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BACKGROUND: While the prognosis of patients with Ewing sarcoma (EwS) is improving, little is known about the frequency of pain and its risk factors in survivors of EwS. This study aims to analyse the prevalence and risk factors of pain and its predictive value for recurrence. PATIENTS AND METHODS: In patients with remission after treatment of EwS, frequency and characteristics of pain within the first 5 years of follow up were assessed retrospectively. RESULTS: Of 80 patients, 37 (46%) presented with at least one episode of pain. Chronic pain (>3 months) was observed in 10 patients (13%). Experience of at least one episode of pain was associated with prior combined local treatment (surgery and radiation compared to surgery alone; odds ratio [OR] 5.83, 95% confidence interval [CI] 1.43-34.9, P = .007). A total of 59 episodes of pain were observed, including 47 acute and 12 chronic episodes. Lower limb pain accounted for 46% (27/59) of all episodes of pain, and was associated with primary tumour of the pelvis or lower extremity (OR 4.29, 95% CI 1.18-18.21, P = .025), which represented 64% (51/80) of all EwS. The positive predictive value of pain for recurrence was only 12%. CONCLUSION: Pain is a common problem in survivors of EwS, which mostly affects the lower extremity, and should be regularly assessed. Interventions to reduce pain may be particularly important in patients with combined local treatment with surgery and radiation, who seem to be at considerably increased risk for pain. Patients presenting with pain should be examined for recurrence.
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Protocolos de Quimioterapia Combinada Antineoplásica/efectos adversos , Neoplasias Óseas/tratamiento farmacológico , Dolor en Cáncer/patología , Supervivientes de Cáncer/estadística & datos numéricos , Recurrencia Local de Neoplasia/diagnóstico , Sarcoma de Ewing/tratamiento farmacológico , Adolescente , Neoplasias Óseas/patología , Dolor en Cáncer/inducido químicamente , Femenino , Estudios de Seguimiento , Alemania/epidemiología , Humanos , Estudios Longitudinales , Masculino , Recurrencia Local de Neoplasia/epidemiología , Prevalencia , Pronóstico , Estudios Retrospectivos , Sarcoma de Ewing/patologíaRESUMEN
Corynebacterium (C.) diphtheriae is one of the two etiological pathogens for human diphtheria with significant morbidity and mortality. Recently, members of its biovar Belfanti have been described as two novel species, C. belfantii and C. rouxii. The most important virulence factor and also the premise to cause diphtheria is the isolate's capacity to encode and express the diphtheria toxin (DT). In contrast to C. ulcerans, which represents a potentially zoonotic pathogen, C. diphtheriae (incl. the novel deduced species) has almost exclusively been found to comprise a human pathogen. We here report three rare cases of C. rouxii isolation from dogs suffering from disseminated poly-bacterial exsudative to purulent dermatitis and a traumatic labial defect, respectively. The isolates were identified as C. diphtheriae based on commercial biochemistry and matrix-assisted laser desorption/ionisation-time of flight mass spectrometry (MALDI-TOF MS) analysis. However, recently described specific spectral peaks were highly similar to spectra of C. rouxii, which was confirmed by whole genome sequencing. Further investigations of the dog isolates for the presence of DT by tox gene qPCR revealed negative results. The findings from this study point out that skin infections in companion animals can be colonized by uncommon and so believed human specific pathogens, thereby resembling the clinical signs of cutaneous diphtheria.
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Infecciones por Corynebacterium , Corynebacterium diphtheriae , Difteria , Enfermedades de los Perros , Úlcera Cutánea , Animales , Corynebacterium/genética , Infecciones por Corynebacterium/veterinaria , Corynebacterium diphtheriae/genética , Difteria/veterinaria , Toxina Diftérica , Enfermedades de los Perros/microbiología , Perros , Úlcera Cutánea/microbiología , Úlcera Cutánea/veterinaria , Secuenciación Completa del GenomaRESUMEN
The carrier Endoplasmic Reticulum Adenylate Transporter1 (ER-ANT1) resides in the endoplasmic reticulum (ER) membrane and acts as an ATP/ADP antiporter. Mutant plants lacking ER-ANT1 exhibit a dwarf phenotype and their seeds contain reduced protein and lipid contents. In this study, we describe a further surprising metabolic peculiarity of the er-ant1 mutants. Interestingly, Gly levels in leaves are immensely enhanced (26×) when compared with that of wild-type plants. Gly accumulation is caused by significantly decreased mitochondrial glycine decarboxylase (GDC) activity. Reduced GDC activity in mutant plants was attributed to oxidative posttranslational protein modification induced by elevated levels of reactive oxygen species (ROS). GDC activity is crucial for photorespiration; accordingly, morphological and physiological defects in er-ant1 plants were nearly completely abolished by application of high environmental CO(2) concentrations. The latter observation demonstrates that the absence of ER-ANT1 activity mainly affects photorespiration (maybe solely GDC), whereas basic cellular metabolism remains largely unchanged. Since ER-ANT1 homologs are restricted to higher plants, it is tempting to speculate that this carrier fulfils a plant-specific function directly or indirectly controlling cellular ROS production. The observation that ER-ANT1 activity is associated with cellular ROS levels reveals an unexpected and critical physiological connection between the ER and other organelles in plants.
