As this family exhibits a great diversity in shell morphology but extremely homogeneous anatomical traits, the systematics of Lymnaeidae has long been controversial. Using the most complete dataset to date, we examined phylogenetic relationships among 50 taxa of this family using a supermatrix approach  involving both Maximum Likelihood and Bayesian Inference.Lymnaeidae snails play a prominent role in the transmission of helminths, mainly trematodes of medical and veterinary importance . We found that intermediate hosts of F. gigantica cluster within one deep clade, while intermediate hosts of F. hepatica are widely spread across the phylogeny. In addition, chromosome number seems to have evolved from n = 18 to n = 17 and n = 16.Our phylogenetic analysis demonstrates the existence of three deep clades of Lymnaeidae representing the main geographic origin of species . This phylogeny allowed us to discuss on potential biological invasions and map important characters, such as, the susceptibility to infection by Our study contributes to deepen our understanding of Lymnaeidae phylogeny by both sampling at worldwide scale and combining information from various genes (supermatrix approach). This phylogeny provides insights into the evolutionary relationships among genera and species and demonstrates that the nomenclature of most genera in the Lymnaeidae does not reflect evolutionary relationships. This study highlights the importance of performing basic studies in systematics to guide epidemiological control programs. The Lymnaeidae, Physidae and Planorbidae comprise ~90% of the Basommatophoran species Radix sperding to ), on theribed in  with no Recent studies have suggested that the Lymnaeidae contains approximately 100 species , meaningL. diaphana in a phylogenetic analysis and suggests that this species is sister to the North American Stagnicola and L. occulta (= C. occulta) (clade C1a), although with low support (0.54 BP and 0.71 PP). Baker L. diaphaG. truncatula-like) in the C1b clade cluster together. The only exception is L. cousini which clearly belongs to this clade  but is morphologically different from all other species in terms of both shell morphology and the anatomy of reproductive organs. Its phylogenetic position suggests the ancestor of clade C1b had a G. truncatula-like morphology and the L. cousini morphology is a derived character. The inclusion of G. truncatula within this clade is unambiguous and agrees with previous results 18]. It dposed by .Radix species. According to this tree, Austropeplea is a polyphyletic genus, in agreement with results obtained by  and andRadixained by . Remigioent with , 11], Radix speent with , sister nonymous . Their snonymous ,55. Alteent with , and migRadix clade diversified and then expanded towards Eurasia and Africa. This is also true of G. truncatula, a branch of the American clade that invaded the Old World. In general, the Lymnaeidae morphology has evolved slowly and most species within clades are similar: small-shelled turriform, G. truncatula-like in the American clade; large and high-spired shells in the Eurasian Lymnaea; and large, rounded or ovate shells in the Indo-Pacific clade, especially in Radix. A few branches, including O. glabra, and L. cousini, have evolved distinctive morphologies that differ from all other lymnaeids. Note, however, that limpet-shaped species  were not analyzed here, and it is not possible to discuss their phylogenetic position and morphological evolution. If they constitute a separate clade, say C4, our hypothesis of slow morphological evolution would be valid.In summary (and ignoring recent introductions) we have three old centers of diversification in the Lymnaeidae family: America, Eurasia and the Indo-Pacific region. In the latter, the Austropeplea with 16 chromosome pairs was thought to be the most "primitive" form, followed by Radix with 17 pairs and Stagnicola with 18 pairs. Our molecular phylogeny contradicts this idea. The ancestral state in Lymnaeidae seems to be n = 18, as it is in other Basommatophoran gastropods  56] hypothysidae) . Eighteeested by , either Radix (including L. (R.) natalensis).The nomenclature of genera has been one of the most confusing issues in the Lymnaeidae systematics. Most genus names are not fixed and are based more on phenotypic resemblances than on sound evolutionary and phylogenetic considerations. For instance, a single genus was recognized by , two by Lymnaea is L. stagnalis Linnaeus, 1758; the type species of Stagnicola Jeffreys, 1830 is S. palustris ; and the type species of Omphiscola Rafinesque, 1819 is O. glabra. However, it is clear from our results that these three species belong to the same clade (C2) and that Lymnaea is not a monophyletic group. We propose that species of clade C2 should all be called Lymnaea, according to the principle of priority of the International Code of Zoological Nomenclature (ICZN). By extension, Stagnicola should not be used to name species in clade C1a since the type species belongs to clade C2. Meier-Brook and Bargues [S. emarginata, S. elodes, S. catascopium and L. occulta within a new genus Catascopium, while S. caperata would belong to the genus Hinkleyia Baker, 1928 [Hinkleyia would be the preferable name according to the ICZN. On the other hand, at least four genera names have been used for species of clade C1b: Lymnaea Lamarck, 1799; Galba Schrank, 1803; Fossaria Westerlund, 1885; and Bakerilymnaea. In the light of the present results, it would be preferable to unify nomenclature. According to the ICZN, Lymnaea should be the unified name, but given that the type species belongs to clade C2, Galba could be a more appropriate name. Finally, as said above, Austropeplea Cotton, 1942 is not a monophyletic group, and employing the genus Kutikina Ponder and Waterhouse, 1997 (one species: K. hispida) seems unjustified on the basis of the current phylogeny. This would also be consistent with results of [Bullastra Pfeiffer, 1839 for all species of clade C3a to fit the ICZN.The type species of  Bargues  suggestesults of . It woulAlthough the Lymnaeidae phylogeny presented in Figure G. truncatula Müller, 1774, the main vector of fascioliasis in the Old World. The idea that this species is native to Europe, as it was described from Germany, is widely accepted [G. truncatula originated. This is consistent with its detection in Alaska and the Yukon territory [G. truncatula in North America is broader, but has remained cryptic because it has been confounded with other taxa. Indeed, Hubendick noted that, "it is a matter of some doubt whether Lymnaea humilis in North America is a distinct species or is specifically connected to L. truncatula" (= G. truncatula) [L. humilis in reality correspond to G. truncatula. At least 10 species of Lymnaeidae from North America placed in synonymy of L. humilis by [G. truncatula: L. galbana Say, 1825; L. modicella Say, 1825; L. obrussa Say, 1825; L. parva Lea, 1841; L. exigua Lea, 1841; L. rustica Lea, 1841; L. tazevelliana Wolf, 1869; L. dalli Baker, 1906; L. cyclostoma Walker, 1908; and L. peninsulae Walker, 1908. Unfortunately, neither detailed anatomical descriptions nor molecular data are available in any of these taxa. Sampling and sequencing of morphologically similar North American taxa could shed light on this question.First, one of our most puzzling results concerns the origin of accepted . Howevererritory . It is pncatula) . It coulmilis by , but conmilis by , presentL. occulta (= C. occulta) is the only species from Europe clustering within clade C2 and seems to correspond to a passage from North America to Europe. As stated by [L. occulta is distributed in eastern Germany, Poland, the former Czechoslovakia, the former Yugoslavia, Ukraine, Sweden, and some rivers in the delta of Lake Baikal. It is hypothesized that the species could reach the far east Asia where it could have been confounded with other stagnicoline species because of similarity in shell morphology [L. occulta has its origins in America.Second, as mentioned above, tated by   are found in distant geographic locations is indicative of recent introductions.Finally, the two F. hepatica seems to be different. Our results show that lymnaeid species serving as intermediate hosts of this trematode are widely distributed across the phylogeny  or stenoxenous  . This isy Figure . Basicaly Figure . In conty Figure . Althougtionship . The behG. truncatula and L. occulta (= C. occulta), as well as passages from the Indo-Pacific to Europe and North America for Radix sp. However, sampling and sequencing efforts remain to be done especially in the Palaearctic and Nearctic regions in which the family diversity is thought to be the largest. This would help resolve the weakly supported relationships , determine the pace of morphological evolution, establish taxonomic synonymy, and determine the phylogenetic relevance of poorly known genera . Second, with the exception of Radix (including the African L. (R.) natalensis), genus names in Lymnaeidae do not reflect the phyletic relationships among species. The group taxonomy should be reconsidered to gain some biological meaning. Third, the number of chromosomes in Lymnaeidae has evolved from an ancestral state of 18 pairs to a derived 17 and 16 pairs. Finally, while the intermediate hosts of F. gigantica are basically restricted to clade C3, F. hepatica is able to infect species from all main clades . This suggests that the cosmopolitan distribution of F. hepatica is largely favored by its capacity to infect local lymnaeids, and highlights the importance of the correct identification of intermediate-host species in fascioliasis control programs.At least four conclusions can be drawn from this study. First, combining information from different genes (supermatrix) is a robust approach to reconstruct the evolutionary history of the Lymnaeidae. Our results indicate that members of this family diverged in three deeply-rooted clades corresponding to the geographic origin of species . Our phylogeny allowed us to pinpoint discordances between ancestral and current geographic distributions of some species, potentially indicating more or less recent biological invasions. Transfers from America to Eurasia are suggested for ACC and SHB designed the study. ACC, JSE, PDavid, PJ and JPP sampled snails. ACC and PDurand carried out experiments. ACC and JSE performed analyses. All authors discussed the results and wrote the paper.


