Soft Tissue Infection of Immunocompetent Man with Cat-Derived Globicatella Species.

We report a novel Globicatella species causing extensive soft tissue infection in a man bitten by a stray domestic cat in the United Kingdom. We identified this bacterium by 16S rRNA gene sequencing, whole-genome sequencing, and biochemical profiling and determined antimicrobial drug susceptibility.

Quality of MALDI-TOF mass spectra in routine diagnostics: results from an international external quality assessment including 36 laboratories from 12 countries using 47 challenging bacterial strains.

OBJECTIVES: Matrix assisted laser desorption ionization-time of flight (MALDI-TOF) mass spectrometry (MS) is a widely used method for bacterial species identification. Incomplete databases and mass spectral quality (MSQ) still represent major challenges. Important proxies for MSQ are the number of detected marker masses, reproducibility, and measurement precision. We aimed to assess MSQs across diagnostic laboratories and the potential of simple workflow adaptations to improve it. METHODS: For baseline MSQ assessment, 47 diverse bacterial strains, which are challenging to identify by MALDI-TOF MS, were routinely measured in 36 laboratories from 12 countries, and well-defined MSQ features were used. After an intervention consisting of detailed reported feedback and instructions on how to acquire MALDI-TOF mass spectra, measurements were repeated and MSQs were compared. RESULTS: At baseline, we observed heterogeneous MSQ between the devices, considering the median number of marker masses detected (range = [2-25]), reproducibility between technical replicates (range = [55%-86%]), and measurement error (range = [147 parts per million (ppm)-588 ppm]). As a general trend, the spectral quality was improved after the intervention for devices, which yielded low MSQs in the baseline assessment as follows: for four out of five devices with a high measurement error, the measurement precision was improved (p-values <0.001, paired Wilcoxon test); for six out of ten devices, which detected a low number of marker masses, the number of detected marker masses increased (p-values <0.001, paired Wilcoxon test). DISCUSSION: We have identified simple workflow adaptations, which, to some extent, improve MSQ of poorly performing devices and should be considered by laboratories yielding a low MSQ. Improving MALDI-TOF MSQ in routine diagnostics is essential for increasing the resolution of bacterial identification by MALDI-TOF MS, which is dependent on the reproducible detection of marker masses. The heterogeneity identified in this external quality assessment (EQA) requires further study.

Branding Dynamics for the Esthetic Dentist: Building Your Brand to Build Your Practice.

The objective of this article is to introduce the concept of branding to dentists interested in implementing elective esthetic treatment into their practice. For many, this will serve as an introduction to begin; for others, it can provide a road map for revising and reinforcing a branding program already in place.

Bartonella clarridgeiae infection in a patient with aortic root abscess and endocarditis.

INTRODUCTION: Bartonella species are increasingly recognized as agents of culture-negative endocarditis. However, to date, almost all human cases have been associated with two members of the genus, Bartonella henselae and Bartonella quintana. B. henselae infections are zoonotic, with domestic cats serving as reservoir hosts for the pathogen. Bartonella clarridgeiae also exploits cats as reservoir hosts, but its zoonotic potential is far less established. CASE PRESENTATION: A 34-year-old male presented with palpitations after a history of aortic incompetence. During surgery for an aortic valve replacement, two vegetations were found on the aortic valve. PCR analysis of the vegetation demonstrated the presence of Bartonella species and so the patient was treated post-operatively with ceftriaxone and doxycycline, making a good recovery. Further PCR-based analysis of the patient's aortic vegetation confirmed the presence of B. clarridgeiae . CONCLUSION: This report expands the number of Bartonella species associated with endocarditis and provides clear evidence that B. clarridgeiae should be considered a zoonotic pathogen.

rpoB gene sequencing highlights the prevalence of an E. miricola cluster over other Elizabethkingia species among UK cystic fibrosis patients.

Difficulties in distinguishing species of the Elizabethkingia genus by MALDI-TOF prompted use of rpoB sequencing to investigate species distribution among 44 isolates from cystic fibrosis (CF) patients. Forty-three isolates from 38 patients formed a cluster comprising E. miricola and proposed novel species E. bruuniana sp. nov., the exception clustering with proposed species E. ursingii sp. nov., also part of this wider cluster. All 44 isolates were PCR-positive for urease gene ureG, whereas only one of 23 E. anophelis isolates from non-CF patients was positive, suggesting that this gene is largely associated with the E. miricola cluster. Antibiotic susceptibilities of 12 CF isolates revealed all were resistant to beta-lactams with the exception of piperacillin-tazobactam, and were only susceptible to minocycline and co-trimoxazole. Pulsed-field gel electrophoresis analysis revealed 4 shared strains among 17 CF patients in one pediatric clinic, but epidemiological investigations did not support patient-to-patient transmission except between one sibling pair.