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Adenosina Trifosfato/metabolismo , Antiportadores/metabolismo , Proteínas de Arabidopsis/metabolismo , Retículo Endoplásmico/metabolismo , Mitocondrias/metabolismo , Antiportadores/genética , Arabidopsis/genética , Arabidopsis/metabolismo , Arabidopsis/efectos de la radiación , Proteínas de Arabidopsis/genética , Expresión Génica/efectos de la radiación , Glicina/efectos de los fármacos , Glicina-Deshidrogenasa (Descarboxilante)/genética , Glicina-Deshidrogenasa (Descarboxilante)/metabolismo , Immunoblotting , Proteínas Mitocondriales/genética , Proteínas Mitocondriales/metabolismo , Mutación , Consumo de Oxígeno/genética , Consumo de Oxígeno/efectos de la radiación , Plantas Modificadas Genéticamente , Reacción en Cadena de la Polimerasa de Transcriptasa InversaRESUMEN
The key to success of extraintestinal pathogenic Escherichia coli (ExPEC) to colonize niches outside the intestinal tract and to establish infection is the coordinated action of numerous virulence and fitness factors. Intense research revealed not only an arsenal of unique virulence determinants with specific action, but also the multi-functionality of single elements. Especially iron uptake systems of ExPEC proved to be of prime importance. Apart from iron acquisition they optimize certain virulence properties. Here we analyzed the contribution of the salmochelin siderophore system to the ability of ExPEC to form biofilms. In the same iron limited environment, ExPEC displayed a distinct transcriptional profile of siderophore systems. During biofilm formation the iroN gene coding for the specific receptors of the siderophore salmochelin was highly upregulated. Almost no induction was observed during planctonic growth. Disruption of iroN resulted in a reduction of almost 50% in biofilm production. Efficient biofilm formation was not affected in a salmochelin synthesis mutant. Thus, the contribution of IroN is independent from the ability to produce salmochelin. Enhanced expression of IroN did not increase significantly the capacity to form biofilms in ExPEC. Interestingly, the additional expression of IroN or even the acquisition of the entire salmochelin system was not able to improve biofilm formation in a poor biofilm producer like a laboratory E. coli K12 strain. However, complementation with only IroN in an ExPEC iroA deletion mutant was able to restore biofilm formation. The contribution of IroN to biofilm formation appears to require a certain background found in ExPEC, but not in E. coli K12. This study identified the contribution of IroN to biofilm formation and highlights the multi-functional role of iron uptake systems in ExPEC.
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Biopelículas/crecimiento & desarrollo , Proteínas de Escherichia coli/metabolismo , Escherichia coli/fisiología , Hierro/metabolismo , Receptores de Superficie Celular/metabolismo , Medios de Cultivo/química , Escherichia coli/genética , Proteínas de Escherichia coli/genética , Perfilación de la Expresión Génica , Técnicas de Inactivación de Genes , Humanos , Receptores de Superficie Celular/genéticaRESUMEN
Uropathogenic Escherichia coli (UPEC) are the major cause of urinary tract infections (UTI). These bacteria are equipped with an arsenal of virulence factors, such as siderophores and adhesins enabling UPECs to sufficiently colonize the urinary tract of humans and animals. Such virulence factors manipulate and impair the recognition of UPECs by the host's innate immune system. Among those, factors like the TIR domain containing proteins in E. coli (TcpC) have been described to interfere with the Toll-like receptor 4 signaling cascade. Nevertheless, some UPECs such as strain UTI89 lack TcpC, but also manipulate the innate immune response. By a random mutant-library approach we identified the galU gene of strain UTI89 to be responsible for a reduced immune response of macrophages. Consequently, we created a site directed knockout mutant of the galU gene in strain UTI89. This mutant caused a significantly increased cytokine response when co-incubated with J774A.1 macrophages. This phenotype could be recomplemented in trans by insertion of a galU-expressing plasmid. No differences in the viability of macrophages co-incubated with either the wild-type (WT) or the ΔgalU mutant strain could be observed. Nor could any growth impairment be detected in the ΔgalU mutant compared to WT strain. Hence, the increased cytokine response was not due to differences in the bacterial cytotoxicity or bacterial counts in the assay. Our results also demonstrated a reduction of intracellular counts of UTI89ΔgalU in the infection model. We were able to show a loss of the O-polysaccharide side chain of the ΔgalU mutant LPS. A comparable LPS structure could be generated by the deletion of the waaL gene in the UTI89. This also caused an impaired immune modulation. In contrast, purified LPS was not sufficient to impair cytokine release of macrophages. Moreover, no differences could be detected by applying bacteria inactivated with heat or formalin treatment. From this, we assume that the aberration of the LPS structure caused by the knockout of the galU gene is an important but not the exclusive cause for the loss of UPEC's immune modulating properties.