A common belief is that, unlike today, ancient urban areas developed in a sustainable way within the environmental limits of local natural resources and the ecosystem's capacity to respond. This long-held paradigm is based on a weak knowledge of the processes underpinning the emergence of urban life and the rise of an urban-adapted environment in and beyond city boundaries. Here, we report a 6000-year record of environmental changes around the port city of Akko (Acre), Israel, to analyse ecological processes and patterns stemming from the emergence and growth of urban life. We show that early urban development deeply transformed pre-existing ecosystems, swiftly leading to an urban environment already governed by its own ecological rules and this, since the emergence of the cities. The question of whether or not the emergence of cities13456891011121411112345Here, the emergence of an urban environment as a unique setting was analyzed through the investigation of patterns of pressure and selection on ecosystems on the coastal strip of Israel, the ancient and present economic heart19202118O values on Ashdod Coast233033342021An outstanding feature of the recorded urban environmental history of Akko is that the area rapidly shifted from a densely forested landscape to a shrub-steppe  between versus groundwater; depending on functional pits) for urban growth. Although the 3200 cal yr BP event locally ended at ~2900–2800 cal yr BP, the re-emergence of extensive habitation pattern and trade networks only occurred after ~2650 BP, during the Phoenician/Persian and Early Hellenistic periods, and is clearly marked in the environment  ages of short-lived terrestrial samples  extracted from a continuous core  drilled on the southern flank of Tel Akko , close t samples . All conArchaeological and historical data for urban development derive from 40 years of excavation at the ancient site of Tel Akko (since the work of M. Dothan) and in the “old city”, especially the crusader town (since the surveys of the British Mandate Authorities).A total of 70 samples were prepared for pollen analysis using the standard palynological procedure for clay samples. Pollen grains were counted at ×400 and ×1000 magnification using an Olympus microscope. Pollen frequencies (%) are based on the terrestrial pollen sum excluding local hygrophytes and spores of non-vascular cryptogams. Aquatic taxa frequencies were calculated by adding the local hygrophytes-hydrophytes to the terrestrial pollen sum.Paired group as algorithm and Spearman's Rho  as similarity measure. The pollen-types from each cluster were summed to create nine pollen-derived ecosystems , Kernel density-2D, Matrix, and Principal Components Analysis (PCA). The CA is a technique for hierarchical clustering, finding ranked groupings in multivariate data sets. Here, it is based on pollen-type time-series (presence/absence and abundance). The CA  was usedosystems .Gaussian as a basis function and three units as radius  correspond to the main loadings in the PCA, explaining most of the variance for the PCA-Axis 1 ordination of the data, which accounts for +0.777 of total inertia. Natural ecosystems  are loaded in positive values, whereas negative values correspond to urban-adapted ecosystems . The PCA-Axis 1 scores have been plotted on a linear age-scale to determine the main changes in the Akko core . A simulation of each vegetation assemblage is obtained and compared to the pollen data. Acceptable scenarios (according to MCMC rules)i.e. a strong underestimation of the winter temperatures, because several tree taxa cannot be unequivocally assigned to cool or warm conifer plant typesThe climatic data  used areD.K., E.V.C., C.M. and M.A. had the idea for the research. D.K. and E.V.C. performed the research, undertook the data collection, and led the statistical analysis. J.G. led the climate reconstruction. D.K., E.V.C., C.M., J.G., D.Z., I.S., T.O. and M.A. contributed to the analysis and interpretation of the results. D.K. wrote the first draft of the paper and all authors contributed to writing the manuscript.Supplementary Figures and Tables


In this article, the authors acknowledged Dr Sarah Cleaveland and Dr Craig Packer as having provided detailed comments on earlier versions of the manuscript. Although the authors sent a revised version to Dr Cleaveland mentioning her name in the acknowledgments before submission to PLoS ONE and received no correspondence suggesting any objection, they accept that she did not formally agree to be included in the Acknowledgments.Similarly, the authors did not seek Dr Packer's formal agreement for him to be listed in the Acknowledgements. As a result, they would like to issue this Correction in order to remove the reference to Drs. Cleaveland and Packer and revise the text under the Acknowledgments to:"We thank TANAPA, TAWIRI and the Tanzania Commission for Science and Technology for providing permission to conduct the long-term study in the Serengeti. We would like to thank all who have contributed to the fieldwork of the Serengeti Cheetah Project. Numerous people and organisations in Tanzania and Kenya have provided much needed logistical support over the years, including B. Allen, O. Newman, A. Barrett, J. Driessen, J. Jackson, A. Geertsema, P. and L. White and the staff and management of Ndutu Safari Lodge, H. van Lawick and his team, fellow scientists at SWRC, G. and M. Russell and M. Borner and the staff at FZS. We thank John Harwood for initial discussions, as well as Tim Coulson and Dominique Pontier for providing detailed comments on earlier versions of this manuscript. The authors also gratefully acknowledge the many organisations that have funded the project, principally The Howard G. Buffett Foundation, Wildlife Conservation Society, Frankfurt Zoological Society (FZS), National Geographic Society, Leverhulme Trust, Royal Society UK, Times Christmas Appeal 1998, Messerli Foundation and People's Trust for Endangered Species."


Anopheles darlingi is the major malaria vector in countries located in the Amazon region. Anopheles aquasalis and Anopheles albitarsis s.l. are also proven vectors in this region. Anopheles nuneztovari s.l. and Anopheles triannulatus s.l. were found infected with Plasmodium vivax; however, their status as vectors is not yet well defined. Knowledge of susceptibility of Amazon anopheline populations to Plasmodium infection is necessary to better understand their vector capacity. Laboratory colonization of An. darlingi, the main Amazon vector, has proven to be difficult and presently An. aquasalis is the only available autonomous colony.An. darlingi, An. albitarsis s.l., An. nuneztovari s.l. and An. triannulatus s.l. were collected in the field and reared until adult stage. Adults of An. aquasalis were obtained from a well-established colony. Mosquitoes were blood-fed using a membrane-feeding device containing infected blood from malarial patients.Larvae of Anopheles species was evaluated by the impact variance of the following parameters: (a) parasitaemia density; (b) blood serum inactivation of the infective bloodmeal; (c) influence of gametocyte number on infection rates and number of oocysts. The goal of this work was to compare the susceptibility to P. vivax of four field-collected Anopheles species with colonized An. aquasalis.The infection of the distinct Anopheles species tested were susceptible to P. vivax infection, nevertheless the proportion of infected mosquitoes and the infection intensity measured by oocyst number varied significantly among species. Inactivation of the blood serum prior to mosquito feeding increased infection rates in An. darlingi and An. triannulatus s.l., but was diminished in An. albitarsis s.l. and An. aquasalis. There was a positive correlation between gametocyte density and the infection rate in all tests  but varied among the mosquito species. Anopheles albitarsis s.l., An. aquasalis and An. nuneztovari s.l. had higher infection rates than An. darlingi.All Anopheles species, as well as colonized An. aquasalis are susceptible to experimental P. vivax infections by membrane feeding assays. Anopheles darlingi, An. albitarsis s.l. and An. aquasalis are very susceptible to P. vivax infection. However, colonized An. aquasalis mosquitoes showed the higher infection intensity represented by infection rate and oocyst numbers. This study is the first to characterize experimental development of Plasmodium infections in Amazon Anopheles vectors and also to endorse that P. vivax infection of colonized An. aquasalis is a feasible laboratory model.All field-collected  Malaria is an infectious disease with major impact on global public health and economy. Currently, malaria threatens almost one third of the world population in 103 tropical countries, resulting in 207 million of clinical cases and 627,000 deaths in 2012 [Plasmodium are transmitted by anopheline mosquitoes. Plasmodium falciparum and Plasmodium vivax are the main species-affecting humans. Actually, since the mid-1990s, P. vivax has become the predominant New World malaria species, as it has expanded its range throughout South America [P. vivax[P. vivax malaria is generally considered to be relatively benign, there are numerous reports of complex cases with severe clinical complications and deaths [Human malaria parasites in the genus [P. vivax. The cond deaths -10.Plasmodium life cycle is the infection of mosquito vectors. Among the 33 Anopheles mosquito species described from the Brazilian Amazon region, Anopheles darlingi is considered to be the main malaria vector. Other anopheline species can be considered secondary or occasional malaria vectors because of their population density, anthropophilic behavior and natural infectivity across their geographic distributions [Anopheles albitarsis s.l., An. nuneztovari s.l. and An. triannulatus s.l. are commonly collected in the Amazon, and they have been observed infected with P. vivax and P. falciparum, but their role as malaria vectors has not yet been elucidated [Anopheles aquasalis is distributed predominantly along the Atlantic coast because of its tolerance to saltwater environment and has been found naturally infected by P. vivax[One keystone stage in the ibutions -14. Anopucidated -25. Anop P. vivax,26-28 inAn. darlingi has been associated with malaria transmission in Bolivia, Colombia, French Guiana, Guyana, Peru, Suriname, and Venezuela [An. albitarsis s.l. in Venezuela [An. nuneztovari s.l. in Venezuela [An. triannulatus s.l. in Venezuela [An. aquasalis in Trinidad [P. vivax[Outside the Brazilian Amazon, enezuela ,29; An. enezuela ; An. nunenezuela , Peru [3enezuela  and Coloenezuela ,33; An. enezuela  and Peruenezuela ; and, AnTrinidad , Guyana Trinidad  and Vene[P. vivax.Plasmodium spp. starts when mosquitoes ingest gametocytes, the parasite sexual stage, during the blood meal taken through the skin of infected vertebrate hosts. Inside the mosquito alimentary tract, fusion between male and female gametocytes produces motile ookinetes, which traverse the mosquito midgut epithelium to form oocysts [Plasmodium establishment in a susceptible vector and this parameter is used to determine the infection rate of a mosquito population [The life cycle of  oocysts ,38. The pulation ,40.Plasmodium oocysts, is an important parameter for defining vector competence and thus, a key indicator in the description of malaria dynamics and transmission biology in a given geographic region. Indeed, infection rates in mosquito vectors are related to gametocyte survival, viability and success of fertilization, and finally, midgut invasion by the resulting ookinetes. However, not all gametocytes that are ingested by susceptible mosquito vectors reach the ookinete stage [Plasmodium in the mosquito, sporozoites are released from the oocyst and go on to invade