ABSTRACT
BACKGROUND: Patient-reported outcomes (PROs) play a crucial role in assessing rheumatic diseases, offering insights into disease evaluation and treatment efficacy. This study focuses on PRO assessment in large vessel vasculitides, including Takayasu Arteritis and Giant Cell Arteritis (GCA). METHODS: We retrospectively analyzed routine data from patients treated at our rheumatology clinic over a 10-year span. Patient and physician-rated global disease activity scale (G-DAS) scores, measured on a numeric rating scale (0-10 points), were collected at each visit. Clinical variables like age, sex, body mass index (BMI), disease duration, lab values, pain perception, and questionnaire responses were recorded. Linear regression and generalized additive linear regression (GAM analysis) examined associations between PROs and these factors. RESULTS: The study included 138 patients, primarily diagnosed with GCA (94.4%). Mean follow-up was 2.5 years (0-7.7). Patient and physician G-DAS exhibited a moderate correlation (Pearson R 0.19, CI 0.14-0.24, p < 0.001). Higher patient G-DAS correlated with younger age (CI -3.4 - -1.5, p < 0.001), increased pain (CI 3.5-4, p < 0.001), functional limitations (HAQ, CI 0.5-0.6, p < 0.001), reduced physical (CI 2.3-2.7, p ≤ 0.001) and psychological well-being (CI 2.1-2.5, p < 0.001), and higher BMI (CI 1.3-2.4, p < 0.001). Physician G-DAS correlated with Birmingham Vasculitis Activity Score (V3.0; R 0.42, p 0.046) and were significantly linked to serum CRP elevations (ß = 0.04, CI 0.0-0.08, p 0.028). CONCLUSIONS: These findings underscore the need to integrate PRO measures into vasculitis disease management strategies, enhancing the understanding of disease activity from the patient's perspective.
Subject(s)
Giant Cell Arteritis , Takayasu Arteritis , Humans , Retrospective Studies , Giant Cell Arteritis/diagnosis , Takayasu Arteritis/diagnosis , Patient Reported Outcome Measures , Ambulatory Care FacilitiesABSTRACT
The microbiome of freshwater sponges is rarely studied, and not a single novel bacterial species has been isolated and subsequently characterized from a freshwater sponge to date. A previous study showed that 14.4% of the microbiome from Ephydatia fluviatilis belong to the phylum Planctomycetes. Therefore, we sampled an Ephydatia sponge from a freshwater lake and employed enrichment techniques targeting bacteria from the phylum Planctomycetes. The obtained strain spb1T was subject to genomic and phenomic characterization and found to represent a novel planctomycetal species proposed as Planctopirus ephydatiae sp. nov. (DSM 106606â¯=â¯CECT 9866). In the process of differentiating spb1T from its next relative Planctopirus limnophila DSM 3776T, we identified and characterized the first phage - Planctopirus phage vB_PlimS_J1 - infecting planctomycetes that was only mentioned anecdotally before. Interestingly, classical chemotaxonomic methods would have failed to distinguish Planctopirus ephydatiae strain spb1T from Planctopirus limnophila DSM 3776T. Our findings demonstrate and underpin the need for whole genome-based taxonomy to detect and differentiate planctomycetal species.
Subject(s)
Phylogeny , Planctomycetales/classification , Porifera/microbiology , Animals , Fresh Water , Microbiota , Planctomycetales/isolation & purificationABSTRACT
After a minor blow to the neck from the handlebars of a bike, a 5-year-old boy developed a massive subcutaneous emphysema with respiratory distress. Orotracheal intubation was performed. A computed tomography (CT) scan of the neck and thorax showed a pneumomediastinum and a bilateral pneumothorax. No injury to the large airways was identified. The patient was stabilized by insertion of chest tubes and controlled ventilation. The endoscopic examination of the trachea revealed a tear of the pars membranacea, which was successfully treated conservatively. The specific features of the injury and the airway management are discussed based on a review of the current literature.
