Oswaldocruzia franciscoensis n. sp. (Nematoda: Molineidae) in Leptodactylus macrosternum Miranda-Ribeiro, 1926 (Anura: Leptodactylidae) from Caatinga morphoclimatic domain, Brazil: morphological and molecular characterisation.

A new species of Oswaldocruzia Travassos, 1917 (Nematoda, Molineidae), parasite of Leptodactylus macrosternum Miranda-Ribeiro, 1926 (Anura: Leptodactylidae), from Caatinga morphoclimatic domain, Brazil, is described based on morphological and molecular data. Oswaldocruzia franciscoensis n. sp. is characterised by an anterior extremity with a cephalic vesicle divided into two portions, a body covered by cuticular longitudinal ridges, and cervical alae. Males of the new species have caudal bursa of type I with a 2-1-2 pattern, spicules divided into a shoe, bifurcated fork, and blade with two unequal branches, in which the longer branch bifurcates at its distal portion end and the smaller branch with three distal processes, each with distal bifurcations. Females have didelphic and amphidelphic uteri, an ovijector divided into vestibule, anterior and posterior sphincters, and anterior and posterior infundibula. The new species differs from its Neotropical congeners that have caudal bursa of type I, based on the presence of cervical alae and by having a spicular blade distally divided into two unequal branches, with the longer branch bifurcating at its distal portion and smaller branch with three distal processes, each distally bifurcated. The partial 18S rDNA sequence generated for Oswaldocruzia franciscoensis n. sp. is the first of a representative belonging to this genus in the Neotropical region.

Morphological description and molecular characterization of Ancyracanthus electrophori n. sp. (Gnathostomatoidea: Gnathostomatidae): a new nematode parasitic in the electric eel Electrophorus varii (Gymnotiformes: Gymnotidae), from the Brazilian Amazon.

A new species of Ancyracanthus, parasite of the electric eel Electrophorus varii, in the Brazilian Amazon, is described based on morphological and molecular characterization. Ancyracanthus electrophori n. sp. differs from the two congeners namely, Ancyracanthus pinnatifidus and Ancyracanthus schubarti, based on the structure of cephalic appendages, number and arrangement of caudal papillae in males, vulva very close to anus in females, eggs with smoothly mamillated shell, host taxon and geographical origin. Moreover, the new species is the first in the genus to be described with thorny cuticular rings and to be observed with the use of scanning electron microscopy (SEM). The morphology of A. pinnatifidus and A. schubarti is still poorly-known and should be revised in details; however, the separation between them and the new species was clear. Genetic characterization based on 28S rDNA and cytochrome c oxidase subunit I (cox1) mtDNA partial sequences, performed for the first time in Acyracanthus, along with phylogenetic reconstructions using both genetic markers, placed Ancyracanthus electrophori n. sp. in a suggestive basal position within Gnathostomatidae. Phylogenetic reconstructions using cox1 sequences also suggested lack of monophyly in the genera Gnathostoma and Spiroxys and, consequently, in the subfamilies Gnathostominae and Spiroxyinae. However, such results are preliminary. With the first genetic characterization and observations using SEM in Ancyracanthus, resulting in the discovery of a new species and in the expansion of the geographical occurrence of the genus to Amazonian fish, an important step towards a better understanding of these nematodes has been taken.

Co-occurrence of Eutrombicula alfreddugesi and Oswaldofilaria chabaudi in Tropidurus torquatus and first report of microfilariae in the chigger mite: possible evidence of a lifecycle pathway?

While much attention has been paid to vector-borne filariasis, diseases that threaten millions of people in tropical and subtropical countries, the literature on host-parasite associations and transmission strategies of filarial nematodes in wildlife is scarce. Here, we report the co-occurrence of chigger mites (Eutrombicula alfreddugesi) and onchocercid nematodes (Oswaldofilaria chabaudi) parasitizing the lizard Tropidurus torquatus in the State of Minas Gerais, Brazil. Examination of chiggers established, for the first time, the occurrence of microfilariae in trombiculid mites (Trombiculidae). These larvae were morphologically similar to those recovered from adult females of O. chabaudi. The current evidence suggests that chiggers do not play a role in the transmission of filarioid nematodes, but rather act as accidental or dead-end hosts. Nevertheless, considering the polyphagous nature of trombiculid mites, similar to blood-sucking insects involved in the transmission of several infectious diseases, further studies may shed light on the potential role of chiggers as vectors of filarioids.

