Interspecific hybridization and inoculation with Pantoea eucalypti improve forage performance of Lotus crop species under alkaline stress.

This study was designed to elucidate the physiological responses of three Lotus forage accessions to alkaline stress, and the influence of inoculating with Pantoea eucalypti endophyte strain on alkaline stress mitigation. A diploid L. corniculatus (Lc) accession, L. tenuis (Lt), and the interspecific hybrid Lt × Lc obtained from these two parental lines were exposed to alkaline stress (pH 8.2). Both Lt and the Lt × Lc hybrid are alkaline-tolerant compared to Lc, based on observations that dry mass was not reduced under stress, and there were no chlorosis symptoms on leaf blades. In all three Lotus accessions, Fe2+ concentration under stress decreased in aerial parts and simultaneously increased in roots. Inoculation with P. eucalypti considerably increased Fe2+ content in shoots of all three Lotus forage species under alkaline treatment. Photochemical efficiency of PSII was affected in Lc accession only when exposed to alkaline treatment. However, when cultivated under alkalinity with inoculation, plants recovered and had photosynthetic parameters equivalent to those in the control treatment. Together, the results highlight the importance of inoculation with P. eucalypti, which contributes significantly to mitigating alkaline stress. All results provide useful information for improving alkaline tolerance traits of Lotus forage species and their interspecific hybrids.

Contrasting response of two Lotus corniculatus L. accessions to combined waterlogging-saline stress.

Waterlogging and salinity impair crop growth and productivity worldwide, with their combined effects being larger than the additive effects of the two stresses separately. Here, a common forage tetraploid Lotus corniculatus (cv. San Gabriel) and a diploid L. corniculatus accession, collected from a coastal area with high frequency of waterlogging-saline stress events, were evaluated for tolerance to waterlogging, salinity and these two stresses combined. We hypothesize that, due to its environmental niche, the diploid accession would show better adaptation to combined waterlogging-saline stress compared to the tetraploid L. corniculatus. Plants were evaluated under control conditions, waterlogging, salinity and a combined waterlogging-saline treatment for 33 days. Shoot and root growth were assessed, together with chlorophyll fluorescence and gas exchange measurements. Results showed that salinity and waterlogging effects were more severe for the tetraploid accession, with a larger effect being observed under the combined stress condition. Concentrations of Na+ , Cl- and K+ were measured in apical and basal leaves, and in roots. A larger accumulation of Na+ and Cl- was observed under both saline and combined stress treatments for the tetraploid L. corniculatus, for which ion toxicity effects were evident. The expression of CLC gene, coding for a Cl- transporter, was only increased in diploid L. corniculatus plants in response to the combined stress condition, suggesting that ion compartmentalization mechanisms were induced in this accession. Thus, this recently characterized L. corniculatus could be used for the introduction of new tolerance traits in other Lotus species used as forage.

Parent-Infant Interaction during the First Year of Life in Infants at High Risk for Cerebral Palsy: A Systematic Review of the Literature.

Introduction: Perinatal adverse events put neonates at high risk for short and long-term disabilities, including cerebral palsy (CP). The most recent guidelines about early intervention in infants with brain damage have emphasized the importance of family involvement from the very first phases of development. Early parent-infant interactions are pivotal in promoting infant cognitive and social developmental trajectories. However, little is known about the extent to which severe adverse perinatal events can affect the quality of early parent-infant interactions. Patients and Methods: We systematically searched five databases (PubMed, PsycINFO, EMBASE, CINAHL, and Cochrane Library) for the publications assessing parent-infant interactions in infants at high neurological risk within 1 year of age. Articles were selected if they involved direct comparison between high-risk populations and healthy controls or low-risk populations, and if quantitative or semiquantitative tools were used to assess the parent-infant interaction. Measures of parent-infant interaction included infant interactive behaviors, parental interactive behaviors, and dyadic interactive patterns. Results: The search yielded 18 publications that met the inclusion criteria. The articles represent a high level of heterogeneity in terms of infant neurological risk, infant age, and tools assessing interactive behaviors. Both infant and maternal behaviors within the investigated interactive exchanges were reported to be compromised, leading to subsequent overall impairment of the dyadic patterns. Conclusion: While the studies reviewed here provide general and important information, the review did not yield a clear picture of early dyadic interactions in high-risk infant populations. Further observational studies are warranted in order to provide a more accurate knowledge of the early dyadic exchanges between infants at high neurological risk and their parents, as they might provide a critical opportunity for early family centered habilitative interventions.

