1,721,107 research outputs found
Serological evidence of Midichloria mitochondrii circulation in humans parasitized by I. ricinus in Germany and development of a marker for tick bite
Full text linkThe tick Ixodes ricinus transmits several microorganisms of medical and veterinary importance. Midichloria mitochondrii (order Rickettsiales; family Midichloriaceae) is an intracellular symbiont present in the ovaries and salivary glands of 100% of adult I. ricinus females (Sassera et al., 2008) and is transmitted to the vertebrate host during the tick bite (Mariconti et al., 2012; Bazzocchi et al., 2013) despite its infective role is not demonstrated.
In this experimental study, a total of 324 human from different areas of Germany were analysed in order to investigate the seropositivity against the flagellar FliD protein of M. mitochondrii using an ELISA approach. Fifty sera were collected from patients living in non-endemic areas and used as negative controls while 274 sera were obtained from subjects exposed to tick bite and suffering from several symptoms referred to tick borne diseases and collected at the BCA clinic in Augsburg.
Since the positivity for the spirochete Borrelia burgdorferi transmitted by I. ricinus is indicative for the tick bite, we considered also this additional information.
The obtained results showed that 82 out of 274 sera were positive to M. mitochondrii and 42 out of 175 sera of subjects negative to B. burgdorferi were positive to M. mitochondrii to confirm the good property of FliD protein as a tick bite marker. However, the high number (133 out of 274) of subjects parasitized by I. ricinus but negative to both bacteria prompted us to detect new more suitable I. ricinus/M. mitochondrii antigenic proteins to use as markers for tick bite. For this purpose, three I. ricinus proteins and one surface protein of M. mitochondrii were selected and 12 synthetic peptides were designed in order to set-up a new ELISA test for investigating the exposure of humans and animals to this tick species
Going Beyond Counting First Authors in Author Co-citation Analysis
Full text linkThe present study examines one of the fundamental aspects of author co-citation analysis (ACA) - the way co-citation
counts are defined. Co-citation counting provides the data on which all subsequent statistical analyses and mappings
are based, and we compare ACA results based on two different types of co-citation counting - the traditional type that
only counts the first one among a cited work's authors on the one hand and a non-traditional type that takes into
account the first 5 authors of a cited work on the other hand. Results indicate that the picture produced through this non-traditional author co-citation counting contains more coherent author groups and is therefore considerably clearer. However, this picture represents fewer specialties in the research field being studied than that produced through the traditional first-author co-citation counting when the same number of top-ranked authors is selected and analyzed. Reasons for these effects are discussed
Production and characterization of a polyclonal antibody against the bovine prion-like doppel protein
No full textPRODUCTION AND CHARACTERIZATION OF A POLYCLONAL ANTIBODY AGAINST THE BOVINE PRION-LIKE DOPPEL PROTEIN
Marco Rondena, Fabrizio Ceciliani, Chiara Bazzocchi, Saverio Paltrinieri
Dipartimento di Patologia Animale, Igiene e Sanità Pubblica Veterinaria – Milano
AIM OF THE STUDY: Doppel (Dpl) is a prion-like protein recently identified in mammalian. The sequence is notably homologous to that of PrP. Dpl is associated to neurodegeneration when ectopically expressed in transgenic mouse. The aim of the study is to raise and characterize a polyclonal antibody (pAb) against bovine Dpl to study the tissue distribution of this protein. MATERIAL AND METHODS. A synthetic peptide spanning from residues 67 to 81 of bovine Dpl sequence was synthetized. Using this peptide, a pAb (Dpl 67-81), was raised in rabbit and purified by means of Protein A affinity chromatography. In order to test the specificity of Dpl 67-81, bovine Dpl was cloned and overexpressed in E.coli in the expression vector PQE30 and purified following methal chelate affinity and bovine PrP was purified by means of cation-exchange chromatography and cobalt-chelate affinity chromatography. Homogenate of tissues were tested for the presence of Dpl after SDS-PAGE and Western Blotting. Blots were visualized using ECL. Immunohistochemistry on cryostatic sections using the avidin-biotin complex methods was performed. RESULTS: Dpl 67-81 strongly react against rDpl (titre =1:20000) and not against bovine PrP. Dpl was detected in testis, spleen, limphnodes by both immunoblotting and immunohistochemistry. In particular endothelial and stromal cells showed a granular cytoplasmic positivity. CONCLUSIONS: Dpl 67-81 can detect the expression of Dpl in tissues using both Western Blotting and immunohistochemistry. Moreover, it does not crossreact with PrP, and therefore it could be also used in experiments of functional blocking of Dpl in order to study its function and its relationship with PrP
Batteri della famiglia Midichloriaceae (ordine Rickettsiales): possibili patogeni emergenti per i vertebrati?
