Dynamics of Mycoplasma bovis in Dutch dairy herds during acute clinical outbreaks.

Mycoplasma bovis (M. bovis) can cause serious illness in cattle, presenting as arthritis and mastitis in dairy cows and pneumonia, arthritis and otitis media in calves. This study aimed to provide insight into the dynamics of M. bovis within dairy herds, experiencing an acute outbreak in dairy cows. Twenty farms were followed with laboratory testing of suspected dairy cows. Each outbreak farm was sampled five times, at 2-3 week intervals, sampling blood and milk and conjunctival fluid from clinically suspected dairy cows and healthy animals from three different age groups: dairy cows, young stock (7-24 months) and calves (1-6 months). Additionally, bulk tank milk was sampled every visit and environmental samples were taken on the first and last visits. The presence of M. bovis was tested by evaluating antibody titres in blood, bacterial DNA in conjunctival fluid and environmental samples and viable bacteria in milk samples. All data were analysed using logistic regression models, corrected for repeated sampling and within-herd correlation. Sixty percent (12/20) of the herds showed a combination of arthritis and mastitis, while other herds experienced only clinically mastitis (3/20) or arthritis (5/20). From the time an outbreak was confirmed, M. bovis infection was not only present in dairy cows, but also in young stock and calves (80% of the farms). Laboratory tests also confirmed the presence of M. bovis in healthy animals. The M. bovis PCR levels of calves and young stock were highly correlated at all visits (rtotal = 0.81, P < 0.01). Furthermore, M. bovis was present in the environment of the animals. At the end of the 3-month study period, none of the 20 clinical outbreak farms were M. bovis-'negative', based on laboratory testing, although hardly any clinical cases were observed at that time.

Comparing the curative efficacy of topical treatment with thiamphenicol and oxytetracycline on digital dermatitis lesions in dairy cattle.

The efficacy of two topically applied antibiotics for the treatment of painful ulcerative stage of bovine digital dermatitis (BDD) lesions was compared in a clinical trial conducted on five dairy farms in the Netherlands during the autumn of 2015. A total of 109 cows with an ulcerative (M2) stage of BDD were randomly appointed a treatment with an antibiotic-based spray. One treatment contained thiamphenicol as active ingredient (TAF). The other treatment had oxytetracycline as active ingredient (ENG). The experimental unit for this study was the hind claw with the presence of an ulcerative BDD lesion. On day 0, claws with ulcerative BDD lesions were trimmed, cleaned, photographed and thereafter treated randomly either with TAF or ENG. Cure was defined as the transition of an ulcerative lesion into a non-painful chronic (M4) or into a healed (M0) stage of BDD at day 28 post-treatment. The cure rate at day 28 of M2 BDD lesions treated with TAF was 89 per cent (95 per cent CI 0.78 to 0.94), and for ENG 75 per cent (95 per cent CI 0.67 to 0.86). So the difference in cure rate was 14 per cent (95 per cent CI 0.00 to 0.27), which was statistically significant. The P value in this experiment is very close to 0.05 indicating that the effect is quite small. If a two-sided test would be used, the small significant effect, in this experiment, will disappear. Overall, the significant better curative effect of TAF on BDD M2 lesions was small, compared with ENG.

One fungus, which genes? Development and assessment of universal primers for potential secondary fungal DNA barcodes.

