Optical mapping compendium of structural variants across global cattle breeds.

Structural variants (SV) have been linked to important bovine disease phenotypes, but due to the difficulty of their accurate detection with standard sequencing approaches, their role in shaping important traits across cattle breeds is largely unexplored. Optical mapping is an alternative approach for mapping SVs that has been shown to have higher sensitivity than DNA sequencing approaches. The aim of this project was to use optical mapping to develop a high-quality database of structural variation across cattle breeds from different geographical regions, to enable further study of SVs in cattle. To do this we generated 100X Bionano optical mapping data for 18 cattle of nine different ancestries, three continents and both cattle sub-species. In total we identified 13,457 SVs, of which 1,200 putatively overlap coding regions. This resource provides a high-quality set of optical mapping-based SV calls that can be used across studies, from validating DNA sequencing-based SV calls to prioritising candidate functional variants in genetic association studies and expanding our understanding of the role of SVs in cattle evolution.

A cattle graph genome incorporating global breed diversity.

Despite only 8% of cattle being found in Europe, European breeds dominate current genetic resources. This adversely impacts cattle research in other important global cattle breeds, especially those from Africa for which genomic resources are particularly limited, despite their disproportionate importance to the continent's economies. To mitigate this issue, we have generated assemblies of African breeds, which have been integrated with genomic data for 294 diverse cattle into a graph genome that incorporates global cattle diversity. We illustrate how this more representative reference assembly contains an extra 116.1 Mb (4.2%) of sequence absent from the current Hereford sequence and consequently inaccessible to current studies. We further demonstrate how using this graph genome increases read mapping rates, reduces allelic biases and improves the agreement of structural variant calling with independent optical mapping data. Consequently, we present an improved, more representative, reference assembly that will improve global cattle research.

Immunisation of mice with Mycoplasma hyopneumoniae antigens P37, P42, P46 and P95 delivered as recombinant subunit or DNA vaccines.

Porcine enzootic pneumonia (PEP), which is caused by the fastidious bacterium Mycoplasma hyopneumoniae, is one of the most economically important diseases in the pig industry worldwide. Commercial bacterins provide only partial protection; therefore, the development of more efficient vaccines against PEP is necessary. In this study, the cellular and humoral immune responses elicited by DNA and recombinant subunit vaccines based on the P37, P42, P46 and P95 antigens of M. hyopneumoniae were evaluated after the intramuscular inoculation of BALB/c mice. The expression of the cytokines INFγ, TNFα and IL1 was evaluated by real-time RT-PCR in splenocytes from vaccinated mice. All antigens delivered as subunit vaccines, especially P42 and P95, and the pcDNA3/P46 DNA vaccine were able to elicit strong immune responses. These vaccines induced cellular immune responses and the production of antibodies able to react with native M. hyopneumoniae proteins. Because both cellular and humoral immune responses were induced, P42 and P95 are promising candidates for a recombinant subunit vaccine and P46 is a promising candidate for a DNA vaccine against PEP.

Production and characterization of recombinant transmembrane proteins from Mycoplasma hyopneumoniae.

Mycoplasma hyopneumoniae is the etiological agent of swine enzootic pneumonia (EP), a chronic respiratory disease which causes significant economic losses to the swine industry worldwide. More efficient strategies for controlling this disease are necessary. In this study, we cloned17 genes coding for transmembrane proteins from M. hyopneumoniae, among which six were successfully expressed in Escherichia coli and had their immunogenic and antigenic properties evaluated. All proteins were immunogenic in mice and sera from naturally infected pigs reacted with the recombinant proteins, suggesting that they are expressed during infection. These antigens may contribute for the development of new recombinant vaccines and diagnostic tests against EP.

Paraoxonase 1 (PON1) gene variants are not associated with clopidogrel response.

