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1.  Draft Genome Sequence of the First Human Isolate of the Ruminant Pathogen Mycoplasma capricolum subsp. capricolum 
Genome Announcements  2015;3(3):e00583-15.
Mycoplasma capricolum subsp. capricolum is a well-known pathogen of small ruminants. A recent human case of septicemia involving this agent raised the question of its potential pathogenicity to humans. We present the first draft genome sequence of a human Mycoplasma capricolum subsp. capricolum isolate.
PMCID: PMC4472885  PMID: 26089408
2.  Evaluation of Specific Metabolic Rates of Major Organs and Tissues: Comparison Between Nonobese and Obese Women 
Obesity (Silver Spring, Md.)  2011;20(1):95-100.
Elia (1992) identified the specific resting metabolic rates (Ki) of major organs and tissues in young adults with normal weight: 200 for liver, 240 for brain, 440 for heart and kidneys, 13 for skeletal muscle, 4.5 for adipose tissue and 12 for residual mass (all units in kcal/kg per day). The aim of the present study was to assess the applicability of Elia’s Ki values for obese adults. A sample of young women (n = 80) was divided into two groups, nonobese (BMI <29.9 kg/m2) and obese (BMI 30.0–43.2 kg/m2). This study was based on the mechanistic model: REE = Σ (Ki × Ti), where REE is whole-body resting energy expenditure measured by indirect calorimetry and Ti is the mass of individual organs and tissues measured by magnetic resonance imaging. For each organ/tissue, the corresponding Elia’s Ki value was analyzed respectively for nonobese and obese groups by using stepwise univariate regression analysis. Elia’s Ki values were within the range of 95% confidence intervals (CIs) in the nonobese group. However, Elia’s Ki values were outside the right boundaries of 95% CIs in the obese group and a corresponding obesity-adjusted coefficient was calculated as 0.98, indicating that Elia’s values overestimate Ki by 2.0% in obese adults. Obesity-adjusted Ki values were 196 for liver, 235 for brain, 431 for heart and kidneys, 12.7 for skeletal muscle, 4.4 for adipose tissue, and 11.8 for residual mass. In conclusion, although Elia’s Ki values were validated in nonobese women, obesity-adjustments are appropriate for application in obese women.
PMCID: PMC3319030  PMID: 21836642
3.  Evaluation of Specific Metabolic Rates of Major Organs and Tissues: Comparison Between Men and Women 
American Journal of Human Biology  2010;23(3):333-338.
The specific resting metabolic rates (Ki, in kcal/kg per day) of major organs and tissues in the Reference Man were suggested in 1992 by Elia: 200 for liver, 240 for brain, 440 for heart and kidneys, 13 for skeletal muscle, 4.5 for adipose tissue and 12 for the residual mass. However, it is unknown whether gender influences the Ki values. The aim of the present study was to compare the Ki values observed in non-elderly non-obese men to the corresponding values in women.
Elia’s Ki values were evaluated based on a mechanistic model: REE = Σ(Ki × Ti), where REE is whole-body resting energy expenditure measured by indirect calorimetry and Ti is the mass of major organs and tissues measured by magnetic resonance imaging. Marginal 95% confidence intervals (CIs) for the model-estimated Ki values were calculated by stepwise univariate regression analysis. Subjects were non-elderly (age 20 - 49 yrs) non-obese (BMI 18.5 - 29.9 kg/m2) men (n = 49) and women (n = 57).
The measured REE (REEm) and the mass of major organs and skeletal muscle were all greater in the men than in women. The predicted REE by Elia’s Ki values were correlated with REEm in men (r = 0.87) and women (r = 0.86, both P <0.001). Elia’s Ki values were within the range of 95% CIs for both men and women groups, revealing that gender adjustment is not necessary.
Elia’s proposed adult Ki values are valid in both non-elderly non-obese men and women. Further studies are needed to explore the potential influences of age and obesity on Ki values in humans.