The aim of this study was to assess potential candidate gene regions and corresponding universal primer pairs as secondary DNA barcodes for the fungal kingdom, additional to ITS rDNA as primary barcode. Amplification efficiencies of 14 (partially) universal primer pairs targeting eight genetic markers were tested across > 1 500 species (1 931 strains or specimens) and the outcomes of almost twenty thousand (19 577) polymerase chain reactions were evaluated. We tested several well-known primer pairs that amplify: i) sections of the nuclear ribosomal RNA gene large subunit (D1-D2 domains of 26/28S); ii) the complete internal transcribed spacer region (ITS1/2); iii) partial ß -tubulin II (TUB2); iv) γ-actin (ACT); v) translation elongation factor 1-α (TEF1α); and vi) the second largest subunit of RNA-polymerase II (partial RPB2, section 5-6). Their PCR efficiencies were compared with novel candidate primers corresponding to: i) the fungal-specific translation elongation factor 3 (TEF3); ii) a small ribosomal protein necessary for t-RNA docking; iii) the 60S L10 (L1) RP; iv) DNA topoisomerase I (TOPI); v) phosphoglycerate kinase (PGK); vi) hypothetical protein LNS2; and vii) alternative sections of TEF1α. Results showed that several gene sections are accessible to universal primers (or primers universal for phyla) yielding a single PCR-product. Barcode gap and multi-dimensional scaling analyses revealed that some of the tested candidate markers have universal properties providing adequate infra- and inter-specific variation that make them attractive barcodes for species identification. Among these gene sections, a novel high fidelity primer pair for TEF1α, already widely used as a phylogenetic marker in mycology, has potential as a supplementary DNA barcode with superior resolution to ITS. Both TOPI and PGK show promise for the Ascomycota, while TOPI and LNS2 are attractive for the Pucciniomycotina, for which universal primers for ribosomal subunits often fail.

The NLstart2run study: Incidence and risk factors of running-related injuries in novice runners.

Running is a popular form of physical activity, despite of the high incidence of running-related injuries (RRIs). Because of methodological issues, the etiology of RRIs remains unclear. Therefore, the purposes of the study were to assess the incidence of RRIs and to identify risk factors for RRIs in a large group of novice runners. In total, 1696 runners of a 6-week supervised "Start to Run" program were included in the NLstart2run study. All participants were aged between 18 and 65, completed a baseline questionnaire that covered potential risk factors, and completed at least one running diary. RRIs were registered during the program with a weekly running log. An RRI was defined as a musculo-skeletal complaint of the lower extremity or back attributed to running and hampering running ability for three consecutive training sessions. During the running program, 10.9% of the runners sustained an RRI. The multivariable Cox regression analysis showed that a higher age, higher BMI, previous musculo-skeletal complaints not attributed to sports and no previous running experience were related to RRI. These findings indicate that many novice runners participating in a short-term running program suffer from RRIs. Therefore, the identified risk factors should be considered for screening and prevention purposes.

Cognitive precursors of arithmetic development in primary school children with cerebral palsy.

The aim of this study was to examine the development of arithmetic performance and its cognitive precursors in children with CP from 7 till 9 years of age. Previous research has shown that children with CP are generally delayed in arithmetic performance compared to their typically developing peers. In children with CP, the developmental trajectory of the ability to solve addition- and subtraction tasks has, however, rarely been studied, as well as the cognitive factors affecting this trajectory. Sixty children (M=7.2 years, SD=.23 months at study entry) with CP participated in this study. Standardized tests were administered to assess arithmetic performance, word decoding skills, non-verbal intelligence, and working memory. The results showed that the ability to solve addition- and subtraction tasks increased over a two year period. Word decoding skills were positively related to the initial status of arithmetic performance. In addition, non-verbal intelligence and working memory were associated with the initial status and growth rate of arithmetic performance from 7 till 9 years of age. The current study highlights the importance of non-verbal intelligence and working memory to the development of arithmetic performance of children with CP.

[Factor structure of the German version of the BIS/BAS Scales in a population-based sample]. / Faktorenstruktur der deutschsprachigen version der BIS/BAS-Skalen in einer bevölkerungsstichprobe.

BACKGROUND: The Behavioural Inhibition System/Behavioural Activation System Scale (BIS/BAS-Scales) developed by Carver and White 1 is a self-rating instrument to assess the dispositional sensitivity to punishment and reward. The present work aims to examine the factor structure of the German version of the BIS/BAS-Scales. MATERIAL AND METHODS: In a large German population-based sample (n = 1881) the model fit of several factor models was tested by using confirmatory factor analyses. RESULTS: The best model fit was found for the 5-factor model with two BIS (anxiety, fear) and three BAS (drive, reward responsiveness, fun seeking) scales, whereas the BIS-fear, the BAS-reward responsiveness, and the BAS-fun seeking subscales showed low internal consistency. The BIS/BAS scales were negatively correlated with age, and women reported higher BIS subscale scores than men. CONCLUSION: Confirmatory factor analyses suggest a 5-factor model. However, due to the low internal reliability of some of the subscales the use of this model is questionable.

