The increasing threat to European forests from the invasive foliar pine pathogen, Lecanosticta acicola.

European forests are threatened by increasing numbers of invasive pests and pathogens. Over the past century, Lecanosticta acicola, a foliar pathogen predominantly of Pinus spp., has expanded its range globally, and is increasing in impact. Lecanosticta acicola causes brown spot needle blight, resulting in premature defoliation, reduced growth, and mortality in some hosts. Originating from southern regions of North American, it devastated forests in the USA's southern states in the early twentieth century, and in 1942 was discovered in Spain. Derived from Euphresco project 'Brownspotrisk,' this study aimed to establish the current distribution of Lecanosticta species, and assess the risks of L. acicola to European forests. Pathogen reports from the literature, and new/ unpublished survey data were combined into an open-access geo-database (http://www.portalofforestpathology.com), and used to visualise the pathogen's range, infer its climatic tolerance, and update its host range. Lecanosticta species have now been recorded in 44 countries, mostly in the northern hemisphere. The type species, L. acicola, has increased its range in recent years, and is present in 24 out of the 26 European countries where data were available. Other species of Lecanosticta are largely restricted to Mexico and Central America, and recently Colombia. The geo-database records demonstrate that L. acicola tolerates a wide range of climates across the northern hemisphere, and indicate its potential to colonise Pinus spp. forests across large swathes of the Europe. Preliminary analyses suggest L. acicola could affect 62% of global Pinus species area by the end of this century, under climate change predictions. Although its host range appears slightly narrower than the similar Dothistroma species, Lecanosticta species were recorded on 70 host taxa, mostly Pinus spp., but including, Cedrus and Picea spp. Twenty-three, including species of critical ecological, environmental and economic significance in Europe, are highly susceptible to L. acicola, suffering heavy defoliation and sometimes mortality. Variation in apparent susceptibility between reports could reflect variation between regions in the hosts' genetic make-up, but could also reflect the significant variation in L. acicola populations and lineages found across Europe. This study served to highlight significant gaps in our understanding of the pathogen's behaviour. Lecanosticta acicola has recently been downgraded from an A1 quarantine pest to a regulated non quarantine pathogen, and is now widely distributed across Europe. With a need to consider disease management, this study also explored global BSNB strategies, and used Case Studies to summarise the tactics employed to date in Europe.

Tracking the invasion: dispersal of Hymenoscyphus fraxineus airborne inoculum at different scales.

Ash dieback is caused by an invasive pathogen Hymenoscyphus fraxineus, which emerged in Europe in the 1990s and jeopardizes the management of ash stands. Although the biological cycle of the pathogen is well understood, its dispersal patterns via airborne spores remain poorly described. We investigated the seasonal and spatial patterns of dispersal in France using both a passive spore-trapping method coupled with a real-time PCR assay and reports of ash dieback based on symptom observations. Spores detection varies from year to year with a detection ability of 30-47% depending on meteorological conditions, which affect both production of inoculum and efficiency of the trapping. Nevertheless, our results are consistent and we showed that the sporulation peak occurred from June to August and that spores were detected up to 50-100 km ahead of the disease front, proving the presence of the pathogen before any observation of symptoms. The spore dispersal gradient was steep, most of inoculum remaining within 50 m of infected ashes. Two dispersal kernels were fitted using Bayesian methods to estimate the mean dispersal distance of H. fraxineus from inoculum sources. The estimated mean distances of dispersal, either local or regional scale, were 1.4 km and 2.6 km, respectively, the best fitting kernel being the inverse power-law. This information may help to design disease management strategies.

[Anti-NEP and anti-PLA2R antibodies in membranous nephropathy: an update]. / Anticorps anti-EPN et anti-PLA2R dans les glomerulopathies extramembraneuses: le point en 2014.