Tritium and radiocarbon levels in the Rhône river delta and along the French Mediterranean coastline.

The Rhône is characterised by a heavy concentration of nuclear-based industries including nuclear power stations and nuclear sites housing civilian and military facilities. Here, we report the results of a four-year survey (2010-2013) of tritium and radiocarbon levels in a variety of matrices within the Rhône delta and along the French Mediterranean coastline. The aim of the study is to create a spatial reference framework of environmental levels of these two radionuclides, which are the most prevalent in radioactive effluents from nuclear power stations. Although both tritium and radiocarbon levels in the samples analysed are very low and can only be detected using ultra-sensitive analytical techniques, they clearly show the influence of the tritium and radiocarbon discharges carried by the Rhône plume along the Mediterranean coast. The tritium content of suspended matter and sediments of the Rhône is a special case, which shows elevated tritium values not seen in other French rivers with similar nuclear facilities. The north-south spatial distribution of this tritium anomaly shows that these trace values are at their highest in the upper Rhône, close to the Swiss border and upstream of Creys Malville, the northernmost nuclear power station on the Rhône. This points to a legacy of past tritium releases by the watchmaking industry. A dedicated study would be needed to clearly identify the source and the exact nature of this contamination.

Evaluation of mean echogenicity of tendons and ligaments of the metacarpal region in neonatal foals: A preliminary study.

The aims of this research were to evaluate mean echogenicity (ME) of the deep and superficial digital flexor tendons (DDFT and SDFT), the interosseous muscle (IM), and the accessory ligament of the deep digital flexor tendon (ALDDFT) of the metacarpal region in neonatal foals, and determine the effect of sex, side and body weight on this quantitative ultrasonographic evaluation. Thirteen orthopedically sound neonatal foals were examined. Four areas of study (1A, 1B, 2A, 2B) were identified. Transverse scans of the DDFT, SDFT, IM and ALDDFT were obtained, recorded, and analyzed. The most echogenic structures were the ALDDFT and DDFT, while the SDFT was significantly less echogenic than all other structures (P<0.05). No influence of sex, forelimb, or body weight was observed. The echogenicity of the tenodesmic structures of foals partially overlapped that reported in the metacarpal region in adult horses, except for IM.

Gross placental morphology and foal serum biochemistry as predictors of foal health.

The aim of this study was to verify if changes in blood glucose, creatinine, urea, and fibrinogen concentrations evaluated at birth reflect gross placenta abnormalities, and are useful to identify foals that suffered from placental dysfunction. A total of 92 mares were included in the present study: 68 delivered healthy foals and they were included in group 1; 24 delivered sick foals and they were included in group 2. In group 2, foals' clinical diagnoses included perinatal asphyxia syndrome (PAS; n = 20) and prematurity and/or dysmaturity (n = 4). The proportion of sick foals was greater when placental abnormalities were observed (χ(2) [1, n = 89] = 5.00; P = 0.025). Serum creatinine concentration at birth was higher in sick than in healthy foals (P = 0.003), and blood glucose concentrations at birth was smaller in sick than in healthy foals (P = 0.007). No difference was found in blood chemistry results between survivors and nonsurvivors of group 2. Serum creatinine concentration was higher in foals born from grossly abnormal than in foals born from grossly normal placenta (P = 0.029), and it was higher in foals affected by PAS (311.17 µmol/L) than in healthy foals (238.24 µmol/L) (P = 0.004). In a clinical setting, serum creatinine and blood glucose concentrations should be evaluated at birth, particularly in foals born from grossly abnormal placenta. The association of clinical and laboratory data could be particularly important to promptly identify and treat foals with a higher risk to develop PAS.

[Effectiveness of a new cervicometer in evaluating the risk of preterm delivery]. / Validità di un nuovo cervicometro per la valutazione del rischio di parto pretermine.

AIM: The ultrasound measurement of the cervix is the gold standard for assessing the risk of preterm delivery. We compared the accuracy of this technique with measurements obtained by the cervicometer Cervilenz®. The purpose was to show that this device can be used as a screening tool, low cost, in the diagnosis of preterm labor. METHODS: Fifty patients were included in the study at gestational age between 21+0 and 26 +0 or weeks. The study was blinded between the two performers utilizing the cervicometer and the transvaginal ultrasound. RESULTS: The study showed 100% concordance and a correlation between the two techniques of 0.94. CONCLUSION: The cervicometer proved to be a reliable and easy to use device to detect a normal or short cervix, with contained healthcare costs.