PMCID: PMC3139779  PMID: 21484913
Gender; Magnetic resonance imaging; Organ mass; Stepwise univariate regression analysis; Tissue mass
4.  The Origin of the ‘Mycoplasma mycoides Cluster’ Coincides with Domestication of Ruminants 
PLoS ONE  2012;7(4):e36150.
The ‘Mycoplasma mycoides cluster’ comprises the ruminant pathogens Mycoplasma mycoides subsp. mycoides the causative agent of contagious bovine pleuropneumonia (CBPP), Mycoplasma capricolum subsp. capripneumoniae the agent of contagious caprine pleuropneumonia (CCPP), Mycoplasma capricolum subsp. capricolum, Mycoplasma leachii and Mycoplasma mycoides subsp. capri. CBPP and CCPP are major livestock diseases and impact the agricultural sector especially in developing countries through reduced food-supply and international trade restrictions. In addition, these diseases are a threat to disease-free countries. We used a multilocus sequence typing (MLST) approach to gain insights into the demographic history of and phylogenetic relationships among the members of the ‘M. mycoides cluster’. We collected partial sequences from seven housekeeping genes representing a total of 3,816 base pairs from 118 strains within this cluster, and five strains isolated from wild Caprinae. Strikingly, the origin of the ‘M. mycoides cluster’ dates to about 10,000 years ago, suggesting that the establishment and spread of the cluster coincided with livestock domestication. In addition, we show that hybridization and recombination may be important factors in the evolutionary history of the cluster.
PMCID: PMC3338596  PMID: 22558362
5.  A Novel Rapid DNA Microarray Assay Enables Identification of 37 Mycoplasma Species and Highlights Multiple Mycoplasma Infections 
PLoS ONE  2012;7(3):e33237.
Mycoplasmas comprise a conglomerate of pathogens and commensals occurring in humans and animals. The genus Mycoplasma alone contains more than 120 species at present, and new members are continuously being discovered. Therefore, it seems promising to use a single highly parallel detection assay rather than develop separate tests for each individual species. In this study, we have designed a DNA microarray carrying 70 oligonucleotide probes derived from the 23S rRNA gene and 86 probes from the tuf gene target regions. Following a PCR amplification and biotinylation step, hybridization on the array was shown to specifically identify 31 Mycoplasma spp., as well as 3 Acholeplasma spp. and 3 Ureaplasma spp. Members of the Mycoplasma mycoides cluster can be recognized at subgroup level. This procedure enables parallel detection of Mollicutes spp. occurring in humans, animals or cell culture, from mono- and multiple infections, in a single run. The main advantages of the microarray assay include ease of operation, rapidity, high information content, and affordability. The new test's analytical sensitivity is equivalent to that of real-time PCR and allows examination of field samples without the need for culture. When 60 field samples from ruminants and birds previously analyzed by denaturing-gradient gel electrophoresis (DGGE) were tested by the microarray assay both tests identified the same agent in 98.3% of the cases. Notably, microarray testing revealed an unexpectedly high proportion (35%) of multiple mycoplasma infections, i.e., substantially more than DGGE (15%). Two of the samples were found to contain four different Mycoplasma spp. This phenomenon deserves more attention, particularly its implications for epidemiology and treatment.
PMCID: PMC3315567  PMID: 22479374
6.  Evaluation of Ultra-Low Dose CT in the Diagnosis of Pediatric-Like Fractures Using an Experimental Animal Study 
Korean Journal of Radiology  2012;13(2):165-173.
The aim of this prospective study was to evaluate the performance of ultra-low dose CT for the diagnosis of pediatric-like fractures and ascertain the lowest dose level sufficient for diagnostics.
Materials and Methods
Fifty-one bones of young pig cadavers were artificially fractured and subsequently examined by using a 64 multi-detector CT with 36 various dose levels down to a dose comparable with that of X-rays. Two pediatric radiologists analysed the CT scans according to the presence or absence of a fracture, determination of the fracture type and the displacement as well as the diagnostic certainty. For each dose protocol, a success rate for the correct determination of the above-mentioned CT analyses was calculated. A success rate of at least 95% was considered sufficient for diagnostics.