A common functional variant in paraoxonase 1 (PON1), Q192R, was recently reported to be a major determinant of clopidogrel response. This variant was genotyped in 566 participants of the Amish Pharmacogenomics of Anti-Platelet Intervention (PAPI) study and in 227 percutaneous coronary intervention (PCI) patients. Serum paraoxonase activity was measured in a subset of 79 PAPI participants. PON1 Q192R was not associated with pre- or post-clopidogrel platelet aggregation in the PAPI study (P = 0.16 and P = 0.21, respectively) or the PCI cohort (P = 0.47 and P = 0.91, respectively). The Q192 allele was not associated with cardiovascular events (hazard ratio (HR) 0.46, 95% confidence interval (CI) 0.20-1.06; P = 0.07). No correlation was observed between paraoxonase activity and post-clopidogrel platelet aggregation (r(2) < 0.01, P = 0.78). None of 49 additional PON1 variants evaluated was associated with post-clopidogrel platelet aggregation. These findings do not support a role for PON1 as a determinant of clopidogrel response.

Molar substitution and C2/C6 ratio of hydroxyethyl starch: influence on blood coagulation.

BACKGROUND: Development of hydroxyethyl starches (HES) with a low impact on blood coagulation but a long intravascular persistence is of clinical interest. A previous in vitro study showed that low substituted high molecular weight HES does not compromise blood coagulation more than medium molecular weight HES. In the present study we assessed the individual effects on blood coagulation of molar substitution and C2/C6 ratio of a high molecular weight HES. METHODS: Blood was obtained from 30 healthy patients undergoing elective surgery and mixed with six high molecular weight (700 kDa) HES solutions differing in their molar substitution (0.42 and 0.51) and C2/C6 ratio (2.7, 7 and 14) to achieve 20, 40 and 60% dilution. Blood coagulation was assessed by Thrombelastograph analysis (TEG) and plasma coagulation tests. Data were compared using a three-way analysis of variance model with repeated measures on the three factors. RESULTS: Higher molar substitution compromised blood coagulation most (for all TEG parameters, P<0.05). The lowest C2/C6 ratio was associated with the lowest effect on blood coagulation; r (P<0.001), angle alpha (P=0.003) and coagulation index (P<0.001). No effect on k and maximum amplitude was observed (P for both >0.50). The higher molar substitution was associated with a lesser increase in PT (P=0.007) and a greater decrease in factor VIII (P=0.010). PTT, functional and antigenic von Willebrand factors were not significantly influenced by molar substitution (P for all >0.20). No significant differences between solutions with the same molar substitution but different C2/C6 ratios were found in plasma coagulation parameters (P for all >0.05). CONCLUSIONS: TEG analysis indicates that high molecular HES with a molar substitution of 0.42 and a C2/C6 ratio of 2.7 has the lowest effect on in vitro human blood coagulation.

Molecular weight of hydroxyethyl starch: is there an effect on blood coagulation and pharmacokinetics?

BACKGROUND: The development of hydroxyethyl starches (HES) with low impact on blood coagulation but higher volume effect compared with the currently used HES solutions is of clinical interest. We hypothesized that high molecular weight, low-substituted HES might possess these properties. METHODS: Thirty pigs were infused with three different HES solutions (20 ml kg(-1)) with the same degree of molar substitution (0.42) but different molecular weights (130, 500 and 900 kDa). Serial blood samples were taken over 24 h and blood coagulation was assessed by Thromboelastograph analysis and analysis of plasma coagulation. In addition, plasma concentration and in vivo molecular weight were determined and pharmacokinetic data were computed based on a two-compartment model. RESULTS: Thromboelastograph analysis and plasma coagulation tests did not reveal a more pronounced alteration of blood coagulation with HES 500 and HES 900 compared with HES 130. In contrast, HES 500 and HES 900 had a greater area under the plasma concentration-time curve [1542 (142) g min litre(-1), P<0.001, 1701 (321) g min litre(-1), P<0.001] than HES 130 [1156 (223) g min litre(-1)] and alpha half life (t(alpha)(1/2)) was longer for HES 500 [53.8 (8.6) min, P<0.01] and HES 900 [57.1 (12.3) min, P<0.01] than for HES 130 [39.9 (10.7) min]. Beta half life (t(beta)(1/2)), however, was similar for all three types of HES [from 332 (100) to 381 (63) min]. CONCLUSIONS: In low-substituted HES, molecular weight is not a key factor in compromising blood coagulation. The longer initial intravascular persistence of high molecular weight low-substituted HES might result in a longer lasting volume effect.

Investigation of concurrent outbreaks of gastroenteritis and typhoid fever following a party on a floating restaurant, France, March 1998.