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Proteínas de Escherichia coli/metabolismo , Interacciones Huésped-Patógeno , Evasión Inmune , Macrófagos/inmunología , Factor de Necrosis Tumoral alfa/metabolismo , UTP-Glucosa-1-Fosfato Uridililtransferasa/metabolismo , Escherichia coli Uropatógena/inmunología , Animales , Línea Celular , Proteínas de Escherichia coli/genética , Eliminación de Gen , Prueba de Complementación Genética , Ratones , UTP-Glucosa-1-Fosfato Uridililtransferasa/genética , Escherichia coli Uropatógena/genéticaRESUMEN
Prosthetic lipoyl groups are required for the function of several essential multienzyme complexes, such as pyruvate dehydrogenase (PDH), α-ketoglutarate dehydrogenase (KGDH), and the glycine cleavage system (glycine decarboxylase [GDC]). How these proteins are lipoylated has been extensively studied in prokaryotes and yeast (Saccharomyces cerevisiae), but little is known for plants. We earlier reported that mitochondrial fatty acid synthesis by ketoacyl-acyl carrier protein synthase is not vital for protein lipoylation in Arabidopsis (Arabidopsis thaliana) and does not play a significant role in roots. Here, we identify Arabidopsis lipoate-protein ligase (AtLPLA) as an essential mitochondrial enzyme that uses octanoyl-nucleoside monophosphate and possibly other donor substrates for the octanoylation of mitochondrial PDH-E2 and GDC H-protein; it shows no reactivity with bacterial and possibly plant KGDH-E2. The octanoate-activating enzyme is unknown, but we assume that it uses octanoyl moieties provided by mitochondrial ß-oxidation. AtLPLA is essential for the octanoylation of PDH-E2, whereas GDC H-protein can optionally also be octanoylated by octanoyltransferase (LIP2) using octanoyl chains provided by mitochondrial ketoacyl-acyl carrier protein synthase to meet the high lipoate requirement of leaf mesophyll mitochondria. Similar to protein lipoylation in yeast, LIP2 likely also transfers octanoyl groups attached to the H-protein to KGDH-E2 but not to PDH-E2, which is exclusively octanoylated by LPLA. We suggest that LPLA and LIP2 together provide a basal protein lipoylation network to plants that is similar to that in other eukaryotes.
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Pathogenicity of Yersinia pseudotuberculosis is determined by an arsenal of virulence factors. Particularly, the Yersinia outer proteins (Yops) and the Type III secretion system (T3SS) encoded on the pYV virulence plasmid are required for Yersinia pathogenicity. A specific group of Y. pseudotuberculosis, responsible for the clinical syndrome described as Far East scarlet-like fever (FESLF), is known to have an altered virulence gene cluster. Far East strains cause unique clinical symptoms for which the pYV virulence plasmid plays apparently a rather secondary role. Here, we characterize a previously unknown protein of Y. pseudotuberculosis serotype I strains (TcpYI) which can be found particularly among the FESLF strain group. The TcpYI protein shares considerable sequence homology to members of the Toll/IL-1 receptor family. Bacterial TIR domain containing proteins (Tcps) interact with the innate immune system by TIR-TIR interactions and subvert host defenses via individual, multifaceted mechanisms. In terms of virulence, it appears that the TcpYI protein of Y. pseudotuberculosis displays its own virulence phenotype compared to the previously characterized bacterial Tcps. Our results clearly demonstrate that TcpYI increases the intracellular survival of the respective strains in vitro. Furthermore, we show here that the intracellular survival benefit of the wild-type strain correlates with an increase in tcpYI gene expression inside murine macrophages. In support of this, we found that TcpYI enhances the survival inside the spleens of mice in a mouse model of peritonitis. Our results may point toward involvement of the TcpYI protein in inhibition of phagocytosis, particularly in distinct Y. pseudotuberculosis strains of the FESLF strain group where the pYV virulence plasmid is absent.