Profile of endoparasites in dairy cattle in the microregion of São João del-Rei, state of Minas Gerais, Brazil / [Perfil de endoparasitas em bovinos leiteiros na microrregião de São João del-Rei, Minas Gerais, Brasil]

The present study provides the first evaluation of the endoparasite profile in dairy cattle from the microregion of São João del-Rei, Minas Gerais, Brazil, including seasonal variations in parasitic infections. Fecal samples were collected directly from rectum of 123 dairy cows and 160 calves on 15 farms. These were kept in insulated boxes and sent for immediate analysis. Using eggs per gram of feces (EPG) we found the following prevalences: Strongyloidea 43.5%, Moniezia sp. 7.6% and both Strongyloides sp. and Trichuris spp. 2.2%, in dairy cows. In calves, the prevalence was Strongyloidea 50.9%, Moniezia sp. 5.6% and Strongyloides sp. 1.9%. The third-stage larvae recovered from coprocultures from cows and calves consisted mainly of Cooperia spp., Haemonchus spp. and Ostertagia spp. EPG counts were significantly higher in calves. Regarding seasonality, the EPG counts and prevalence of Moniezia sp. tended to be higher in the dry period, whereas those of Strongyloidea were higher in the rainy season. Evidence suggests that parasite control and prophylaxis were inadequately managed on the farms studied here. The present parasitological profile correlates with those found by other similar studies, indicating that the main problem is still the inadequate management of parasite control and prophylaxis by farmers.(AU)

O presente estudo representa a primeira avaliação do perfil de endoparasitos em bovinos leiteiros da microrregião de São João del-Rei, Minas Gerais, Brasil, e das variações sazonais das infecções parasitárias. Foram avaliadas amostras fecais de 123 vacas e 160 bezerros de 15 propriedades rurais, coletadas diretamente do reto, mantidas em caixas térmicas isoladas e analisadas imediatamente. A contagem de ovos por grama de fezes (OPG) revelou prevalências de 43,5% de Strongyloidea, 7,6% de Moniezia sp. e 2,2% de Strongyloides sp. e Trichuris spp. em vacas leiteiras. As prevalências de OPG em bezerros foram 50,9% de Strongyloidea, 5,6% de Moniezia sp. e 1,9% de Strongyloides sp. As larvas do terceiro estádio recuperadas de coproculturas de vacas e bezerros foram principalmente de Cooperia spp., Haemonchus spp. e Ostertagia spp. OPG dos parasitos foram significativamente maiores em bezerros. Em relação à sazonalidade, OPG e a prevalência de Moniezia sp. tenderam a ser maiores durante a estação seca, enquanto OPG de Strongyloidea foram maiores na estação chuvosa. Com base nos resultados, é possível concluir que o manejo antiparasitário nas propriedades estudadas não é realizado de maneira plenamente satisfatória. O presente perfil parasitológico se assemelha a outros encontrados em estudos semelhantes, indicando que o maior problema ainda é o manejo antiparasitário inadequado realizado nas propriedades de bovinos leiteiros.(AU)

Modeling microsolvation clusters with electronic-structure calculations guided by analytical potentials and predictive machine learning techniques.