Subject(s)
Neck Injuries/diagnostic imaging , Subcutaneous Emphysema/therapy , Airway Management , Child, Preschool , Endoscopy , Humans , Intubation, Intratracheal , Male , Neck Injuries/therapyABSTRACT
In the last few years, the Biophysics Working Group of the Institute of Aerospace Medicine of the German Aerospace Center (DLR) started the development of a small low power consumption radiation detector system for the measurement of the absorbed dose to be applied in various environments, such as onboard aircraft, in space, and also as a demonstration tool for students. These so called DLR M-42 detectors are based on an electronics design, which can easily be adjusted to the user- and mission-requirements. M-42 systems were already applied for measurements in airplanes, during two MAPHEUS (Materialphysikalische Experimente unter Schwerelosigkeit) rocket missions, and are currently prepared for long term balloon experiments. In addition, they will be part of the dosimetry suite of the upcoming Matroshka AstroRad Radiation Experiment on the NASA Artemis I mission. This paper gives an overview of the design and the testing of the DLR M-42 systems and provides highlighted results from the MAPHEUS campaigns where the detectors were tested for the first time under space flight conditions. Results clearly show that the system design enables independent measurements starting upon rocket launch due to the built-in accelerometer sensors and provides data for the relevant 6 min of µ-gravity as given for the MAPHEUS missions. These 6 min of the µ-gravity environment at altitudes between 100 and 240 km lead to a total absorbed dose of 1.21 ± 0.15 µGy being equivalent to half a day of radiation background measured with the M-42 in the laboratory at DLR, Cologne, Germany.
ABSTRACT
Enzootic pneumonia incurs major economic losses to pork production globally. The primary pathogen and causative agent, Mycoplasma hyopneumoniae, colonises ciliated epithelium and disrupts mucociliary function predisposing the upper respiratory tract to secondary pathogens. Alleviation of disease is reliant on antibiotics, vaccination, and sound animal husbandry, but none are effective at eliminating M. hyopneumoniae from large production systems. Sustainable pork production systems strive to lower reliance on antibiotics but lack of a detailed understanding of the pathobiology of M. hyopneumoniae has curtailed efforts to develop effective mitigation strategies. M. hyopneumoniae is considered an extracellular pathogen. Here we show that M. hyopneumoniae associates with integrin ß1 on the surface of epithelial cells via interactions with surface-bound fibronectin and initiates signalling events that stimulate pathogen uptake into clathrin-coated vesicles (CCVs) and caveosomes. These early events allow M. hyopneumoniae to exploit an intracellular lifestyle by commandeering the endosomal pathway. Specifically, we show: (i) using a modified gentamicin protection assay that approximately 8% of M. hyopneumoniae cells reside intracellularly; (ii) integrin ß1 expression specifically co-localises with the deposition of fibronectin precisely where M. hyopneumoniae cells assemble extracellularly; (iii) anti-integrin ß1 antibodies block entry of M. hyopneumoniae into porcine cells; and (iv) M. hyopneumoniae survives phagolysosomal fusion, and resides within recycling endosomes that are trafficked to the cell membrane. Our data creates a paradigm shift by challenging the long-held view that M. hyopneumoniae is a strict extracellular pathogen and calls for in vivo studies to determine if M. hyopneumoniae can traffic to extrapulmonary sites in commercially-reared pigs.