the salivary glands. Once the salivary glands become infected with sporozoites, the mosquito is infectious to humans during the next blood meal [In the field studies, the infection rate, i.e., the amount of individuals in a mosquito population that carry well-developed te stage -43. Factte stage -48. To cood meal -51.Anopheles only about 10% are important as vectors of human malaria. There is a multitude of both ecological and genetic determinants that influence vector competence, both among species and even at the level of geographic populations within a single species [Plasmodium infection among the putative vectors of malaria in the Amazon have never been fully and carefully considered until this study.It is well known that among the over 400 species of mosquitoes in the genus  species -45,47. DAnopheles gambiae, the major vector in several African countries, is the most well studied mosquito, including its interaction with human and murine Plasmodium species causative agents of malaria [An. darlingi, the major Amazon vector, has proven to be difficult as well as other anopheline species from the region, and presently there is only An. aquasalis as an available autonomous colony. Anopheles aquasalis has been reared in laboratory as free mating since 1995 [Mosquito vectors of malaria from Africa and Asia have been well established in colonies and are feasible to maintain in laboratory. For example the  malaria ,53. Distnce 1995 -56 and rnce 1995 -59.P. vivax of colonized An. aquasalis with four groups of field-collected Anopheles species. This study was focused on vector infection rates as defined by the presence, quantification of oocysts and proportion (percentage) of infected individuals in these mosquito populations, following exposure to blood obtained from infected human patients. In addition, P. vivax infection in the vector was correlated with gametocyte numbers present in the circulating blood of infective human hosts. In laboratory studies, the infection rate is a critical part of the determination of vector competence. It is important to study P. vivax infection of New World vectors due to the huge gap regarding the knowledge comparing other vector-parasite pairs from the Old World.The goal of this work was to compare the susceptibility to P. vivax malaria infection diagnosed by blood smears were invited to participate in the study. Volunteers signed informed consent documents as blood sample donors. About 3 ml of blood samples were collected by venipuncture from volunteers and placed into a sterile lithium heparinate vacutainer tube. After blood collection, all patients were treated at the Fundação de Medicina Tropical Dr Heitor Vieira Dourado (located in the city of Manaus) or in the health posts from the region of Manaus where they were diagnosed, following ethical procedures determined by the Brazilian Health Ministry. This study was approved by the Brazilian National Ethics Committee Board .Adult volunteers (ages >18 years) residents from the region of Manaus  with P. vivax parasites. Sexual (gametocyte) and asexual stages counting per 500 leukocytes were performed.Thick and thin blood smears from malarial patients were prepared by Giemsa staining method and examined under light microscope x100 oil immersion lens to confirm the presence of An. darlingi, An. albitarsis s.l., An. nuneztovari s.l. and An. triannulatus s.l., as described elsewhere [Anopheles aquasalis mosquitoes originated from a colony established in 1995 [ad libitum fed 10% sugar solution and kept in laboratory conditions at 26-28°C and 70-80% RH (relative humidity). Three- to five-days old adult females were used in all experiments. Pinned voucher specimens were deposited at the Biological Collection at the Instituto Leônidas e Maria Deane .Mosquito larvae were collected during one year at different breeding sites near the city of Manaus, capital of Amazonas State, Brazil: Puraquequara Road ; Brasileirinho Road . Larvae were reared in the insectary as described elsewhere . Emergedlsewhere ,61. Fiel in 1995  were reaP. vivax patients for a period of 45 to 90 minutes via membrane feeding assay (glass device covered with Parafilm®). Blood was held at 37-39°C through a hose system connected to a thermal bath. The second group was treated in similar way but with inactivated-blood serum (IBS). The P. vivax infective blood samples were centrifuged for 15 minutes at 2,000 g and the serum removed and heated for 1 hour at 56°C. Then, the inactivated serum was added back to the red blood cells containing parasites, suspended and offered to the mosquitoes. After the infective blood meals only fully engorged mosquitoes were transferred to rearing containers and maintained in the insectary as described above for the development of infection.Adult mosquitoes were sugar starved overnight prior to infection via membrane feeding assay. Individuals from each of the five species were separated in two experimental groups. One group was offered whole blood (WB) from Five to eight days after infective blood meal, midguts from the experimentally infected mosquitoes were dissected in phosphate buffered saline, stained with 2% commercial Mercurochrome (Merbromin), placed under a coverglass and examined for the presence of oocysts. The number of oocysts on the mosquito midgut was recorded.Anopheles species to P. vivax was evaluated by the presence and the number of oocysts in the midguts. The population infection rates were calculated by dividing the number of infected mosquitoes (those with one or more oocysts) by the number of dissected mosquitoes. G tests were used to compare the frequency of infection among all the studied mosquito species conjointly, as well as pairwise comparisons between each pair of species. The Kruskal-Wallis test evaluates differences in the number of oocysts between the infected mosquito species. Only positively infected mosquitoes were used for this last analysis. Conover-Inman test, a posteriori, was used for comparison of the number of oocysts among all studied mosquito species [In this study, the blood-feeding rate was calculated as the proportion of female mosquitoes that were fully engorged after a blood meal. The susceptibility of the  species ,63. For P. vivax isolates from malarial patients were used for the different experimental feeding assays and only the fully engorged mosquitoes after blood feeding were analyzed in this study  . Actually, An. aquasalis showed the highest infection rate  followed by An. albitarsis s.l. (44.7%), which were significantly different to all other mosquito species . Anopheles nuneztovari s.l. held the third highest infection rate (24.5%), and it was not statistically different to An. darlingi, which had an infection rate of 18.3% , but was different to An. triannulatus s.l., with only 8.8% of the individuals infected . Anopheles darlingi also had a significantly higher infection rate compared with An. triannulatus s.l.   and showed statistical difference when compared with all the other species. Anopheles darlingi  was followed by An. albitarsis s.l.  and An. nuneztovari s.l. . There was no significant difference in the number of oocysts between An. darlingi, An. albitarsis s.l. and An. nuneztovari s.l. However, An. triannulatus s.l. had significantly less oocysts than all other studied species .The intensity of infection, measured by the numbers of oocysts per infected mosquitoes, fed on WB and IBS varied significantly among species  Figure . The higAn. triannulatus s.l., followed by An. darlingi, An. nuneztovari s.l., An. aquasalis, and An. albitarsis s.l. Approximately 16% of individuals from all five species were infected with 1 to 10 oocysts and only 10% were infected with 11 to 50 oocysts, with the lower number of infected individuals observed in An. triannulatus s.l. (0.4%) and the higher in An. albitarsis s.l. (21.3%). Only An. darlingi and An. aquasalis had more than 50 oocysts, with 1.3 and 7.6% of the individuals, respectively.To outline the distribution of infection intensities, among WB-infected mosquitoes, we organize the oocyst numbers into four groups, as follows: 1–10, 11–50, 51–100, and more than 100 oocysts. A comparison of the distribution of these groups among individuals within species is illustrated in Figure Anopheles species as a whole group, the inactivation of factors present in blood serum was not of significant importance to change the infection rate . However, the pattern of the infection rate differed among IBS-infected mosquito species. While the serum inactivation did not cause changes in infection rates on An. aquasalis  and An. nuneztovari s.l. , serum inactivation resulted in a 53% increase in infection rates in An. darlingi , and 87% increase in An. triannulatus s.l. . Inversely, the infection rate for An. albitarsis s.l. decreased by 55%   and An. nuneztovari s.l. (Mann–Whitney U = 0.466 p = 0.495). On the other hand, the inactivation slightly increased the oocyst formation on An. triannulatus s.l.  and decreased in An. darlingi  and An. albitarsis s.l.  . There was no difference in oocyst numbers formed on P. vivax isolates from malarial patients were used for An. darlingi feeding experiments, 13 for An. albitarsis s.l., 17 for An. nuneztovari s.l., 20 for An. triannulatus s.l. and 29 for An. aquasalis  and An. aquasalis  infection rates increased with an increase in the number of gametocytes. For An. nuneztovari s.l., An. triannulatus s.l. and An. albitarsis s.l.  there was no correlation between those two variables. Also, among infected species, An. aquasalis  and An. darlingi  showed a positive correlation between the number of gametocytes and the number of oocysts formed. The data for all other species did not exhibit a correlation between the quantity of gametocytes and the quantity of oocysts (p > 0.05).A total of 17 Anopheles species. Experimental infection of mosquito vectors can involve direct feeding on the skin of patients or offering blood meal through a membrane-feeding device. Ethical preference tends to lean towards the use of membrane-feeding experiments, in order to minimize the human factor interaction. Most experimental results with Amazon outbred Anopheles species have used direct skin feeding on gametocytaemic malarial patients [P. vivax-gametocytaemic patients resulted in An. darlingi infection rates between 22 and 23% with a mean oocyst load per infected midgut of 11.5 with a range of 1–175 [Anopheles aquasalis and An. albitarsis s.l. had a significantly higher infection rate than An. darlingi, considered the main malaria vector in the Brazilian Amazon, and all three species showed high quantity of oocysts, being that the highest one was found in An. aquasalis. These results confirm that the membrane-feeding assay is as efficient as direct feeding on human skin when it comes to the study of Plasmodium infection on mosquito vectors.The number of field mosquito specimens available for experimental infection was limited by the seasonality of the species and the malaria transmission peaks in the Amazonas region near to Manaus city. A membrane-feeding assay was used to infect patients ,27,64. Tof 1–175 ,66. AnopAnopheles darlingi in Amazon area is more abundant during the late wet season and early dry season, while the other species are more abundant during the early dry season [An. darlingi was in high abundance, An. albitarsis s.l. and An. nuneztovari s.l. were in low abundance. When the abundance of An. darlingi was low, An. albitarsis s.l. and An. nuneztovari s.l. abundance increased. This dynamics in species succession is a very important factor for maintenance of malaria transmission, and can present difficulties for malaria control in this region due to susceptibility of all studied species to P. vivax infection [.y season ,67. Siminfection .An. darlingi, An. albitarsis