We propose a new methodology to study, at the density functional theory (DFT) level, the clusters resulting from the microsolvation of alkali-metal ions with rare-gas atoms. The workflow begins with a global optimization search to generate a pool of low-energy minimum structures for different cluster sizes. This is achieved by employing an analytical potential energy surface (PES) and an evolutionary algorithm (EA). The next main stage of the methodology is devoted to establish an adequate DFT approach to treat the microsolvation system, through a systematic benchmark study involving several combinations of functionals and basis sets, in order to characterize the global minimum structures of the smaller clusters. In the next stage, we apply machine learning (ML) classification algorithms to predict how the low-energy minima of the analytical PES map to the DFT ones. An early and accurate detection of likely DFT local minima is extremely important to guide the choice of the most promising low-energy minima of large clusters to be re-optimized at the DFT level of theory. In this work, the methodology was applied to the Li+Krn (n = 2-14 and 16) microsolvation clusters for which the most competitive DFT approach was found to be the B3LYP-D3/aug-pcseg-1. Additionally, the ML classifier was able to accurately predict most of the solutions to be re-optimized at the DFT level of theory, thereby greatly enhancing the efficiency of the process and allowing its applicability to larger clusters.

Diaphanocephalus galeatus (Nematoda: Diaphanocephalidae), a poorly known parasite of lizards: redescription, first genetic characterization and a revision of its congeners from Brazil.

Diaphanocephalus galeatus collected from the small intestine of the lizard Dracaena paraguayensis in the Pantanal wetlands, State of Mato Grosso do Sul, Brazil, is redescribed. Genetic characterization and observations using scanning electron microscopy (SEM) were performed for the first time. The vouchers of D. galeatus and the type specimens of its congeners, deposited in the Coleção Helmintológica do Instituto Oswaldo Cruz (CHIOC), were consulted. Light and SEM observations revealed several undescribed features of D. galeatus, i.e. structure of the cephalic end and of the buccal capsule, position and morphology of deirids, presence of phasmids in females and presence of unpaired papilla on the membranous projection that covers the genital cone in males. After observation of the specimens deposited in the helminthological collection, D. jacuruxi is considered a synonym of D. galeatus, and D. diesingi, despite its incomplete description, is tentatively retained as valid due to the poor condition of the type material. The results also indicated low host specificity of D. galeatus, contradicting previous assertions. Genetic comparisons using patristic distances and phylogenetic trees generated from sequences of the 28S rRNA nuclear gene indicated that D. galeatus is closer to the taxa within Ancylostomatoidea and Strongyloidea than any lineage of Metastrongyloidea or Trichostrongyloidea. However, most of the nodal supports were low. Based on the genetic and morphological characterization, the validity of D. galeatus was confirmed. These data may serve for further comparative approaches for different populations of the parasite, from different hosts in different geographical areas, mitigating taxonomic confusions.

A new species of Raphidascaris (Nematoda: Raphidascarididae) infecting the fish Gymnogeophagus balzanii (Cichlidae) from the Pantanal wetlands, Brazil and a taxonomic update of the subgenera of Raphidascaris based on molecular phylogeny and morphology.

Raphidascaris (Sprentascaris) andersoni n. sp. (Nematoda: Raphidascarididae) collected in the intestine of the humphead cichlid Gymnogeophagus balzanii (Perugia) from the Pantanal wetlands, State of Mato Grosso do Sul (Brazil) is described and genetically characterized. The new species differs from its congeners mainly by having a conspicuous papilla-like formation slightly anterior to the cloacal aperture. Furthermore, males of R. (S.) lanfrediae and R. (S.) mahnerti have caudal alae, and R. (S.) hypostomi and R. (S.) pimelodi lack lateral alae, whereas in the new species caudal alae are absent and lateral alae present. The remaining congeners, namely, R. (S.) marano and R. (S.) saltaensis differ from Raphidascaris (Sprentascaris) andersoni n. sp. mainly because males have three pairs of postcloacal papillae (vs five pairs). In the phylogenetic reconstructions, using three nuclear genetic markers (18S, ITS1-5.8S-ITS2 and 28S rDNA) and one mitochondrial (cox1 mtDNA), the new species was separated from other representatives of Raphidascarididae, and the absence of monophyly in Hysterothylacium and Raphidascaroides was confirmed. Moreover, the subgenera Sprentascaris and Ichthyascaris appeared to be monophyletic. Therefore, even though Raphidascaris (Raphidascaris) was apparently not monophyletic, the subgenera of Raphidascaris should be re-erected as valid genera. The updated diagnoses of Ichthyascaris, Raphidascaris and Sprentascaris are given. The present study represents the first parasitological survey in G. balzanii.