A retrospective cohort study was carried out to investigate concurrent outbreaks of gastroenteritis and typhoid fever that occurred among guests of a supper on a floating restaurant in France in March 1998. A total of 133 guests (attack rate = 90%) reported gastroenteritis within 12 days of the supper. Twenty-seven guests developed typhoid fever (attack rate = 18%) of whom 15 were confirmed by stool or blood culture. All patients with typhoid fever had had an initial gastroenteritis. The results suggest that the same food items served during the supper, chicken and rice, were the vehicles of both gastroenteritis and typhoid fever, but the authors could not determine the specific source of infection. Initial gastroenteritis has been described as a clinical manifestation of typhoid fever but whether or not these two syndromes (gastroenteritis and typhoid fever) were due to the same etiology remains unclear in this outbreak.

NO reduces PMN adhesion to human vascular endothelial cells due to downregulation of ICAM-1 mRNA and surface expression.

Reperfusion damage is largely due to the adherence of polymorphonuclear leukocytes to the endothelium initiated by adhesion molecule upregulation. The reduced endothelial nitric oxide release during ischemia may be involved in the upregulation of intercellular adhesion molecule 1. In this study, we tested if nitric oxide donors suppress polymorphonuclear leukocyte adherence to activated endothelial cells by inhibition of the intercellular adhesion molecule 1 surface expression. Confluent human umbilical vein endothelial cells were stimulated with tumor necrosis factor alpha (300 U/mL) after preincubation with increasing concentrations of the nitric oxide donors CAS 1609 (0.005-5 mM/L) and 3-(4-morpholinyl)-sydnonimine (0.01-1 mM/L). Intercellular adhesion molecule 1 surface expression was measured in a cell surface enzyme-linked immunosorbent assay, intercellular adhesion molecule 1 mRNA by Northern analysis. Human saphenous vein endothelial cells were transfected with the inducible nitric oxide synthase gene and stimulated with tumor necrosis factor alpha (300 U/mL). Fluorescein green-labeled polymorphonuclear leukocytes adhering to activated human umbilical vein endothelial cells/human saphenous vein endothelial cells were quantified by epifluorescent microscopy. The intercellular adhesion molecule 1 surface expression of activated human umbilical vein endothelial cells/human saphenous vein endothelial cells was significantly diminished to 40 to 60% of the maximum after treatment with CAS 1609, 3-(4-morpholinyl)-sydnonimine, or transfection with the inducible nitric oxide synthase gene. Intercellular adhesion molecule 1 mRNA was diminished by CAS 1609 and 3-(4-morpholinyl)-sydnonimine in the same manner. The functional relevance of our data was shown by reduction of polymorphonuclear leukocyte adherence to activated human umbilical vein endothelial cells/human saphenous vein endothelial cells following treatment with CAS 1609 and 3-(4-morpholinyl)-sydnonimine or transfection with inducible nitric oxide synthase. Tumor necrosis factor-induced polymorphonuclear leukocyte adherence was abolished by blocking antibody against intercellular adhesion molecule 1. Thus, exogenous or endogenous substitution of nitric oxide diminishes the expression of endothelial intercellular adhesion molecule 1 and its mRNA following tumor necrosis factor alpha stimulation. This results in a reduced polymorphonuclear leukocyte adherence to activated endothelium.

Malaria chemoprophylaxis of 3,446 French travelers departing from Paris to eight tropical countries.

BACKGROUND: Each year more and more French travelers are visiting areas where malaria is endemic. The aim of this study was to assess prophylactic regimens used by French travelers and to determine whether they meet current published recommendations. METHODS: This 12 month transversal study (May 1, 1995 to April 31, 1996) was conducted in embarkment lounges of Roissy Charles de Gaulle Airport to eight "tropical" destinations. RESULTS: 3,446 French travelers were enrolled. Twenty two and three-fifths percent of travelers had not sought any advice. The percentages of travelers staying less than 3 months (n = 2899) at risk of malaria (i.e., using none or inadequate chemoprophylaxis) were, according to the destination: Brazil (20%), Gabon (83%), Ivory Coast (26%), Kenya (43%), Madagascar (39%), Thailand (22%), Venezuela (41%) and Vietnam (8%). The suitability of the prophylaxis according to the information source for travelers staying less than 3 months varied as follows: specialist physician (OR = 1), travel agent (OR = 1.01, CI = 0.9 - 1. 1), occupational physician (OR = 1.13, CI = 0.6 - 2.1), GP (OR = 1. 58, CI = 1.1 - 2.3), none (OR = 1.95, CI = 1.3 - 2.9), friends (OR = 3, CI = 1.8 - 5) and pharmacist (OR = 3.94, CI = 2.1 - 7.5). Suitability of prophylaxis also varied according to the type of trip: organized tour (OR = 1), business trip (OR = 1.04, CI = 0.8 - 1.4), adventure tourism (OR = 2.1, CI = 1.6 - 2.9) and visit to family or friends (OR = 2.3, CI = 1.7 - 3.1). CONCLUSIONS: This study shows that the quality of advice on antimalarial chemoprophylaxis varies markedly according to the source, and that nearly one in three French travelers (29.3 %, 850/2899) to tropical areas is at risk of malaria.

