Extra hepatic expression of the acute phase protein alpha 1-acid glycoprotein in normal bovine tissues

Combined quantitative (real-time) polymerase chain reaction and immunohistochemistry were used to evaluate the expression of the minor acute phase protein alpha 1-acid glycoprotein (AGP, orosomucoid) in bovine extrahepatic tissues. AGP was produced mainly in the salivary glands and spleen, whereas minor expression was detected in all other tissues sampled, including lung, lymph nodes, uterus, ovary, kidney and tongue. The findings were consistent with immunohistochemical results. In view of the immunomodulatory and direct antibacterial activity of AGP, its expression in the salivary glands may signal an involvement in the regulation of the local immunity, even in non-pathological conditions.     

Proline and light as quality enhancers of rocket (Eruca sativa Miller) grown under saline conditions

Stress conditions experienced during growth may affect plant responses during post-harvest storage and eventually determine the overall quality of commercial products. In this context, we hypothesized that foliar applications of proline during the growth cycle and light exposure during post-harvest storage could be two important modulators of yield and quality parameters of rocket plants exposed to NaCl stress. Dry matter percentage increased upon NaCl treatment. However, fresh weight loss during storage did not change over time as a consequence of salt stress. High salinity (100 mM NaCl) moderately reduced both leaf nitrate (14%) and nitrite (3%) contents. Lipophilic (LAC) and hydrophilic (HAC) antioxidant activities also decreased by 10% at the highest salinization (average of two growth cycles). In contrast, during storage, LAC decreased whereas HAC increased. Proline applications reduced the leaf nitrate content during storage by 16%, increased carotenoids and chlorophyll contents in salinized plants and also increased the ascorbate leaf concentration in both salinized and non-salinzed plants. Light storage enhanced fresh weight loss in contrast to dark storage. However the exposure to light reduced leaf nitrate levels by 7% (average of two growth cycles) and contributed to maintain high leaf ascorbate concentrations over time.     

Transgenic fertile soybean plants derived from somatic embryos transformed via the combined DNA-free particle bombardment and <Emphasis Type="Italic">Agrobacterium</Emphasis> system

An Agrobacterium-mediated transformation procedure for soybean [Glycine max L. Merrill] proliferating somatic embryos is here described. The Agrobacterium tumefaciens LBA4404 strain harboring pTOK233, pCAMBIA1390-olp or pH7WG2Dwrky plasmids was used to mediate gene transfer into the plant genome. Prior to Agrobacterium inoculation, proliferative soybean embryogenic clusters were microwounded by DNA-free tungsten particle bombardment. Three independent transformation experiments were performed. In Experiment I, 26 transgenic plants were obtained from a unique clone of cv Bragg, while 580 plants were recovered from 105 clones of cv IAS5. In Experiment II, a single hygromycin-resistant clone of cv BRSMG68 Vencedora was recovered and gave rise to five plants. In Experiment III, 19 plants of cv Bragg and 48 plants of IAS5 were recovered, representing five and 14 independent transformation events, respectively. PCR and Southern analyses confirmed the transgenes’ integration into plant genomes. Transgenic plants were fertile. They flowered, set pods and seeds. Transgene segregation in two T₁ progenies fits the Mendelian pattern (3:1 transgenic:non-transgenic plants). This is the first report of transgenic fertile soybean plants obtained from somatic embryogenic tissues transformed by the system that combines DNA-free particle bombardment and Agrobacterium.     

Solid olive residues: insight into their phenolic composition

Solid olive residues (SOR) are byproducts of the olive-milling process, but they have an increasing importance in the pharmaceutical industry due to their rich content of biophenols. Such compounds are studied widely for their antioxidant and antimicrobial activities, but there is a lack of information about their quantitative recovery. This research highlighted the key role played both by the selection of the cultivar and by the degree of olive fruit ripening on the phenolic content on the SOR. The extraction methods were selected to reach the best quantitative results mainly using a safe food solvent. In light of the results the Soxhlet extraction with ethanol could be proposed as preferential for a higher recovery of verbascoside and its analogues.     

Pre-unfolding resonant oscillations of single green fluorescent protein molecules

Fluorescence spectroscopy of a green fluorescent protein mutant at single-molecule resolution has revealed a remarkable oscillatory behavior that can also be driven by applied fields. We show that immediately before unfolding, several periodic oscillations among the chemical substates of the protein chromophore occur. We also show that applied alternating electric or acoustic fields, when tuned to the protein characteristic frequencies, give rise to strong resonance effects.     

