Specific proteome changes in platelets from individuals with GATA1-, GFI1B-, and RUNX1-linked bleeding disorders.

Mutations in the transcription factors GATA binding factor 1 (GATA1), growth factor independence 1B (GFI1B), and Runt-related transcription factor 1 (RUNX1) cause familial platelet and bleeding disorders. Mutant platelets exhibit common abnormalities including an α-granule reduction resulting in a grayish appearance in blood smears. This suggests that similar pathways are deregulated by different transcription factor mutations. To identify common factors, full platelet proteomes from 11 individuals with mutant GATA1R216Q, GFI1BQ287*, RUNX1Q154Rfs, or RUNX1TD2-6 and 28 healthy controls were examined by label-free quantitative mass spectrometry. In total, 2875 platelet proteins were reliably quantified. Clustering analysis of more than 300 differentially expressed proteins revealed profound differences between cases and controls. Among cases, 44 of 143 significantly downregulated proteins were assigned to platelet function, hemostasis, and granule biology, in line with platelet dysfunction and bleedings. Remarkably, none of these proteins were significantly diminished in all affected cases. Similarly, no proteins were commonly overrepresented in all affected cases compared with controls. These data indicate that the studied transcription factor mutations alter platelet proteomes in distinct largely nonoverlapping manners. This work provides the quantitative landscape of proteins that affect platelet function when deregulated by mutated transcription factors in inherited bleeding disorders.

[Oral and maxillofacial surgeons' satisfaction of the use of procedural sedation and/or analgesia (PSA) in the clinic]. / Tevredenheid van mka-chirurgen over gebruik procedurele sedatie in de kliniek.

Fear of orofacial interventions is a very common problem. Procedural sedation and/or analgesia (PSA) offers added value in reducing anxiety, pain and discomfort. At the Radboud University Medical Centre (Nijmegen, the Netherlands) PSA is conducted under the guidance of physician assistants, resulting in a high degree of patient satisfaction. In this study, the 115 surgeons' satisfaction with PSA procedures for outpatient orofacial surgical interventions was evaluated by means of questionnaires. The study's conclusion is that oral and maxillofacial surgeons are highly satisfied when PSA is administered to very anxious patients. The collaboration with the department of anaestesiology ensures that the outpatient interventions can be performed safely and efficiently by the oral and maxillofacial surgeon.

[Patient satisfaction with procedural sedation in oral and -maxillofacial surgery]. / Patiënttevredenheid met procedurele sedatie in de mka-chirurgie.

Fear of treatment by an oral and maxillofacial (OMF) surgeon or the dentist is a widespread and deep-rooted phenomenon. A patient can be taught to cope with dental phobia by means of psychological, cognitive behavioural therapy, but in cases of invasive oral treatment, such therapy may be insufficiently effective. For these invasive procedures, sedation serves as an alternative treatment option. The department of Oral and Maxillofacial Surgery at the Radboud University Medical Centre in Nijmegen, the Netherlands, offers the possibility to utilise intravenous sedation with midazolam (Dormicum®) and remifentanil (Ultiva®). This results in a reduction in the experience of axiety and pain, which can lead to an anterograde amnesia for the invasive treatment. Procedural sedation and analgesia (PSA) for OMF surgery demonstrate a high level of patient satisfaction, and therefore offer a good alternative treatment to local and general anaesthesia.

Mantle Sources of Recent Anatolian Intraplate Magmatism: A Regional Plume or Local Tectonic Origin?