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Macrófagos/microbiología , Viabilidad Microbiana , Factores de Virulencia/genética , Yersiniosis/microbiología , Yersinia pseudotuberculosis/inmunología , Yersinia pseudotuberculosis/fisiología , Animales , Modelos Animales de Enfermedad , Femenino , Ratones , Ratones Endogámicos BALB C , Ratones Endogámicos C57BL , Peritonitis/microbiología , Bazo/microbiología , Factores de Virulencia/metabolismo , Yersinia pseudotuberculosis/genética , Yersinia pseudotuberculosis/aislamiento & purificaciónRESUMEN
Trueperella pecoris was described as a new species of the genus Trueperella in 2021 and might be pathogenic to various animal species. However, the lack of a suitable diagnostic test system stands in the way of epidemiological surveys to clarify possible causalities. In this study, a Loop-mediated Isothermal Amplification (LAMP) assay was developed and validated that was highly specific for T. pecoris. The assay provided an analytical sensitivity of 0.5 pg/25 µL and showed 100% inclusivity and exclusivity for 11 target and 33 non-target strains, respectively. Three different DNA extraction methods were evaluated to select the most LAMP-compatible method for cell disruption in pure and complex samples. Using an on-site applicable single-buffer DNA extraction with additional heating, the cell-based detection limit was 2.3 CFU/reaction. Finally, the LAMP assay was validated by means of artificially contaminated porcine lung tissue samples in which minimal microbial loads between 6.54 and 8.37 × 103 CFU per swab sample were detectable. The LAMP assay established in this study represents a suitable diagnostic procedure for identifying T. pecoris in clinical specimens and will help to collect epidemiological data on the pathogenicity of this species.
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Actinomycetaceae , Animales , Porcinos , Bioensayo , Membrana Celular , CalefacciónRESUMEN
Sarcoptic mange was detected in five free-ranging raccoon dogs (Nyctereutes procyonoides) in the federal state of Schleswig-Holstein, Germany, during a health assessment study of invasive species, including raccoon dogs, carried out between 2021 and 2022. Four raccoon dogs showed severe lesions, including extensive alopecia with thickening and hyperpigmentation of the skin (lichenification). The fifth animal was less affected, showing only thinning of the hair coat in multiple body locations. Skin scrapings were performed and confirmed the presence of Sarcoptes scabiei. Histopathology of the skin revealed diffuse epidermal hyperplasia and hyperkeratosis, mild eosinophilic dermatitis, and varying amounts of intralesional mites. Staphylococcus pseudintermedius and Corynebacterium auriscanis were detected in the skin samples of the affected animals, indicating a secondary bacterial infection. The source of sarcoptic mange remains unclear; interspecies transmission via direct or indirect contact seems likely. Raccoon dogs are therefore a potential vector for sarcoptic mange, and their behaviour could contribute to disease spread and persistence.
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Recently, the first plant nucleoside hydrolase, NSH1 (former designation URH1), was identified at the molecular level. This enzyme's highest hydrolysis capacity is for uridine, thereby balancing pyrimidine salvage and catabolism. NSH1 was found to be less efficient in the hydrolysis of further nucleosides. However, it remained unclear whether purine nucleosides are processed by NSH1. Moreover, the biochemical and physiological functions of further NSH isoforms in Arabidopsis has not been analyzed. Here we show that NSH1 is also able to hydrolyze xanthosine with high efficiency, and thus represents the leading activity in purine and pyrimidine breakdown in a cell. A knockout mutant for NSH1 showed symptoms of accelerated senescence, accompanied by marked accumulation of uridine and xanthosine under conditions of prolonged darkness. The closest, so far uncharacterized, homolog of NSH1, NSH2, was found to act during the late phase of senescence and may support inosine breakdown. NSH3, another NSH isoform, surprisingly functions as an extracellular, purine-specific hydrolase that is involved in degradation of extracellular nucleosides and may participate in wound and pathogen responses.