s.l., An. nuneztovari s.l., and An. triannulatus s.l. field populations, and the laboratory-colonized An. aquasalis are susceptible to P. vivax under laboratory conditions. All of the studied species might be sporadic competent vectors in nature, although there was a generalized high proportion of uninfected mosquitoes. However, infection rates were much higher than those reported in nature for the five species examined. As determined by the ELISA technique, based on the use of species-specific anti-sporozoite monoclonal antibodies, Amazonian mosquito populations had different P. vivax infection rates: An. darlingi ranged from 0.3 to 9.3%; An. albitarsis s.l. from 0.4 to 5.2%; An. nuneztovari s.l. from 0.3 to 1.1%; An. triannulatus s.l. 0.2% and An. aquasalis from 0.3 to 1.3% [P. vivax genotypes, immune factors in patient sera, and host response mechanisms, all of which could alter gametocyte infectivity [Here, it was demonstrated that  to 1.3% ,27,69-71ectivity ,72-74.An. darlingi is known to be strongly anthropophilic and endophilic, and population of this species occurs through the year. Anopheles albitarsis s.l. was shown to be a very susceptible species to P. vivax, and these results agree with results obtained in other Brazilian Amazonian states of Roraima, Pará, Amapá and Rondônia [An. albitarsis s.l. a dubious malaria vector because of the low number of oocysts, zoophilic behavior and seasonal distribution [An. darlingi and An. albitarsis s.l., An. aquasalis showed a high susceptibility to P. vivax. The high level of susceptibility of An. aquasalis to P. vivax indicates the value of using this species in studying New World malarial parasite vector-interaction. Unlike An. darlingi, An. aquasalis is well adapted to colonization in the laboratory.The number of oocysts has little importance in malaria epidemiology since most infected mosquitoes found in nature only possesses a few oocysts . The degRondônia ,16,19,70ribution . Like AnAnopheles nuneztovari s.l. also showed a high infection rate, although not statistically different to An. darlingi, it was significantly different to An. albitarsis s.l.. Oocyst numbers in An. nuneztovari s.l. were lower than in An. darlingi and An. albitarsis s.l., although the mean number of oocysts was not significantly different between these species. Anopheles nuneztovari s.l. is considered an important malaria vector in some South American countries [An. nuneztovari s.l. as a vector of malaria, which could be considered a risk depending on its density in a given area. In the Manaus area for example, An. nuneztovari s.l. is probably an important species for malaria transmission, because of its high feeding and infection rates observed in laboratory. The population of this species was found in high densities in localities around Manaus where An. darlingi is also abundant [An. triannulatus s.l. became infected with P. vivax in the experiments described here, this species had significantly the lowest infection rates and mean number of oocysts compared with the other species studied. These observations reinforce the conclusion that An. triannulatus s.l. is not an important malaria vector in the Amazon region [ountries ,31; howeountries ,23,24. Tabundant . On the n region ,16,44,68An. albitarsis s.l. and An. aquasalis, mosquito infection rates were increased after blood serum inactivation. Blood serum factors have been shown to influence the ability of P. vivax gametocytes to infect mosquitoes in experiments in which patient plasma was replaced with P. vivax-naïve sera or plasma [Plasmodium infection in An. albitarsis s.l. and An. aquasalis may be more strongly associated with intrinsic factors related to the mosquito’s own immune system, which could respond more effectively to destroy Plasmodium infections. Future studies should include evaluation of P. vivax strain variability on mosquito susceptibility and both intra- and interspecific variation in mosquito immune responses to Plasmodium infection.Except for r plasma ,66. InfeAn. darlingi and An. aquasalis had a positive correlation between the quantity of gametocytes and the other two variables. However, according to Klein and collaborators there was no correlation between the number of P. falciparum gametocytes and the mean number of oocysts formed in An. darlingi, although, in general, low numbers of circulating gametocytes resulted in few infected mosquitoes [P. vivax transmission to mosquitoes [The results also indicate that the quantity of gametocytes are positively correlated with the infection rate and the number of oocysts formed, but when results were analyzed by individual species only squitoes . Distincsquitoes .An. darlingi, which has not been colonized after several efforts by distinct research groups . Only An. aquasalis, the main vector in coastal Brazil, has been established and maintained in the laboratory. This study established baseline data for key transmission parameters showing that in laboratory, P. vivax infection of colonized An. aquasalis had an infection rate of 44.8% with a mean oocyst count of 29 per infected individual. It is possible now to begin using this system to explore mosquito immune response to P. vivax infection [Development of novel malaria control strategies includes methods aimed at disrupting parasite development in the mosquito vector. Studying natural vector-parasite interactions, as opposed to model systems, is critical to the development of strategies that can ultimately be used in the field. These studies are made difficult in cases where mosquito vectors cannot be colonized in the laboratory. Such is the case with many of the vectors in South America, including the main vector nfection -59.The authors declare that they have no competing interests.CMRV, FACP, RS, PFPP, NFCS, WPT, and MVGL conceived and designed the experiments; CMRV, KMC, RS, FACP, EVS, and JBPL: field collection and laboratory procedures; CMRV and TJI: data analysis; CMRV, PFPP, TJI, WMM, FACP, WPT, MVGL, and NFCS: data interpretation and manuscript preparation. This manuscript is a part of the PhD thesis developed by CMRV and supervised by PFPP. PFPP is a senior visiting researcher at the Foundation of the Tropical Medicine Dr. Heitor Vieira Dourado in Manaus (FMT-HVD). PFPP, NFCS and MVGL are productivity fellows of the Brazilian Council for Scientific and Technological Development (CNPq). All authors read and approved the final version of this article.


Professor Prem Narain Saxena was born on October 15, 1925 in a respected family of a village Bithrichainpur of Bareilly. It was a joint orthodox family village. From village, he rose to a peak of excellence in medical science career.Dr. Saxena was the first child of his parents Mr. Raghunandan Prasad and Mrs. Saraswati Devi, apart from three other sisters. He married Ms Rajeshwari Saxena, MA (LT), a working lady but latterly left the job and remained as housewife.Dr. P. N. Saxena took early education first in his hometown village Bithrichainpur. He then shifted to Dehradun at an early age of 6 to 7 years to take admission in the first standard, as his father was posted already in Dehradun as ‘Accountant’ in the Department of Irrigation. Dr. Saxena was a very brilliant student since school days despite of malnutrition, rickets and typhoid. He topped with first position both in matriculation from AP Mission School and Intermediate from DAV College of Dehradun. In the very first attempt, he qualified medical entrance and did MBBS with 10th position from the King George Medical College, Lucknow  in 1947. He was keen in medical research and hence after getting MD degree in Medicine in 1950, he again did MD in Pharmacology in 1952 from the King George Medical College, Lucknow. In addition, he completed his PhD from the Patna University, Patna, in 1963 and DSc from the Aligarh Muslim University, Aligarh, in 1978.[He joined the Department of Pharmacology of Jawaharlal Nehru Medical College, Aligarh Muslim University, Aligarh, on April 14, 1964 as Professor and Founder Chairman, Department of Pharmacology. He served the Department as Head till September 30, 1985 and in view of his outstanding merits, he was reemployed by the University for two years, that is, till September 30, 1987. Soon after retirement, he was selected as emeritus scientist of ICMR, New Delhi, for a period from September 1, 1987 till September 14, 1990. On January 31, 1991, the University appointed him as ‘Professor Emeritus.’ He was among the few scientists in the university who was offered this ‘Professor Emeritus.’He is nationally and internationally known as a leading Neuropharmacologist as he worked mostly in the field of neuroscience. He was awarded Fellowship of the Rockefeller Foundation Fellow in USA during 1960-61; Commonwealth Medical Fellowship and then Wellcome Research Fellowship in England during 1970s. In London at the National Institute for Medical Research, he worked on mechanism of action of Pyrogen and in the field of thermoregulation with Wilhelm Feldberg (1900 – 1993), a renownProfessor P. N. Saxena has to his credit, approximately 145 published research papers. He wrote Hospital Formulary in 1969 and a book-cum-manual for practical pharmacy and experimental pharmacology laboratory. After serving the university as professor and head of the department of pharmacology, he was then retired from active service on September 30, 1987. In the university, he also worked as Dean, Faculty of Medicine, In-charge, Animal House and on few occasions served as acting Principal of Ajmal Khan Tibbiya College and acting Vice Chancellor of the University. However, despite of his retirement from active service, he continued his research activities as Professor Emeritus in the Department of Pharmacology till late December 1994. During those days, his wife was also suffering from cancer and hence could not manage to visit the department regularly and finally discontinued all his academic services in 1995. She died on March 19, 1996.He had been the founding member of many academic bodies such as Indian Pharmacological Society, Association of Physiologists and Pharmacologists of India, Indian Medical Association, Indian Academy of Neurosciences and Indian Association for the Advancement of Medical Education in India. Indian National Science Academy (INSA) elected him as Fellow (FNA) in 1989.Dr. Saxena was very simple, kindhearted and pious person. He used laboratory animals  very gently and was very kind towards all the staff of the medical college.Dr. P. N. Saxena succumbed to death due to severe second stroke on November 29, 1999. He was admitted to KK Hospital, Aligarh before death. He was fond of Urdu and Persian poetry and also maintained a diary, where he used to write his favourite couplets of Urdu poetry. His mother tongue was Urdu. He had a good friends’ circle in academics. To name a few, Professors R. D. Myers, C. von Euler, W. Feldberg, W. J. Lang, A. Herxheimer, Richard J. Schain, K. P. Bhargava, M. B. Waller, Hakim Syed Zillur Rahman, W. R. Martin, Eleanor Zaimis, J. N. Sinha, B. N. Dhawan were the most closest colleagues.Coincidentally, both of his parents Mr. Raghunandan Prasad and Mrs. Saraswati Devi died in Aligarh due to paralytic attack. Dr. Saxena invited them in Aligarh to stay with his family and children. Dr. Saxena had two daughters and one son.Wilhelm Feldberg assisted many research workers who came to England as a part of their Commonwealth Medical Fellowship and Wellcome Research Fellowship. Under this Fellowships, Professor P. N. Saxena, during 1970s, got many papers published together with Wilhelm Feldberg while their vocation at National Institute for Medical Research, Mill Hill, London. Most of the reprints of their research papers are preserved in the Library of Ibn Sina Academy of Medieval Medicine and Sciences, Aligarh, India.