The relationship between emotional experience with dyslexia and work self-efficacy among adults with dyslexia.

This study responds to a call for more research on working adults with dyslexia investigating how employment-related factors affect and relate to one another. Two important work-related factors are self-efficacy and emotional experience with dyslexia. Work self-efficacy is viewed one of the most vital intrapersonal capacities in the work environment. It can be viewed as a subjective indicator of work success and is conceptualized as a multidimensional construct. Research speaks to a unique emotional experience stemming from living with an often misunderstood and stereotyped learning difference. The participants were 173 working adults with dyslexia (average age = 43.5 years, females = 56.6%) who participated in a web-based survey. Hierarchical multiple regression analyses revealed that more negative or uncomfortable emotions emanating from living with dyslexia predicted lower levels of total work self-efficacy, work attributes, work competency, and work anxiety over and beyond background contextual variables. Implications of these findings are discussed.

Sphingobacterium spiritivorum infection in a patient with end stage renal disease on haemodialysis.

BACKGROUND: Sphingobacterium spiritivorum is a microorganism that is ubiquitously found in the environment. However, it is rarely isolated from human clinical specimens. There are few reports to date of Sphingobacterium spiritivorum causing disease in humans. CASE REPORT: We describe a case of Sphingobacterium spiritivorum infection in a patient on haemodialysis, which to our knowledge, has not been described before. Further testing revealed this strain was sensitive to multiple antimicrobials. CONCLUSION: Despite interrupted courses of several antibiotics, our patient clinically made a good recovery and continued to receive haemodialysis.

Case report: Actinomycetoma caused by Nocardia aobensis from Lao PDR with favourable outcome after short-term antibiotic treatment.

BACKGROUND: Mycetoma is a neglected, chronic, localized, progressively destructive, granulomatous infection caused either by fungi (eumycetoma) or by aerobic actinomycetes (actinomycetoma). It is characterized by a triad of painless subcutaneous mass, multiple sinuses and discharge containing grains. Mycetoma commonly affects young men aged between 20 and 40 years with low socioeconomic status, particularly farmers and herdsmen. METHODOLOGY/PRINCIPAL FINDINGS: A 30 year-old male farmer from an ethnic minority in Phin District, Savannakhet Province, Lao PDR (Laos) developed a painless swelling with multiple draining sinuses of his right foot over a period of approximately 3 years. X-ray of the right foot showed osteolysis of tarsals and metatarsals. Aerobic culture of sinus discharge yielded large numbers of Staphylococcus aureus and a slow growing Gram-positive rod. The organism was subsequently identified as Nocardia aobensis by 16S ribosomal RNA gene sequencing. The patient received antimicrobial treatment with amikacin and trimethoprim-sulfamethoxazole according to consensus treatment guidelines. Although slight improvement was noted the patient left the hospital after 14 days and did not take any more antibiotics. Over the following 22 weeks the swelling of his foot subsequently diminished together with healing of discharging sinuses. CONCLUSION: This is the first published case of Actinomycetoma caused by Nocardia aobensis and the second case of Actinomycetoma from Laos. A treatment course of only 14 days with amikacin and trimethoprim-sulfamethoxazole was apparently sufficient to cure the infection, although long-term treatment up to one year is currently recommended. Treatment trials or prospective descriptions of outcome for actinomycetoma should investigate treatment efficacy for the different members of Actinomycetales, particularly Nocardia spp., with short-term and long-term treatment courses.

Actinomycetoma in SE Asia: the first case from Laos and a review of the literature.

BACKGROUND: Mycetoma is a chronic, localized, slowly progressing infection of the cutaneous and subcutaneous tissues caused either by fungi (eumycetoma or implantation mycosis) or by aerobic actinomycetes (actinomycetoma). It is acquired by traumatic implantation, most commonly in the tropics and subtropics, especially in rural agricultural communities. Although well recognized elsewhere in Asia, it has not been reported from the Lao People's Democratic Republic (Laos). CASE PRESENTATION: A 30 year-old female elementary school teacher and rice farmer from northeast Laos was admitted to Mahosot Hospital, Vientiane, with a massive growth on her left foot, without a history of trauma. The swelling had progressed slowly but painlessly over 5 years and multiple draining sinuses had developed. Ten days before admission the foot had increased considerably in size and became very painful, with multiple sinuses and discharge, preventing her from walking. Gram stain and bacterial culture of tissue biopsies revealed a branching filamentous Gram-positive bacterium that was subsequently identified as Actinomadura madurae by 16S rRNA gene amplification and sequencing. She was treated with long-term co-trimoxazole and multiple 3-week cycles of amikacin with a good therapeutic response. CONCLUSION: We report the first patient with actinomycetoma from Laos. The disease should be considered in the differential diagnosis of chronic skin and bone infections in patients from rural SE Asia.