Increased platelet sensitivity toward platelet inhibitors during physical exercise in patients with coronary artery disease.

Generalized atherosclerosis and coronary artery disease (CAD) are associated with endothelial dysfunction and during acute myocardial ischemia platelet activation has been reported. Activated platelets exert activated fibrinogen receptors (GP IIb/IIIa) and express CD 62p being regarded as reliable marker for platelet activation. Patients with angiographically proven CAD performed a bicycle exercise test until the onset of angina or ST-segment depression. We studied the ischemia-induced alterations in fibrinogen binding to activated platelet GP IIb/IIIa receptors and CD 62p expression. Therefore, the basal fibrinogen binding to GP IIb/IIIa and CD 62p expression and the thrombin-concentration for half-maximal platelet activation before and after exercise testing were determined. Additionally, inhibition of thrombin-induced platelet activation by increasing concentrations of the prostacyclin-analog iloprost and the NO-donor SIN-1 was examined. In patients with CAD, a significantly reduced basal activation and a highly significant reduction in sensitivity towards thrombin was measured. The thrombin-induced expression of GP IIb/IIIa and CD 62p was significantly diminished in patients with CAD after physical exercise and their platelets were significantly more sensitive towards the inhibitory effects of iloprost and SIN-1. These data demonstrate a significant reduction in platelet activation in response to physical exercise in patients with CAD and advanced atherosclerosis. Despite exercise induced myocardial ischemia as evidenced by angina and ECG-changes, the platelets are not generally activated, as it could be expected. Thus, patients with myocardial ischemia experienced a reduced platelet activity and enhanced sensitivity towards prostacyclin (PGI2) and nitric oxide, probably due to an augmented release of endogenous platelet inhibitory mediators.

A multicentre serosurvey on diphtheria immunity in a French population of 1004 subjects.

Diphtheria immunity was determined in serum specimens obtained in 1994 from 1004 subjects seen in emergency departments of three distant French cities. An enzyme immunoassay was used to measure serum diphtheria antitoxin concentrations according to the following criteria: (a) antitoxin < 0.01 IU/ml: susceptibility, (b) 0.01-0.09 IU/ml: basic protection, (c) > or = 0.10 IU/ml: full protection. Among these patients, 20.3% were fully susceptible to diphtheria, 30.3% had basic but doubtful protection and only 49.4% were fully protected. Protection was different by age-groups: 73.5% of the subjects under 40 years of age, 46% between 40 and 65 and 33% over 65 were fully protected. Protection decreased with increasing age (p < 0.001)and was greater for men than women after 40 years of age (p < 0.001). The results of this exploratory study indicate that the enhancement of diphtheria immunity by boosters in adult population should be reconsidered in France as well as in many industrialized countries.

MR of cerebral malaria.

In three cases of cerebral malaria, MR imaging disclosed either cortical infarcts (one case) or hyperintense areas of white matter (two cases) on T2-weighted and fluid-attenuated inversion-recovery sequences. These white matter abnormalities were, in one case, sharply limited, symmetrical, hyperintense, and unenhanced; in the other case, they were diffuse, hyperintense, and had a more limited focus. The diffuse hyperintensity was probably due to edema, whereas focal lesions were probably associated with gliosis.

Health hazards in international tourists visiting Paris in August: a five-year retrospective epidemiologic survey.