Membranous nephropathy (MN) is the most common cause for nephrotic syndrome in adults and occurs as an idiopathic (primary) or secondary disease. Since the early 2000's, substantial advances have been made in the understanding of the molecular bases of MN. The neutral endopeptidase (NEP) and the receptor for secretory phospholipase A2 (PLA2R) have been identified as target antigens for circulating and deposited antibodies in allo-immune neonatal and adult " idiopathic " MN, respectively. These antibodies recognize specific antigens of podocytes, precipitate as subepithelial immune complexes and activate complement leading to proteinuria. Anti-PLA2R antibodies are of particular clinical importance. Indeed, they are detected in approximately 70% of primary MN in adults, demonstrating that MN actually is an autoimmune condition specific to the kidney. In Europeans, genome-wide studies have shown an association between alleles of PLA2R1 and HLA DQA1 (class II genes of tissue histocompatibility complex) genes and idiopathic MN. Newly developed diagnostic tests detecting circulating anti-PLA2R antibody and PLA2R antigen in glomerular deposits have induced a change in paradigm in the diagnostic approach of idiopathic MN. Measurement of circulating anti-PLA2R antibody is also very useful for the monitoring of MN activity. However, the mechanisms responsible for the formation of anti-PLA2R antibodies as well as those involved in the progression of MN to end-stage renal disease remain to be defined.

Evidence for homoploid speciation in Phytophthora alni supports taxonomic reclassification in this species complex.

Alder decline has been a problem along European watercourses since the early 1990s. Hybridization was identified as the main cause of this emerging disease. Indeed, the causal agent, a soil-borne pathogen named Phytophthora alni subsp. alni (Paa) is the result of interspecific hybridization between two taxa, Phytophthora alni subsp. multiformis (Pam) and Phytophthora alni subsp. uniformis (Pau), initially identified as subspecies of Paa. The aim of this work was to characterize the ploidy level within the P. alni complex that is presently poorly understood. For that, we used two complementary approaches for a set of 31 isolates of Paa, Pam and Pau: (i) quantification of allele copy number of three single-copy nuclear genes using allele-specific real-time PCR and (ii) comparison of the genome size estimated by flow cytometry. Relative quantification of alleles of the three single-copy genes showed that the copy number of a given allele in Paa was systematically half that of its parents Pau or Pam. Moreover, DNA content estimated by flow cytometry in Paa was equal to half the sum of those in Pam and Pau. Our results therefore suggest that the hybrid Paa is an allotriploid species, containing half of the genome of each of its parents Pam and Pau, which in turn are considered to be allotetraploid and diploid, respectively. Paa thus results from a homoploid speciation process. Based on published data and on results from this study, a new formal taxonomic name is proposed for the three taxa Paa, Pam and Pau which are raised to species status and renamed P. ×alni, P. ×multiformis and P. uniformis, respectively.

[Experimental nephrology unit: overview and research projects]. / Unité de néphrologie expérimentale facultaire: etat des lieux et projets d'avenir.

After a short historical background of the Laboratory, the main research topics--renal toxicology, physiopathology of renal interstitial fibrosis and hormonology--are described in the perspective of a partnership between research clinicians and full time scientists. National as well as international scientific collaborations underline the need of combining expertises, stimulating also the training of youngest colleagues to the experimental approach of their future discipline.

Control of the duration of urinary catheterization: impact on catheter-associated urinary tract infection.

Catheter-associated urinary tract infection (CAUTI) is one of the most common types of nosocomial infection. Reducing the duration of catheterization may be a key intervention in CAUTI prevention. We carried out a prospective time-sequence, non-randomized intervention study in five departments of our university hospital. We wished to determine the impact of daily reminders from nurses to physicians to remove unnecessary urinary catheters four days after insertion. The intervention significantly decreased the duration of catheterization in two out of five departments. The frequency of late CAUTI (LCAUTI) among catheterized patients in all five departments decreased from 10.6 to 1.1 per 100 patients (P = 0.003) and the incidence of LCAUTI decreased from 12.3 to 1.8 per 1000 catheter-days (P = 0.03). Logistic regression analysis showed that duration of catheterization and iterative catheter changes were associated with LCAUTI. This study demonstrates that a simple measure can reduce the frequency of LCAUTI.

First Report of Phytophthora ramorum on Ornamental Plants in France.