All but the lowest dose levels were sufficient to identify the fracture. Only the two lowest dose levels were insufficient to detect the fracture type. All dose levels were adequate for the identification of the displacement. The lowest dose level sufficient for diagnostics was 120 kVp, 11 mAs, and pitch 1.5, with a CTDIvol of 10% of a standard dose and an effective dose three times as large as that of X-rays.
Ultra-low dose CT provides the feasibility of a significant dose reduction, still allowing sufficient diagnostics of pediatric-like fractures.
PMCID: PMC3303899  PMID: 22438683
Ultra-low dose CT; Fractures in children; Skeletal-appendicular; Experimental study
7.  Serological testing of cattle experimentally infected with Mycoplasma mycoides subsp. mycoides Small Colony using four different tests reveals a variety of seroconversion patterns 
To study the specific antibody response to infection with Mycoplasma mycoides subsp. mycoides Small Colony (MmmSC), the agent of Contagious Bovine Pleuropneumonia (CBPP), we examined three panels of sera collected during three experimental infection trials in African cattle. The methods used included an in-house complement fixation test (CFT), a commercially available CFT, a competitive antibody ELISA (cELISA) and the immunoblotting test (IBT). In addition, lung tissue samples were examined by culture.
A total of 89% (51/59) of all experimentally infected animals tested positive on at least one of the serological tests throughout the trial. The specific antibody titres to the MmmSC infection became positive first by CFT (6 to 9 days post infection [dpi]), followed by IBT (9 to 13 dpi) and cELISA (13 to 16 dpi). Individual animals were found to display remarkably distinct seroconversion patterns, which allowed their classification into i) early high responders, ii) late high responders, and iii) low responders. In accordance with other studies, none of the present serological tests was capable of detecting all CBPP infected animals.
Comparison of the assays' performance in terms of sensitivity and specificity raises serious questions as to their reliability for identification of infected individuals in the field. In view of these limitations, a combination of CFT and cELISA can markedly improve CBPP diagnosis at single-animal level.
PMCID: PMC3377920  PMID: 22098816
8.  Assessment of a novel multiplex real-time PCR assay for the detection of the CBPP agent Mycoplasma mycoides subsp. mycoides SC through experimental infection in cattle 
Mycoplasma mycoides subsp. mycoides SC is the pathogenic agent of contagious bovine pleuropneumonia (CBPP), the most important disease of cattle in Africa causing significant economic losses. The re-emergence of CBPP in Europe in the 1980s and 1990s illustrates that it is still a threat also to countries that have successfully eradicated the disease in the past. Nowadays, probe-based real-time PCR techniques are among the most advanced tools for a reliable identification and a sensitive detection of many pathogens, but only few protocols have been published so far for CBPP diagnosis. Therefore we developed a novel TaqMan®-based real-time PCR assay comprising the amplification of two independent targets (MSC_0136 and MSC_1046) and an internal exogenous amplification control in a multiplex reaction and evaluated its diagnostic performance with clinical samples.
The assays detected 49 MmmSC strains from diverse temporal and geographical origin, but did not amplify DNA from 82 isolates of 20 non-target species confirming a specificity of 100%. The detection limit was determined to be 10 fg DNA per reaction for the MSC_0136 assay and 100 fg per reaction for the MSC_1046 assay corresponding to 8 and 80 genome equivalents, respectively. The diagnostic performance of the assay was evaluated with clinical samples from 19 experimentally infected cattle and from 20 cattle without CBPP and compared to those of cultivation and a conventional PCR protocol. The two rt-PCR tests proved to be the most sensitive methods and identified all 19 infected animals. The different sample types used were not equally suitable for MmmSC detection. While 94.7% of lung samples from the infected cohort were positively tested in the MSC_0136 assay, only 81% of pulmonal lymph nodes, 31% of mediastinal lymph nodes and 25% of pleural fluid samples gave a positive result.