In the Funding section, the grant number from the Institutional Research Concept Grant was listed incorrectly. The correct grant number is: RVO: 61388971.http://www.gacr.cz/; [^] Synlab (http://www.synlab.cz), [^] the Academy of Sciences of the Czech Republic , http://www.gaav.cz/; [^] the Czech Ministry of Education, Youth and Sports (2B06155 and LC07017), http://www.msmt.cz; [^] and by Institutional Research Concept Grant RVO: 61388971, http://www.isvav.cz. [^] The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. The complete, correct Funding statement is: This work was supported by grants from the Czech Science Foundation ,


This special issue reflects on multiple factors/mechanisms involved in the pathogenesis of ALD. Alcoholic liver injury is known to cause a broad range of liver abnormalities. Alcohol is primarily metabolized in the hepatocyte leading to increased secretion of inflammatory mediators, which, in turn, activate and/or influence the response of the nonparenchymal cells (NPCs)  and subsequently control the degree of liver injury.This issue overviews general aspects of ALD, such as oxidative stress and inflammation (A. Ambade and P. Mandrekar) as well as the molecular aspects of these events, including the role of ethanol-metabolizing enzymes, ADH , and CYP2E1  in the development of alcohol-induced liver injury. In addition, signaling mechanisms induced by alcohol are examined in the paper by L. N. Gerjevic et al. As a separate pathogenic aspect, the role of microRNA in ALD is analyzed in the paper by S. Bala and G. Szabo.in vitro studies of C. A. Casey et al. and B. McVicker et al., respectively.The consequences of alcohol-related liver injury, such as impairment of receptor-mediated endocytosis and lipid droplet accumulation, are presented in experimental One of the mechanisms that affects various liver cell types and affect disease progression is impairment of methylation reactions. In our special issue, the role of impaired methylation in pathogenesis of steatohepatitis as well as treatment modalities with promethylating agent, betaine, is discussed in the paper by C. H. Halsted and V. Medici and K. Kharbanda et al.Liver also serves as an immune organ and accommodates a wide variety of cells, including immune cells. The latter consists of dendritic cells (DCs), natural killer (NK) cells, and lymphocytes, which are present in normal livers. Selective recruitment and retention of certain immune populations occurs during diverse liver diseases, and these cells play a critical role in the development and resolution of liver inflammation, remodeling, and destruction and actively participate in immune defense. The role of adaptive immunity in ALD development is overviewed by E. Albano. Also, the role of stem cells in ALD treatment is discussed in the paper by M. Pai et al. Finally, to underline the role of alcohol in progression of chronic infections (HCV), we included the paper by M. Neuman et al. which reflects on the markers of inflammation and fibrosis in alcoholic hepatitis and hepatitis C.Each paper received external blind review in addition to our reviews.This special issue covers exciting new areas of ALD pathogenesis and treatment and is strongly recommended for the clinicians and basic scientists involved in alcohol research.Natalia OsnaNatalia OsnaKusum KharbandaKusum KharbandaLaura SchrumLaura SchrumAngela DolganiucAngela Dolganiuc


We investigated the course of exacerbations in the published COSMIC study that investigated the effects of 1-year withdrawal of fluticasone after a 3-month run-in treatment period with salmeterol/fluticasone in patients with COPDIn 373 patients, we evaluated diary cards for symptoms, Peak Expiratory Flow (PEF), and salbutamol use and assessed their course during exacerbations.There were 492 exacerbations in 224 patients. The level of symptoms of cough, sputum, dyspnea and nocturnal awakening steadily increased from 2 weeks prior to exacerbation, with a sharp rise during the last week. Symptoms of cough, sputum, and dyspnea reverted to baseline values at different rates , whereas symptoms of nocturnal awakening were still increased after eight weeks. The course of symptoms was similar around a first and second exacerbation. Increases in symptoms and salbutamol use and decreases in PEF were associated with a higher risk to develop an exacerbation, but with moderate predictive values, the areas under the receiver operating curves ranging from 0.63 to 0.70.Exacerbations of COPD are associated with increased symptoms that persist for weeks and the course is very similar between a first and second exacerbation. COPD exacerbations are preceded by increased symptoms and salbutamol use and lower PEF, yet predictive values are too low to warrant daily use in clinical practice. Goals in the management COPD are to prevent and control symptoms, to reduce the frequency and severity of exacerbations, and to improve health status and exercise tolerance and prevent disease progression. In partStudies in asthma have shown that exacerbations resolve generally within 2 weeks with respect to symptoms, salbutamol use and peak expiratory flow (PEF) values. Data onAccording to the GOLD  guidelines, the goals of clinical control in patients with COPD include both health status  and clinical goals (prevention of disease progression and minimization of symptoms). The Clinical COPD questionnaire (CCQ) is the first practical instrument to be used for routine evaluation of clinical control concerning patients with COPD in general practice,12. A woCOPD and Seretide: a Multi-Center Intervention and Characterization) population with moderate to severe COPD, it has been shown that there are no significant differences with respect to the number of moderate to severe exacerbations experienced by patients in whom inhaled steroids were withdrawn after a run-in period with fixed dose of fluticasone (500 μg) and salmeterol (50 μg), versus those who continued this treatment[In a previous report on the COSMIC  in this group of COPD patients. It furthermore assesses the similarity of the course of symptoms around a first and second exacerbation. Information on the intra-individual variability of the course of symptoms around an exacerbation is important, since we wished to assess whether a change in symptoms can predict the occurrence of a moderate or a severe COPD exacerbation.Inclusion and exclusion criteria, methods for lung function measurements and design of the COSMIC study have been reported previously. In shorPatients recorded use of rescue medication  over the last 24 hours in daily paper record cards. If the patient’s condition worsened and a course of oral corticosteroids  was indicated based on the judgment of the research physician or general practitioner, the exacerbation was defined as moderate. If hospitalization was required at the discretion of the clinician, the exacerbation was considered severe. The time of onset and end of exacerbation was based on clinical judgment according to the research physician at each site.Every morning patients recorded severity scores of symptoms over the previous 24 hours on shortness of breath, cough, sputum production and night-time sleep disturbance (breathlessness 0 (none) to 4 (breathless at rest); cough and sputum production 0 (none) to 3 (severe); night-time sleep disturbance due to respiratory symptoms 0 (none) to 4 . Patients recorded their daily morning PEF throughout the run-in and the treatment period, prior to the inhalation of COPD (study) medication. The highest of three values was used for analysis.The Clinical COPD Questionnaire (CCQ) was administered to assess health status, a higher score meaning greater impairment. It is a 10-item, self administered questionnaire that can be completed in less than 2 minutes. Items are divided into three domains: symptom, functional state and mental state. Patients are required to respond to each item on a seven-point Likert scale where 0= asymptomatic/no limitation and 6 = extremely symptomatic/total limitation. The final score in the mean of all ten items and scores for the three domains can be calculated separately if required.1 (%predicted), age, gender, smoking status and CCQ. Mixed model Anova (SAS PROC MIXED) was used to evaluate associations between these factors and the log-transformed durations of both the moderate and severe exacerbations. Profiles of all longitudinal diary card data were used to characterize exacerbations. Only patients with a proper scoring of diaries in each of both periods were evaluated in these analyses, i.e. at least 50 percent of diary days. In addition patients were required not to have a new exacerbation in the 8-week period after the onset of the exacerbation. We analyzed the diary card data to investigate whether the increase in total or separate symptom scores could predict the occurrence of an exacerbation at each day of follow-up as follows. The increase of the mean symptom score was calculated for each patient separately during the week before the day of onset of the exacerbation, i.e. days 1 to −6, as compared to the mean symptom score during the week covering days −8 to −14. These increases were calculated for all patients during each day of follow-up, irrespective of whether the patient had experienced an exacerbation or not. So in fact a time-window of 14 days width of 2 successive weeks is used and this window is moved one day along time step by step, resulting at each step in a difference of 2 moving averages of symptom scores. For example, if a patient has a mean score of 10 points during days 101 to 107 and this was 13 points during days 108 to 114, then the value of the calculated increase at day 115 equals +3 points. The information at all follow-up days can be combined using Cox-regression, with the calculated increases of symptom scores per patient at each day of follow-up as a time-dependent covariate. With this method, the daily risk of an exacerbation is quantified in relation to the observed longitudinal changes in symptom scores. Regarding prediction of an exacerbation, only data to the first exacerbation were analyzed. The same method with moving averages was used to investigate the predictive value of changes in daily PEF measurements and salbutamol use. Receiver operating characteristic curves (ROC-curves) were constructed to evaluate the predictive value of the changes in total or separate symptom scores, PEF or salbutamol use as explained above. An area under the Receiver Operating Curve (AUC) of 0.8 or greater is generally considered to indicate a good predictor. A two-sided p-value below 0.05 was considered to be significant in all analyses.Poisson-regression was used to evaluate the relation between the annual exacerbation rate and the factors FEV1 % predicted and smoking status, showed that the annual moderate to severe exacerbation rate was 1.6 per patient year in the salmeterol group and 1.3 per patient year in the salmeterol/fluticasone group : 0.9-1.5; p=0.15). When considering moderate and severe exacerbations separately, no significant difference was observed between the treatment arms as well.Baseline characteristics of the 373 patients enrolled are presented in Table1 % predicted, showed that the number of pack-years smoking was the only significant predictor for the duration of a severe exacerbation. With every 5 pack-years smoking, the mean duration of a severe exacerbation increased 1.15 times. There were no significant effects of any of these parameters on the duration of a moderate exacerbation. Furthermore, the CCQ total score at baseline did not significantly contribute to the duration of moderate and/or severe exacerbations. The same applied to the separate domains of CCQ.There were 492 moderate or severe exacerbations in a total of 224 COPD patients, median duration of these 492 exacerbations being 10 days (range 2–119 days). The duration of a severe exacerbation was on average 46% longer than a moderate one ; no significant differences existed between the treatment arms. Mixed model Anova, allowing for gender, age, current and pack-years smoking, treatment and FEVth and 7th week after the exacerbation respectively compared to the weekly average scores during stable disease, i.e. the fourth week prior to the onset of the exacerbation , the pattern being similar for separate symptoms of cough, sputum, shortness of breath and night-time awakening . Adding baseline CCQ values to this Poisson regression model showed that the CCQ significantly contributed to the exacerbation rate. When adding each domain , the functional domain remained in the Poisson-model with a highly significant contribution (p=0.001). The factor gender became of borderline significance (p=0.08), whereas FEV1 % predicted remained significant (p=0.019).We reported previously that femThe averages of the symptom scores in two subsequent weeks were compared to assess whether an exacerbation is pending. Although there were individuals who had a marked increase in symptoms during the week preceding an exacerbation, a large inter-individual variability was observed Figure. Table3Our study shows that exacerbations of COPD are associated with increased symptoms that persist for weeks. Especially shortness of breath and nocturnal awakening due to respiratory symptoms persist for many weeks after the resolution of the acute phase of an exacerbation (7 and 8 weeks respectively). In addition, we demonstrate that the course of symptoms is very similar between a first and second exacerbation. Finally, we show that exacerbations are preceded by lower functional status, increased symptoms and salbutamol use and lower PEF yet predictive values are too low to warrant their use for clinical practice.1 and FVC) and found that there was only a limited fall in lung function during an exacerbation, compatible with our findings. Additionally, they followed the course of an exacerbation with diary cards and reported that the median time to recovery of symptoms was 7 days with an interquartile range of 4 to 14 days. However, the study did not report on the course of individual symptoms. In that respect our findings are new and show that some symptoms remit more rapidly than others. Nocturnal awakening due to respiratory symptoms was especially lingering on for many weeks after an exacerbation. This is in agreement with the findings by Partridge and colleagues that COPD patients indicate the morning as the time when their symptoms are most severe[This is in contrast to observations in asthma where symptoms during an exacerbation remit within 7–10 days after treatment with prednisone. Especiat severe. Whethert severe. This clInterestingly, the mean course of the increase and decrease in symptoms around the onset of a moderate exacerbation was very similar at the first and the second exacerbation. It thus appears that, despite a large inter-individual difference in the course of an exacerbation, the overall course of a first and second moderate exacerbation is quite similar in one and the same individual. As one can anticipate, the mean values of symptom scores were higher in the period before a severe exacerbation than before the first and second moderate exacerbations. Similarly, salbutamol use was significantly higher in the period before onset of a severe exacerbation and PEF values were significantly lower. Unfortunately, we had insufficient numbers of patients to compare the course of a severe and moderate exacerbation within the same individuals. It remains to be clarified which factors determine a severe exacerbation that requires hospitalization and one that can be managed in the home situation. It may well be, as Donaldson and coworkers suggested that sevWe sought for factors in diary cards that would predict an exacerbation in the near future. With each increase in total and separate symptom scores or salbutamol use per week, or decrease in PEF the risk of an exacerbation increased. Although the relative risks were high with weekly increases in reports, the overall predictive value of these changes was still rather limited in view of the moderate values of areas under the ROC curves. Whichever cut-off level one chooses for the increase of the various parameters, the rate of false positive findings remains high and therefore the specificity is low for acceptable values of the sensitivity Figure. The lat1[1 %predicted to be associated with an increased number of exacerbations, a finding compatible with data in the literature[A study that divided patients into frequent and infrequent exacerbators showed that those with frequent exacerbations had a faster decline in FEV1. Moreove1,17. In lterature,17. InteThere are some limitations to our study. First, there were relatively few severe exacerbations, probably reflecting the fact that patients were treated and followed accurately in a randomized controlled trial which may have influenced our results. In addition, we cannot rule out the possibility that regular treatment with salmeterol with or without added fluticasone, may have blunted the changes in PEF which recovered very quickly after a COPD exacerbation, whereas this as not the case for the perception of symptoms.In conclusion, this prospective study in moderate to severe COPD patients shows that symptoms persist for several weeks after an exacerbation, suggesting that the underlying pathophysiology is not resolved with a two-week course of oral corticosteroids and/or antibiotics. This may have implications for a better understanding why exacerbations, which may have a systemic effect on COPD as well, may affect the outcome of COPD. In addition, we show that the course of symptoms is very similar around a first and second exacerbation. Finally, COPD exacerbations can be predicted by an increase in respiratory symptoms, salbutamol use and decrease in PEF, yet the predictive values of these parameters are too low to warrant their daily use in clinical practice.1: Forced expiratory volume in one second; FVC: Forced vital capacity; GOLD: Global obstructive lung disease; COSMIC: COPD and Seretide: a Multi-Center Intervention and Characterization; ROC: Receiver operating curve.AUC: Area under the curve; COPD: Chronic obstructive pulmonary disease; CCQ: Control of COPD questionnaire; PEF: Peak expiratory flow; FEVM. van den Berge has received research grants from GlaxoSmithKline, E.F.M. Wouters, D.S. Postma and T van der Molen have been consultants for, and received research grants from, GlaxoSmithKline. W.C.J. Hop is a regular statistical consultant for GlaxoSmithKline. J.A. van Noord, J.P.H.M. Creemers, A.J.M. Schreurs received research grants from GlaxoSmithKline.MvdB reviewed and interpreted the results and wrote the manuscript. EFMW and DSP designed the study, analyzed the data, reviewed and interpreted the results and wrote the manuscript. WCJH performed the statistical analysis of the data and wrote the manuscript. JAvN, JPHMC and AJMS reviewed and interpreted the results. ThvdM reviewed and interpreted the results and wrote the manuscript. All authors have access to all data in the study and held final responsibility for the decision to submit for publication. All authors read and approved the final manuscript.COSMIC investigatorsDr. R. Aalbers, Dr. F. Beaumont, Dr. W. Boersma, Prof. Dr. J. Bogaard, M. Bunnik, J. Creemers, W. Dalinghaus, C. de Graaff, Dr. J. de Jong, Dr. P. de Jong, D. de Munck, D. de Vries, I. de Vries, W. den Hertog, H. Dik, E. Dubois, M. Eland, W. Evers, S. Gans, W. Geraedts, Dr. H. Heijerman, A. Hendriks, Dr. Ho, Dr. B. Hol, J. Kersbergen, H. Los, P. Luursema, B. Pannekoek, Dr. W. Pieters, R. Quanjel, E. Quanjel-Wisselo, R. Rammeloo, J. Retera, Dr. A. Roldaan, Dr. A. Rudolphus, L. Sala, N. Schlösser, Dr. A. Schols, Dr. A. Schreurs, Dr. J. Simons, A. Sips, Dr. F. Smeenk, W. Strankinga, I. Utama, F. van Beek, H. van de Woude, P. van den Berg, Dr. J. van den Bosch, J. van den Bosch, W. van der Brink, Dr. F. van den Elshout, Dr. B. van der Bruggen- Bogaarts, J. van der Zeijden, A. van Harreveld, Dr. A. van Keimpema, Dr. J. van Noord, H. van Pagée, R. van Snippenburg, P. van Spiegel, Dr. J. Verbraecken, J. Westbroek; The Netherlands.This study shows that COPD exacerbations can be predicted by an increase in respiratory symptoms, salbutamol use and decrease in Peak Expiratory Flow (PEF) values, but the predictive value of these parameters is low. Remarkably, symptoms persist for up to 8 weeks after a COPD exacerbation. Whether this reflects incomplete resolution of the pathophysiology underlying exacerbations remains to be elucidated.The study sponsor, GlaxoSmithKline, was involved in the study design, together with the principal investigators, in the collection and analysis of data, which were made freely available to the investigators, and in the decision to submit the paper for publication.


Botanicals are accepted worldwide as medicinal agents and nutraceuticals. Extensive scientific investigations have been performed over the past 200 years which have resulted in the evolution of botanical utilization. This special issue highlights some of this research in 7 review articles and in 24 original research articles.Review Articles. W. Jiratchariyakul and G. B. Mahady wrote about the botanical status and popular herbs in EU, US, and Thailand. M. Miroddi et al. updated the market and the regulatory of botanical products in EU and US. Traditional Chinese and Indian Medicines (TCM and TIM) play an important role in Asian countries. Z. Wang et al. presented and discussed the role of TCM in the treatment of epidemic type II diabetes mellitus. M. M. Pandey et al. reported the use of TIM as a nutritional supplement in malnutrition. Y. Kamisah et al. presented the chemoprevention and antioxidation of Parkia speciosa. A. P. Bartolome et al. reviewed laboratory evidence of Bidens pilosa. T.-P. Huynh et al. discussed the promising botanical compounds for prevention and treatment of eye diseases.Research Articles. New biological and pharmacological activities of botanicals are reported. They included the enhancement of learning, memory and antistress of Acanthopanax trifoliatus , the sedative effect of Ziziphus mauritiana , the cardioprotective effect of Phyllanthus emblica , the cytoprotection and antioxidative stress of Citrus maxima , anti-influenza viral activity of Momordica charantia , antioxidation from Nypa fruticans , Nigella glandulifera , TIM , Herba Cynomorii . The alleviation of metabolic disorder of Citrus ichangensis , and anticancer activity of Vitex agnus-castus .Zingiber zerumbet , phenyl butenoid dimmer from Zingiber cassumunar , and girinimbine from Murraya koenigii . Biochanin A, the major isoflavone from Trifolium pratense, prevented the bone loss in the ovariectomized rat . Quercetin isolated from Caesalpinia mimosoides had neuroprotective effect  and inhibited eosinophile (M. K. Asano Sakai-Kashiwahara). C. Li et al. reported new anti-inflammatory triterpenoids from Illicium difengpi.The mechanisms of anticancer action are deeply investigated with the botanical compounds, zerumbone from Gynura procumbens” .Besides the herbal activities, the cohort study of TCM, Si-Wu-Tang, in postpartum women was performed to evaluate the health benefits. The effectiveness of the modern herbal drug was carried out using the double-blind randomized controlled clinical trial, as shown under the title “Antiherpetic effects of Moringa oleifera  and Pueraria tuberosa . In addition, the arsenic accumulation in Zingiberaceous rhizomes was reported .The research on the quality assessment of the botanicals was also presented in this issue. It included the HPLC analysis of Weena JiratchariyakulWeena JiratchariyakulLudger BeerhuesLudger BeerhuesGail B. MahadyGail B. MahadyTanawan KummalueTanawan KummalueMolvibha VongsakulMolvibha Vongsakul


