Description of the first-stage juveniles of Xiphinema cretense and X. herakliense - Distribution of Xiphinema and Longidorus species in olive orchards and grapevines in Crete, Greece

The occurrence of nematodes of the family Longidoridae was investigated in soil samples collected from cultivated and wild olives and grapevines in Crete. The first-stage juveniles of Xiphinema cretense and X. herakliense are described for the first time. The species X. israeliae, X. cretense, X. herakliense and Longidorus pseudoelongatus, previously recorded exclusively from olives in Crete, are herein reported in the rhizosphere of grapevines. Also L. iranicus is reported for the first time in cultivated olive, while X. italiae and L. closelongatus are reported for the first time in wild olive in Crete. Data on the occurrence of phytoparasitic nematode species in cultivated olives, wild olives and grapevines are updated with those previously published.This research was supported by grant KBBE 219262 ArimNET-ERANET FP7 2012-2015 Project PESTOLIVE ‘Contribution of olive history for the management of soilborne parasites in the Mediterranean basin’ from Hellenic Agricultural Organization-DEMETER and Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria (INIA), grant AGR-136 from ‘Consejería de Economía, Innvovación y Ciencia’ from Junta de Andalucía, and Union Europea, Fondo Europeo de Desarrollo regional, “Una manera de hacer Europa”. I.G. Birmpilis and E. Nasiou were employed by the ARIMNET-PESTOL-IVE project.Peer reviewe

ISO-66, a novel inhibitor of macrophage migration inhibitory factor, shows efficacy in melanoma and colon cancer models

Macrophage migration inhibitory factor (MIF) is a pleiotropic pro-inflammatory cytokine, which possesses a contributing role in cancer progression and metastasis and, thus, is now considered a promising anticancer drug target. Many MIF-inactivating strategies have proven successful in delaying cancer growth. Here, we report on the synthesis of ISO-66, a novel, highly stable, small-molecule MIF inhibitor, an analog of ISO-1 with improved characteristics. The MIF:ISO-66 co-crystal structure demonstrated that ISO-66 ligates the tautomerase active site of MIF, which has previously been shown to play an important role in its biological functions. In vitro, ISO-66 enhanced specific and non-specific anticancer immune responses, whereas prolonged administration of ISO-66 in mice with established syngeneic melanoma or colon cancer was non-toxic and resulted in a significant decrease in tumor burden. Subsequent ex vivo analysis of mouse splenocytes revealed that the observed decrease in tumor growth rates was likely mediated by the selective in vivo expansion of antitumor-reactive effector cells induced by ISO-66. Compared to other MIF-inactivating strategies employed in vivo, the anticancer activity of ISO-66 is demonstrated to be of equal or better efficacy. Our findings suggest that targeting MIF, via highly specific and stable compounds, such as ISO-66, may be effective for cancer treatment and stimulation of anticancer immune responses

Presence of Longidoridae (Xiphinema and Longidorus) nematodes in grapevine, cultivated olive and wild olive in Crete

Trabajo presentado en el XVI Panhellenic Entomological Congress, celebrado en Heraklion del 20 al 23 de octubre de 2015.The presence of the nematodes Xiphinema and Longidorus (Longidoridae) was studied in the rhizosphere of grapevine, cultivated olive and wild olive. Soil samples from grapevines were brought in the lab by farmers for investigation while samples from olives were collected in the framework of ARIMNET-PESTOLIVE project Morphological and morphometric studies combined with molecular techniques were applied for nematode identification. Five species of Longidorus and seven species Xiphinema were found in totaL Longidorus closelongatus, L moesicus, and L pseudoelongatus were found in grapevine and _olive with the two last species recorded for first time in Greece. L orientalis was found only in grapevine something that consists a new record for Greece and an unusual record for Europe. L cretensis, which was first described in Crete in 2001 and has never been reported elsewhere, was found in both crops. Xiphinema pachtaicum and X italiae which are widespread along the Mediterranean basin, were found in both crops. X simile, was found in grapevine and consists a new record for the country. X index, beside in grapevine where is quite common, was found ata low percentage (1 ,5%) in cultivated olive. Even though this nematode is not a virus vector in olive, it could be potentially damaging in grapevines adjacent to cultivated olives. X israeliae was found in cultivated and wild ol ives; its presence consists the second record worldwide, after the original identification of the species in IsraeL Furthermore, two new species were described for first time, X cretense n.sp in cultivated olive and X herak/iense n.sp in both wild and cultivated olive. All the nematodes mentioned above, were found in the rhizosphere of olive but it is not confirmed whether olives or/and weeds are their hosts. Except Xindex, no other species has ever been reported as a virus vector. The most important finding is the presence of X israeliae and the two new species X cretense and X herakliense; a study on their biology and their potential to be virus vectors consists a quite interesting resea rch area.N

