Abscisic Acid Sprays Significantly Increase Yield per Plant in Vineyard-Grown Wine Grape (<Emphasis Type="Italic">Vitis vinifera</Emphasis> L.) cv. Cabernet Sauvignon Through Increased Berry Set with No Negative Effects on Anthocyanin Content and Total Polyphenol Index of Both Juice and Wine

In many cultivars of Vitis vinifera periods of mild water stress during ripening are thought to increase grape quality for winemaking, even though yields may be negatively affected. Because abscisic acid (ABA) is involved in the signaling of water stress in plants, we examine the effects of the ABA signal being given without the concomitant water stress. ABA at 250 mg l⁻¹ was sprayed weekly or biweekly from bud-burst until harvest onto the leaves of vineyard-grown plants of cv. Cabernet Sauvignon. For ABA-treated plants berry yield per bunch and per plant was significantly increased (1.5- to 2.0-fold) across three consecutive harvests (2005 through 2007). Number of berries per bunch and per plant was the primary basis for the significant crop increases, although bunches per plant also tended to increase (1.1- to 1.3-fold) across all three harvests. Other parameters assessed included number of internodes, shoot length, leaf area, leaf water potential at midday, photosynthesis, and stomatal conductance. These parameters showed no significant change with ABA treatment, although shoot length tended to be reduced, as was leaf area relative to control plants. The significantly increased fruit yields were thus accomplished without accompanying increases in leaf photosynthesis and leaf areas. Juice at harvest had equal levels of sugars (Brix) and somewhat higher levels of anthocyanins and total polyphenols relative to control values. The two latter trends continued for the resultant wine across two vintage years. In conclusion, three seasons of experimental trials have demonstrated that ABA application can significantly enhance yield per plant in the field-grown grape (cv. Cabernet Sauvignon) by favoring increased berry set without diminishing the quality of the fruit for winemaking use.     

Multiple mechanisms generate HLA class I altered phenotypes in laryngeal carcinomas: high frequency of HLA haplotype loss associated with loss of heterozygosity in chromosome region 6p21

Major histocompatibility complex (MHC) class I loss or downregulation in cancer cells is a major immune escape route used by a large variety of human tumors to evade anti-tumor immune responses mediated by cytotoxic T lymphocytes. Multiple mechanisms are responsible for such HLA class I alterations. However, the precise frequency of these molecular defects has not been clearly determined in tumors derived from specific tissues. To analyze such defects we aim to define the major HLA class I-altered phenotypes in different tumor types. In this paper we report on HLA class I expression in 70 laryngeal carcinomas. We used immunohistological techniques with a highly selective panel of anti-HLA monoclonal antibodies (mAb), and polymerase chain reaction (PCR) microsatellite amplification of previously selected microsatellite markers (STR) located in chromosome 6 and 15. DNA was obtained from microdissected tumor tissues and surrounding stroma to define the loss of heterozygosity (LOH) associated with chromosome 6p21. Our results showed that LOH in chromosome 6 produced HLA haplotype loss (phenotype II) in 36% of the tumors. In addition, HLA class I total loss (phenotype I) was found in 11%; HLA A or B locus downregulation (phenotype III) was detected in 20%; and HLA class I allelic loss (phenotype IV) in 10% of all cases. We sometimes observed two or more associated mechanisms in the same HLA-altered phenotype, such as LOH and HLA total loss in phenotype I. In only 23% of tumors it was not possible to identify any HLA class I alteration. We conclude that the combination of immunohistological techniques and molecular analysis of tumor DNA obtained from microdissected tumor tissues provides a means for the first time of determining the actual frequency of the major HLA class I-altered phenotypes in laryngeal carcinomas.     

Higher accumulation of F1-V fusion recombinant protein in plants after induction of protein body formation

Improving foreign protein accumulation is crucial for enhancing the commercial success of plant-based production systems since product yields have a major influence on process economics. Cereal grain evolved to store large amounts of proteins in tightly organized aggregates. In maize, γ-Zein is the major storage protein synthesized by the rough endoplasmic reticulum (ER) and stored in specialized organelles called protein bodies (PB). Zera^® (γ-Zein ER-accumulating domain) is the N-terminal proline-rich domain of γ-zein that is sufficient to induce the assembly of PB formation. Fusion of the Zera^® domain to proteins of interest results in assembly of dense PB-like, ER-derived organelles, containing high concentration of recombinant protein. Our main goal was to increase recombinant protein accumulation in plants in order to enhance the efficiency of orally-delivered plant-made vaccines. It is well known that oral vaccination requires substantially higher doses than parental formulations. As a part of a project to develop a plant-made plague vaccine, we expressed our model antigen, the Yersinia pestis F1-V antigen fusion protein, with and without a fused Zera^® domain. We demonstrated that Zera^®-F1-V protein accumulation was at least 3× higher than F1-V alone when expressed in three different host plant systems: Ncotiana benthamiana, Medicago sativa (alfalfa) and Nicotiana tabacum NT1 cells. We confirmed the feasibility of using Zera^® technology to induce protein body formation in non-seed tissues. Zera^® expression and accumulation did not affect plant development and growth. These results confirmed the potential exploitation of Zera^® technology to substantially increase the accumulation of value-added proteins in plants.     