We present an extensive study of rehomogenized olivine-hosted melt inclusions, olivine phenocrysts, and chromian spinel inclusions to explore the link between geodynamic conditions and the origin and composition of Pliocene-Quaternary intraplate magmatism in Anatolia at Kula, Ceyhan-Osmaniye, and Karacadag. Exceptional compositional variability of these products reveals early and incomplete mixing of distinct parental melts in each volcanic center, reflecting asthenospheric and lithospheric mantle sources. The studied primitive magmas consist of (1) two variably enriched ocean island basalt (OIB)-type melts in Kula; (2) both OIB-type and plume mid-ocean ridge basalt (P-MORB)-like melts beneath Toprakkale and Üçtepeler (Ceyhan-Osmaniye); and (3) two variably enriched OIB-type melts beneath Karacadag. Estimated conditions of primary melt generation are 23-9 kbar, 75-30 km, and 1415-1215 °C for Kula; 28-19 kbar, 90-65 km, and 1430-1350 °C for Toprakkale; 23-18 kbar, 75-60 km, and 1400-1355 °C for Üçtepeler; and 35-27 kbar, 115-90 km, and 1530-1455 °C for Karacadag, the deepest levels of which correspond to the depth of the lithosphere-asthenosphere boundary in all regions. Although magma ascent was likely facilitated by local deformation structures, recent Anatolian intraplate magmatism seems to be triggered by large-scale mantle flow that also affects the wider Arabian and North African regions. We infer that these volcanics form part of a much wider Arabian-North African intraplate volcanic province, which was able to invade the Anatolian upper plate through slab gaps.

Colonization of Campylobacter spp. in broiler chickens and laying hens reared in tropical climates with low-biosecurity housing.

The onset and prevalence of Campylobacter colonization in broilers and layers at commercial farms with low biosecurity in tropical climates were tested. Despite the presence of positive animals at the same farms, the broiler flocks tested negative until, on average, 21 days. Prelaying flocks showed a higher prevalence than laying flocks.

Clostridium difficile in Dutch animals: their presence, characteristics and similarities with human isolates.

The presence and characteristics of Clostridium difficile were investigated in 839 faecal samples from seven different animal species in the Netherlands. The number of positive samples ranged from 3.4% (cattle) to 25.0% (dogs). Twenty-two different PCR ribotypes were identified. Among 96 isolates, 53% harboured toxin genes. All C. difficile isolates from pigs, cattle and poultry were toxinogenic, whereas the majority of isolates from pet animals consisted of non-toxinogenic PCR ribotypes 010 and 039. Ribotype 012 was most prevalent in cattle and ribotype 078 in pigs. No predominant ribotypes were present in horse and poultry samples. Overall, PCR ribotypes 012, 014 and 078 were the most frequently recovered toxinogenic ribotypes from animal samples. Comparison with human isolates from the Dutch Reference Laboratory for C. difficile at Leiden University Medical Centre (LUMC) showed that these types were also recovered from human hospitalized patients in 2009/2010, encompassing 0.8%, 11.4% and 9.8% of all isolates, respectively. Application of multiple-locus variable-number tandem-repeat analysis indicated a genotypic relation of animal and human ribotype 078 strains, but a clear genotypic distinction for ribotypes 012 and 014. We conclude that toxinogenic C. difficile PCR ribotypes found in animals correspond to PCR ribotypes associated with human disease in hospitalized patients in the Netherlands. Contrary to PCR ribotype 078, significant genetic differences were observed between animal and human PCR ribotype 012 and 014 isolates.

Enterococcus hirae-associated endocarditis outbreaks in broiler flocks: clinical and pathological characteristics and molecular epidemiology.