Antonina graminis Maskell (Hemiptera: Pseudococcidae), by the encyrtid wasp Neodusmetia sangwani is considered a textbook example of classical biological control. However, recent evidence suggests that A. graminis is abundant in the southeastern United States and no recent surveys have been conducted to determine the status of N. sangwani or other A. graminis parasites. A survey was conducted and it was found that N. sangwani was uncommon overall, occurring at only 20 percent of survey sites. In addition, N. sangwani exhibited a patchy geographic distribution. Possible causes for these results are that N. sangwani has not dispersed widely since its introduction, or that the imported fire ant, Solenopsis invicta, is interfering with biological control. These results suggest that a reevaluation of the efficacy of biological control may be necessary. The survey also found two other encyrtid wasps utilizing A. graminis as a host. One, Acerophagus sp., is apparently native and was nearly as frequent as N. sangwani, while the other, Pseudectroma sp., is apparently introduced and relatively rare.Control of the rhodesgrass mealybug,  Nezara viridula , in the southeastern United States , Xanthoencyrtus phragmitis, and Boucekiella antoninae from France, and Neodusmetia sangwani, from India. Anagyrus antoninae was introduced into Texas in 1949 and Florida in 1954, P. europaea was introduced into Texas in 1954–1955, and Florida in 1959, X. phragmitis and B. antoninae were introduced into Texas in 1954–1955, and N. sangwani was introduced into Florida in 1957, and Texas in 1959  was targeted because it is an important preferred host plant of A. graminis in rangeland systems  and/or crabgrass (Digitana sp.). After collecting, A. graminis were removed from plants, counted, and placed into a rearing chamber at 30°C as outlined for the Texas survey. Parasitoids that emerged from both surveys were counted, cleared, placed into 90% ethanol, and identified . For most sites, field estimates of a minimum of approximately 100 entified . ParasitA. graminis at five (38.5%) of 13 sites, primarily in the southern half of the state  of 16 sites, two in Louisiana, and two in Florida. Two parasitoid species, N. sangwani and Acerophagus sp. emerged at one site in Florida (Fl–1), while only Acerophagus sp., emerged from the remaining three sites, two in Louisiana  and one site in Florida (FL–5)  . The per to 16.8 . PoolingA. graminis by N. sangwani is viewed as an important success of biological control, there is surprisingly little quantitative data on the frequency of either the host or parasitoid since shortly after initiation of the biological control program. In this study no quantitative data on the frequency of A. graminis is provided, however, it appeared common and often abundant at the study sites, which is consistent with reports from other recent studies in the southeastern United States .A second possible reason why d debris . A recengraminis . WhetherA. graminis was parasitized nearly as frequently by a species of Acerophagus as it was by N. sangwani. Because Acerophagus is native to North America and the Caribbean (Acerophagus sp. at survey sites was nearly equal to that of N. sangwani and is another indicator that N. sangwani is not providing widespread control of A. graminis. If so, the native parasitoid may be an important additional tool in the biological control of the mealybug.aribbean , it is lPseudectroma sp. was found at only a single site, the discovery that it was a parasite of A. graminis is of interest. While P. europaea was one of the early parasitoids introduced, it was not believed to become established and has not been found in any subsequent survey. Indeed, it is likely that Pseudectroma sp. is not P. europaea. A Pseudectroma sp. identified as not being P. europaea, and presumably from Asia, was found utilizing A. graminis as a host in Florida surveys from 1975–1991 (N. sangwani (A. graminis is unclear but deserves further study.While 975–1991 . It was sangwani . The oriA. graminis from the parasitoid N. sangwani may not be as widespread or effective as when first released. This could occur because N. sangwani has failed to disperse widely, or because A. graminis is protected from parasitoids by S. invicta. Other causes are also possible. Further study would be valuable in exploring these causes and in determining the generality of our results. Because A. graminis is an important invasive species with a history of significant negative impacts on agriculture, it is important to determine whether its biological control remains effective or whether a new program may be warranted.The surveys involved a number of sites over a large range, and they suggest that the biological control of


Colpoclypeus florus  (Hymenoptera: Eulophidae). C. florus, a species introduced from Europe, requires medium to large host larvae late in autumn on which to overwinter, and few leafroller species display this biology. Over the four years, five potential C. florus hosts were collected, including: Ancylis comptana (Froelich), Xenotemna pallorana (Robinson), and Syndemis sp. (Tortricidae), Filatima sp. (Gelechiidae), and Caloptilia burgessiellia (Zeller) (Gracillariidae). Of these, A. comptana, Syndemis sp., and Filatima sp. have been confirmed as overwintering hosts for C. florus. During the four years, the Syndemis sp. was rare and observed at only one location feeding on redosier dogwood, Cornus sericea L.  although, at this location, many of the larvae collected were parasitized by C. florus. Filatima sp. was common in the Yakima valley feeding on balsam poplar, Populus balsamifera L. ssp. trichocarpa (Torr. & Gray ex Hook) Brayshaw  but was rarely parasitized. A. comptana, however, was collected at many locations in central Washington and was frequently found as an overwintering host for C. florus. A. comptana was found feeding on two Rosaceae: Wood's rose, Rosa woodsii Lindl., and strawberry, Fragaria ananassa Duchesne . Based on the number of host larvae collected, A. comptana appears to be the primary overwintering host for C. florus in Washington. Introduction of A. comptana populations to near-orchard habitats may facilitate biological control of leafrollers that are orchard pests.Thirty sites of managed and native habitats were surveyed for leafrollers (Lepidoptera: Tortricidae) in the apple producing region of central Washington State and northern Oregon from September through November 1997–2000 to discover species that supported overwintering by the parasitoid  Choristoneura rosaceana and Pandemis pyrusana are important pests of pome fruits, especially apples, in Central Washington , was discovered attacking leafrollers in unsprayed apple orchards of central Washington  shington . Leafrolomonella . In 1992shington . The popshington  or from shington . C. florn Europe  and has shington , 2000.C. florus has been reported to overwinter in the larval or pupal stage on suitably sized leafroller hosts  or as eggs (Archips rosanus)   and woulrosanus) . C. florneration .C. florus populations to seasonally disappear from orchards  and bitterbrush, Purshia tridentata (Pursh) de Canolle  and more lush and diverse riparian habitats dominated by willow  and balsam poplar  Brayshaw ) at lower elevations and redosier dogwood, Cornus sericea L.  and chokecherry, Prunus virginiana L. at higher elevations. During the summer and fall, much of the vegetation dies, except in riparian areas or among xeric adapted plants such as sage and bitterbrush, or in the weeds along orchard margins where irrigation water may spread. The search was focused in riparian areas, although collections were also done in orchard margins and the shrub-steppe habitats. Additionally, commercial alfalfa fields were sampled specifically for Xenotemna pallorana (Robinson) (Lepidoptera: Tortricidae).Agricultural and non-agricultural habitats in central Washington were sampled for leafrollers during September, October, and November from 1997 to 2000. Most locations were in areas that ablished , and mosAt all sites, all plant types were examined for evidence of leafroller infestation and, particularly, the presence of leafroller larvae. In addition to perennial species, the many herbaceous species present including the grasses (Poaceae) were examined. Leafroller infestations were characterized by the presence of foliage that had been rolled or folded and was held in this position by silk. Where evidence of leafroller feeding was observed, a more intensive search was conducted, and leafroller larvae were collected. Samples of > 25 individuals were collected whenever possible; collections of < 25 individuals occurred when that was all that could be located after about 1 h of searching.C. florus in the United States , near apple orchards or feral apples. In 2000, sampling was restricted to a gelechiid associated with balsam poplar near Yakima, WA.In 1997, samples were collected in the Wenatchee area of central Washington, the site of the first discovery of d States . In 1998X. pallorana, artificial leafroller diet  with tight fitting lids for rearing at 22° C and a 14:10 L:D photoperiod to evaluate parasitism. When needed, larvae were provided with additional food, which consisted of foliage from the host on which they were collected, or in the case of ler diet . In 1998C. florus were difficult to locate in most habitats in September and October of all years. Other than small diapausing (unsuitable) larvae of C. rosaceana and P. pyrusana, only five species of potential leafroller hosts were collected after September 1 in any year. These included Ancylis comptona (Froelich), X. pallorana, and Syndemis sp. (Tortricidae), Filatima sp. (Gelechiidae), and Caloptilia burgessiellia (Zeller) (Gracillariidae). Syndemis sp. was the only leafroller that was easily recognizable as large enough to support parasitism by C. florus; the stages of the other four leafrollers all seemed to be smaller than the suitable sizes of leafrollers typically parasitized by C. florus. A. comptana was the most common of these leafrollers and, once identified, could be collected from roses in numerous locations in the Columbia river basin near Wenatchee and the Kittitas, Wenatchee and Yakima valleys  . Syndemis sp. was rare, with only 14 individuals collected over the three years of the survey. All Syndemis sp. larvae were collected at one location feeding on redosier dogwood, despite sampling that host plant in numerous locations . C. florus were also discovered diapausing on A. comptana feeding on Wood's rose at Squilchuck 1 in 1997. Additional observations of C. florus diapausing on A. comptana were made on roses at 3 sites in 1998 and 7 sites in 1999 and in strawberries at one site in 1999 . This was a low level of parasitism of Filatima sp. compared to the very high parasitism of sentinel P. pyrusana (100%) in nearby apples and parasitism of A. comptona  in an adjacent rose patch. The small number of A. comptana collected for this sample was due to its scarcity in this area. Many other rose patches were sampled in this general area  during that day, and no other A. comptana were collected, nor was additional evidence of larval feeding observed. In 2000, > 200 Filatima sp. were collected, and no parasitism by C. florus was observed, despite very high parasitism of sentinel P. pyrusana (> 95%) in nearby orchards. No parasitism of X. pallorana or C. burgesselia by C. florus was observed during this survey.In 1999, diapausing A. comptana, Syndemis sp., and Filatima sp., that were observed to overwinter in potentially suitable stages for C. florus were observed with diapausing C. florus in the field. However, only A. comptana appeared to be a numerically important overwintering host. Parasitism of A. comptana by C. florus varied widely from site to site ranging from 0 to 44%   .Syndemis sp. was only observed at one location and in small numbers, thus this host is unlikely to play an important role in the population dynamics of C. florus. However, despite its rarity, C. florus larvae diapausing on Syndemis sp. were collected each fall. Parasitism of Filatima sp. was much more difficult to observe. Despite the ease of locating larvae of this species, Filatima sp. appears to not be a particularly suitable host for C. florus. It is smaller than A. comptana and may rarely reach a size large enough for C. florus to parasitize. C. burgessiellia does not appear to be a suitable host for C. florus as there were no recoveries with parasitism by C. florus. The third tortricid collected, X. pallorana, is very suitable for parasitism by C. florus , and its size appears too small to support overwintering by C. florus. However, the overwintering instars of X. pallorana have not been definitively determined.. florus , but it C. florus has been reported to be the most important parasitoid of leafrollers in Europe . A host of C. florus in Europe -Anagrus spp. system in California vineyards. Habitats containing alternate hosts for overwintering by Anagrus spp., including blackberry patches and prune plantings that are in proximity to vineyards can lead to reduction of pest leafhopper populations in grapes . Future studies will evaluate the potential of creating near orchard habitats containing roses and strawberries and introducing A. comptana to increase the local density of C. florus and subsequent parasitism of leafrollers in orchards, particularly in the spring.