Rotylenchus cretensis n. sp. and R. cypriensis Antoniou 1980 (Nematoda: Hoplolaimidae) recovered from the rhizosphere of olive at Crete (Greece) with a molecular phylogeny of the genus

A new and a known species of Rotylenchus from Crete, each collected from the rhizosphere of olive, are described and illustrated based on morphological, morphometric and molecular studies. Rotylenchus cretensis n. sp. is characterised by having a truncate lip region with irregular longitudinal striation, lateral field areolated only in pharynx region, stylet length less than 41 μm, vulva located at 53–59 %, conoid-rounded to dorsally convex-conoid female tail with 7–11 annuli and specific D2-D3 of 28S, ITS, and partial 18S rRNA. Rotylenchus cretensis n. sp. appears to be similar to R. urmiaensis, having overlapping character ranges. Rotylenchus cretensis is also similar to R. striaticeps from which has got shorter stylet length (35.5–41.5 vs 39–50 μm), female tail shape (conoid-rounded to dorsally convex-conoid vs rounded), shorter spicules length (34–40 vs 41–50 μm), and phasmid position (varying from at level to three annuli posterior to the anus vs at level to seven annuli anterior to anus). Multivariate analysis of variance performed on R. cretensis n. sp., as well as two populations of R. striaticeps and R. urmiaensis from Iran on combined morphometric characters data set revealed a significant effect by species populations, indicating that closely morphological and morphometrically species differed significantly from each other. Additionally, analysis of variance showed that morphometric characters such as lip region width, distance from phasmid to tail tip (PHS) or a ratio can be used as key morphometric characters for species differentiation in this group. The results of phylogenetic analyses based on D2-D3 region, ITS of rRNA, and the partial 18S sequences confirmed the close molecular relationships between R. cretensis n. sp., R. urmiaensis and R. striaticeps, but clearly support the proposal of R. cretensis n. sp. as a new species. Also, the recognition of this cryptic species within Rotylenchus shows that the biodiversity of these nematodes is still not fully understood and requires further studies. Additionally, R. cypriensis is described and molecularly analysed for first time in Greece.The present research was partially funded by grant KBBE 219262 ArimNET-ERANET FP7 2012–2015 Project PESTOLIVE ‘Contribution of olive history for the management of soilborne parasites in the Mediterranean basin’ from Hellenic Agricultural Organization-DEMETER and Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria (INIA), grant AGR-136 from ‘Consejería de Economía, Innvovación y Ciencia’ from Junta de Andalucía, and Union Europea, Fondo Europeo de Desarrollo regional, ‘Una manera de hacer Europa’. I. G. Birmpilis was employed by the ARIMNET-PESTOLIVE project.Peer reviewe

Prevalence and molecular diversity of reniform nematodes of the genus Rotylenchulus (Nematoda: Rotylenchulinae) in the Mediterranean Basin

The reniform nematodes of the genus Rotylenchulus are semi-endoparasites of numerous herbaceous and woody plant roots and distributed in regions with Mediterranean, subtropical and tropical climates. In this study, we provide morphological and molecular characterisation of three out of 11 valid species of the genus Rotylenchulus: R. macrodoratus, R. macrosoma, and R. reniformis from Greece (Crete), Italy and Spain. The overall prevalence of reniform nematodes in wild and cultivated olives in Greece, Italy, and Spain was 11.5%, 19.0% and 0.6%, respectively. In Greece, R. macrodoratus and R. macrosoma were detected in cultivated olive with a prevalence of 8.2% and 6.2%, respectively, but none of them were found in wild olive. This is the first report of R. macrosoma in Greece. Only one reniform nematode species was detected in olive from Italy and Spain, viz. R. macrodoratus and R. macrosoma, respectively. The parasitism of R. macrosoma on hazelnut in northern Spain was also confirmed for the first time. This study demonstrates that R. macrodoratus and R. macrosoma have two distinct rRNA gene types in their genomes, specifically the two types of D2-D3 for R. macrosoma and R. macrodoratus, the two types of ITS for R. macrodoratus and the testing of the ITS variability in other R. macrosoma populations in different countries. Rotylenchulus macrosoma from Greece and Spain showed differences in nucleotide sequences in the ITS region and D2-D3 of 28S rRNA gene.This research was supported by grant P12-AGR 1486 from ‘Consejería de Economía, Innvovación y Ciencia’ from Junta de Andalucía, and Union Europea, Fondo Europeo de Desarrollo regional, ‘Una manera de hacer Europa’, grant KBBE 219262 ArimNET-ERANET FP7 2012-2015 Project PESTOLIVE ‘Contribution of olive history for the management of soilborne parasites in the Mediterranean basin’ from Hellenic Agricultural Organization-DEMETER and Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria (INIA), and AGL-2012-37521 from ‘Ministerio de Economía y Competitividad’ of Spain. I. G. Birmpilis was employed by the ARIMNET-PESTOLIVE project.Peer reviewe