BACKGROUND: Enterococcus hirae-associated endocarditis, characterized by a peak in mortality during the second week of the grow-out, and occasionally lameness, was diagnosed at Dutch broiler farms. OBJECTIVES: Field cases were studied to increase knowledge on clinical and pathological characteristics, pathogenesis and epidemiology of these infections. ANIMALS AND METHODS: In total, 1266 birds of 25 flocks from 12 farms were examined. Post-mortem examinations, bacteriology, histopathology, PCR and DNA fingerprinting was carried out. Six flocks were followed longitudinally (n = 1017 birds). RESULTS: Average mortality was 4.1% for the entire grow-out, of which 36% was attributed to endocarditis. Fibrinous thromboendocarditis of the right atrioventricular (AV) valve was found in 24% of hearts, compared to 7% and 4% with lesions of left and both AV valves, respectively. Thrombotic lesions were found in 24% (n = 432) of lungs, but only in larger branches of the Arteria pulmonalis. Occasionally, thrombi were found in the Arteria ischiadica externa and in liver and brain vessels. Enterococcus was cultured from 54% (n = 176) of heart and in 75% (n = 28), 62% (n = 106) and 31% (n = 16) of liver, bone marrow and lung samples, respectively. Further identification, using the Rapid ID Strep 32 API system and a PCR targeting mur-2 and mur-2(ed) genes was carried out on a subset of Enterococcus positive isolates (n = 65): both techniques identified the isolates as Enterococcus hirae. Pulsed-field gel electrophoresis did not indicate evidence of clonality between farms and flocks. CONCLUSIONS: The relevance of these findings for pathogenesis and epidemiology of E. hirae infections is discussed. CLINICAL IMPORTANCE. This study may facilitate diagnosis of field cases and may contribute to the design of further research and development of control measures.

Breakthrough Saprochaete capitata infections in patients receiving echinocandins: case report and review of the literature.

We report a rare case of a fatal Saprochaete capitata breakthrough infection in a patient with acute myeloid leukemia receiving empirical caspofungin therapy. S. capitata is an uncommon, yet emerging cause of invasive infections, especially in patients with haematological malignancies. Blood cultures from our patient yielded S. capitata which was found to be resistant, in vitro, to caspofungin. We consecutively reviewed all published cases of breakthrough infections caused by S. capitata in patients receiving echinocandins. S. capitata should be considered in those patients who remain febrile or who develop invasive mould infections while under echinocandin therapy.

[Multiresistant Brachyspira hyodysenteriae in a Dutch sow herd]. / Multiresistentie bij Brachyspira hyodysenteriae-isolaten op een varkensvermeerderingsbedrijf in Nederland.

This case study describes the isolation ofa multiresistant strain ofBrachyspira hyodysenteriae in April 2007 in a Dutch sow herd with recurrent diarrhoea. Examination of faecal samples taken from 7-month-old breeding gilts with diarrhoea revealed the presence of resistance against tiamulin, lincomycin, tylosin, doxycycline, and tylvalosin (the active substance in Aivlosin) in four of five samples. Tiamulin resistance has not been reported in The Netherlands before. The repeated use of tiamulin on the affected farm was assumed to be the main cause of the development of resistance to the drug. The farmer was advised to adopt a medication strategy and to implement management practices that would prevent an ongoing cycle of infection on the farm. It is important that the Dutch swine industry appreciates that tiamulin-resistant strains of B. hyodysenteriae may be found on other farms as well. The appropriate and prudent use of antibiotics is essential in order to prevent the development of resistance against the last option left to cure B. hyodysenteriae infections: valnemulin.

Bacteriophage therapy to reduce salmonella colonization of broiler chickens.

Acute enteric infections caused by salmonellas remain a major public health burden worldwide. Poultry, particularly chickens, are known to be the main reservoir for this zoonotic pathogen. Although some progress has been made in reducing Salmonella colonization of broiler chickens by using biosecurity and antimicrobials, it still remains a considerable problem. The use of host-specific bacteriophages as a biocontrol is one possible intervention by which Salmonella colonization could be reduced. A total of 232 Salmonella bacteriophages were isolated from poultry farms, abattoirs, and wastewater in 2004 and 2005. Three phages exhibiting the broadest host ranges against Salmonella enterica serotypes Enteritidis, Hadar, and Typhimurium were characterized further by determining their morphology and lytic activity in vitro. These phages were then administered in antacid suspension to birds experimentally colonized with specific Salmonella host strains. The first phage reduced S. enterica serotype Enteritidis cecal colonization by > or = 4.2 log10 CFU within 24 h compared with controls. Administration of the second phage reduced S. enterica serotype Typhimurium by > or = 2.19 log10 CFU within 24 h. The third bacteriophage was ineffective at reducing S. enterica serotype Hadar colonization. Bacteriophage resistance occurred at a frequency commensurate with the titer of phage being administered, with larger phage titers resulting in a greater proportion of resistant salmonellas. The selection of appropriate bacteriophages and optimization of both the timing and method of phage delivery are key factors in the successful phage-mediated control of salmonellas in broiler chickens.