Subject(s)
Epithelial Cells/microbiology , Mycoplasma hyopneumoniae/pathogenicity , Pneumonia of Swine, Mycoplasmal/microbiology , Animals , Cell Membrane/metabolism , Cell Membrane/microbiology , Endosomes/metabolism , Endosomes/microbiology , Epithelial Cells/metabolism , Fibronectins/metabolism , Integrin beta1/metabolism , Pneumonia of Swine, Mycoplasmal/metabolism , SwineABSTRACT
Microfluidic synthesis techniques can offer improvement over batch syntheses which are currently used for radiopharmaceutical production. These improvements are, for example, better mixing of reactants, more efficient energy transfer, less radiolysis, faster reaction optimization, and overall improved reaction control. However, scale-up challenges hinder the routine clinical use, so the main advantage is currently the ability to optimize reactions rapidly and with low reactant consumption. Translating those results to clinical systems could be done based on calculations, if kinetic constants and diffusion coefficients were known. This study describes a microfluidic system with which it was possible to determine the kinetic association rate constants for the formation of [177Lu]Lu-DOTA-TATE under conditions currently used for clinical production. The kinetic rate constants showed a temperature dependence that followed the Arrhenius equation, allowing the determination of Arrhenius parameters for a Lu-DOTA conjugate (A = 1.24 ± 0.05 × 1019 M-1 s-1, EA = 109.5 ± 0.1 × 103 J mol-1) for the first time. The required reaction time for the formation of [177Lu]Lu-DOTA-TATE (99% yield) at 80 °C was 44 s in a microfluidic channel (100 µm). Simulations done with COMSOL Multiphysics® indicated that processing clinical amounts (3 mL reaction solution) in less than 12 min is possible in a micro- or milli-fluidic system, if the diameter of the reaction channel is increased to over 500 µm. These results show that a continuous, microfluidic system can become a viable alternative to the conventional, batch-wise radiolabelling technique.
ABSTRACT
While the oral health-related quality of life (OHRQoL) is known to be reduced in patients with cleft lip and palate (CLP), its inter-dependency with the soft tissue characteristics of the CLP area remains unclear. This study aimed to evaluate the soft tissue characteristics in the treated cleft area in order to investigate whether gingival esthetics correlate with OHRQoL. Thirty-six patients with unilateral or bilateral CLP (46 cleft areas) were investigated after secondary/tertiary alveolar bone grafting and orthodontic/prosthetic implant treatment using an adapted score to rate gingival esthetics (clinical esthetic score, CES). The patient's OHRQoL was determined using the German short version of the Oral Health Impact Profile questionnaire (OHIP-G14). The results showed a significantly better rating in patients with their own teeth in situ (12.05±1.10) than in patients with implants (6.95±4.78) or prosthetics (4.00±3.58). The best OHRQoL values were achieved by patients with their own teeth integrated into the cleft area (1.32±2.31), followed by patients with implants (2.33±2.33) and prosthetics (3.75±5.87). A significant (P=0.017) correlation was found between OHIP-G14 and CES scores, suggesting an increased OHRQoL in cases with higher oral esthetics in the cleft area. The therapeutic strategy contributes to both gingival esthetics and OHRQoL. The patient's subjective perception of OHRQoL can be attributed to objective gingival esthetic ratings.
Subject(s)
Cleft Lip/surgery , Cleft Palate/surgery , Esthetics, Dental , Gingiva/anatomy & histology , Oral Health , Quality of Life , Adolescent , Adult , Female , Humans , Male , Plastic Surgery Procedures , Surveys and QuestionnairesABSTRACT
BACKGROUND: Neopterin is produced by activated macrophages upon stimulation with interferon-γ (IFN-γ) and thus, elevated neopterin concentrations in patients indicate cellular inate immune response. Most studies in patients with malignant diseases found an association between higher neopterin concentrations and reduced survival and impaired prognosis. Nevertheless, neopterin is not a classical tumor marker since it is not produced by the cancer cells themselves. PATIENTS AND METHODS: In a study conducted by the Austrian Gynecologic Oncology Group (AGO) in 114 patients with ovarian cystadenomas and 223 patients with invasive ovarian cancer, patients' urinary neopterin was determined before and after primary therapy. The relevance of neopterin in long-term median follow-up was assessed. RESULTS: Elevated levels (cut-off 250 µmol/mol creatinine) were found less frequently in women with benign ovarian cystadenomas (24%) than in patients with malignant disease (58%). After 10 years, only 57% of ovarian cancer patients with elevated urinary neopterin levels survived without disease progression following primary therapy when compared with 86% of women with normal levels (P < 0.001). Along with residual tumor, FIGO stage, age and histological type, neopterin was significantly associated with overall survival (OS) and progression-free survival (PFS). The median PFS was 52 and 12 months and the median OS was 81 and 24 months for patients with normal and elevated neopterin, respectively, P < 0.001. In a multivariate Cox regression analysis, only residual tumor, neopterin and age were independently associated with OS, while only residual tumor was predictive for PFS. Thirty patients with early-stage invasive ovarian cancer (FIGO I and II) were analyzed separately. Of 3 patients with elevated neopterin, 2 died of disease in contrast to 2 out of 27 patients with normal neopterin (P = 0.004). CONCLUSION: In ovarian cancer, the negative impact of elevated urinary neopterin levels indicates a detrimental effect of cancer-associated inflammatory reaction.