BACKGROUND: Travel-related illnesses have been studied in visitors to developing countries, but no studies have examined the incidence of health problems in visitors to developed countries. METHODS: 4, 093 foreign tourists visiting Paris in August and attending to emergency medical care for acute health problems were included in an epidemiological survey conducted over 5 consecutive years. The objective was to determine what types of acute health problems occur in a foreign tourist population and to estimate the incidence of the main health hazards. RESULTS: Gastroenteritis represented the main cause of medical care in that population (from 14.5-21.9%) followed by traumatology, ENT problem, viral syndrome and dermatology which represented altogether 60-64% of all medical problems. Two factors were related to the distribution of diseases observed: age and nationality. The monthly incidence of gastroenteritis was estimated to be between 1.33 to 2.92 per 10,000 visitors, and the overall incidence of health problems between 8 to 10 per 10,000. CONCLUSIONS: Even if the incidence rate of gastroenteritis is low compared with developing countries, further studies are needed to support the hypothesis that gastroenteritis could be attributed to sanitary conditions in some restaurants of the French capital.

[Emergency consultations of foreign tourists in Paris in the month of August. 5 years of prospective surveillance (1992-1996)]. / Consultations d'urgence des touristes étrangers à Paris au mois d'août. Cinq années de surveillance prospective (1992-1996).

For five consecutive years, five major Parisian institutions in charge of emergencies have participated in a prospective collection of medical data for foreign patients visiting Paris in August; 4093 subjects have been studied. Gastroenteritis represented the main cause in calling on emergency medical care (14.5 to 21.9%), followed by traumatology, ear-nose-throat problems, syndromes labelled as viral, skin problems: these five categories represented 60 to 64% of all the serious problems encountered by tourists. The statistical frequency of different causes in calling on emergency care varied significantly according to two variables: the tourists' age and nationality. The incidence of gastroenteritis is estimated at between 13 and 30 per 100,000 visitors and the incidence of pathological problems taken all together--at 80 to 100 per 100,000.

Prostacyclin receptor desensitization is a reversible phenomenon in human platelets.

BACKGROUND: Long-term exposure of platelets to endogenous or exogenous prostacyclin or its analogues might result in desensitization of the platelet prostacyclin receptor in vitro and in vivo accompanied by a loss in receptor density on the platelet surface and a reduced sensitivity toward the inhibitory effects of prostacyclins. However, the reversibility of this process in platelets has not yet been investigated. METHODS AND RESULTS: Human platelets desensitized by the chemically stable prostacyclin analogue iloprost showed a significant reduction in [3H]-iloprost binding sites that was reversed by saponin permeabilization. This indicates functionally active internalized prostacyclin receptors. To assess whether the internalized prostacyclin receptors recycle to the cell surface after withdrawal of the agonist, iloprost sensitivity and prostacyclin receptor binding properties of iloprost (30 nmol/L, 2 hours) desensitized platelets incubated in iloprost-free autologous plasma were investigated. While desensitized platelets showed a significant increase in IC50 for iloprost inhibition of thrombin-induced platelet aggregation, serotonin release, and p-selectin expression and a reduced iloprost-stimulated cAMP formation, platelet iloprost sensitivity was restored 3 hours after iloprost withdrawal. In addition, the significant reduction in Bmax and the increase in K(D) of prostacyclin receptors in desensitized platelets as revealed by [3H]-iloprost binding studies also returned to the initial values. CONCLUSIONS: These results indicate that prostacyclin receptors internalized during short-term desensitization are not degraded but can be recycled rapidly to the platelet surface in a functionally active form after withdrawal of the agonist.

Stability of essential drugs in the field: results of a study conducted over a two-year period in Burkina Faso.

To evaluate the stability of essential drugs stored in realistic tropical conditions, we have carried out a two-year prospective study in western Burkina Faso. Twenty-seven essential drugs were stored in a rural site and a urban one where temperature and hygrometry were recorded daily. Samples of each drug were taken for further analysis to the World Health Organization Collaborative Center for the Study of Stability of Drugs in Nantes, France every three months. Quantitative analysis showed that the majority of samples suffered no significant loss of their active ingredient. In contrast, ampicillin, erythromycin, sulfaguanidine, injectable furosemide, penicillin G, trimethoprim, and chloroquine showed more than a 10% quantitative loss of their active ingredient. Thus, it is not recommended that these essential drugs be stored for more than one year in a tropical climate.