Integration of TGF-beta and Ras/MAPK signaling through p53 phosphorylation

During development and tissue homeostasis, cells must integrate different signals. We investigated how cell behavior is controlled by the combined activity of transforming growth factor-beta (TGF-beta) and receptor tyrosine kinase (RTK) signaling, whose integration mechanism is unknown. We find that RTK/Ras/MAPK (mitogen-activated protein kinase) activity induces p53 N-terminal phosphorylation, enabling the interaction of p53 with the TGF-beta-activated Smads. This mechanism confines mesoderm specification in Xenopus embryos and promotes TGF-beta cytostasis in human cells. These data indicate a mechanism to allow extracellular cues to specify the TGF-beta gene-expression program.     

Splice variants of the alpha subunit of the G protein Gs activate both adenylyl cyclase and calcium channels

Signal transducing guanine nucleotide binding (G) proteins are heterotrimers with different alpha subunits that confer specificity for interactions with receptors and effectors. Eight to ten such G proteins couple a large number of receptors for hormones and neurotransmitters to at least eight different effectors. Although one G protein can interact with several receptors, a given G protein was thought to interact with but one effector. The recent finding that voltage-gated calcium channels are stimulated by purified Gs, which stimulates adenylyl cyclase, challenged this concept. However, purified Gs may have four distinct alpha-subunit polypeptides, produced by alternative splicing of messenger RNA. By using recombinant DNA techniques, three of the splice variants were synthesized in Escherichia coli and each variant was shown to stimulate both adenylyl cyclase and calcium channels. Thus, a single G protein alpha subunit may regulate more than one effector function.     

Efficacy of a single milbemycin oxime administration in combination with praziquantel against experimentally induced heartworm (Dirofilaria immitis) infection in cats

The efficacy of a combination of milbemycin oxime and praziquantel in preventing the establishment of experimentally induced heartworm (Dirofilaria immitis) infection was investigated in a study involving 24 young domestic short-hair cats. The animals were inoculated with 50 infective larvae on day 0. Subsequently they were divided into two groups of 12 animals each. The animals in group 1 were treated once with medicated tablets containing 4 mg milbemycin (minimum dose 2 mg/kg body weight) and 10 mg praziquantel (MILBEMAX) on day 30 after infection. Cats in group 2 received placebo tablets on the same day. On day 183 post-infection a blood sample was taken from each animal before euthanasia and necropsy. The blood samples were tested for the presence of microfilariae and the necropsied animals were examined for the presence of adult worms. Microfilariae were not found in any of the investigated cats. No heartworms were found in the animals in group 1 (treated with medicated tablets). Out of the 12 placebo-treated cats 1 was heartworm-free, whereas all the others were found to be infected with 1-3 adult heartworms.     

Morphological evaluation of olive germplasm present in Tuscany region

Undesirable characteristic of rapeseed oil is a relatively high level of linolenic acid (18:3), which is easily oxidized leading to rancidity and a shortened shelf life of the oil. Previous attempts to reduce linolenic acid levels in rapeseed oil through breeding have been impaired by complex genetics and strong environmental sensitivity of this trait. Therefore, our objective was to develop molecular markers for low linolenic acid that could facilitate the breeding of low linolenic rapeseed. Bulked segregant analysis was employed to identify two RAPD markers associated with 18:3 in a doubled haploid population segregating for linolenic and erucic acid levels. Based on analysis of individual DH lines, the markers RM350 and RM574, representing two independent loci, accounted for a total of 39% of the genetic variability in this population. This marker RM350 alone accounted for 25% genetic variation for this trait with no evidence of recombination. Significant interlocus interaction found between the markers RM350 and RM574 suggested that epistasis was involved in the genetic control of 18:3 level in this population. Another marker designated as RM322, which was independent of the other two, was found significantly associated with the erucic acid level and oil content. RAPD markers identified in this study should be a useful tool for the early detection of low linolenic, or low or high erucic acid genotypes in rapeseed breeding programs based on doubled haploids. This revised version was published online in July 2006 with corrections to the Cover Date.     

Composition and Utilization of Barley Pearling By‐Products for Making Functional Pastas Rich in Dietary Fiber and β‐Glucans