Subject(s)
Biomarkers, Tumor/urine , Immunity, Innate/drug effects , Neopterin/urine , Ovarian Neoplasms/urine , Adult , Aged , Austria , Disease-Free Survival , Female , Humans , Interferon-gamma/administration & dosage , Interferon-gamma/urine , Middle Aged , Neoplasm Staging , Neoplasm, Residual/pathology , Neoplasm, Residual/urine , Ovarian Neoplasms/drug therapy , Ovarian Neoplasms/pathologyABSTRACT
BACKGROUND: In recent years interactions between surgical treatment of oral carcinoma and incidence of anxiety or depression have become a subject of discussions. This prospective study is a comparison between the extent of loss of speech intelligibility and presence of depressive symptoms or anxiety as a result of oral carcinoma. MATERIAL AND METHODS: One year after surgical therapy for oral carcinoma, 90 patients of an average age of 60±12 years were examined. Their speech intelligibility degree was measured using standardized automatic speech recognition (word recognition rate, WR). Symptoms of anxiety and depression were detected by use of HAD-Scales (HADS). Next to the relationship between WR and HADS other influential variables related to WR and HADS were statistically evaluated. RESULTS: The WR average was 53.2±17.2. Female WR was better than male. The difference between tumor classifications T1 and T4 compared to the WR reached statistical significance. Significant differences were detected between WR and "tumor localization", "graft donor site", "graft morphology", "tongue motility", and "tracheostoma" groups. There was a relationship between tongue motility and graft morphology, graft donor site and tumor localization. HAD-Scores in the mean were elevated: HADS-Total=43.3%, HADS-A=43.3% und HADS-D= 51.1%. WR correlates with HADS-D-Subscale, but not with HADS-A-Subscale. CONCLUSION: Communication disorders as a result of neoplasmic orofacial surgery may be related to extent of the treatment and to affective impairments. This should receive attention in the concept of rehabilitation.
Subject(s)
Anxiety Disorders , Mouth Neoplasms/surgery , Speech Intelligibility , Aged , Anxiety , Female , Humans , Male , Middle Aged , Prospective StudiesABSTRACT
The control of Johne's disease requires the identification of Mycobacterium avium ssp. paratuberculosis (MAP)-positive herds. Boot swabs and liquid manure samples have been suggested as an easy-to-use alternative to sampling individual animals in order to diagnose subclinical Johne's disease at the herd level, but there is a need to evaluate performance of this approach in the field. Using a logistic regression model, this study aimed to calculate the threshold level of the apparent within-herd prevalence as determined by individual faecal culture, thus allowing the detection of whether a herd is MAP positive. A total of 77 boot swabs and 75 liquid manure samples were taken from 19 certified negative and 58 positive dairy herds. Faecal culture, three different polymerase chain reaction (PCR) methods and the combination of faecal culture with PCR were applied in order to detect MAP. For 50% probability of detection, a within-herd prevalence threshold of 1·5% was calculated for testing both matrices simultaneously by faecal culture and PCR, with the threshold increased to 4·0% for 90% probability of detection. The results encourage the use of boot swabs or liquid manure samples, or a combination both, for identifying MAP-positive herds and, to a certain extent, for monitoring certified Johne's disease-negative cattle herds.