Redescription and genetic characterization of Cystidicoloides vaucheri, including first description of male and current status on the phylogeny of Cystidicolidae (Nematoda: Habronematoidea).

Cystidicoloides vaucheri collected in the stomach of the redtail catfish Phractocephalus hemioliopterus from River Acre, State of Acre, Brazil is redescribed, including the first description of males and the first genetic characterization based upon 18S and 28S genes of the rRNA. Newly collected females were biometrically smaller than those reported in the original description, but similar morphology shared by the two samples revealed that they belong to the same species. Scanning electron micrographs showed the accurate structure of the cephalic region, described here in detail. Furthermore, the morphology of males completed the specific diagnosis, strengthening the validity of the species. The three other congeners differ from C. vaucheri mainly as follows: in C. dlouhyi the area rugosa is absent, the cephalic structures in C. fischeri are completely distinct, and in both species the spicules have membranous outgrowths, absent in C. vaucheri. Despite the dubious generic assignment of C. izecksohni, it differs from C. vaucheri in several biometrical and morphological features. Because of data availability, only sequences of the 18S were used for phylogenetic reconstructions. Results showed that the genus Ascarophis and the families Cystidicolidae and Physalopteridae are not monophyletic. Cystidicoloides vaucheri formed an independent branch clustering with representatives of Cystidicolidae, confirming its validity. The inclusion of Salmonema and Spinitectus within Cystidicolidae should be reviewed, since they formed an assemblage with species from Rhabdochonidae. In fact, current classification of some taxa belonging to Habronematoidea, Physalopteroidea and Thelazioidea need to be re-evaluated, mainly based on molecular data from different genes.

A global optimization perspective on molecular clusters.

Although there is a long history behind the idea of chemical structure, this is a key concept that continues to challenge chemists. Chemical structure is fundamental to understanding most of the properties of matter and its knowledge for complex systems requires the use of state-of-the-art techniques, either experimental or theoretical. From the theoretical view point, one needs to establish the interaction potential among the atoms or molecules of the system, which contains all the information regarding the energy landscape, and employ optimization algorithms to discover the relevant stationary points. In particular, global optimization methods are of major importance to search for the low-energy structures of molecular aggregates. We review the application of global optimization techniques to several molecular clusters; some new results are also reported. Emphasis is given to evolutionary algorithms and their application in the study of the microsolvation of alkali-metal and Ca2+ ions with various types of solvents.This article is part of the themed issue 'Theoretical and computational studies of non-equilibrium and non-statistical dynamics in the gas phase, in the condensed phase and at interfaces'.

Helminth parasites of South American fishes: current status and characterization as a model for studies of biodiversity.

The South American subcontinent supports one of the world's most diverse and commercially very important ichthyofauna. In this context, the study of South American fish parasites is of increased relevance in understanding their key roles in ecosystems, regulating the abundance or density of host populations, stabilizing food webs and structuring host communities. It is hard to estimate the number of fish parasites in South America. The number of fish species studied for parasites is still low (less than 10%), although the total number of host-parasite associations (HPAs) found in the present study was 3971. Monogeneans, with 835 species (1123 HPAs, 28.5%), and trematodes, with 662 species (1127 HPAs, 30.9%), are the more diverse groups. Data gathered from the literature are useful to roughly estimate species richness of helminths from South American fish, even though there are some associated problems: the reliability of information depends on accurate species identification; the lack of knowledge about life cycles; the increasing number of discoveries of cryptic species and the geographically biased number of studies. Therefore, the closest true estimations of species diversity and distribution will rely on further studies combining both molecular and morphological approaches with ecological data such as host specificity, geographical distribution and life-cycle data. Research on biodiversity of fish parasites in South America is influenced by problems such as funding, taxonomic impediments and dispersion of research groups. Increasing collaboration, interchange and research networks in the context of globalization will enable a promising future for fish parasitology in South America.