In April 2002, Phytophthora ramorum was associated with twig blight and brown spots on Rhododendron spp. leaves from a nursery in France. The isolate was identified by its morphological characters on V8 agar: slow growth, deciduous and semipapillate sporangia, and abundant production of large chlamydospores (3). The identification was confirmed by ITS rDNA sequencing. During 2002, P. ramorum was also isolated from diseased Viburnum tinus and V. × bodnantense plants exhibiting symptoms of wilting and stem base discoloration. Subsequently, repeated surveys for P. ramorum were carried out in nurseries and areas surrounding nurseries throughout France. Since 2004, a large range of known hosts were investigated in approximately 2,000 nurseries and 200 other sites each year. P. ramorum was detected exclusively in nurseries at 29 locations in 2002, 9 in 2003, 23 in 2004, 17 in 2005, and 19 in 2006. Rhododendron spp. and occasionally V. tinus were the major hosts. In addition, the pathogen was detected for the first time on Pieris japonica in two nurseries in 2005 and on Camellia sp. in one nursery in 2006 from plants exhibiting leaf and twig blight. In both cases, P. ramorum had already been detected on Rhododendron spp. in the same nurseries. Most of the infected plants were found in northwestern France (Bretagne and Pays-de-la-Loire), or came from this region, which is the main rhododendron-growing area in France. In some cases, plants were imported from Belgium or the Netherlands. P. ramorum was also detected in a nursery in soil close to diseased Rhododendron spp. plants and pond water used for irrigation by using a combination of baiting with Rhododendron spp. leaves and PCR assay with species-specific primers (1). Overall, approximately 1% of the investigated nurseries were found positive each year, and this ratio was quite stable from 2004 to 2006. To date, P. ramorum has not been detected outside of nurseries, although many surveys were conducted on the west coast of France where the risk is considered to be high because of a favorable mild and humid climate and the presence of suitable hosts. In addition, 78 isolates of P. ramorum collected between 2002 and 2004 on Rhododendron spp. and V. tinus were found to be of A1 mating type based on pairings with P. cryptogea A1 and A2 mating types (2). References: (1) K. J. Hayden et al. Phytopathology 94:1075, 2004. (2) S. Werres and B. Zielke J. Plant Dis. Prot. 110:129, 2003. (3) S. Werres et al. Mycol. Res. 105:1155, 2001.

Risk factors for the phytophthora-induced decline of alder in northeastern france.

ABSTRACT A lethal disease of common alder caused by Phytophthora alni, a new hybrid pathogen, has been spreading in Europe since the early 1990s. In 2004, we conducted an epidemiological survey in northeastern France to determine disease frequency and to investigate the impact of environmental factors on disease prevalence. Seventy-eight plots in the Rhin-Meuse basin were investigated. The survey was structured to enable critical examination of the possible impact of nitrogen pollution of the river water on disease prevalence. P. alni-induced alder decline was common throughout northeastern France. Altogether, disease was found in 80% of the plots containing alder, with 16% of all the alders affected. Striking differences existed between watercourse types. Lower proportions of diseased alders were found in watercourse types with rapid water flow, such as mountain streams of the Vosges and piedmont or watercourses on steep calcareous slopes, than in the slow watercourses of the low-lying valleys of the calcareous plateaus and of the clayey plains. Disease prevalence was not related to the total oxidized nitrogen concentration of the water. However, prevalence increased with the mean summer temperature of the river water and where clayey soils were found in the river banks. The results of this work can be used for the assessment of P. alni-induced alder decline risks in affected European countries and in areas where the disease could be introduced.

Orofacial granulomas after injection of cosmetic fillers. Histopathologic and clinical study of 11 cases.

BACKGROUND: Purposely, cosmetic injections in orofacial tissues of various resorbable, biodegradable, or permanent fillers may be followed by development of foreign-body granulomas. The aim of this article is to contribute to the histologic identification of the filler material. METHODS: Histologic and clinical features of 11 cases of granulomas on orofacial fillers are described. RESULTS: Only 3/11 patients knew the nature of the injected product. Four histologic patterns were found: (i) Artecoll, (ii) Dermalive, and (iii) New-Fill granuloma, all three of the classic giant cell granuloma type, differing in respect of foreign particles; and (iv) Liquid Silicone granuloma, which featured a cystic and macrophagic type. Information was often missing or misleading, patients or practitioners being reluctant to give the details. CONCLUSION: Increasing demand for orofacial tissue augmentation makes pathologists face new, and sometimes, puzzling granuloma types. Identification of the foreign product might be required for therapeutic or medico-legal reasons.

