1H, 13C and 15N resonance assignments of the complement control protein modules of the complement component C7

Human C7 is one of four homologous complement proteins that self-assemble on the nascent activation-specific fragment, C5b, thus forming the cytolytic membrane attack complex (MAC). In addition to the conserved modular core of the MAC/perforin protein family, C7 has four C-terminal domains comprising a pair of complement control protein modules (CCPs) preceding two Factor-I like modules (FIMs). It is proposed that the C7-CCPs might serve as a molecular arm for delivery of C7-FIMs to their binding site on C5b. Here we present the NMR chemical shift assignments for the C7-CCPs produced as a 14-kDa recombinant protein. Based upon triple-resonance experiments, 98 and 94 % of the backbone and side-chain (¹H, ¹³C and ¹⁵N) assignments, respectively, have been completed. The chemical shifts and assignments have been deposited in the BioMagResBank database under accession number 18530.     

Comparison of Fibronectin and Collagen in Supporting the Isolation and Expansion of Endothelial Progenitor Cells from Human Adult Peripheral Blood

Background

Endothelial colony-forming cells (ECFCs), are circulating endothelial progenitor cells increasingly studied in various diseases because of their potential for clinical translation. Experimental procedures for their ex vivo culture still lack standardization. In particular two different extracellular matrix proteins, either fibronectin or collagen, are commonly used by different Authors for coating plastic plates, both allowing to obtain cells that have all the features of ECFCs. However, possible differences in the impact of each substrate on ECFCs have not been analysed, so far. Therefore, in this study we investigated whether fibronectin and collagen may differentially affect ECFC cultures.

Methodology/Principal Findings

ECFCs were isolated and cultured from peripheral blood mononuclear cells of healthy donors. The impact of fibronectin compared with collagen as the only variable of the experimental procedure was analysed separately in the phase of isolation of ECFC colonies and in the following phase of cell expansion. In the isolation phase, although similar frequencies of colonies were obtained on the two substrates, ECFC colonies appeared some days earlier when mononuclear cells were seeded on fibronectin rather than collagen. In the expansion phase, ECFCs cultured on collagen showed a longer lifespan and higher cell yields compared with ECFCs cultured on fibronectin, possibly related to the higher levels of IL-6 and IL-8 measured in their supernatants. ECFCs cultured on both substrates showed similar immunophenotype and ability for in vitro tube formation.

Conclusions/Significance

Overall, the results of this study indicate that, although both fibronectin and collagen efficiently sustain ECFC cultures, each of them brings some advantages within individual steps of the entire process. We suggest that colony isolation performed on fibronectin followed by cell expansion performed on collagen may represent a novel and the most efficient strategy to obtain ECFCs from adult peripheral blood samples.     

New Insight into Wolbachia Epidemiology: Its Varying Incidence During the Host Life Cycle Can Alter Bacteria Spread

Wolbachia is an obligate endosymbiont whose spread depends mainly on its capacity to alter host reproduction by, for instance, cytoplasmic incompatibility. Several mathematical models have been developed to explain the dynamics of bacterial spread, because of its applied interest. However, some aspects of the host’s and bacterium’s biology have not been considered in modelling: for instance, changes in Wolbachia proportions during the host’s life cycle have been observed in several species, including Drosophila sp., Nasonia sp. and Aedes sp. (Diptera), but also in the grasshopper Chorthippus parallelus (Orthoptera), the species studied in this article. These changes influence the proportion of incompatible crosses and, consequently, infection prevalence in subsequent generations. In this paper, we are interested in ascertaining whether these changes in the infection proportions during the host’s life cycle can influence the dynamics of the spread of these bacteria. We have examined its consequences using a mathematical model to predict the evolution of Wolbachia infection frequencies. The simulations were validated by experimental field data from C. parallelus. The main outcome is that those changes above mentioned might affect long-term infection spread, with possible consequences for the current distribution of Wolbachia and the way it affects its host’s reproduction.     

l-Arginine induces protein aggregation and transformation of supramolecular structures of the aggregates