Subject(s)
Cattle Diseases/epidemiology , Mycobacterium avium subsp. paratuberculosis/isolation & purification , Paratuberculosis/epidemiology , Animals , Cattle , Cattle Diseases/microbiology , Female , Germany/epidemiology , Logistic Models , Manure/microbiology , Paratuberculosis/microbiology , Prevalence , Real-Time Polymerase Chain Reaction/veterinarySubject(s)
Biomarkers, Tumor/genetics , Leukemia-Lymphoma, Adult T-Cell/classification , Mutation/genetics , Precursor T-Cell Lymphoblastic Leukemia-Lymphoma/classification , Adolescent , Adult , Child , Cohort Studies , Female , Humans , Leukemia-Lymphoma, Adult T-Cell/genetics , Leukemia-Lymphoma, Adult T-Cell/pathology , Male , Neoplasm Staging , Precursor T-Cell Lymphoblastic Leukemia-Lymphoma/genetics , Precursor T-Cell Lymphoblastic Leukemia-Lymphoma/pathology , Prognosis , Risk AssessmentABSTRACT
Bacteria of many species are able to invade and colonize solid tumours in mice. We have focused on Salmonella enterica serovar Typhimurium. Detailed analysis revealed that such tumour-invading Salmonella form biofilms, thus providing a versatile in vivo test system for studying bacterial phenotypes and host-pathogen interactions. It appears that biofilm formation by S. typhimurium is induced as a defence against the immune system of the host, and in particular against neutrophils. Further, we extended our work to the clinically more relevant biofilm infection by Pseudomonas aeruginosa. The induction of P. aeruginosa biofilms in neoplastic tissue appears to be elicited as a reaction against the immune system. Reconstitution experiments reveal that T cells are responsible for biofilm induction. Isogenic mutants that are no longer able to form biofilms can be used for comparison studies to determine antimicrobial resistance, especially therapeutic efficacy against P. aeruginosa located in biofilms.
Subject(s)
Biofilms/growth & development , Host-Pathogen Interactions , Neoplasms, Experimental/microbiology , Pseudomonas aeruginosa/growth & development , Salmonella typhimurium/growth & development , Animals , Antibiosis , Mice , Neutrophils/microbiology , Phagocytosis , Pseudomonas aeruginosa/immunology , Salmonella Infections, Animal/microbiology , Salmonella typhimurium/immunology , T-Lymphocytes/microbiologyABSTRACT
AIMS: Improvement of clinical diagnostics of idiopathic giant cell myocarditis (IGCM) and cardiac sarcoidosis (CS), two frequently fatal human myocardial diseases. Currently, IGCM and CS are diagnosed based on differential patterns of inflammatory cell infiltration and non-caseating granulomas in histological sections of endomyocardial biopsies (EMBs), after heart explantation or postmortem. We report on a method for improved differential diagnosis by myocardial gene expression profiling in EMBs. METHODS AND RESULTS: We examined gene expression profiles in EMBs from 10 patients with histopathologically proven IGCM, 10 with CS, 18 with active myocarditis (MCA), and 80 inflammation-free control subjects by quantitative RT-QPCR. We identified distinct differential profiles that allowed a clear discrimination of tissues harbouring giant cells (IGCS, CS) from those with MCA or inflammation-free controls. The expression levels of genes coding for cytokines or chemokines (CCL20, IFNB1, IL6, IL17D; P < 0.05), cellular receptors (ADIPOR2, CCR5, CCR6, TLR4, TLR8; P < 0.05), and proteins involved in the mitochondrial energy metabolism (CPT1, CYB, DHODH; P < 0.05) were deregulated in 2- to 300-fold, respectively. Bioinformatic analyses and correlation of the gene expression data with immunohistochemical findings provided novel information regarding the differential cellular and molecular pathomechanisms in IGCM, CS, and MCA. CONCLUSION: Myocardial gene expression profiling is a reliable method to predict the presence of multinuclear giant cells in the myocardium, even without a direct histological proof, in single small EMB sections, and thus to reduce the risk of sampling errors. This profiling also facilitates the discrimination between IGCM and CS, as two different clinical entities that require immediate and tailored differential therapy.