Chain length-dependent association of distamycin-type oligopeptides with A·T and G·C pairs in polydeoxynucleotide duplexes

Different binding affinities of various distamycin analogs including the deformylated derivative with poly(dA-dC)·poly(dG-dT) were investigated using CD measurements. The inhibitory effect of distamycins on the DNAase I cleavage activity of DNA duplexes strongly supports the binding data. The base specificity of the ligand interaction with duplex DNA depends on the chain length of distamycin analogs. Netropsin, distamycin-2 and the deformylated distamycin-3 show no binding to dG·dC containing sequences at moderate ionic strength and are classified as highly dA·dT specific. In contrast distamycin having three, four or five methylpyrrolecarboxamide groups also forms more or less stable complexes with dG·dC-containing duplexes. These ligands possess a lower basepair specificity. The correlation between binding behavior and oligopeptide structure shows that presence of the number of hydrogen acceptor and donor sites determines the basepair and sequence specificity. The additional interaction with dG·dC pairs becomes essential when the number of hydrogen acceptor sites exceeds n = 3.     

A new small mammal assemblage from the M013 Terre Rosse fissure filling (Gargano,South-Eastern Italy)

The taxonomic study of the small mammal assemblage from fissure M013, sampled by a team of the University of Torino during the 2005–2009 excavations in the Dell’Erba Quarry (Apricena, Foggia) is presented. The assemblage includes the Echinosoricinae Apulogalerix cf. pusillus and the Crocidosoricinae ?Lartetium cf. dehmi; the Myomiminae species Stertomys simplex and Stertomys lyrifer; a new genus and species of Cricetodontinae and a single specimen of Hattomys cf. nazarii. Muridae include Mikrotia cf. parva, Mikrotia sp. 1 and a new genus and species of Murinae, phylogenetically related to Mikrotia. The occurrence of the new Murinae, the new Cricetodontinae, the two species of glirids and the Crocidosoricinae, as well as the absence of Apodemus and Prolagus, indicates M013 as the oldest Gargano's faunal assemblage known to date, despite the occurrence of Hattomys cf. nazarii, Mikrotia cf. parva and Mikrotia sp. 1, which most probably results from infiltrations from younger fissure fillings. The M013 assemblage is an absolute novelty for the Abruzzo-Apulian Palaeobioprovince, opening new perspectives for the timing and mode of dispersal of the forerunners of the Gargano fauna.     

Glutamate metabolism in cerebral mitochondria after ischemia and post‐ischemic recovery during aging: relationships with brain energy metabolism

Glutamate is involved in cerebral ischemic injury, but its role has not been completely clarified and studies are required to understand how to minimize its detrimental effects, contemporarily boosting the positive ones. In fact, glutamate is not only a neurotransmitter, but primarily a key metabolite for brain bioenergetics. Thus, we investigated the relationships between glutamate and brain energy metabolism in an in vivo model of complete cerebral ischemia of 15 min and during post‐ischemic recovery after 1, 24, 48, 72, and 96 h in 1‐year‐old adult and 2‐year‐old aged rats. The maximum rates (V _max) of glutamate dehydrogenase (GlDH ), glutamate‐oxaloacetate transaminase, and glutamate‐pyruvate transaminase were assayed in somatic mitochondria (FM ) and in intra‐synaptic ‘Light’ mitochondria and intra‐synaptic ‘Heavy’ mitochondria ones purified from cerebral cortex, distinguishing post‐ and pre‐synaptic compartments. During ischemia, none of the enzymes were modified in adult animals. In aged ones, glutamate‐oxaloacetate transaminase was increased in FM and GlDH in intra‐synaptic ‘Heavy’ mitochondria, stimulating glutamate catabolism. During post‐ischemic recovery, FM did not show modifications at both ages while, in intra‐synaptic mitochondria of adult animals, glutamate catabolism was increased after 1 h of recirculation and decreased after 48 and 72 h, whereas it remained decreased up to 96 h in aged rats. These results, with those previously published about Krebs’ cycle and Electron Transport Chain (Villa et al ., [2013] Neurochem. Int . 63, 765–781), demonstrate that: (i) V _max of energy‐linked enzymes are different in the various cerebral mitochondria, which (ii) respond differently to ischemia and post‐ischemic recovery, also (iii) with respect to aging.