Morphological and molecular characterization of cucullanid nematodes including Cucullanus opisthoporus n. sp. in freshwater fish from the Brazilian Amazon.

Molecular and morphological characterization of two species of Cucullanidae from freshwater fish in Brazil are provided, one of which represented an undescribed taxon. Cucullanus opisthoporus n. sp. was collected in Cichla melaniae from River Xingu, State of Pará, and in C. pinima from River Jamarí, State of Rondônia. Nematodes referable to Cucullanus grandistomis were collected in Oxydoras niger from River Xingu. The new species has an appendage in the tail tip, ventrally covered by small spines, which is an exclusive feature of Cucullanus tucunarensis. However, C. tucunarensis differs from C. opisthoporus n. sp. based upon the relative position of deirids and the excretory pore, which are more posterior from the oesophageal end in the new species. Observations of C. tucunarensis type specimens also revealed features that were wrongly or not reported in the original description. Type specimens of C. grandistomis were observed, although they were poorly preserved. After evaluation of newly collected specimens of C. grandistomis, features unreported in the original description were observed for the first time, including the presence of an intestinal caecum. Thus, C. grandistomis was transferred to Dichelyne. Sequences of the 18S and 28S rRNA genes revealed high genetic similarity between the specimens of C. opisthoporus n. sp. from the two different hosts as well as their genetic distance from Dichelyne grandistomis n. comb. Phylogenetic reconstructions using representatives of Cucullanidae suggested the artificiality of the current morphological system adopted to separate the genera, since most genera were not monophyletic, although the availability of genetic data is still fragmented.

Improved evolutionary algorithm for the global optimization of clusters with competing attractive and repulsive interactions.

We propose improvements to our evolutionary algorithm (EA) [J. M. C. Marques and F. B. Pereira, J. Mol. Liq. 210, 51 (2015)] in order to avoid dissociative solutions in the global optimization of clusters with competing attractive and repulsive interactions. The improved EA outperforms the original version of the method for charged colloidal clusters in the size range 3 ≤ N ≤ 25, which is a very stringent test for global optimization algorithms. While the Bernal spiral is the global minimum for clusters in the interval 13 ≤ N ≤ 18, the lowest-energy structure is a peculiar, so-called beaded-necklace, motif for 19 ≤ N ≤ 25. We have also applied the method for larger sizes and unusual quasi-linear and branched clusters arise as low-energy structures.

The relationship between nematode infections and ontogeny and diet of the lizard Tropidurus torquatus (Wied, 1820) (Squamata: Tropiduridae) from the Atlantic Rainforest in south-eastern Brazil.

The aim of the present study was to investigate the relationship between nematode infection and the ontogeny and diet of the lizard Tropidurus torquatus from a rocky outcrop in the state of Minas Gerais, south-eastern Brazil. Eighty-nine of 110 lizards examined (81.9%) harboured nematodes. Two nematode species were identified, namely, Physaloptera lutzi in the stomach and Parapharyngodon bainae in the intestine, with prevalence values of 67.3 and 60.0%, respectively. The lizard diet was composed mainly of ants, other hymenopterans, beetles and flowers of the species Centrosema coriaceum (Fabaceae). Host body size was positively correlated with nematode abundance, with adults more heavily parasitized than juveniles. The consumption of C. coriaceum had a negative effect on the abundance of both nematode species, suggesting that this plant may possess anthelmintic properties. The probability of a higher worm burden in adult hosts is likely linked with a longer exposure time to infective stages. Beetles, ants and hymenopterans appear to be the main intermediate hosts for P. lutzi. In general, ontogeny and diet composition determine the structure of the helminth community in this species of lizard.

An evolutionary algorithm for the global optimization of molecular clusters: application to water, benzene, and benzene cation.