A pith necrosis caused by Xanthomonas perforans on tomato plants

Pith necrosis is a common disease of tomato in Europe, mainly caused by Pseudomonas corrugata and other soil-borne species of Pseudomonas. During 2011–2012 a survey was conducted in soil-grown tomato crops in southeastern Sicily (Italy). Plants showed pith necrosis, brown discolouration of the vascular tissues, leaf chlorosis and sometimes wilting of leaves. Thirty bacterial isolates from symptomatic tissues, forming colonies on NA and KB, were identified by morphological, biochemical and physiological tests. Among them, seven isolates were analyzed for their 16S rDNA and 16S–23S spacer region sequence that resulted in 99 % identity to that of the Xanthomonas perforans type strain (GenBank accession number GQ46173over 2.085 bp.). Additional sequences of fusA, gapA, gltA, gyrB, lacF, and lepA from one selected isolate were 100% identical to sequences of the Xanthomonas perforans type strain. X. perforans local isolates showed similar genomic patterns with REP-PCR and fAFLP, and were clearly distinguished from other Xanthomonas spp. type strains. In stem-inoculation assays, bacteria isolated from symptomatic tomato plants identified as P. fluorescens, P. putida, P. marginalis, P. citronellolis, P. straminea, and Pantoea agglomerans induced discolouration of vascular tissues, while Pectobacterium carotovorum subsp. atrosepticum isolates induced soft rot. Conversely, the isolates here identified as Xanthomonas perforans were able to induce pith necrosis, vascular discolouration, longitudinal splits and external lesions on stems. This report of X. perforans causing pith necrosis on tomato represents a potentially serious problem that may limit the productivity of tomato crops.     

Antimicrobial activity and cytotoxicity of Cirsium tenoreanum

The methanol and ethyl acetate extracts of the aerial parts of Cirsium tenoreanum, have been evaluated for their phytochemical constituents and antibacterial and cytotoxic properties. Three flavonoids, apigenin, quercetin-3-O-galactoside and kaempferol-3-O-ramnoside, were isolated. The ethyl acetate extract showed a good antiproliferative activity.     

Endotoxin Structures in the Psychrophiles Psychromonas marina and Psychrobacter cryohalolentis Contain Distinctive Acyl Features

Lipid A is the essential component of endotoxin (Gram-negative lipopolysaccharide), a potent immunostimulatory compound. As the outer surface of the outer membrane, the details of lipid A structure are crucial not only to bacterial pathogenesis but also to membrane integrity. This work characterizes the structure of lipid A in two psychrophiles, Psychromonas marina and Psychrobacter cryohalolentis, and also two mesophiles to which they are related using MALDI-TOF MS and fatty acid methyl ester (FAME) GC-MS. P. marina lipid A is strikingly similar to that of Escherichia coli in organization and total acyl size, but incorporates an unusual doubly unsaturated tetradecadienoyl acyl residue. P. cryohalolentis also shows structural organization similar to a closely related mesophile, Acinetobacter baumannii, however it has generally shorter acyl constituents and shows many acyl variants differing by single methylene (-CH₂-) units, a characteristic it shares with the one previously reported psychrotolerant lipid A structure. This work is the first detailed structural characterization of lipid A from an obligate psychrophile and the second from a psychrotolerant species. It reveals distinctive structural features of psychrophilic lipid A in comparison to that of related mesophiles which suggest constitutive adaptations to maintain outer membrane fluidity in cold environments.     

Toxigenic Fusarium species and Mycotoxins Associated with Head Blight in Small-Grain Cereals in Europe

The Fusarium species predominantly found associated with Fusarium head blight (FHB) in wheat and other small-grain cereals all over Europe are F. graminearum, F. avenaceum and F. culmorum. Among the less frequently encountered species are several others which are less pathogenic or opportunistic, but also toxigenic. These include F. poae, F. cerealis F. equiseti F. sporotrichioides F. tricinctum and, to a lesser extent, F. acuminatum F. subglutinans F. solani F. oxysporum F. verticillioides F. semitectum and F. proliferatum. The species profile of FHB is due to several factors, primarily climatic conditions, particularly rain and the temperature at flowering stage, but also agronomic factors, such as soil cultivation, nitrogen fertilization, fungicides, crop rotation, and host genotype. The most frequently encountered Fusarium mycotoxins in FHB in Europe has proved to be deoxynivalenol and zearalenone produced by F. graminearum and F. culmorum with the former more common in southern (warmer) and the latter in northern (colder) European areas. Nivalenol was usually found associated with deoxynivalenol and its derivatives (mono-acetyldeoxynivalenols), together with fusarenone-X, formed by F. graminearum F. cerealis F. culmorum and, in northern areas, by F. poae. Moreover, from central to northern European countries, moniliformin has been consistently reported, as a consequence of the widespread distribution of F. avenaceum whereas the occurrence of T-2 toxin derivatives, such as T-2 toxin and HT-2 toxin, and diacetoxyscirpenol have been recorded in conjunction with sporadic epidemics of F. sporotrichioides and F. poae. Finally, beauvericin and various enniatins have recently been found in Finnish wheat colonized by F.avenaceum and F. poae.