No full textLa famiglia Midichloriaceae è una famiglia batterica il cui nome deriva da Midichloria mitochondrii, simbionte intramitocondriale presente nell’ovario della zecca Ixodes ricinus (Montagna et al., 2013). Le Midichloriaceae sono comparabili, per numero di specie e per complessità, alle altre due famiglie dell’ordine Rickettsiales (Rickettsiaceae ed Anaplasmataceae), di cui fanno parte diversi batteri intracellulari patogeni per l’uomo ed altri vertebrati. In seguito alla scoperta di M. mitochondrii in I. ricinus e di batteri a esso correlati in altre specie di zecca dura, altri batteri della famiglia Midichloriaceae sono stati osservati in organismi molto diversi tra loro, spaziando da artropodi parassiti (ad esempio, pulci e cimici dei letti) e organismi dei phyla Porifera e Cnidaria, fino ad arrivare a organismi acquatici quali protisti appartenenti a specie di ciliati e ad amebe patogene del genere Acanthamoeba (Montagna et al., 2013). L’associazione di alcune midichloriaceae con organismi parassiti fa supporre un loro possibile coinvolgimento nel ruolo patogeno svolto dall’ospite parassita. Questa ipotesi è in primo luogo supportata da dati molecolari e sierologici riguardanti M. mitochondrii in I. ricinus: è infatti noto che questo batterio, presente anche nelle ghiandole salivari e nel rostro della zecca, viene trasmesso all’ospite vertebrato durante il pasto di sangue (Mariconti et al., 2012; Bazzocchi et al., 2013), anche se il suo destino, una volta inoculato nell’ospite, è tutt’ora ignoto. Altre linee di evidenza riguardanti l’infettività di batteri ascritti alla famiglia Midichloriaceae sono riscontrabili nell’ipotesi di un loro coinvolgimento nello sviluppo di patologie quali la red mark syndrome/strawberry disease nella trota iridea (Onchorynchus mykiss) (Cafiso et al., 2015) e di un possibile ruolo come coadiuvante nello sviluppo delle cheratiti oculari da Ancanthamoeba spp. Tuttavia al momento la potenziale patogenicità di alcuni membri della famiglia Midichloriaceae per l’uomo e altri vertebrati deve ancora essere dimostrata. Ad esempio, in soggetti parassitati da zecca che mostrano aspetti clinici attribuibili a rickettsiosi o ehrlichiosi, ma che mancano di titolo anticorpale contro microrganismi trasmessi tipicamente con il morso da zecca, le infezioni dovrebbero essere re-investigate sulla base di segni derivanti da infezioni correlate a Midichloriaceae. In conclusione, quindi, diversi aspetti suggeriscono il possibile ruolo di alcuni membri appartenenti alla famiglia Midichloriaceae come agenti infettivi/patogeni per i vertebrati
Attivazione dei granulociti neutrofili in presenza di wolbachia surface protein (WSP): osservazioni preliminari
No full textTransmission of members of the “Candidatus midichloriaceae“ family to vertebrates and possibile involvement in disease pathogenesis
No full text“Candidatus Midichloriaceae” (order Rickettsiales) is a family that encompasses obligate intracellular bacteria present in a wide range of hosts, from parasitic arthropods (ticks, fleas, bedbugs) to aquatic animals (e.g. sponges and corals) and protists, including pathogenic amoebae (genus Acanthamoeba). In addition, multiple studies have detected evidence of the presence of “Candidatus Midichloriaceae” in vertebrates, suggesting a capacity to cause infections, with possible pathogenic effects. Two members of the family that are described as symbionts of ticks, i.e. “Candidatus Midichloria mitochondrii” and “Candidatus Lariskella arthropodarum” have also been found in the blood of humans and other mammals, using molecular and serological methods. Consequently, it has been hypothesized that these arthropods could transmit the microorganisms during the blood meal, as is known to happen with pathogenic members of the order Rickettsiales; for example, ticks can be vectors of rickettsiosis or ehrlichiosis. Indeed, clinical cases compatible with these pathologies in humans or animals parasitized by ticks, but characterized by the absence of antibody titers for known pathogenic rickettsiae, could be reinvestigated for the presence of “Candidatus Midichloriaceae”. A third member of the family, a “Candidatus Midichloria”-related organism, has repeatedly been found in association with red mark syndrome in the rainbow trout Oncorhynchus mykiss. Although the etiology of this economically relevant disease is currently unknown, multiple independent studies suggest that this member of “Candidatus Midichloriaceae” is the most probable causative agent
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