Subject(s)
Cardiomyopathies/diagnosis , Gene Expression Profiling/methods , Sarcoidosis/diagnosis , DNA, Complementary/metabolism , Female , Humans , Immunohistochemistry , Male , Middle Aged , Myocarditis/diagnosis , Retrospective StudiesABSTRACT
Streptococcus pneumoniae is not only a commensal of the nasopharyngeal epithelium, but may also cause life-threatening diseases. Immune-electron microscopy studies revealed that the bacterial glycolytic enzyme, phosphoglycerate kinase (PGK), is localised on the pneumococcal surface of both capsulated and non-capsulated strains and colocalises with plasminogen. Since pneumococci may concentrate host plasminogen (PLG) together with its activators on the bacterial cell surface to facilitate the formation of plasmin, the involvement of PGK in this process was studied. Specific binding of human or murine PLG to strain-independent PGK was documented, and surface plasmon resonance analyses indicated a high affinity interaction with the kringle domains 1-4 of PLG. Crystal structure determination of pneumococcal PGK together with peptide array analysis revealed localisation of PLG-binding site in the N-terminal region and provided structural motifs for the interaction with PLG. Based on structural analysis data, a potential interaction of PGK with tissue plasminogen activator (tPA) was proposed and experimentally confirmed by binding studies, plasmin activity assays and thrombus degradation analyses.
Subject(s)
Bacterial Proteins/metabolism , Phosphoglycerate Kinase/metabolism , Plasminogen/metabolism , Streptococcus pneumoniae/physiology , Tissue Plasminogen Activator/metabolism , Animals , Bacterial Proteins/genetics , Cloning, Molecular , Crystallography, X-Ray , Fibrinolysin/metabolism , Humans , Laryngeal Mucosa/microbiology , Mice , Nasal Mucosa/microbiology , Phosphoglycerate Kinase/genetics , Protein Binding , Protein Conformation , Protein Interaction Domains and Motifs/genetics , Protein Transport , Surface Plasmon ResonanceABSTRACT
Suilysin is a pore-forming cholesterol-dependent cytolysin secreted by Streptococcus suis (S. suis), an important swine and zoonotic pathogen. The role of suilysin in S. suis host-cell interaction is still unclear. We found a higher adherence and invasion rate of an unencapsulated sly-positive strain in comparison to its isogenic sly-negative mutant. Electron microscopy revealed that formation of membrane ruffles accompanying invasion of the sly-positive strain was abolished in the sly-negative mutant. Inhibition experiments showed that the actin cytoskeleton was involved in suilysin-mediated effects. Point-mutation of the domain putatively responsible for macropore-formation resulted in abolished hemolytic and cytolysin activity, but had no effect on S. suis host cell association. Concluding, our results indicate that subcytolytic suilysin promotes S. suis association with epithelial cells.