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Proteínas de Arabidopsis/metabolismo , Arabidopsis/enzimología , N-Glicosil Hidrolasas/metabolismo , Purinas/metabolismo , Arabidopsis/genética , Proteínas de Arabidopsis/genética , Técnicas de Inactivación de Genes , Mutagénesis Insercional , Mutación , N-Glicosil Hidrolasas/genética , Plantas Modificadas Genéticamente/enzimología , Plantas Modificadas Genéticamente/genética , ARN de Planta/genética , Ribonucleósidos/metabolismo , XantinasRESUMEN
Infections due to extraintestinal pathogenic E. coli (ExPEC) are very common in humans as well as in animals. In humans ExPEC infections include urinary tract infections (UTI), septicemia, and wound infections, which result in significant morbidity, mortality, and substantial healthcare costs. In view of the increasing number of ExPEC infections caused by more and more resistant strains, effective prevention would be desirable. Given the rising treatment costs, a vaccine may be cost-effective in selected patient groups, such as women with recurrent UTI, patients with neurologic disorders impairing bladder function and men with prostate hyperplasia. Previous vaccine studies used single target proteins or whole inactivated ExPEC cells. Here, we describe a vaccine system for oral application based on artificial multiple subunit vaccine proteins. Those multi-epitope proteins are composed of predicted epitopes derived from ExPEC virulence-associated proteins. As ExPEC are known to form intracellular biofilms in the urothelium and can also resist killing by non-activated macrophages, T-cell responses are supposed to be an important measure to counteract these stages of ExPEC during infection. Therefore, a live bacterial antigen delivery system based upon the Salmonella type-III secretion system (T3SS) was used in this study to directly deliver the vaccine proteins into the cytoplasm of the host cells. Epitope-rich domains of the proteins FyuA, IroN, ChuA, IreA, Iha, and Usp were expressed in an attenuated Salmonella enterica serovar Typhimurium strain and translocated into target cells for extended periods of time inducing a strong T-cell response. No significant antibody titre increase against the secreted vaccine proteins could be detected in vaginal wash or serum. Despite that, one of the vaccine proteins was able to significantly reduce bacterial load in the challenge model of intraperitoneal sepsis. This study shows that a vaccine encompassing distinct epitopes of virulence-associated ExPEC proteins (i) can be applied for a T3SS-dependent vaccination strategy, (ii) elicits T-cell responses and (iii) confers protection after a single application.
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Anticuerpos Antibacterianos/inmunología , Sistemas de Secreción Bacterianos , Infecciones por Escherichia coli/prevención & control , Vacunas contra Escherichia coli/administración & dosificación , Escherichia coli/inmunología , Administración Oral , Animales , Carga Bacteriana , Sistemas de Secreción Bacterianos/inmunología , Epítopos/genética , Epítopos/inmunología , Infecciones por Escherichia coli/inmunología , Infecciones por Escherichia coli/microbiología , Proteínas de Escherichia coli/genética , Proteínas de Escherichia coli/inmunología , Proteínas de Escherichia coli/aislamiento & purificación , Femenino , Cabras , Humanos , Masculino , Ratones , Estructura Terciaria de Proteína , Salmonella/genética , Salmonella/metabolismo , Sepsis/inmunología , Sepsis/microbiología , Sepsis/prevención & control , Vacunación , Vacunas de Subunidad/administración & dosificación , Vacunas Sintéticas/administración & dosificación , VirulenciaRESUMEN
Horizontal gene transfer is a key step in the evolution of bacterial pathogens. Besides phages and plasmids, pathogenicity islands (PAIs) are subjected to horizontal transfer. The transfer mechanisms of PAIs within a certain bacterial species or between different species are still not well understood. This study is focused on the High-Pathogenicity Island (HPI), which is a PAI widely spread among extraintestinal pathogenic Escherichia coli and serves as a model for horizontal transfer of PAIs in general. We applied a phylogenetic approach using multilocus sequence typing on HPI-positive and -negative natural E. coli isolates representative of the species diversity to infer the mechanism of horizontal HPI transfer within the E. coli species. In each strain, the partial nucleotide sequences of 6 HPI-encoded genes and 6 housekeeping genes of the genomic backbone, as well as DNA fragments immediately upstream and downstream of the HPI were compared. This revealed that the HPI is not solely vertically transmitted, but that recombination of large DNA fragments beyond the HPI plays a major role in the spread of the HPI within E. coli species. In support of the results of the phylogenetic analyses, we experimentally demonstrated that HPI can be transferred between different E. coli strains by F-plasmid mediated mobilization. Sequencing of the chromosomal DNA regions immediately upstream and downstream of the HPI in the recipient strain indicated that the HPI was transferred and integrated together with HPI-flanking DNA regions of the donor strain. The results of this study demonstrate for the first time that conjugative transfer and homologous DNA recombination play a major role in horizontal transfer of a pathogenicity island within the species E. coli.