Utility of real-time amplification of selected 16S rRNA gene sequences as a tool for detection and identification of microbial signatures directly from clinical samples.

The potential of incorporating a real-time PCR for amplification and detection of 16S rRNA gene signatures directly from clinical samples was assessed as a tool for diagnostics. Universal PCR primers spanning short variable regions (~500 bp) were optimized for real-time PCR and tested in comparison with a longer fragment (~1400 bp) generated from block-based amplification. Real-time PCR had improved sensitivity of detection (8% increase), decreased amplification time and simplified downstream processing. The real-time PCR primers also offered an improvement in detection of bacteria from samples that demonstrated inhibition with the block-based primers and in the resolution of mixed-sequence traces. In addition to testing primer sensitivity, the effect of amplifying and sequencing two different variable regions of the 16S rRNA gene on organism identification was compared. By amplifying and sequencing a shorter variable region, the number of species that were identified to the species level was reduced by 18%.

Dyslexic entrepreneurs: the incidence; their coping strategies and their business skills.

This comparative study explores the incidence of dyslexia in entrepreneurs, corporate managers and the general population. It examines the suggestion that dyslexic entrepreneurs develop coping strategies to manage their weaknesses, which are subsequently of benefit in the new venture creation process. Results of this study suggest that there is a significantly higher incidence of dyslexia in entrepreneurs than in the corporate management and general US and UK populations and some of the strategies they adopt to overcome dyslexia (such as delegation of tasks) may be useful in business. The study was undertaken in two parts. First, entrepreneurs and corporate managers completed an online questionnaire, which combined questions about their company, their management or leadership role and their business skills together with questions that were designed to explore the likely incidence of dyslexia. A follow-up study that made use of a semi-structured questionnaire explored business issues and educational experience in more depth with those who had been diagnosed as dyslexic and those who did not have any history of dyslexia or any other learning difficulty.

Beating-heart percutaneous mitral valve repair using a transcatheter endovascular suturing device in an animal model.

BACKGROUND: The edge-to-edge (Alfieri) technique for mitral valve repair is a versatile method of treating mitral insufficiency. Because of its simplicity, it has been applied in minimally invasive surgery, and recently, in the design of endovascular closed-heart devices. AIM: The purpose of this study was to evaluate the acute in-vivo safety and feasibility of a novel percutaneous mitral valve repair system based on Alfieri technique in an animal model. METHODS: Under general anesthesia, 11 pigs (90-100 kgs), underwent percutaneous Alfeiri procedure. The right femoral vein was punctured and the mitral valve was approached via a standard transeptal puncture. Combined intracardiac echo and fluoroscopic guidance was used. The procedure included: the positioning of a guide catheter for multiple access to the left atrium and for directing devices; the use of a therapy device to capture the free edge of the mitral valve leaflets using vacuum, and to deliver the suture to the valve and finally the fixation with a Nitinol suture clip, and trimming of the suture with a fastener catheter. RESULTS: Leaflet capture, suture placement, and suture-clip deployment was successful in all 11 animals. There were no acute cardiac or access site complications. Procedural time (from wire in left atrium to completion of the procedure was 18 +/- 9 min (range 9-30 min). Blood loss was 67 +/- 44 ml (range 0-125 ml). A double orifice configuration was visible by echocardiography at the end of the procedure in all animals. CONCLUSION: This acute animal study demonstrated the feasibility of a beating heart percutaneous Alfieri procedure in a non-diseased porcine valve using an endovascular suturing device to safely access the mitral valve, place a stitch through the mitral valve leaflets, and deploy a suture-clip that reproduces the surgical technique. Clinical application of this device in humans needs to be evaluated.

Flagellin gene sequence evolution in Salmonella.

Salmonella exhibits 70 serologically distinct flagellins, used internationally to diagnose and track infections. The terminal sequences of flagellin protein subunits are conserved in a range of bacteria and are here used as evolutionary markers to reveal how new serotypes arise. Terminal sequences of flagellins that exhibit factors g or m (G-group) were distinct from other Salmonella antigens (Non-G-group) and cluster more closely with Escherichia coli. It is postulated that G-group flagellins were inherited from a common ancestor of E. coli and Salmonella and that these antigens were among the original set in Salmonella. Sequence differences at the 5' termini may prevent recombination between co-infecting strains. Evidence of increased variation of flagellin in rare biphasic G-group serotypes suggests that the presence of a second flagellin locus allows mutation of the G-group flagellin. FljB probably arose from a single duplication of a Non-G gene, since which synonymous mutations resulted in the fljB-specific sequence at the 5' termini.

