Seed cryopreservation of Fraxinus angustifolia Vahl

Fraxinus angustifolia is used for afforestation and for production of both valuable timber and manna, a substance with pharmacological applications. The aim of this research was to establish the optimized condition for cryopreservation of F. angustifolia seeds, in order to protect the germplasm of this species. Germination percentage and mean time of germination of non-treated seeds (control) and liquid nitrogen-treated seeds were established in water or in 10-6 M gibberellic acid (GA3). The seeds could be cryopreserved with 3% of moisture content (MC) and germinated easily in water (70.0 ± 5.0%), while seeds cryopreserved with 6% MC showed a physiological dormancy. This state could be efficiently removed by addition of GA3, which increased germination to 76.0 ± 2.4%. Liquid nitrogen did not impair embryo viability, allowing a successful cryopreservation of seeds. The results are particularly useful for both germplasm conservation programs and in narrow leaved ash nurseries.Keywords: Fraxinus angustifolia, germination, germplasm conservation, seed cryopreservationAfrican Journal of Biotechnology Vol. 12(16), pp. 1930-193

Boosting the MHC class II-restricted tumor antigen presentation to CD4+ T helper cells: A critical issue for triggering protective immunity and re-orienting the tumor microenvironment toward an anti-tumor state

6noopenAlthough the existence of an immune response against tumor cells is well documented, the fact that tumors take off in cancer patients indicates that neoplastic cells can circumvent this response. Over the years many investigators have described strategies to rescue the anti-tumor immune response with the aim of creating specific and long-lasting protection against the disease. When exported to human clinical settings, these strategies have revealed in most cases a very limited, if any, positive outcome. We believe that the failure is mostly due to the inadequate triggering of the CD4+ T helper (TH) cell arm of the adaptive immunity, as TH cells are necessary to trigger all the immune effector mechanisms required to eliminate tumor cells. In this review, we focus on novel strategies that by stimulating MHC class II-restricted activation of TH cells generate a specific and persistent adaptive immunity against the tumor. This point is of critical importance for both preventive and therapeutic anti-tumor vaccination protocols, because adaptive immunity with its capacity to produce specific, long-lasting protection and memory responses is indeed the final goal of vaccination. We will discuss data from our as well as other laboratories which strongly suggest that triggering a specific and persistent anti-tumor CD4+ TH cell response stably modify not only the tumor microenvironment but also tumor-dependent extratumor microenvironments by eliminating and/or reducing the blood-derived tumor infiltrating cells that may have a pro-tumor growth function such as regulatory CD4+/CD25+ T cells and myeloid-derived-suppressor cells. Within this frame, therefore, we believe that the establishment of a pro-tumor environment is not the cause but simply the consequence of the tumor strategy to primarily counteract components of the adaptive cellular immunity, particularly TH lymphocytes.openAccolla, R.S.; Lombardo, L.; Abdallah, R.; Raval, G.; Forlani, G.; Tosi, G.Accolla, Roberto; Lombardo, L.; Abdallah, R.; Raval, G.; Forlani, Greta; Tosi, Giovann

Biotechnology for microbial monitoring of indoor cultural heritage environments

An integrated approach for the characterization of bioaerosol was employed in different sites (that include hypogeal and semi-confined areas), characterized by great cultural/artistic interest besides peculiar architectural structures, thermo- hygrometric and lighting parameters. These typologies of indoor environments preserve several artworks like mural paintings, stone-works, paper or parchments that are susceptible of microbial colonization. The presence of fungal spores and low air change can induce both potentially effects to human health (users/operators) or biodegradation of historical-artistic manufacts. We perform bioaerosol sampling by a portable sampler (Sartorius MD8), equipped with gelatin filters and non-invasive sampling (Nylon membrane or sterile swab) is carried out on works of art surface. Microbial consortia is revealed and characterized by Optical, Scanning Electron and Confocal Laser Scanning Microscopy (OM, SEM, CLSM), in vitro culture and molecular analysis (PCR, sequencing, sequence analysis). The inter-disciplinary approach applied in this study, represents a valuable contribution for the proper planning of both direct and/or indirect biological growth control and for the conservative restoration procedure (1, 2). This work was supported by Research Project It@cha - Italian Technologies for Advanced application in Cultural Heritage Assets, grant PON 01_00625 “Ricerca e Competitività” 2007-2013. (1) Palla F. et al. (2006) Characterization of bacterial community in indoor environment. Heritage, Weathering and Conservation, 1: 361-365. (2) Palla F. et al. (2010) Microscopy and molecular biology techniques for the study biocenosis diversity in semi-confined environments. Conservation Science in Cultural Heritage, 10: 185-194

Real-Life effects of benralizumab on exacerbation number and lung hyperinflation in atopic patients with severe eosinophilic asthma.