We have developed an evolutionary algorithm (EA) for the global minimum search of molecular clusters. The EA is able to discover all the putative global minima of water clusters up to (H(2)O)(20) and benzene clusters up to (C(6)H(6))(30). Then, the EA was applied to search for the global minima structures of (C(6)H(6))(n)(+) with n = 2-20, some of which were theoretically studied for the first time. Our results for n = 2-6 are consistent with previous theoretical work that uses a similar interaction potential. Excluding the very symmetric global minimum structure for n = 9, the growth pattern of (C(6)H(6))(n)(+) with n ≥ 7 involves the (C(6)H(6))(2)(+) dimer motif, which is placed off-center in the cluster. Such observation indicates that potentials commonly used in the literature for (C(6)H(6))(n)(+) cannot reproduce the icosahedral-type packing suggested by the available experimental data.

How different are two chemical structures?

We extend the scope of a recent method for superimposing two molecules ( J. Chem. Phys. 2009, 131, 124126-1-124126-10 ) to include the identification of chiral structures. This methodology is tested by applying it to several organic molecules and water clusters that were subjected to geometry optimization. The accuracy of four simpler, non-superimposing approaches is then analyzed by comparing pairs of structures for argon and water clusters. The structures considered in this work were obtained by a Markovian walk in the coordinate space. First, a random geometry is generated, and then, the iterative application of a mutation operator ensures the creation of increasingly dissimilar structures. The discriminating power of the non-superimposing approaches is tested by comparing the corresponding dissimilarity measures with the root-mean-square distance obtained from the superimposing method. Finally, we showcase the application of those methods to characterize the diversity of solutions in global geometry optimization by evolutionary algorithms.

Red blood cell antigen alloimmunization in liver transplant recipients.

Orthotopic liver transplantation (OLT) is a life-saving procedure for patients with end-stage liver disease. Transfusion support is an important part of OLT. Intraoperative transfusion of large volumes of blood products is recognized to be a poor prognostic factor, probably due to the negative effects of blood transfusions, such as transfusion reactions, infectious contamination of blood products, or immune modulation of the transfused patient. The aim of this study was to evaluate the frequency of alloimmunization and its specificity to red blood cell (RBC) antigens among patients undergoing OLT. We identified 74 RBC alloantibodies in 70 (23%) patients when the indirect antiglobulin test (IAT) was performed. The most common RBC alloantibodies were against Rh system antigens. The majority (41.9%) were directed against the E antigen. Despite the ethnic heterogeneity of our population there were no cases of intravascular hemolysis. The incidence of alloimmunization (23%) was slightly higher among patients than in the literature, most probably as a consequence of our ethnic heterogeneity.

Oswaldofiaria chabaudi n. sp. (Nematoda: Onchocercidae) from a South American tropidurid lizard (Squamata: Iguania) with an update on Oswaldofilariinae.

A new species of Oswaldofilaria is described from Tropidurus torquatus (Tropiduridae: Iguania); its prevalence at the rocky study area at Juiz de Fora, Minas Gerais, Brazil, was approximately 30% and its mean intensity 3.13 +/- 2.51. Oswaldofilaria chaboudi n. sp. is distinct from the thirteen Oswaldofilaria species known in Australia, Africa and South-America in having the following characteristics: oesophagus medium-sized, left spicule 1 mm long and high spicular ratio (about 5), tail extremity ornated in both sexes with a bifurcated projection, and tooth-like structures near phasmids in the female. A long left spicule and high spicular ratio are convergent derived characters also found in a parasite of Australian crocodilians, O. kanbaya, and in several species of the closely related genus Befilaria, such as the Central American B. puertoricensis from polychrotids. Oswaldofilaria in South America is represented by eight species. Within these, a primitive group that is parasitic in Iguanidae, Polychrotidae (Iguania) and Crocodylidae and that possesses a long oesophagus is recognised, together with two distinct derived lines: three species with numerous, aligned precloacal papillae, parasitic in Teiidae (Laterato) and Scincidae (Scincomorpha), and O. chabaudi n. sp., in which this character is absent. Tropidurids (Tropiduris and Plica) had previously been reported in the host range of two oswaldofilarine genera, Oswaldofilaria and Piratuba, and their parasites assigned to known species described from other groups of lizards.