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Evolución Biológica , Escherichia coli/patogenicidad , Transferencia de Gen Horizontal , Islas Genómicas/genética , Secuencia de Bases , Conjugación Genética/genética , Escherichia coli/genética , Filogenia , Recombinación Genética , Virulencia/genéticaRESUMEN
BACKGROUND AND OBJECTIVES: One of the most difficult problems in tumor surgery is the treatment of pelvic tumors, particularly those in the periacetabular region. This retrospective study serves to analyze clinical and functional outcome of the new surgical technique of hip transposition. PATIENTS AND METHODS: Sixty-two patients were analyzed (chondrosarcoma 15, Ewing's sarcoma or PNET 26, osteosarcoma 14, synovial cell sarcoma 2, others 5) and clinical, oncological, and functional outcome were evaluated. RESULTS: The surgical procedure focused on hip transpositions Type I (n = 9), Type IIa using the MUTARS attachment tube (n = 36), and Type IIb using the MUTARS attachment tube with a proximal femur endoprosthesis and bipolar cup (n = 17). In 53 patients (85.5%), wide margins were achieved. The 1- and 5-year overall survival rates were 96.4% and 80.5%, respectively. The 5-year (10 years) overall survival was 82.1% (82.1%) for Ewing's sarcoma, 60.1% (40.6%) for osteosarcoma, and 92.3% (46.2%) for chondrosarcoma. Wound healing problems were observed in 14 cases, deep infection in 20 cases. The mean Musculoskeletal Tumor Society (MSTS) score was 62% (18.6 Pts ± 3.73). CONCLUSION: Hip transposition can be considered as an easy and safe surgical procedure to employ in order to achieve good functional outcome in periacetabular tumors.
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Acetábulo/cirugía , Artroplastia de Reemplazo de Cadera/métodos , Recuperación del Miembro , Neoplasias Pélvicas/cirugía , Adolescente , Adulto , Niño , Femenino , Humanos , Masculino , Persona de Mediana Edad , Estudios Retrospectivos , Resultado del TratamientoRESUMEN
Infections due to extraintestinal pathogenic Escherichia coli (ExPEC) are common in humans and animals and include urinary tract infections (from uropathogenic E. coli [UPEC]), septicemia, and wound infections. These infections result in significant morbidity and mortality and in high health care costs. In view of the increasing number of ExPEC infections and the ever-growing antibiotic resistance capability of ExPEC isolates, preventive measures such as an effective vaccine against ExPEC are desirable. An ExPEC vaccine may be cost-effective for select patient groups. Previous vaccine candidates consisted of single target proteins or whole ExPEC cells. Here we describe a subunit vaccine against ExPEC which is based on immunodominant epitopes of the virulence-associated ExPEC proteins FyuA, IroN, ChuA, IreA, Iha, and Usp. Using a novel approach of computer-aided design, two completely artificial genes were created, both encoding eight peptide domains derived from these ExPEC proteins. The recombinant expression of these two genes resulted in a protein vaccine directed against ExPEC but not against commensal E. coli of the gut flora. In mice, the vaccine was highly immunogenic, eliciting both strong humoral and cellular immune responses. Nasal application resulted in high secretory immunoglobulin A (sIgA) production, which was detectable on the mucosal surface of the urogenital tract. Finally, it conveyed protection, as shown by a significant reduction of bacterial load in a mouse model of ExPEC peritonitis. This study provides evidence that a novel vaccine design encompassing distinct epitopes of virulence-associated ExPEC proteins may represent a means for providing a protective and pathogen-specific vaccine.
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Infecciones por Escherichia coli/prevención & control , Proteínas de Escherichia coli/inmunología , Vacunas contra Escherichia coli/inmunología , Administración Intranasal , Animales , Anticuerpos Antibacterianos/sangre , Epítopos/genética , Epítopos/inmunología , Infecciones por Escherichia coli/inmunología , Proteínas de Escherichia coli/administración & dosificación , Proteínas de Escherichia coli/genética , Vacunas contra Escherichia coli/administración & dosificación , Vacunas contra Escherichia coli/genética , Femenino , Inmunidad Mucosa , Inmunoglobulina A Secretora/análisis , Inmunoglobulina A Secretora/inmunología , Linfocitos/inmunología , Ratones , Ratones Endogámicos BALB C , Peritonitis/inmunología , Peritonitis/prevención & control , Vacunas de Subunidad/administración & dosificación , Vacunas de Subunidad/genética , Vacunas de Subunidad/inmunología , Vacunas Sintéticas/administración & dosificación , Vacunas Sintéticas/genética , Vacunas Sintéticas/inmunologíaRESUMEN
Extraintestinal pathogenic Escherichia coli (ExPECs) possess an armament of virulence factors to colonize and infect the host, such as adhesins, toxins, capsules and iron-uptake systems. Recently, we could identify a novel virulence factor of ExPECs that interferes with the innate immune response of the host by interrupting the NF-κB signaling pathway. This protein named TcpC shows considerable homology to motifs of the Tir domain of Toll-like receptors. Here we demonstrate that the tcpC gene is widely distributed among clinical ExPEC isolates with almost half of the E. coli strains from patients suffering pyelonephritis shown to be tcpC positive as compared to only 8% in commensal isolates. However, this gene is only present in phylogenetic group B2 strains. Interestingly, the tcpC gene is strongly associated with presence of the high-pathogenicity island (HPI). The phylogenetic history of the tcpC gene, in the E. coli reference collection (ECOR) and other well-defined E. coli strains, compared to the phylogenetic histories of the HPI and the strains, showed that the tcpC gene (i) is scattered among various B2 subgroups with specific O-types, (ii) has a phylogeny incongruent with the strain phylogeny, but (iii) congruent with the HPI phylogenetic history. This, together with the strong conservation of the tcpC gene, indicates a very recent introduction of this virulence factor into E. coli by horizontal gene transfer which occurred "en bloc" with the HPI at one major hot spot of recombination in the E. coli genome. The present data provide evidence for a strong impact of homologous recombination events in the spread of the TcpC virulence trait among E. coli.