Partial Resistance to Melampsora larici-populina Leaf Rust in Hybrid Poplars: Genetic Variability in Inoculated Excised Leaf Disk Bioassay and Relationship with Complete Resistance.

ABSTRACT Inoculated excised leaf disk technique allows decomposition of poplar partial resistance to Melampsora larici-populina leaf rust into key epidemiological components such as latent period (LP), uredinia number (UN), uredinia size (US), and spore production (SP) for a given M. larici-populina strain under controlled environmental conditions. Three hundred thirty-six genotypes from an interspecific Populus deltoides x P. trichocarpa F(1) progeny segregating for complete resistance to M. larici-populina strain 93ID6 were inoculated with M. larici-populina strain 93CV1. This strain was able to infect the whole family, except few probable recombinants. LP, final UN, and final US after one infectious cycle proved to be relevant complementary descriptors of partial resistance. Area under the disease progress curve and other parameters of uredinia appearance dynamics did not yield additional information. Indirect assessment of SP by US scoring was reliable and easy to access compared with direct spore counting. UN was the only trait for which a doubling of the inoculum pressure level had a significant effect, leading to greater differentiation between genotypes. Consistent with previous studies is the clear relationship between presence of complete resistance against M. larici-populina strain 93ID6 and higher partial resistance to M. larici-populina strain 93CV1 (32% longer LP, 76% smaller UN, and 34% smaller US). In the subpopulation compatible with 93ID6, bimodal distribution of genotypic means for US suggested implication of a major gene inherited from the P. trichocarpa parent. Residual variation was noted for the three epidemiological components, suggesting that additional genes might condition these quantitative traits.

Solid state and solution studies of lanthanide(III) complexes of cyclohexanetriols, models of the coordination sites found in sugars.

This report covers studies in trivalent lanthanide complexation by two simple cyclohexanetriols that are models of the two coordination sites found in sugars and derivatives. Several complexes of trivalent lanthanide ions with cis,cis-1,3,5-trihydroxycyclohexane (L(1)()) and cis,cis-1,2,3-trihydroxycyclohexane (L(2)()) have been characterized in the solid state, and some of them have been studied in organic solutions. With L(1)(), Ln(L)(2) complexes are obtained when crystallization is performed from acetonitrile solutions whatever the nature of the salt (nitrate or triflate) [Ln(L(1)())(2)(NO(3))(2)](NO(3)) (Ln = Pr, Nd); [Ln(L(1)())(2)(NO(3))H(2)O](NO(3))(2) (Ln = Eu, Ho, Yb); [Ln(L(1)())(2)(OTf)(2)(H(2)O)](OTf) (Ln = Nd, Eu). Lanthanum nitrate itself gives a mixed complex [La(L(1)())(2)(NO(3))(2)][LaL(1)()(NO(3))(4)] from acetonitrile solution while [La(L(1)())(2)(NO(3))(2)](NO(3)) is obtained using dimethoxyethane as reaction solvent and crystallization medium. With L(2)(), Ln(L)(2) complexes have also been crystallized from methanol solution [Ln(L(2)())(2)(NO(3))(2)]NO(3), (Ln = Pr, Nd, Eu). Single-crystal X-ray diffraction analyses are reported for these complexes. Complex formation in solution has been studied for several triflate salts (La, Pr, Nd, Eu, and Yb) with L(1 )()and L(2)(), respectively in acetonitrile and in methanol. In contrast to the solid state, both structures Ln(L) and Ln(L)(2) equilibrate in solution, as was demonstrated by low-temperature (1)H NMR and electrospray ionization mass spectrometry experiments. Competing experiments in complexing abilities of L(1)() and L(2)() with trivalent lanthanide cations have shown that only L(2)() exhibits a small selectivity (Nd > Pr > Yb > La > Eu) in methanol.