On the use of different potential energy functions in rare-gas cluster optimization by genetic algorithms: application to argon clusters.

We study the effect of the potential energy function on the global minimum structures of argon clusters arising in the optimization performed by genetic algorithms (GAs). We propose a robust and efficient GA which allows for the calculation of all of the putative global minima of Ar(N) (N = 3-78) clusters modeled with four different potentials. Both energetic and structural properties of such minima are compared among each other and with those previously obtained for the Lennard-Jones function. In addition, the possibility of obtaining global minima of one potential through local optimization over the corresponding cluster geometry given by other potentials was associated with some structural parameters. The influence of the contribution from the three-body (Axilrod-Teller-Muto) triple-dipole potential (including or not a damping function to describe its correct behavior at smaller interatomic distances) has also been analyzed.

Interbilayer lipid mixing induced by the human immunodeficiency virus type-1 fusion peptide on large unilamellar vesicles: the nature of the nonlamellar intermediates.

A peptide corresponding to the 23 N-terminal amino acid residues of the human immunodeficiency virus type-1 (HIV-1) gp41 has the capacity to induce intervesicular lipid mixing in large unilamellar liposomes composed of dioleoylphosphatidylcholine (DOPC), dioleoylphosphatidylethanolamine (DOPE) and cholesterol (CHOL) (molar ratio, 1:1:1). Cryo-transmission electron microscopy (cryo-TEM) of diluted vesicles to which peptides has been externally added reveals a morphology that is compatible with the formation of nonlamellar lipidic aggregates during the time-course of lipid mixing. 31P-nuclear magnetic resonance and 1-(4-trimethylaminophenyl)-6-phenyl-1,3,5-hexatriene (TMADPH) steady-state anisotropy data at equilibrium indicate that the peptide is able to modulate the lipid polymorphism in pelletted membranes by: (i) promoting the thermotropic formation of inverted phases; and (ii) driving the lamellar-to-nonlamellar transition towards the formation of isotropic phases. Therefore, our combined morphological and spectroscopic data reveal the existence of a direct correlation between the ability of the externally added peptide to induce lipid-mixing in dilute liposome samples and its capacity to modulate lipid polymorphism in stacked bilayers.

Phosphatidylinositol-dependent membrane fusion induced by a putative fusogenic sequence of Ebola virus.

The membrane-interacting abilities of three sequences representing the putative fusogenic subdomain of the Ebola virus transmembrane protein have been investigated. In the presence of calcium, the sequence EBO(GE) (GAAIGLAWIPYFGPAAE) efficiently fused unilamellar vesicles composed of phosphatidylcholine, phosphatidylethanolamine, cholesterol, and phosphatidylinositol (molar ratio, 2:1:1:0.5), a mixture that roughly resembles the lipid composition of the hepatocyte plasma membrane. Analysis of the lipid dependence of the process demonstrated that the fusion activity of EBO(GE) was promoted by phosphatidylinositol but not by other acidic phospholipids. In comparison, EBO(EA) (EGAAIGLAWIPYFGPAA) and EBO(EE) (EGAAIGLAWIPYFGPAAE) sequences, which are similar to EBO(GE) except that they bear the negatively charged glutamate residue at the N terminus and at both the N and C termini, respectively, induced fusion to a lesser extent. As revealed by binding experiments, the glutamate residue at the N terminus severely impaired peptide-vesicle interaction. In addition, the fusion-competent EBO(GE) sequence did not associate significantly with vesicles lacking phosphatidylinositol. Tryptophan fluorescence quenching by vesicles containing brominated phospholipids indicated that the EBO(GE) peptide penetrated to the acyl chain level only when the membranes contained phosphatidylinositol. We conclude that binding and further penetration of the Ebola virus putative fusion peptide into membranes might be governed by the nature of the N-terminal residue and by the presence of phosphatidylinositol in the target membrane. Moreover, since insertion of such a peptide leads to membrane destabilization and fusion, the present data would be compatible with the involvement of this sequence in Ebola virus fusion.