Protein misfolding, self-assembly, and aggregation are an essential problem in cell biology, biotechnology, and biomedicine. The protein aggregates are very different morphologically varying from soluble amorphous aggregates to highly ordered amyloid-like fibrils. The objective of this study was to elucidate the role of the amino acid l-arginine (Arg), a widely used suppressor of protein aggregation, in the regulation of transformations of soluble aggregation-prone proteins into supramolecular structures of higher order. However, a striking potential of Arg to govern the initial events in the process of protein aggregation has been revealed under environment conditions where the protein aggregation in its absence was not observed. Using dynamic light scattering we have demonstrated that Arg (10–100 mM) dramatically accelerated the dithiothreitol-induced aggregation of acidic model proteins. The inhibitory effect on the protein aggregation was revealed at higher concentrations of Arg. Using atomic force microscopy it was shown that aggregation of α-lactalbumin from bovine milk induced upon addition of Arg reached a state of formation of supramolecular structures of non-fibrillar species profoundly differing from those of the individual protein in type, size, and shape. The interaction of another positively charged amino acid l-lysine with α-lactalbumin also resulted in profound acceleration of the aggregation process and transformation of supramolecular structures of the aggregates.     

Going going gone: the loss of a reef building gastropod (Mollusca: Caenogastropoda: Vermetidae) in the southeast Mediterranean Sea

A new harvestman species, Rilaena artvinensis sp. n. (Opiliones: Phalangiidae), is described and illustrated based on males collected in north-eastern Anatolia, Turkey. The new species is similar to R. buresi (?ilhavý, 1965 ?ilhavý, V. (1965): Die Weberknechte der Unterordnung Eupnoi aus Bulgarien; zugleich eine Revision europäischer Gattungen der Unterfamilien Oligolophinae und Phalangiinae (Arachnoidea, Opilionidea). Acta Entomologica Bohemoslovaca, 62, 369–406. [Google Scholar]) and R. lenkoranica Snegovaya, 2007 Snegovaya, N. Y. (2007): Two new harvestman species from Lenkoran, Azerbaijan (Arachnida: Opiliones: Phalangiidae). Bulletin of the British Arachnological Society, 14, 88–92. [Google Scholar]. The differences between these species are elaborated.

http://www.zoobank.org/urn:lsid:zoobank.org:pub:A53452BB-A85D-4BE8-8A6B-D0FC3421AFE3     

New alien insect pests to Portugal on urban ornamental plants and additional data on recently introduced species

The spread of invasive pests is an important topic in pest control and early detection is crucial for containment of invasive aliens. In this study the results of recent surveys on alien insects of ornamental urban plants in Portugal are reported. Overall, 13 alien species associated with allochthonous ornamental plants were recorded. Seven of these pests are new to the Portuguese fauna: Tinocallis kahawaluokalani (Kirkaldy); Illinoia liriodendri (Monell); Appendiseta robiniae (Gillette) (Hemiptera: Aphididae); Acizzia jamatonica (Kuwayama) (Hemiptera: Psyllidae); Dasineura gleditchiae (Osten Sacken); Obolodiplosis robiniae (Haldeman) (Diptera: Cecidomyiidae); and Phytoliriomyza jacarandae Steyskal & Spencer (Diptera: Agromyzidae). Additional distribution data are also provided for other recently introduced alien pests in Portugal: Myzocallis (Lineomyzocallis) walshii (Monell) (Hemiptera: Aphididae); Platycorypha nigrivirga Burckhardt; Glycaspis brimblecombei Moore (Hemiptera: Psyllidae); Calophya schini Tuthill (Hemiptera: Calophyidae); Corythucha ciliata (Say) (Hemiptera: Tingidae); and Lantanophaga pusillidactylus (Walker) (Lepidoptera: Pterophoridae). Three species in particular (Illinoia liriodendri, Obolodiplosis robiniae and Phytoliriomyza jacarandae) were not previously found in the Iberian Peninsula. Details on current distribution, host plants and biological remarks are given for each species. Possible origin areas, pathways and impacts are also discussed.     