Role of complement and platelet-activating factor in the stimulation of phagocytosis and reactive oxygen species production during haemodialysis.

BACKGROUND: Neutrophil phagocytic functions have been studied extensively in haemodialysis (HD) patients; however, results are contradictory and the mechanisms that modulate phagocytosis and oxidative burst during dialysis are not completely understood. METHODS: The present study investigated neutrophil functions in a selected population of patients before and during clinical dialysis with cuprophane, and polyacrylonitrile (AN69) membranes. We measured phagocytosis of Escherichia coli and intracellular hydrogen peroxide (H2O2) production by flow cytometry in whole blood. RESULTS: Before dialysis, neutrophils from HD patients showed normal phagocytic capability and H2O2 formation. Phagocytosis of FITC-E. coli was significantly stimulated in cuprophane but not AN69-treated patients. Spontaneous and stimulated H2O2 production was enhanced with both cuprophane and AN69 membranes. We then investigated in vitro the role of complement and platelet-activating factor (PAF) in the activation of neutrophils. Incubation of whole blood with C5a increased phagocytosis but not H2O2 production. On the contrary, the addition of synthetic PAF showed a markedly stimulated H2O2 production without increase in phagocytosis. Moreover, during dialysis with formaldehyde-reused cuprophane, complement activation was abolished and phagocytosis was no longer enhanced, while the stimulation of H2O2 production persisted. In addition, we also excluded a particular role of the membrane itself in the activation of neutrophils. CONCLUSION: We demonstrated that in a selected population of HD patients, neutrophils exhibit normal phagocytic capability and normal intracellular H2O2 production. During dialysis, the stimulation of phagocytosis observed with cuprophane is complement dependent, whereas the enhanced H2O2 production observed with both cuprophane and AN69 membranes might be related to PAF production.

Role of the cholinergic pathway in mediating platelet-activating factor-induced pulmonary dysfunctions in unsedated calves.

The purpose of this study was to investigate the possible role of the cholinergic pathway in mediating platelet-activating factor (PAF)-induced pulmonary dysfunctions in unsedated calves. In a placebo group, PAF infusion challenge induced significant dysfunctions in the pattern of breathing [a significant increase in respiratory rate (RR) and a significant decrease in tidal volume (VT)], the mechanics of breathing [a significant increase in total lung resistance (RL) and a significant decrease in dynamic lung compliance (CLdyn)] and gas exchange, whereas in atropine pre-treated calves, PAF infusion challenge induced a significant increase in RR, VT and HR and a significant decrease in CLdyn. The RL increase was prevented by atropine pre-treatment. On the basis of our findings, we suggest that, in cattle, the PAF-induced pattern of breathing dysfunctions and the diffuse bronchoconstriction and microvascular leakage of small airways are not mediated through the cholinergic pathway. By contrast, our data suggest that PAF-induced bronchoconstriction of upper airways is at least partly mediated through muscarinic receptors.

Efficacy of oral immunotherapy on respiratory infections in hemodialysis patients: a double-blind, placebo-controlled study.

BACKGROUND: Hemodialysis (HD) patients suffer from several immune defects that make them prone to develop bacterial infections, in particular respiratory tract infections (RTIs). PATIENTS AND METHODS: As previous studies have shown that oral immunotherapy with an immunomodulating bacterial extract (IBE) is effective against RTIs, we decided to test its efficacy and safety in HD patients during a double-blind placebo-controlled prospective study. 40 HD patients with a documented history of RTIs in the previous year were treated for 24 weeks of the endemic season with one capsule daily of IBE (n = 21) or placebo (PL, n = 19). Clinical examinations, measurements of Mac-1 and gp150.95 on circulating phagocytes and routine laboratory evaluations were performed at week 0, 4, 12 and 24. Patients were also examined at each dialysis session allowing an accurate recording of any infectious episode, its treatment and of any untoward effect. RESULTS: During the last period of the study (weeks 13-24), IBE significantly reduced the number of patients with RTIs and consequently of antibiotic treatment courses as compared to PL (p = 0.018), whereas no difference was detected between IBE and PL during periods I (weeks 0-4) and II (weeks 5-12). There was no difference between IBE and PL for other, non respiratory infections. IBE was associated at several time points with an increased expression on phagocytes of adhesion molecules involved in phagocytosis (Mac-1 and gp150.95). However, the expression of these molecules was not predictive for the occurrence of RTI. IBE was on the whole as well tolerated as PL, 7 patients presented side effects (5 IBE, 2 PL, NS) which led to drop-out in 4 cases (3 IBE, 1 PL). No serious side effect was recorded, gastrointestinal upset being the most prevalent type. CONCLUSION: The results of this study indicate that immunomodulation with selected bacterial extracts constitutes a promising approach for the prevention of bacterial airway infections in groups at risk, such as HD patients.