Immunogenic Cell Death, DAMPs and Prothymosin α as a Putative Anticancer Immune Response Biomarker

The new and increasingly studied concept of immunogenic cell death (ICD) revealed a previously unknown perspective of the various regulated cell death (RCD) modalities, elucidating their immunogenic properties and rendering obsolete the notion that immune stimulation is solely the outcome of necrosis. A distinct characteristic of ICD is the release of danger-associated molecular patterns (DAMPs) by dying and/or dead cells. Thus, several members of the DAMP family, such as the well-characterized heat shock proteins (HSPs) HSP70 and HSP90, the high-mobility group box 1 protein and calreticulin, and the thymic polypeptide prothymosin α (proTα) and its immunoreactive fragment proTα(100–109), are being studied as potential diagnostic tools and/or possible therapeutic agents. Here, we present the basic aspects and mechanisms of both ICD and other immunogenic RCD forms; denote the role of DAMPs in ICD; and further exploit the relevance of human proTα and proTα(100–109) in ICD, highlighting their possible clinical applications. Furthermore, we present the preliminary results of our in vitro studies, which show a direct correlation between the concentration of proTα/proTα(100–109) and the levels of cancer cell apoptosis, induced by anticancer agents and γ-radiation. © 2022 by the authors. Licensee MDPI, Basel, Switzerland

Immunogenic Cell Death, DAMPs and Prothymosin α as a Putative Anticancer Immune Response Biomarker

The new and increasingly studied concept of immunogenic cell death (ICD) revealed a previously unknown perspective of the various regulated cell death (RCD) modalities, elucidating their immunogenic properties and rendering obsolete the notion that immune stimulation is solely the outcome of necrosis. A distinct characteristic of ICD is the release of danger-associated molecular patterns (DAMPs) by dying and/or dead cells. Thus, several members of the DAMP family, such as the well-characterized heat shock proteins (HSPs) HSP70 and HSP90, the high-mobility group box 1 protein and calreticulin, and the thymic polypeptide prothymosin α (proTα) and its immunoreactive fragment proTα(100–109), are being studied as potential diagnostic tools and/or possible therapeutic agents. Here, we present the basic aspects and mechanisms of both ICD and other immunogenic RCD forms; denote the role of DAMPs in ICD; and further exploit the relevance of human proTα and proTα(100–109) in ICD, highlighting their possible clinical applications. Furthermore, we present the preliminary results of our in vitro studies, which show a direct correlation between the concentration of proTα/proTα(100–109) and the levels of cancer cell apoptosis, induced by anticancer agents and γ-radiation

Prothymosin α and Its C-Terminal Immunoreactive Decapeptide Show no Evidence of Acute Toxicity: A Preliminary in Silico, in Vitro and in Vivo Investigation.

BACKGROUND: Members of the α-thymosin family have long been studied for their immunostimulating properties. Among them, the danger-associated molecular patterns (DAMPs) prothymosin α (proTα) and its C-terminal decapeptide proTα(100-109) have been shown to act as immunomodulators in vitro, due to their ability to promote T helper type 1 (Th1) responses. Recently, we verified these findings in vivo, showing that both proTα and proTα(100-109) enhance antitumor-reactive T cell-mediated responses. METHODS: In view of the eventual use of proTα and proTα(100-109) in humans, we investigated their safety profile in silico, in human leukocytes and cancer cells lines in vitro, and in immunocompetent mice in vivo, in comparison to the proTα derivative thymosin alpha 1 (Τα1), a 28-mer peptide extensively studied for its safety in clinical trials. RESULTS: In silico prediction via computational tools showed that all three peptide sequences likely are non-toxic or do not induce allergic regions. In vitro, proTα, proTα(100-109) and Tα1 did not affect the viability of human cancer cell lines and healthy donor-derived leukocytes, did not promote apoptosis or alter cell cycle distribution. Furthermore, mice injected with proTα, proTα(100-109) and Tα1 at doses equivalent to the suggested dose regimen of Tα1 in humans, did not show signs of acute toxicity, whereas proTα and proTα(100-109) increased the levels of proinflammatory and Th1-type cytokines in their peripheral blood. CONCLUSION: Our preliminary findings suggest that proTα and proTα(100-109), even at high concentrations, are non-toxic in vitro and in an acute toxicity model in vivo; moreover, we show that the two peptides retain their immunomodulatory properties in vivo and, eventually, could be considered for therapeutic use in humans