[Meaningful 'romming' (routine outcome monitoring): seeking a balance between userfriendliness and clinical relevance]. / Zinvol 'rommen': zoeken naar een evenwicht tussen gebruiksvriendelijkheid en klinische relevantie.

BACKGROUND: In the course of the last few years several ROM systems have been developed in which the measurements and feedback are restricted to the therapy outcome in terms of symptom reduction. In these ROM systems the emphasis is on the user-friendliness and on the ease with which the measurements can be implemented in clinical practice. However, research shows that many therapists and clients consider that the measurement of symptom reduction is too simplistic. AIM: To develop a ROM system that is therapeutically more relevant. METHOD: We describe the development and implementation of the Leuven Systematic Case study Protocol. RESULTS: In this multidimensional and multimodal monitoring system the emphasis is on the therapeutic relevance of 'romming:' not simply in terms of symptom reduction, but also in terms of processes of change and growth-orientated processes. In addition, the monitoring system takes into account the unique characteristics of client and therapist and the synergy existing between them. CONCLUSION: Not only do therapists have to learn to deal with the practical aspects of 'romming', they also need to be trained and supported so that they can deal adequately with a more explicit evaluation and feedback 'culture'.

Development of non-verbal intellectual capacity in school-age children with cerebral palsy.

BACKGROUND: Children with cerebral palsy (CP) are at greater risk for a limited intellectual development than typically developing children. Little information is available which children with CP are most at risk. This study aimed to describe the development of non-verbal intellectual capacity of school-age children with CP and to examine the association between the development of non-verbal intellectual capacity and the severity of CP. METHODS: A longitudinal analysis in a cohort study was performed with a clinic-based sample of children with CP. Forty-two children were assessed at 5, 6 and 7 years of age, and 49 children were assessed at 7, 8 and 9 years of age. Non-verbal intellectual capacity was assessed by Raven's Coloured Progressive Matrices (RCPM). Severity of CP was classified by the Gross Motor Function Classification System, type of motor impairment and limb distribution. manova for repeated measurements was used to analyse time effects and time × group effects on both RCPM raw scores and RCPM intelligence quotient scores. RESULTS: The development of non-verbal intellectual capacity was characterised by a statistically significant increase in RCPM raw scores but no significant change in RCPM intelligence quotient scores. The development of RCPM raw scores was significantly associated with the severity of CP. Children with higher levels of gross motor functioning and children with spastic CP showed greater increase in raw scores than children with lower levels of gross motor functioning and children with dyskinetic CP. CONCLUSIONS: Children with CP aged between 5 and 9 years show different developmental trajectories for non-verbal intellectual capacity, which are associated with the severity of CP. The development of non-verbal intellectual capacity in children with less severe CP seems to resemble that of typically developing children, while children with more severe CP show a limited intellectual development compared to typically developing children.

Anionic and cationic drug secretion in the isolated perfused rat kidney after neonatal surgical induction of ureteric obstruction.