[Seroprevalence of anti-human parvovirus B19 antibodies in a sample of blood donors in Galicia]. / Seroprevalencia de anticuerpos antiparvovirus B19 en una muestra de donantes de sangre en Galicia.

BACKGROUND: To determine the seroprevalence of antibodies to human parvovirus B19 (PVB19) in a series of 92 blood donors in Galicia. METHODS: Antibodies of the immunoglobulin G (IgG) class were investigated in serum samples from 44 females and 48 males aged 18-63 years (mean: 29.40; SD: 10.81) using a commercial ELISA system consisting of highly specific recombinant antigen. RESULTS: Nine positive cases (9.78%; CI 95%: 4.76-17.98%) were detected, including six females and three males with a mean age slightly higher as compared to all blood donors group. The prevalence of antibodies against PVB19 increased with age. No sex or age differences in the positive results were observed. CONCLUSIONS: These data indicate a PVB19 seroprevalence lower that in previous studies in several countries and in other areas in our country; since published references are limited in this community, complementary studies to establish the importance of this virus infection are justified.

Direct activation of human peritoneal mesothelial cells by heat-killed microorganisms.

OBJECTIVE: The aim of the study was to determine if human peritoneal mesothelial cells (HPMCs) can be activated directly by bacterial products contained in preparations of heat-killed Escherichia coli and staphylococci. SUMMARY BACKGROUND DATA: It has been shown recently that cytokine-activated HPMCs produce the inflammatory mediators, interleukin-1, interleukin-6, interleukin-8, and macrophage chemotactic protein-1. Studies concerning the effects of bacterial products on HPMCs are scarce and have not yielded conclusive results. METHODS: Growth-arrested HPMC monolayers were prepared from cell suspensions obtained by enzymatic disaggregation of small pieces of omentum. They were incubated for 24 hours with heat-killed E. coli (ATCC 25922), heat-killed staphylococci (ATCC 25933), or E. coli lipopolysaccharide, and the release of various cytokines in the culture media was measured by radioimmunoassays or enzyme-linked immunosorbent assays. Results were expressed as mean +/- standard error of the mean in picograms per milliliter of supernatant and analyzed with the Wilcoxon test; p values of less than 0.05 were considered significant. RESULTS: Baseline production of interleukin-6, interleukin-8, the chemokine "regulated upon activation, normal T cell expressed and secreted" (RANTES), and macrophage chemotactic protein-1 varied widely from one omental preparation to the other. E. coli increased the release of these mediators: from 1206 +/- 316 pg/mL to 8480 +/- 2189 pg/mL for interleukin-6, from 285 +/- 58 pg/mL to 3164 +/- 1053 pg/mL for interleukin-8, from 7 +/- 5 pg/mL to 684 +/- 264 pg/mL for RANTES, and from 2212 +/- 346 pg/mL to 7726 +/- 1473 pg/mL for macrophage chemotactic protein-1. Heat-killed staphylococci did not alter significantly the production of RANTES or macrophage chemotactic protein-1 but increased the production of the two other cytokines from 1325 +/- 389 pg/mL to 2206 +/- 523 pg/mL for interleukin-6 and from 318 +/- 70 pg/mL to 819 +/- 265 pg/mL for interleukin-8. CONCLUSIONS: The authors' results show that HPMCs are able to react to a direct stimulation with heat-killed microbes. They suggest that HPMCs, as well as resident macrophages, participate actively in the initiation and possibly in the modulation of intraperitonen inflammatory reactions.