Pearling by‐products and the pearled products of two commercial stocks of hulled barley, pearled according to an industrial process consisting of five consecutive pearling steps, were analyzed for β‐glucans, dietary fiber (total, soluble, and insoluble), protein, lipid, ash, and digestible carbohydrate. The data showed that the pearling flour fractions, abraded in the fourth and fifth hullers, contained interesting amounts of β‐glucans (3.9–5.1% db) from a nutritional point of view. These fractions were subsequently enriched in β‐glucans using a milling‐sieving process to double β‐glucan content (9.1–10.5% db). Functional pastas, enriched with β‐glucans and dietary fiber, were produced by substituting 50% of standard durum wheat semolina with β‐glucan‐enriched barley flour fractions. Although darker than durum wheat pasta, these pastas had good cooking qualities with regard to stickiness, bulkiness, firmness, and total organic matter released in rinsing water. The dietary fiber (13.1–16.1% wb) and β‐glucan (4.3–5.0% wb) contents in the barley pastas were much higher than in the control (4.0 and 0.3% wb, respectively). These values amply meet the FDA requirements of 5 g of dietary fiber and 0.75 g of β‐glucans per serving (56 g in the United States and 80 g in Italy). At present, the FDA has authorized the health claim “may reduce the risk of heart disease” for food containing β‐glucans from oat and psyllium only.     

Clarifying the identity of the main ellagitannin in the fruit of the strawberry, Fragaria vesca and Fragaria ananassa Duch

Although the composition of strawberry fruit has been extensively studied, especially for the most abundant phenolic compounds, agrimoniin has never been univocally identified as one of the most abundant phenolic compounds in the fruit. In this study agrimoniin was isolated in the fruit of Fragaria vesca and its structure characterized. Furthermore, its presence was definitively established to be the main ellagitannin in both F. vesca and Fragaria ananassa D. fruit. The presence of sanguiin H-6 and lambertianin C as minor compounds was confirmed in both F. vesca and F. ananassa D. samples. For the first time here is reported the full NMR assignments for agrimoniin. These data should represent a point of reference for NMR analysis of this and other structurally related ellagitannins. Finally, the establishment of an HPLC protocol for separation provided information making it possible to avoid confusion with sanguiin H-6, the main ellagitannin in Rubus species, which is also present in strawberries but at a much lower concentration.     

Complexing the Marine Sesquiterpene Euplotin C by Means of Cyclodextrin-Based Nanosponges: A Preliminary Investigation

Euplotin C is a sesquiterpene of marine origin endowed with significant anti-microbial and anti-tumor properties. Despite the promising functional profile, its progress as a novel drug candidate has failed so far, due to its scarce solubility and poor stability in aqueous media, such as biological fluids. Therefore, overcoming these limits is an intriguing challenge for the scientific community. In this work, we synthesized β-cyclodextrin-based nanosponges and investigated their use as colloidal carriers for stably complex euplotin C. Results obtained proved the ability of the carrier to include the natural compound, showing remarkable values of both loading efficiency and capacity. Moreover, it also allowed us to preserve the chemical structure of the loaded compound, which was recovered unaltered once extracted from the complex. Therefore, the use of β-cyclodextrin-based nanosponges represents a viable option to vehiculate euplotin C, thus opening up its possible use as pharmacologically active compound.     

Contribution of the xenograft bone plate-screw system in lumbar transpedicular stabilization of dogs: an in-vitro study

We performed biomechanical comparison of a xenograft bone plate-screw (XBPS) system for achieving cadaveric lumbar transpedicular stabilization (TS) in dogs. Twenty dogs'' cadaveric L_2-4 lumbar specimens were harvested and their muscles were removed, but the discs and ligaments were left intact. These specimens were separated to four groups: the L_2-4 intact group as control (group I, n = 5), the L₃ laminectomy and bilateral facetectomy group (LBF) (group II, n = 5), the LBF plus TS with metal plate-screw group (group III, n = 5) and the LBF plus TS with XBPS group (group IV, n = 5). Five kinds of biomechanical tests were applied to the specimens: flexion, extension, left-right bending and rotation. The averages of the 16 stiffness values were calculated and then these were statistically analyzed. The statistical results show that the XBPS system contributes spinal stability and this system can be a good choice for achieving TS.     

Glue barriers reduce earwig damage on apricots in north-western Italy

The European earwig, Forficula auricularia L. (Dermaptera: Forficulidae), is a well-known species that is cosmopolitan and present throughout Europe. Due to its omnivorous feeding behaviour, this species can act as a generalist predator, preying on several top fruit pests, but also as a pest causing shallow gouges or holes in stone and soft fruits such as apricots, strawberries, raspberries or blackberries. In Piedmont (NW Italy), significant fruit damage has been observed lately in apricot orchards where earwigs fed on ripening fruits and made a considerable part of the produce unmarketable. In this study, we sampled earwig populations in three apricot orchards in Piedmont and tested the effectiveness of glue barriers applied to the tree trunks in reducing both earwig density in the canopy and fruit damage. The arboreal glues Rampastop® and Vebicolla® were tested both in the field and laboratory trials. Glue barriers demonstrated to be effective control measures, significantly reducing earwig abundance in the canopy and fruit damage. Rampastop® gave better results on old trees with a very rough and cracked bark, since in that case Vebicolla® could not perfectly bond with the trunk.