OBJECTIVE: To study the pathophysiological changes of renal tubular drug transport mechanisms in congenital renal obstruction, by developing a model for perfusing the isolated kidney (IPK) after neonatal surgical induction of partial ureteric obstruction in Hanover Wistar rats. MATERIAL AND METHODS: Moderately severe obstruction of the right kidney of male rats was created by burying a segment of the right ureter under the psoas fascia at 5-7 days after birth. Different fluorescent substrates for renal organic anion and cation drug transport systems were added to the IPK, and the concentration of these substances with time analysed in perfusate and urine. RESULTS: The reproducibility in all groups of the glomerular filtration rate (GFR) and drug excretion was remarkably good. GFR was significantly lower in obstructed kidneys than in unobstructed kidneys. 123Rhodamine, a marker for organic cation and P-glycoprotein transport, had a significantly lower maximum excretion rate in the obstructed than in unobstructed kidneys. Renal fractional clearance (123rhodamine clearance corrected for diminished GFR) was also significantly lower in obstructed kidneys. There was no significant difference in maximum excretion (absolute and corrected GFR) for Lucifer Yellow, a marker for sodium-dependent organic anion transport. The maximum excretion rate of calcein, a marker for sodium-independent organic anion transport, was significantly lower in the obstructed than in the unobstructed kidneys, but significantly higher after correcting for reduced GFR. CONCLUSION: The IPK is a good model for studying the effect of neonatal renal obstruction on tubular drug transport. These results show that organic anion and cation transport mechanisms are affected differently by obstruction.

The pattern of metastasis of human melanoma to the central nervous system is not influenced by integrin alpha(v)beta(3) expression.

We investigated the effect of integrin alpha(v)beta(3) expression on the metastatic pattern of human melanoma cells in the central nervous system (CNS). For this purpose, we developed a hematogenous CNS melanoma metastasis model in nude mice using a modified internal carotid artery infusion technique. This protocol revealed 2 different patterns of CNS metastasis. The integrin alpha(v)beta(3)-expressing melanoma lines Mel57 and Zkr nearly exclusively produced metastases in the brain parenchyma, whereas cells of the BLM and MV3 lines, devoid of integrin alpha(v)beta(3) expression, preferentially metastasized to dura mater and leptomeninges. Treatment with hyaluronidase to obtain single BLM cell suspensions did not influence the metastatic pattern, indicating that this was not simply the result of entrapment of tumor cell aggregates in large-sized leptomeningeal vessels. The role of integrin alpha(v)beta(3) expression in the process of metastasis was tested by transfection of BLM, but did not lead to an altered pattern of metastasis. We did observe, however, slower growth of the transfected tumors, although the in vitro growth rate was unaltered, indicating a reduction in tumorigenicity. We conclude from our findings that CNS metastasis of melanoma cells in the mouse xenograft model occurs in at least 2 different but very reproducible patterns. Although it is predicted that adhesion of tumor cells to endothelial cells plays a role in this phenomenon, tumor cell integrin alpha(v)beta(3) expression per se does not explain the difference in metastatic behavior in the CNS. We assume that other, as yet unknown factors, must be involved.

Two structurally closely related polypeptides encoded by 14-S mRNA isolated from rat lens.

14-S mRNA from rat lens codes for two subunits of alpha-crystallin, A2 (Mr 20 000) and AIns (Mr 24 000, previously referred to as alphaX). Structural relationship between both translation products has been proved by immunoprecipitation with antisera directed against the different crystallin classes. Competition immunoprecipitation showed that the 14-S mRNA translation products are precipitated by common antibodies, specific for the A subunit of alpha-crystallin. Two-dimensional gel electrophoresis and peptide analysis provided further evidence that the 24 000-Mr polypeptide, synthesized in vitro under direction of 14-S mRNA, is identical with native alphaAIns. Although the structures of alphaA2 and alphaAIns are very similar, no precursor-product relationship exists between both 14-S-mRNA-encoded polypeptides.

Isolation and chracterization of rat-lens messenger RNAs. Comparison of lens proteins, synthesized in lens culture and in homologous and heterologous cell-free systems.

Messenger RNA has been isolated from rat lens tissue. The mRNA species which codes for the A2 chain of alpha-crystallin, revealed the same extremely high sedimentation value (14S) as the corresponding messenger from calf lens. However, it has been shown that in rat lens the 14-S messenger preparation directs the synthesis of an additional alpha-crystallin chain, designated as alpha-X with an approximate molecular weight of 24000. For comparative purpose crystallin synthesis has also been studied as well in cultured rat lens cells as in the rat lens cell-free system.