Potential routes of acquisition of Arcobacter species by piglets.

The aim of this study was to determine the prevalence and the transmission routes of Arcobacter spp. in sows and their offspring on a breeding farm. Twelve Arcobacter-positive sows and their litters were studied for this purpose. Analysis of rectal samples showed a high prevalence of Arcobacter spp. among the sows (approximately 42% of the sows carried one or more Arcobacter species). Intermittent excretion of one particular species and shifts in excretion from one species to another were observed in individual animals over time. The detection of Arcobacter spp. in amniotic fluid of the sows and in rectal samples from newborn piglets (ranging from 38.5-83.3% per litter), as well as the high similarity between PFGE profiles of Arcobacter isolates from sows and their respective newborns indicated the existence of an intra-uterine transmission route for Arcobacter spp. Specific antibodies against Arcobacter spp. were detected in colostrum by Western blot. At 2 weeks of age, only a few piglets were positive for Arcobacter. The reappearance of Arcobacter in these piglets at Week 3 and the shift in the Arcobacter species detected (from a prominent presence of A. cryaerophilus at birth to the presence of A. skirrowii and A. butzleri at 3 weeks after birth) showed that a post-natal infection route from their mothers, newcomers or the environment to the piglets existed. Thus, in this manuscript the transmission of Arcobacter spp. (both vertical and horizontal) from carrying sows to their offspring is demonstrated.

[Campylobacter as a venereal disease in cattle]. / Campylobacter als dekinfectie bij rundvee.

Since the introduction of AI, venereal diseases caused by Tritrichomonas fetus and Campylobacter fetus subsp. venerealis haved been eradicated in The Netherlands. Campylobacter fetus subsp. fetus can cause sporadic abortion and early embryonic death. When natural breeding is practised, venereal diseases must be included in the list of differential diagnoses of fertility problems. A case study of a Campylobacter fetus subsp. fetus infection is described.

Global detection and identification of Campylobacter fetus subsp. venerealis.

Bovine genital campylobacteriosis caused by Campylobacter fetus subsp. venerealis (Cfv) is a genital infection that threatens the cattle industry. Detection and identification of Cfv are key factors in control programmes. Trade regulations should be based on scientifically and internationally accepted methods of detection and identification of Cfv. Such methods are described in the World Organisation for Animal Health (OIE) Manual of Diagnostic Tests and Vaccines for Terrestrial Animals. A study was conducted to determine which methods are in use in OIE Member Countries and to get an overview of new or improved tests. A questionnaire was sent to OIE Member Countries, and 26 out of 166 were returned. Globally, a diversity of methods for the detection and identification of Cfv are in use. The authors conclude that there is a lack of harmonisation that may have consequences for the description of the health status of countries and may lead to disputes with respect to trade regulations.

Comparative study using amplified fragment length polymorphism fingerprinting, PCR genotyping, and phenotyping to differentiate Campylobacter fetus strains isolated from animals.

A collection of Campylobacter fetus strains, including both C. fetus subsp. fetus and C. fetus subsp. venerealis, were phenotypically identified to the subspecies level and genotypically typed by PCR and amplified fragment length polymorphism (AFLP) analysis. Phenotypic subspecies determination methods were unreliable. Genotyping of the strains by PCR and AFLP showed a clear discrimination between the two subspecies.