The developed multiplex rt-PCR assay is recommended as an efficient tool for rapid confirmation of a presumptive CBPP diagnosis in a well-equipped laboratory environment.
PMCID: PMC3170577  PMID: 21838878
9.  Effect of Constitution on Mass of Individual Organs and Their Association with Metabolic Rate in Humans—A Detailed View on Allometric Scaling 
PLoS ONE  2011;6(7):e22732.
Resting energy expenditure (REE)-power relationships result from multiple underlying factors including weight and height. In addition, detailed body composition, including fat free mass (FFM) and its components, skeletal muscle mass and internal organs with high metabolic rates (i.e. brain, heart, liver, kidneys), are major determinants of REE. Since the mass of individual organs scales to height as well as to weight (and, thus, to constitution), the variance in these associations may also add to the variance in REE. Here we address body composition (measured by magnetic resonance imaging) and REE (assessed by indirect calorimetry) in a group of 330 healthy volunteers differing with respect to age (17–78 years), sex (61% female) and BMI (15.9–47.8 kg/m2). Using three dimensional data interpolation we found that the inter-individual variance related to scaling of organ mass to height and weight and, thus, the constitution-related variances in either FFM (model 1) or kidneys, muscle, brain and liver (model 2) explained up to 43% of the inter-individual variance in REE. These data are the first evidence that constitution adds to the complexity of REE. Since organs scale differently as weight as well as height the “fit” of organ masses within constitution should be considered as a further trait.
PMCID: PMC3144246  PMID: 21818376
10.  A minor role of CD4+ T lymphocytes in the control of a primary infection of cattle with Mycoplasma mycoides subsp. mycoides 
Veterinary Research  2011;42(1):77.
Contagious bovine pleuropneumonia (CBPP), caused by Mycoplasma mycoides subsp. mycoides, is an important livestock disease in Africa. The current control measures rely on a vaccine with limited efficacy and occasional severe side effects. Knowledge of the protective arms of immunity involved in this disease will be beneficial for the development of an improved vaccine. In previous studies on cattle infected with M. mycoides subsp. mycoides, a correlation was detected between the levels of mycoplasma-specific IFN-γ-secreting CD4+ T lymphocytes and reduced clinical signs. However, no cause and effect has been established, and the role of such cells and of protective responses acquired during a primary infection is not known.
We investigated the role of CD4+ T lymphocytes in CBPP by comparing disease patterns and post mortem findings between CD4+ T cell depleted and non-depleted cattle. The depletion was carried out using several injections of BoCD4 specific murine monoclonal antibody on day 6 after experimental endotracheal infection with the strain Afadé. All cattle were monitored clinically daily and sacrificed 28-30 days post-infection. Statistically significant but small differences were observed in the mortality rate between the depleted and non-depleted animals. However, no differences in clinical parameters (fever, signs of respiratory distress) and pathological lesions were observed, despite elimination of CD4+ T cells for more than a week. The slightly higher mortality in the depleted group suggests a minor role of CD4+ T cells in control of CBPP.
PMCID: PMC3148206  PMID: 21663697
11.  ‘Functional’ body composition: differentiating between benign and non-benign obesity 
Recent body composition analyses, together with assessments of insulin resistance, aerobic fitness, and intima-media thickness of the common carotid artery, have shown that metabolically-benign obese subjects have a similar BMI, waist circumference, and subcutaneous abdominal fat compared with non-metabolically-benign obese subjects. Research has suggested that 25-30% of the obese population do not need either treatment or prevention of secondary disorders. Therefore, assessment of functional body composition should replace nutritional status-based risk assessments (such as the body mass index) in both metabolic research and clinical decision making. The concept of ‘functional’ body composition gives us a more sophisticated view on nutritional status, metabolism, endocrinology, and diseases. Knowledge of detailed body composition enables characterization of biomedical traits which will give functional evidence relating genetic variants.
PMCID: PMC2948251  PMID: 20948613

Results 1-11 (11)