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Proteínas de Escherichia coli/genética , Escherichia coli/clasificación , Escherichia coli/patogenicidad , Factores de Virulencia/genética , Adulto , Análisis por Conglomerados , ADN Bacteriano/química , ADN Bacteriano/genética , Escherichia coli/genética , Orden Génico , Transferencia de Gen Horizontal , Islas Genómicas , Humanos , Recién Nacido , Datos de Secuencia Molecular , Filogenia , Reacción en Cadena de la Polimerasa , Recombinación Genética , Análisis de Secuencia de ADN , Homología de SecuenciaRESUMEN
This review provides a survey of studies investigating physical activity and exercise interventions in patients during tumor treatment and survivors of childhood cancer. PubMed and Medline databases were searched using relevant terms. References of selected papers were tracked. A total of 28 studies could be identified. Seventeen studies investigated physical activity, 11 studies determined the effect of activity enhancing interventions during and after therapy. Even though most studies showed limitations and results were not consistent, considerably reduced physical activity is highly probable in patients during and after therapy. Studies on interventions provided promising results and revealed challenges to be faced.
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Terapia por Ejercicio , Actividad Motora/fisiología , Neoplasias/rehabilitación , Neoplasias/terapia , Niño , Ensayos Clínicos como Asunto , HumanosRESUMEN
Experimental autoimmune uveitis (EAU) in the Lewis rat has been regarded as an acute and monophasic disease. Uveitis can be induced by immunization with retinal soluble antigen (S-Ag), interphotoreceptor retinoid-binding protein (IRBP) or their peptide derivatives (PDSAg from S-Ag and R14 from IRBP) in CFA as well as by the transfer of activated, antigen-specific T cells. Previously, it has been shown that adoptively transferred, IRBP peptide-specific, but not S-Ag peptide-specific T cells can induce relapsing uveitis in rats. We observed spontaneous recurrences of intra-ocular inflammation even after immunization with R14 in CFA and were able to experimentally re-induce uveitis in rats previously immunized with autoantigen peptide in CFA. The efficiency of re-induction was dependent on the mode of pre-treatment [immunization or adoptive transfer (AT)] as well as on the antigen itself. Primary PDSAg-responses prevented subsequent re-induction of disease much more efficiently than primary R14-mediated EAU. In our model, the suppressive effect of CFA did not play a key role in preventing re-induction or spontaneous relapses. Furthermore, epitope spreading could not be demonstrated as a cause for recurrent inflammation. These data suggest that autoimmune responses with different antigen specificities could underlie similar clinical pictures while being differently regulated, which may help explain the variations in the disease courses in patients and the differential responses to therapeutic modalities.
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Antígenos/inmunología , Proteínas de Unión al Retinol/inmunología , Uveítis/inmunología , Animales , Proliferación Celular , Femenino , Inmunización , Ganglios Linfáticos/citología , Masculino , Ratas , Ratas Endogámicas Lew , Recurrencia , Especificidad del Receptor de Antígeno de Linfocitos T , Factores de Tiempo , Uveítis/patologíaRESUMEN
BACKGROUND: The diagnosis of cancer bears severe implications for pediatric patients. One immense restriction consists in a reduced level of activity due to different factors. Physical activity affects various aspects of development and can be regarded as an essential part of a child's life. In the present study physical activity in patients undergoing cancer therapy was quantified in order to determine the extent of the restriction and to provide baseline information for the assessment of possible interventions. PROCEDURE: Physical activity in 80 patients and 45 healthy children matched for age and gender was measured using the StepWatch 3 Activity Monitor (SAM, OrthoCare Innovations). RESULTS: Pediatric cancer patients, at 2,787 gait cycles (gcs) per day, were significantly less active than their healthy counterparts (8,096 gcs). Patients were significantly more active at home than during inpatient stays (3,185 gcs compared to 1,830 gcs), and patients with bone tumors were less active than those with leukemia regarding both, the amount (1,849 gcs vs. 2,992 gcs) and the intensity of activity. CONCLUSION: The present study quantified an often observed but so far hardly assessable problem. Activity in cancer patients is considerably reduced and patients with bone tumors are at increased risk from the detrimental effects of prolonged inactivity. Both our findings and the accepted fact that activity is an essential element of child development confirm the need for interventions tailored to a patient's needs and abilities during the course of treatment.
