Qualidade da Assistência à Saúde , WashingtonRESUMO
AIMS: Steroidal and non-steroidal anti-inflammatory drugs are used for treatment of peripheral inflammation, but they are not effective in neurodegenerative disorders. Gold compounds are also used to treat peripheral inflammation, but their effects on neuroimmune reactions are unknown. This study investigated the effects of gold compounds on astrocytic cell functions and assessed in vivo distribution of auranofin after its oral administration in mice. MAIN METHODS: Auranofin and three other gold compounds were investigated for their ability to reduce the secretion of pro-inflammatory cytokines and cytotoxins produced by activated human astrocytic cells. Ability of the gold compounds to protect neuronal cells from glial cytotoxins and from oxidative damage induced by hydrogen peroxide was also studied. The in vivo distribution of auranofin was investigated using laser ablation inductively coupled plasma mass spectrometry (LA-ICP-MS). KEY FINDINGS: Auranofin (0.1-5 µM) inhibited the toxicity of stimulated primary human astrocytes and U-373 MG astrocytic cells towards human neuronal cells, but did not inhibit secretion of cytokines. Treatment of neuronal cells with high nanomolar to low micromolar concentrations of auranofin protected them from toxicity induced by hydrogen peroxide and supernatants of stimulated astrocytic cells through the upregulation of heme-oxygenase (HOX)-1. Aurothiomalate, aurothioglucose, and aurothiosulphate were ineffective in the assays used. Auranofin reached low micromolar concentrations in mouse brains following daily oral administration for one week. SIGNIFICANCE: Since auranofin may protect neurons by inhibiting astrocyte toxicity and is also directly neuroprotective, it could be useful in neurodegenerative diseases where activation of astrocytes contributes to the neuronal loss.
Assuntos
Antirreumáticos/farmacologia , Astrócitos/efeitos dos fármacos , Auranofina/farmacologia , Fármacos Neuroprotetores/farmacologia , Animais , Antirreumáticos/farmacocinética , Astrócitos/metabolismo , Auranofina/farmacocinética , Encéfalo/efeitos dos fármacos , Encéfalo/metabolismo , Linhagem Celular , Sobrevivência Celular/efeitos dos fármacos , Citocinas/metabolismo , Relação Dose-Resposta a Droga , Antagonismo de Drogas , Feminino , Heme Oxigenase (Desciclizante)/biossíntese , Humanos , Peróxido de Hidrogênio/farmacologia , Camundongos , Camundongos Endogâmicos C57BL , Oxidantes/farmacologia , Estresse Oxidativo/efeitos dos fármacos , Regulação para CimaRESUMO
Salmonella thin aggregative fimbriae (Tafi; curli) are important in pathogenesis and biofilm formation; however, less is known of their structure and morphogenesis. In the Salmonella agfBAC Tafi operon, the transcription and role of agfC have been elusive. In this study, agfBAC transcripts were detected using a sensitive reverse transcriptase technique. Native AgfC was not detected using polyclonal antibodies generated against purified hexahistidine-tagged AgfC; however, in trans expression revealed that AgfC was localized to the periplasm as a mature form. An isogenic DeltaagfC mutant displayed an abundance of 20 nm fibres, in addition to native Tafi (5-7 nm), and had an increase in cell surface hydrophobicity. Purified 20 nm fibres were depolymerized under exceptionally stringent conditions to release what proved to be AgfA subunits. This revealed that the 20 nm fibres represented a different form of Tafi. The role of AgfC in Tafi assembly was investigated further using an antibody-capture assay of isogenic Deltaagf mutants. A soluble antibody-accessible form of AgfA was captured in wild-type (wt), DeltaagfB and DeltaagfF strains, in support of the extracellular nucleation-precipitation pathway of Tafi assembly, but not in DeltaagfC or DeltaagfE mutants. This indicates that AgfC and AgfE are important for AgfA extracellular assembly, facilitating the synthesis of Tafi.