Ex vivo analysis of pancreatic cancer-infiltrating T lymphocytes reveals that ENO-specific Tregs accumulate in tumor tissue and inhibit Th1/Th17 effector cell functions

Pancreatic cancer (PC) is an aggressive disease with dismal prognosis. Surgical resection is the recommended treatment for long-term survival, but patients with resectable PC are in the minority (with a 5-year survival rate of 20 %). Therefore, development of novel therapeutic strategies, such as anti-PC immunotherapy, is crucial. α-Enolase (ENO1) is an enzyme expressed on the surface of pancreatic cancer cells and is able to promote cell migration and cancer metastasis. The capacity of ENO1 to induce an immune response in PC patients renders it a true tumor-associated antigen. In this study, we characterized the effector functions of ENO1-specific T cells isolated from PC patients, and we specifically evaluated the successful role of intra-tumoral T helper 17 (Th17) cells and the inhibitory role of regulatory T (Tregs) cells in respectively promoting or reducing the cancer-specific immune response. In this ex vivo study, we have demonstrated, for the first time, that ENO1-specific Th17 cells have a specific anti-cancer effector function in PC patients, and that there are decreased levels of these cells in cancer compared to healthy mucosa. Conversely, there are elevated levels of ENO1-specific Tregs in PC patients which lead to inhibition of the antigen-specific effector T cells, thus highlighting a possible role in promoting PC progression. These results may be relevant for the design of novel immunotherapeutic strategies in pancreatic cancer.     

Identification of one novel causative mutation in exon 4 of WFS1 gene in two Italian siblings with classical DIDMOAD syndrome phenotype

The aim of the present paper is to describe a novel missense mutation (G107R) of WFS1 gene that was unexpectedly detected, in two siblings from Southern Italy, outside exon 8; a very unusual finding which has previously been reported only twice in Italian patients with Wolfram syndrome (WS). Although in Spanish pedigrees' WFS1 mutations are frequently located in exon 4, this finding is very infrequent in other pedigrees, particularly in Italian patients. Conclusions: a) our report of two siblings with one novel WSF1 mutation (G107R) expands the molecular spectrum of WS; b) this is the 3rd report of Italian patients harbouring one mutation outside exon 8 and the 2nd with one mutation in exon 4; c) on the basis of the present observations, and literature data we can infer that mutation locations outside exon 8 do not seem to be clearly associated with peculiar phenotype expressions of WFS1 gene.     

Isolated carcinoid tumor metastatic to the thyroid gland: report of a case initially diagnosed by fine needle aspiration cytology

BACKGROUND: Neuroendocrine tumor metastatic to the thyroid gland is rare and may be difficult to differentiate from primary thyroid neuroendocrine tumors, such as medullary thyroid carcinoma (M/ITC). This report describes an unusual case of bronchial carcinoid metastatic to the thyroid diagnosed by fine needle aspiration cytology (FNAC). CASE: A 42-year-old woman with an undiagnosed bronchial carcinoid tumor presented to our clinic with a solitary nodule in the thyroid gland. FNAC of the nodule showed loosely cohesive groups of cuboidal tumor cells with scant, slightly granular cytoplasm; centrally located nuclei with a coarsely granular, salt-and-pepper chromatin pattern and inconspicuous nucleoli. Immunocytochemically the tumor cells were positive for neuron-specific enolase, chromogranin and synaptophysin and negative for thyroglobulin, calcitonin and carcinoembryonic antigen. The cytologic diagnosis of a metastatic neuroendocrine carcinoma was confirmed histologically. CONCLUSION: Metastasis to the thyroid gland may pose a diagnostic problem, particularly with tumors of neuroendocrine origin, as these have similar cytologic features in various organs. The correct preoperative cytologic diagnosis of metastatic carcinoid tumor in patients without a prior history of cancer and differential diagnosis with MTC are crucial because prognosis, workup and treatment are different in each.