Nandina domestica Thunb. were the only ingesta found in the gastrointestinal tract of these birds. Due to their voracious feeding behavior, the birds had eaten toxic doses of N. domestica berries. N. domestica contains cyanide and is one of the few berries readily available at this time of the year in the region. The gross and microscopic findings are consistent with lesions associated with cyanide toxicity. This paper for the first time documents toxicity associated with N. domestica in Cedar Waxwings.Dozens of Cedar Waxwings were found dead in Thomas County, Georgia, USA, in April 2009. Five of these were examined grossly and microscopically. Grossly, all the examined birds had pulmonary, mediastinal, and tracheal hemorrhages. Microscopically, several tissues and organs were diffusely congested and hemorrhagic. Congestion and hemorrhage were marked in the lungs. Intact and partly digested berries of  Bombycilla cedrorum, formerly Ampelis cedrorum) is a member of the family Bombycillidae. It breeds in North America, principally southern half of Canada, and the northern half of the United States [N. domestica forms an excellent backdrop for perennials that disappear in winter. Clusters of the bright red berries of N. domestica last for months on each plant, attracting hungry birds whose food is in short supply during this time of the year and into late spring [N. domestica is not previously reported in Cedar Waxwings. The Cedar Waxwing , and examined by light microscopy. Dozens of Cedar Waxwings were found dead in Thomas County, Georgia, USA, in April 2009. Five of these were necropsied at Tifton Veterinary Diagnostic and Investigational Laboratory of The University of Georgia, College of Veterinary Medicine. The birds were examined grossly and microscopically. For microscopic examination, tissue samples were collected at postmortem and fixed in 10% buffered formalin, processed for routine histopathology, sectioned at 5 N. domestica variably filled the crop (All the examined birds had similar findings on gross and microscopic examinations. Grossly, intact berries of the crop . Ample athe crop . Microscopically, the lungs, liver, kidney, proventriculus, ventriculus, uvea of the eye, heart, the meninges, and brain were diffusely congested. The hemorrhage and congestion were marked in the lungs . The tra Nandina domestica is a native of China and Japan. The species and its dwarf varieties are popular landscape items. The plant has naturalized and invaded habitats in southeastern and other areas of the USA. The bright berries are beloved by birds and attract Cedar Waxwings, mockingbirds, and robins [During winter, the Cedar Waxwings are concentrated in southeastern coastal plains of the USA. They are highly vagile, moving among crops of fruits, including those of ornamental trees and shrubs in suburban areas . Destrucd robins .N. domestica were the sole contents of the gastrointestinal tract of the examined birds. This indicates that the birds had eaten toxic doses of N. domestica berries, one of the few fleshy fruits available in winter and spring in south Georgia. The appetite of the Cedarbird is of so extraordinary nature as to prompt it to devour every fruit or berry that comes in its way . Cedar WNandina domestica berries contain cyanide and other alkaloids [N. domestica, cyanogenesis is the most important intoxication factor [N. domestica. lkaloids , 10. Forn factor . PresumaN. domestica. Because of their voracious feeding behavior, the birds have eaten toxic doses of N. domestica berries for which cyanogenesis is the most important intoxication factor [Cyanogenic plants represent a problem for various range of animals and wildlife, primarily among species that eat rapidly . The gasn factor . Tissue n factor . Hydrogen cyanide is highly poisonous to all animals. Sudden death is often the only presenting sign of acute cyanide poisoning . AlthougN. domestica in Cedar Waxwings. The berries of N. domestica are beloved by other birds such as robins and mockingbirds [In cyanide poisoning, hemorrhages occur commonly in the heart, lungs, and various other organs of affected animals. The characteristic cherry red venous blood seen in acute cyanide poisoning results from the failure of the oxygen-saturated hemoglobin to release oxygen to the tissues because the enzyme cytochrome oxidase is inhibited by cyanide . The groingbirds , indicat


The Federation of European Academies of Medicine (F.E.A.M.), founded in 1993 in Brussels, has the goal of promoting cooperation between national Academies of Medicine. It is the main association functioning as a bridge between the political and administrative authorities of the European Union and the National Medical Academies of Europe, in their role as advisors with concern to medicine and public health. Since the promulgation of its status in the Royal Decree on 31 March, 1995, F.E.A.M. has been given the full rights of an international association with a scientific objective. Its head office is still located in Brussels, in the prestigious Palais des Academies. F.E.A.M. has grouped leading European medical experts for its work, having three main objectives, as stated in their statute: To identify public health issues common to the Member States and to debate them in its conferences organized for the scientific Delegates of its Member Academies;To produce short reports based on an objective and impartial analysis of these issues which are submitted to the European Union and the Member Academies;To facilitate contacts between Member Academies and other organizations, with respect to the activities of the European Union, in matters concerning medicine and public health.th and the 26th of March, in Bucharest. Past conferences were held in Prague, Paris, Brussels, Rome and Lisbon. For the first time in the whole existence of FEAM, representatives from out of Europe, from countries such as China, USA, and Israel have been invited to participate. This year, F.E.A.M. has given the Romanian Academy of Medical Sciences the honor to host and organize the Spring Conference, during the 24This conference had two main objectives, both fulfilled with great success. The first objective, an administrative one, was to elect a new President–to–be for F.E.A.M., who was found in the person of Academician Prof. Dr. Laurentiu Mircea Popescu, President of the Romanian Academy of Medical Sciences. After his election, he will enjoy the Vice–President, President and Past President positions, each for two years. The second objective, an academical one, was to outline the high–end research and opinions in two mainstream domains of medicine: Translational Cardiology, on one part, and Medical Bioethics and Public Health on the other.‘The embryonic epicardium: an essential element of cardiac development’ by Prof. R.M. Chapuli from Spain, ‘The role of the epicardium derived cells in cardiac development and repair’ by Prof. Adriana C. Gittenberger–de Groot from Netherlands, ‘From genes to function: Single nucleotide polymorphisms as markers for cardiac disease’ by Prof. C. deWit from Germany, ‘The role of endogenous carbon monoxide and heme oxygenase system in myocardium’ by Prof. A. Tosaki from Hungary, ‘Regenerative Medicine: a cardiac surgeon's view’ by Prof. S.S.Hu from China ‘Relationships between telocytes and cardiomyocytes during pre– and post–natal life’ by Prof. Maria–Simonetta Faussone–Pellegrini and Prof. D. Bani from Italy, ‘Myocardial telocytes: a new distinct cellular entity’ by Prof. S. Kostin from Germany, ‘Cardiac stem cells, from science to therapy’ by Prof. P. Doevendans from Netherlands, ‘Cardiospheres and tissue engineering for myocardial regeneration: potential for clinical application’ by Prof. G. Frati from Italy, ‘Redox signaling of cardiac stem cells’ by Prof. D. K. Das from S.U.A., ‘Vascular Disease: A Critical Stage in the Vascular Continuum’ by Prof. Maya Simionescu from Romania, ‘Use of Cell and Gene Therapy for Engineering Blood Vessels’  by Prof. M. Flugelman from Israel and ‘Developmental origins of hypertrophic cardiomiopathy phenotypes’ by Prof. F. Cecchi from Italy.Selected speakers from twelve countries and three continents have presented their point of view during the two major scientific sections of the conference. The first section entitled ‘Translational Cardiology: from Molecular and Cellular Cardiology to Bedside’, brought light on the latest scientific breakthroughs of this field. The program included subjects such as A high importance was given to the newly discovered Interstitial ‘Cajal–like’ Cells (ICLCs) telocytes and their importance in Cardiology. Both Prof. G. Frati from Italy and Prof. S. Kostin from Germany focused on the importance of telocytes in Regenerative Cardiology, receiving valuable feedback from Prof. L.M. Popescu. The interactivity of the conference was of great significance, since it directed research towards new grounds, and stands as the fundamental source of new ideas and new perspective in a field that is responsible for the caring of more and more patients worldwide.‘The medical profession in transition: what makes a good doctor?’ by Prof. H.E. Blum from Germany, ‘Reconciling Bioethics with Public Health’ by Sir Peter Lachmann, Prof. from the United Kingdom, ‘Skylla and Kharybdis: stem cell researchers squeezed between advancement of science and self promotion?’ by Prof. Y. T. Konttinen from Finland, ‘Ethical issues in Tissue Engineering and Molecular Medicine’ by Prof. R.E. Horch also from Germany, ‘Bioethics, a basic discipline in public health’  by Prof. A. Knottnerus from The Netherlands, and ‘Bioethics of Patient Cell Reprogramming: In Search for Post JAK2 V617F Events in Blood Cancers’ by Prof. St.N. Constantinescu from Belgium.The second section of the conference focused on Medical Bioethics and Public health, with presentations such as ‘The New Medicine–a general cultural project and a perspective in civilization’ by Prof. C.T. Dragomir from Romania. The final presentation and closing remarks were summarized in an impressive presentation regarding High importance was given to the conference by the Romanian administrative, legislative and academic world, with attendance of Daniel Petru Funeriu the Minister of Education and Culture, Adrian Streinu–Cercel, Secretary of State in the Ministry of Health, Florian Popa, Member of the Romanian Parliament and Rector of the University of Medicine and Pharmacy in Bucharest, Mircea Cinteza, Senator, Stefan Iosif Dragulescu, Deputy and Rector of the University of Medicine in Timisoara and Academician Ion Ababii, Rector of the University of Medicine in Chisinau, as well as other personalities transforming this event into a base–stone for future academical, social and cultural perspectives in Romanian health and education.To quote Acad. Prof. L.M. Popescu in his presentation of the event, we sincerely believe that Bucharest has been the heart of translational cardiology and medical bioethics, even for just a couple of days.