Asunto(s)
Actividad Motora , Neoplasias/fisiopatología , Adolescente , Neoplasias Óseas/fisiopatología , Neoplasias Óseas/terapia , Niño , Preescolar , Marcha , Humanos , Leucemia/fisiopatología , Leucemia/terapia , Neoplasias/terapiaRESUMEN
Purpose Significantly improved survival rates in patients with Ewing sarcoma have raised interest in accessing the quality of long-term survivorship. In this study, subjective and objective measurement tools, preclassified as physical or mental scores, were used to assess clinicofunctional outcome and physical activity after intensive bone tumor treatment. Methods Long-term outcome of 618 survivors from consecutive Ewing sarcoma trials was assessed by the Toronto Extremity Salvage Score, Short-Form Health Survey (SF-36), Brief Symptom Inventory (BSI), and Rosenberg Self-Esteem Scale questionnaires and by the accelerometric StepWatch 3 Activity Monitor. Prospective measurements were correlated retrospectively with standardized primary trial data. Results were compared with 316 nonrandom healthy peers by using effect sizes ( d). Median observation time was 12.9 years from primary diagnosis (range, 3.7 to 31.2 years). Results Absolute subjective scores were moderate to good for survivors. Compared with control subjects, unfavorable outcome was shown on physical Toronto Extremity Salvage Score, SF-36 Physical Component Summary, and BSI-Somatization scales (| d| ≥ 0.50; P < .01), in contrast to SF-36 Mental Component Summary, BSI-Anxiety, BSI-Depression, and Rosenberg Self-Esteem Scale mental scales (| d| ≤ 0.31). Survivors were less active than control subjects, as demonstrated by a step count difference of 1,742 steps per day ( d = -0.43; P < .01); however, on average, the recommended level for an active lifestyle was achieved (≥ 10,000 steps). Location of pelvic tumor was the major inferior disease-specific prognostic factor in physical scores ( P < .01), whereas nondisease-specific inferior factors in questionnaires were older age and female sex ( P < .01). Conclusion Survivors of Ewing sarcoma apparently returned to a normal life with minor limitations. Observed reductions in physical scores should be a focus in future research to optimize treatment strategies to reduce a negative impact on the quality of survivorship.
Asunto(s)
Neoplasias Óseas/fisiopatología , Neoplasias Óseas/psicología , Ejercicio Físico , Sarcoma de Ewing/fisiopatología , Sarcoma de Ewing/psicología , Adolescente , Adulto , Niño , Preescolar , Estudios de Cohortes , Femenino , Humanos , Masculino , Pronóstico , Calidad de Vida , Sobrevivientes , Adulto JovenRESUMEN
In the present in vivo study, the regenerative potential of a new injectable bone substitute (IBS) composed of beta-tricalcium phosphate (ß-TCP) and hyaluronan was tested in a rabbit distal femoral condyle model. To achieve this, 2 defects of 6 mm in diameter and 10 mm in length were drilled into each femur condyle in a total of 12 animals. For each animal, 1 hole was filled with the substitute material, and the other was left empty to serve as the control. After 1, 3, and 6 months, the regenerative process was analyzed by radiography as well as by histological and histomorphometrical analysis. The results revealed that bone tissue formation took place through osteoconductive processes over time, starting from the defect borders to the center. Both the ß-TCP content and the hydrogel support bone tissue growth. The histomorphometrical measurements showed that the amount of bone formation in the experimental group was significantly higher compared with that found in the control group after 3 months (19.51 ± 5.08% vs. 1.96 ± 0.77%, P < .05) and 6 months (4.57 ± 1.56% vs. 0.23 ± 0.21%, P < .05). The application of the IBS gave a restitutio ad integrum result after 6 months and was associated with its nearly complete degradation, in contrast to the results found in the control group. In conclusion, the results of the present study demonstrate that the IBS contributes to sufficient bone regeneration by serving as a scaffold-like structure, combined with its degradation within 6 months.