Background: The humanized monoclonal antibody benralizumab targets the α subunit of the interleukin-5 (IL-5) receptor and the FcγRIIIa receptor expressed by natural killer cells. Through this dual mechanism of action, benralizumab neutralizes the pro-eosinophil functions of IL-5 and promotes eosinophil apoptosis. Objectives and methods: The present real-life study aimed to evaluate, in 22 allergic patients with severe eosinophilic asthma, the effects of benralizumab on asthma exacerbations and lung hyperinflation. Results: In this regard here we show that, after 24 weeks of add-on treatment, benralizumab completely depleted peripheral blood eosinophils (from 810 to 0 cells/μL; p < 0.0001), and significantly decreased both asthma exacerbation number (from 4 to 0; p < 0.0001) and residual volume (from 2720 to 2300 mL; p < 0.01). Moreover, at the same time point (24 weeks) benralizumab also increased pre-bronchodilator FEV1 (from 1295 to 1985 mL; p < 0.0001), FVC (from 2390 to 2974 mL; p < 0.0001), FEF25−75 (from 0.6 to 1.42 L/sec; p < 0.0001), IC (from 1940 to 2460 mL; not significant), and ACT score (from 14.73 to 22.95; p < 0.0001), as well as reduced prednisone intake (from 25 to 0 mg; p < 0.0001). Conclusion: In conclusion, our results suggest that via its anti-eosinophil actions benralizumab improved airflow limitation, lung hyperinflation, and respiratory symptoms, as well as lowered asthma exacerbation rate and abrogated OCS consumption in most patients

Speech outcome in tongue cancer surgery: objective evaluation by acoustic analysis software

BACKGROUND. Cancer of the oral cavity is one of the most common malignancies of which 60% affect the tongue. Carcinoma of the tongue causes significant alterations of the articulatory and swallowing functions. The gold standard of care remains primary surgical resection with or without postoperative adjuvant therapy. Whereas T1 and T2 tongue tumors can be treated with more conservative surgeries, as partial glossectomies, the larger tumors require total and aggressive glossectomies which increase survival, but, on the other hand, they might often make speech, chewing and swallowing impossible. MATERIAL AND METHODS. Our study was performed on a total of 21 patients with Squamous Cell Carcinoma of the tongue who underwent either partial resection or hemiglossectomy. Each subject (either surgical patients or controls) was asked to pronounce the vowels /a/, /e/, /i/, /u/, and all signals were evaluated separately by two operators. Acoustic (F0, jitter, shimmer, NHR) and vowel metric (the ratio F2i/F2u, tVSA, qVSA, FCR) features have been extracted. In order to define the speech intelligibility, all patients were evaluated by two doctors and one speech therapist and all patients received the Speech Handicap Index (SHI) translated into Italian language before recording. RESULTS. No statistically significant variations were observed, regardless of the gender, between controls and surgically resected patients when tumor staging was T1-T2. On the contrary, when patients had to undergo more extensive surgical resection due to the presence of a T3-T4 tumor, a dramatic increase of F2u could be observed. This change, together with a decrease of F2i, led to a highly significant reduction in the F2i/F2u parameter in surgically resected patients as compared to controls. The other parameters which were reduced in a statistically significant manner in T3-T4 surgically resected patients were tVSA and qVSA. Instead, two parameters increased in a statistically significant manner in T3-T4 surgically resected patients: FCR and SHI. Again, none of the above-mentioned parameters was altered in a statistically significant manner in early tumor stage resected patients, regar dless of the gender. CONCLUSION. For the first time, we used a series of newly developed formant parameters, introduced by various authors for the study of the articulatory undershoot of the tongue in various neurodegenerative diseases. The statistical analysis of our results highlighted in an incontrovertible way a strong correlation and significance of each of our parameters F2 / i / / F2 / u /, FCR, tVSA, qVSA, with the entity of the TNM, and therefore of the surgical extension of the resection, and in parallel with the loss of the intelligibility of the speech that proportionally reaches higher values in the advanced stages of the disease as can be deduced from the SHI trend