Subject(s)
Hemolysin Proteins/pharmacology , Host-Pathogen Interactions/drug effects , Streptococcal Infections/microbiology , Streptococcus suis/drug effects , Streptococcus suis/physiology , Swine Diseases/microbiology , Animals , Cell Line , Epithelial Cells/microbiology , Hemolysin Proteins/toxicity , Humans , Mutation , Streptococcus suis/genetics , Streptococcus suis/ultrastructure , SwineABSTRACT
Recent studies have detected erythrovirus genomes in the hearts of cardiomyopathy and cardiac transplant patients. Assessment of the functional status of viruses may provide clinically important information beyond detection of the viral genomes. Here, we report transcriptional activation of cardiotropic erythrovirus to be associated with strongly altered myocardial gene expression in a distinct subgroup of cardiomyopathy patients. Endomyocardial biopsies (EMBs) from 415 consecutive cardiac erythrovirus (B19V)-positive patients with clinically suspected cardiomyopathy were screened for virus-encoded VP1/VP2 mRNA indicating transcriptional activation of the virus, and correlated with cardiac host gene expression patterns in transcriptionally active versus latent infections, and in virus-free control hearts. Transcriptional activity was detected in baseline biopsies of only 66/415 patients (15.9 %) harbouring erythrovirus. At the molecular level, significant differences between cardiac B19V-positive patients with transcriptionally active versus latent virus were revealed by expression profiling of EMBs. Importantly, latent B19V infection was indistinguishable from controls. Genes involved encode proteins of antiviral immune response, B19V receptor complex, and mitochondrial energy metabolism. Thus, functional mapping of erythrovirus allows definition of a subgroup of B19V-infected cardiomyopathy patients characterized by virus-encoded VP1/VP2 transcripts and anomalous host myocardial transcriptomes. Cardiac B19V reactivation from latency, as reported here for the first time, is a key factor required for erythrovirus to induce altered cardiac gene expression in a subgroup of cardiomyopathy patients. Virus genome detection is insufficient to assess pathogenic potential, but additional transcriptional mapping should be incorporated into future pathogenetic and therapeutic studies both in cardiology and transplantation medicine.
Subject(s)
Cardiomyopathies/genetics , Cardiomyopathies/virology , Parvoviridae Infections/virology , Transcriptome , Cardiomyopathies/complications , DNA, Viral/analysis , Female , Humans , Male , Middle Aged , Parvoviridae Infections/complications , Parvoviridae Infections/genetics , Parvovirus B19, Human/genetics , Reverse Transcriptase Polymerase Chain ReactionABSTRACT
Although Streptococcus (S.) canis is known to cause severe infections in dogs and cats and harbors a clear zoonotic potential, knowledge about physiology and pathogenesis is mostly elusive. The arginine deiminase system (ADS) has been described in certain streptococcal species and its role in the establishment of infection has been suggested. In this study we focused on the identification and characterization of the ADS in S. canis. Using genome sequencing and subsequent in silico analysis we identified the ADS of S. canis as a gene cluster composed of seven genes. RT-PCR analysis revealed that the ADS of S. canis is transcribed in four transcriptional units, comprising three monocistronical mRNAs and one operon structure. As a secondary metabolic pathway, the ADS of S. canis is strictly regulated by carbon catabolite repression (CCR) and arginine as demonstrated on transcriptional, translational, and enzymatical level, respectively. Furthermore, growth kinetics with a chemically defined medium clearly showed that arginine, the substrate of the ADS, is essential for the biological fitness of S. canis. Using Immuno-electron microscopy analysis, we observed a surface-exposed localization of the ADS enzymes arginine deiminase (ArcA), ornithine carbamoyltransferase (ArcB), and carbamate kinase (ArcC), respectively, which might suggest the contribution of the ADS to the development of streptococcal infections.