Intracardiac echocardiography for percutaneous mitral valve repair in a swine model.

BACKGROUND: High-quality live imaging assessment of cardiac valves and cardiac anatomy is crucial for the success of percutaneous catheter-based mitral valve (MV) repair techniques. We examined the use of intracardiac echocardiography (ICE) in providing online ICE images necessary for successful perctunaneous MV repair by Alfieri stitch technique-based percutaneous edge-to-edge device in a swine model. METHODS: ICE was performed in 20 healthy adult pigs (90 +/- 8 kg, mean +/- SEM). A 10F ICE catheter was advanced through the left femoral vein and the right jugular vein. Images were obtained from the right atrium, left atrium, right ventricle, and pulmonary artery. Fluoroscopy was used to locate the position of ICE catheter tip in different imaging windows. RESULTS: An echocardiographic protocol was developed for focused visualization of target cardiac structures during the process of percutaneous MV repair. This included visualization of interatrial septum to guide transeptal puncture; to confirm transit of guidewire into the left atrium, across the MV into the left ventricle and across the aortic valve into the aorta; to assist in central positioning of guide catheter in the left atrium above and then across MV leaflets; and to visualize middle scallops of anterior and posterior MV leaflets in short- and long-axis views. Finally, location and orientation of the orifice of the therapy catheter against each MV leaflet was visualized to enable successful capture of MV leaflets, to confirm successful deployment of suture and double orifice (figure of 8" appearance of MV, and finally to confirm central deployment of clip at the site of suture. CONCLUSION: An ICE protocol was developed to visualize serial cardiac structures to guide deployment of suture into the A2-P2 scallops of the MV and to confirm final result before release of clip.

Towards the development of a DNA-sequence based approach to serotyping of Salmonella enterica.

BACKGROUND: The fliC and fljB genes in Salmonella code for the phase 1 (H1) and phase 2 (H2) flagellin respectively, the rfb cluster encodes the majority of enzymes for polysaccharide (O) antigen biosynthesis, together they determine the antigenic profile by which Salmonella are identified. Sequencing and characterisation of fliC was performed in the development of a molecular serotyping technique. RESULTS: FliC sequencing of 106 strains revealed two groups; the g-complex included those exhibiting "g" or "m,t" antigenic factors, and the non-g strains which formed a second more diverse group. Variation in fliC was characterised and sero-specific motifs identified. Furthermore, it was possible to identify differences in certain H antigens that are not detected by traditional serotyping. A rapid short sequencing assay was developed to target serotype-specific sequence motifs in fliC. The assay was evaluated for identification of H1 antigens with a panel of 55 strains. CONCLUSION: FliC sequences were obtained for more than 100 strains comprising 29 different H1 alleles. Unique pyrosequencing profiles corresponding to the H1 component of the serotype were generated reproducibly for the 23 alleles represented in the evaluation panel. Short read sequence assays can now be used to identify fliC alleles in approximately 97% of the 50 medically most important Salmonella in England and Wales. Capability for high throughput testing and automation give these assays considerable advantages over traditional methods.

Fluorescent amplified fragment length polymorphism genotyping of Campylobacter jejuni and Campylobacter coli strains and its relationship with host specificity, serotyping, and phage typing.

Fluorescent amplified fragment length polymorphism (FAFLP) analysis was applied to 276 Campylobacter jejuni strains and 87 Campylobacter coli strains isolated from humans, pigs, cattle, poultry, and retail meats to investigate whether certain FAFLP genotypes of C. jejuni and C. coli are associated with a particular host and to determine the degree of association between FAFLP-defined genotypes and heat-stable serotypes and/or phage types. Within C. coli, the poultry strains clustered separately from those of porcine origin. In contrast, no evidence of host specificity was detected among C. jejuni strains. While C. coli strains show host specificity by FAFLP genotyping, C. jejuni strains that are genotypically similar appear to colonize a range of hosts, rather than being host adapted. Some serotypes and/or phage types (C. jejuni serotype HS18, phage type PT6, and serophage type HS19/PT2 and C. coli HS66, PT2, and HS56/PT2) were the most homogeneous by FAFLP genotyping, while others were more heterogeneous (C. jejuni HS5 and PT39, and C. coli HS24 and PT44) and therefore poor indicators of genetic relatedness between strains. The lack of host specificity in C. jejuni suggests that tracing the source of infection during epidemiological investigations will continue to be difficult. The lack of congruence between some serotypes and/or phage types and FAFLP genotype underlines the need for phenotypic testing to be supplemented by genotyping. This study also demonstrates how, in general, FAFLP generates "anonymous" genetic markers for strain characterization and epidemiological investigation of Campylobacter in the food chain.