MORPHOLOGICAL ANALYSIS OF ANCIENT GRAPE SEEDS FROM A SINK IN THE MIDDLE-AGE TOWN OF PALERMO

The archaeological excavations in Piazza della Vittoria, in the Roman-Middle Age town of Palermo (Sicily) put in light a sink 3.20 m deep and 1 square m. large, partially filled by thin organic sediments. Grape seeds (grape-stones), fish scales and few vertebrate bones have been found in specific strata sealed under a stratum chronologically attributed to Islamic Middle-Age period (a post-quem limit). The finding of well preserved grape seeds is peculiar and their study opens the opportunity to improve the actual knowledge about evolution, cultivation, use and trade of Vitis L. in the Mediterranean area. This preliminary work focuses on morphologic and morphometric analysis of the ancient grape seeds with two aims: i) systematically describe the remains collection and, ii) define seeds typology and a consequent morphotaxonomic attribution. Over 200 seeds have been carefully dry cleaned (soft brush), photographed and analyzed for total breadth (B), total length (L) and length of stalk (LS) parameters, the most efficient for typological attribution (1, 2); Stummer index has been also calculated (1). Apical notch length (AN) has been for the first time evaluated. Measurements on digital images have been performed using ImageJ 1.31 platform; morphological parameters have been assembled in a dedicated database. Descriptive analysis and linear correlations have been performed using SYSTAT 10. Analysis of variance (ANOVA) and Tukey’s HSD (5% level of significance, α = 0.05) have been applied. All the parameters approximate a normal distribution. Major variation has been observed in LS (c.v. = 35.6%) and AN (c.v. = 35.6%), while B and L showed a c.v. of 9.5% and 12.6% respectively. All the analyzed parameters behave as independent variables with the exception of a significant correlation between Stummer index and L (R2 = 0.45; y = 8.17-0.047x with y = L and x = Stummer index). This correlation reveals that Stummer index depends more from the L and not from the B parameter. On the base of LS measures three subgroups have been arbitrarily created in relationship with the LS: LS1 0.90 mm (35 seeds). Analyzing together the LS groups toward AN, we have found a proportional and significant correlation (p = 0.05) between the extremes LS1 and LS3. In the entire collection, Stummer index varies from 55.76 to 100.86; in the LS groups, the range is 68.38-97.87 in LS1, 61.02-100.86 in LS2 and 55.76-81.70 in LS3. A small group (17) of seeds has been excluded for the impossibility to measure the stalk. The analyzed ancient grape seeds show a wide range of variability for all the considered parameters, revealing a polymorphic collection. In general, the seeds have a rounded heart-like shape, with a noticeable pointy stalk and a very invaginated apical notch. This typical shape is more marked in LS3 group. On the base of LS measures, LS1 is ascribable to wild grapevines, while LS2 and LS3 seem to be ascribable neither to wild nor to cultivated autochthonous Vitis. Furthermore, these seeds differ from those already described in other archaeological horizons in Italy (3) and in France (2). The Stummer index varies highly, exceeding the known range of wild Vitis vinifera (4), although values close to 100 have been already found in wild grapevines in Spain (5) and values above 80 have been also described in Extra-European Vitis species (6). A deep evaluation of the sample, including isotopic analysis and aDNA studies, is in progress. 1) T. T. Korenčič, J. Jakše, Z. Korošec-Koruza (2008) Veget. Hist. Archaeobot., 17(Suppl. 1), S93-S102 2) L. Bouby, I. Figueiral, A. Bouchette, N. Rovina, S. Ivorra, T. Lacombe, T. Pastor, S. Picq, P. Marinval, J. F. Terral (2013) PLoS ONE, 8(5), e63195 3) C. Milanesi, F. Antonucci, P. Menesatti, C. Costa, C. Faleri, M. Cresti (2011) Interdisciplinaria Archaeologica – Natural Sciences in Archaeology, II(2), 95-100 4) A. Stummer (1911) Mitt. Anthropol. Gesellschaft Wien, 41, 283-296 5) F. M. De Toda, J. C. Sancha (1999) Am. J. Enol. Vitic., 50(4), 443-446 6) D. Rivera, B. Miralles, C. Obón, E. Carreño, J. A. Palazón (2007) Vitis, 46(4), 158-16