Assuntos
Proteínas de Fímbrias/metabolismo , Fímbrias Bacterianas/metabolismo , Salmonella enteritidis/metabolismo , Proteínas de Fímbrias/genética , Óperon , Salmonella enteritidis/química , Salmonella enteritidis/genética , Transcrição GênicaRESUMO
Salmonella spp. are environmentally persistent pathogens that have served as one of the important models for understanding how bacteria adapt to stressful conditions. However, it remains poorly understood how they survive extreme conditions encountered outside their hosts. Here we show that the rdar morphotype, a multicellular phenotype characterized by fimbria- and cellulose-mediated colony pattern formation, enhances the resistance of Salmonella to desiccation. When colonies were stored on plastic for several months in the absence of exogenous nutrients, survival of wild-type cells was increased compared to mutants deficient in fimbriae and/or cellulose production. Differences between strains were further highlighted upon exposure to sodium hypochlorite, as cellulose-deficient strains were 1,000-fold more susceptible. Measurements of gene expression using luciferase reporters indicated that production of thin aggregative fimbriae (Tafi) may initiate formation of colony surface patterns characteristic of the rdar morphotype. We hypothesize that Tafi play a role in the organization of different components of the extracellular matrix. Conservation of the rdar morphotype among pathogenic S. enterica isolates and the survival advantages that it provides collectively suggest that this phenotype could play a role in the transmission of Salmonella between hosts.
Assuntos
Celulose/metabolismo , Fímbrias Bacterianas/metabolismo , Salmonella typhimurium/fisiologia , Proteínas de Bactérias/genética , Proteínas de Bactérias/fisiologia , Celulose/genética , Dessecação , Fímbrias Bacterianas/genética , Oxidantes/farmacologia , Salmonella typhimurium/efeitos dos fármacos , Salmonella typhimurium/genética , Hipoclorito de Sódio/farmacologia , Fatores de Tempo , Fatores de Transcrição/genética , Fatores de Transcrição/fisiologiaRESUMO
In this study, we show that Salmonella produces an O-antigen capsule coregulated with the fimbria- and cellulose-associated extracellular matrix. Structural analysis of purified Salmonella extracellular polysaccharides yielded predominantly a repeating oligosaccharide unit similar to that of Salmonella enterica serovar Enteritidis lipopolysaccharide O antigen with some modifications. Putative carbohydrate transport and regulatory operons important for capsule assembly and translocation, designated yihU-yshA and yihVW, were identified by screening a random transposon library with immune serum generated to the capsule. The absence of capsule was confirmed by generating various isogenic Deltayih mutants, where yihQ and yihO were shown to be important in capsule assembly and translocation. Luciferase-based expression studies showed that AgfD regulates the yih operons in coordination with extracellular matrix genes coding for thin aggregative fimbriae and cellulose. Although the capsule did not appear to be important for multicellular behavior, we demonstrate that it was important for survival during desiccation stress. Since the yih genes are conserved in salmonellae and the O-antigen capsule was important for environmental persistence, the formation of this surface structure may represent a conserved survival strategy.
Assuntos
Cápsulas Bacterianas/genética , Proteínas de Bactérias/genética , Matriz Extracelular/genética , Regulação Bacteriana da Expressão Gênica , Salmonella/fisiologia , Fatores de Transcrição/genética , Cápsulas Bacterianas/metabolismo , Transporte Biológico , Metabolismo dos Carboidratos , Carboidratos/genética , Celulose/genética , Elementos de DNA Transponíveis , Dessecação , Microbiologia Ambiental , Matriz Extracelular/metabolismo , Fímbrias Bacterianas/genética , Genes Bacterianos/genética , Genes Bacterianos/fisiologia , Mutação , Antígenos O/química , Óperon/genética , Óperon/fisiologia , Salmonella/genética , Salmonella/metabolismoRESUMO
The use of a central venous catheter for long-term intravenous therapy is described. The catheter's history, physical description, and uses are discussed. Also reviewed are complications from use of the catheter, the pharmacist's role in patient teaching, and the procedure for administering medications through the catheter. A listing of drugs administered through the catheter, incompatibility data and patient teaching instructions are also included.