Subject(s)
Hydrolases/genetics , Hydrolases/metabolism , Streptococcus/enzymology , Arginine/metabolism , Base Sequence , Hydrolases/chemistry , Multigene Family , Operon , Ornithine Carbamoyltransferase/metabolism , Phosphotransferases (Carboxyl Group Acceptor)/metabolism , Streptococcal Infections/enzymology , Streptococcal Infections/microbiology , Streptococcus/genetics , Streptococcus/metabolismABSTRACT
A novel Gram-positive, aerobic, actinobacterial strain, CF5/5, was isolated from soil in the Sahara desert, Chad. It grew best at 20-35 °C and at pH 6.0-8.0 and with 0-4 % (w/v) NaCl, forming black-colored colonies. Chemotaxonomic and molecular characteristics of the isolate matched those described for members of the genus Geodermatophilus. The DNA G + C content was 75.9 mol%. The peptidoglycan contained meso-diaminopimelic acid; galactose and xylose were detected as diagnostic sugars. The main phospholipids were diphosphatidylglycerol, phosphatidylcholine, and phosphatidylinositol; MK-9(H(4)) was the dominant menaquinone. The major cellular fatty acids were: iso-C(16:0) and iso-C(15:0). The 16S rRNA gene showed 95.6-98.3 % sequence similarity with the other named members of the genus Geodermatophilus. Based on the polyphasic taxonomy data, the isolate is proposed to represent a novel species, Geodermatophilus saharensis with the type strain CF5/5(T) = DSM 45423 = CCUG 62813 = MTCC 11416.
Subject(s)
Actinomycetales/classification , Phylogeny , Silicon Dioxide , Soil Microbiology , Actinomycetales/chemistry , Actinomycetales/genetics , Actinomycetales/isolation & purification , Actinomycetales/ultrastructure , Cell Wall/chemistry , Chad , DNA, Bacterial/chemistry , DNA, Bacterial/genetics , Microscopy, Electron, Scanning , Molecular Sequence Data , RNA, Ribosomal, 16S/genetics , Species SpecificityABSTRACT
A novel Gram-positive, aerobic, actinobacterial strain, CF5/4(T), was isolated in 2007 during an environmental screening of arid desert soil in Ouré Cassoni, Chad. The isolate grew best in a temperature range of 28-40 °C and at pH 6.0-8.5, with 0-1 % (w/v) NaCl, forming brown-coloured and nearly circular colonies on GYM agar. Chemotaxonomic and molecular characteristics of the isolate matched those described for members of the genus Geodermatophilus. The DNA G + C content of the novel strain was 75.9 mol %. The peptidoglycan contained meso-diaminopimelic acid as diagnostic diaminoacid. The main phospholipids were phosphatidylethanolamine, phosphatidylcholine, phosphatidylinositol, diphosphatidylglycerol and a small amount of phosphatidylglycerol; MK-9(H(4)) was identified as the dominant menaquinone and galactose as diagnostic sugar. The major cellular fatty acids were branched-chain saturated acids: iso-C(15:0) and iso-C(16:0). The 16S rRNA gene showed 96.2-98.3 % sequence identity with the three members of the genus Geodermatophilus: G. obscurus (96.2 %), G. ruber (96.5 %), and G. nigrescens (98.3 %). Based on the chemotaxonomic results, 16S rRNA gene sequence analysis and DNA-DNA hybridization with the type strain of G. nigrescens, the isolate is proposed to represent a novel species, Geodermatophilus arenarius (type strain CF5/4(T) = DSM 45418(T) = MTCC 11413(T) = CCUG 62763(T)).
Subject(s)
Actinomycetales/classification , Actinomycetales/isolation & purification , Geologic Sediments/microbiology , Actinomycetales/cytology , Actinomycetales/genetics , Africa, Northern , Bacterial Typing Techniques , Base Composition , Chad , Desert Climate , Geologic Sediments/chemistry , Phylogeny , RNA, Ribosomal, 16S/genetics , Silicon DioxideABSTRACT
Parvovirus B19 is a frequent virus detected in endomyocardial biopsies of patients with clinically suspected myocarditis or dilated cardiomyopathy (DCM). Viruses often cause a more symptomatic disease with increased tissue injury if they become reactivated. A disease-specific differential expression of microRNAs (miRNAs) has been described in the regulation of replicating viruses. Analyzing patients with latent and reactivated B19V infection, we found 29 differentially regulated miRNAs and, in order to test whether predicted genes are differentially expressed, selected mRNAs were tested by TaqMan-QPCR.