Effects of mesenchymal stem cells isolated from amniotic fluid and platelet-rich plasma gel on severe decubitus ulcers in a septic neonatal foal.

This paper documents the treatment of severe decubitus ulcers with amniotic fluid mesenchymal stem cells and platelets rich plasma (PRP) gel in a septic neonatal foal. The colt needed 25 days of hospitalization: during this period ulcers were treated for 15 days with mesenchymal stem cells (MSCs) plus PRP, PRP gel alone, or aloe gel. Healing was faster using MSCs+PRP, and at 7 months an ulcer treated with aloe gel was still not completely healed.

Autocorrelation of the polarization-dependent loss in fiber routes.

We study the spectral properties of polarization-dependent loss (PDL) in fiber routes with multiple spans and amplifiers. In these systems, PDL stems from a small number of lumped WDM components interconnected by PDL-free fibers, a configuration that is poorly described by the popular model of distributed PDL. We derive transparent and practical analytical expressions for the autocorrelation functions of the PDL vector and of the square PDL magnitude and show that they are strongly affected by the order of the individual PDL elements.

Guiding principles in choosing a therapeutic table for DCI hyperbaric therapy.

Hyperbaric therapy is the basis of treatment for pervasive development disorders. For this reason, the choice of the right therapeutic table for each case is critical. Above all, the delay in recompression time with respect to the first symptoms and to the severity of the case must be considered. In our experience, the use of low-pressure oxygen tables resolves almost all cases if recompression takes place within a short time. When recompression is possible almost immediately, the mechanical effect of reduction on bubble volume due to pressure is of remarkable importance. In these cases, high-pressure tables can be considered. These tables can also be used in severe spinal-cord decompression sickness. The preferred breathing mixture is still disputed. Heliox seems to be favored because it causes fewer problems during the recompression of divers, and above all, because nitrox can cause narcosis and contributes nitrogen. Saturation treatment should be avoided or at least used only in special cases. In cases of arterial gas embolism cerebral injury, it is recommended to start with an initial 6 ATA recompression only if the time between symptom onset and the beginning of recompression is less than a few hours.

GFAP mutations and polymorphisms in 13 unrelated Italian patients affected by Alexander disease.

Alexander disease (AD), a rare neurodegenerative disorder of the central nervous system, is characterized by the accumulation of cytoplasmic protein aggregates (Rosenthal fibers) composed of glial fibrillary acidic protein (GFAP) and small heat-shock proteins within astrocytes. To date, more than 40 different GFAP mutations have been reported in AD. The present study is aimed at the molecular diagnosis of Italian patients suspected to be affected by AD. By analyzing the GFAP gene of 13 unrelated patients (eight with infantile form, two with juvenile form and three with adult form), we found 11 different alleles, including four new ones. Among the novel mutations, three (p.R70Q, p.R73K, and p.R79P) were identified in exon 1 and p.L359P in exon 6. The sequence analysis also detected six different single nucleotide polymorphic variants, including two previously unreported ones, spread throughout non-coding regions (introns 2, 3, 5, 6, and 3'UTR) of the gene. All patients were heterozygous for the mutations, thus confirming their dominant effect.

Radiological consequences of the extreme flooding on the lower course of the Rhone valley (December 2003, south east France).

In early December 2003 unusual weather conditions led to major flooding of the lower Rhone valley. When it floods, the Rhone carries large masses of solid matter in suspension, which potentially includes associated artificial (anthropogenic) radioactive contaminants from soil drainage in the catchment area and from re-uptake of sedimentary matter that has been contaminated with low-level radioactive liquid effluents from almost twenty nuclear facilities situated along the Rhone valley. A sampling campaign was carried out to investigate the level and spread of both sediment mass and associated radioactive contamination across the flooded areas. An attempt was made to assess the radiological consequences of such an extreme event on contamination of the food chain. Our results show that almost 700,000 tons of sediment was transported onto the floodplain, of which 80% were coarse and fine sands. These materials transferred 6660 MBq of 137Cs, 93 MBq of (239+240)Pu, 13 MBq of 238Pu and 204 MBq of 60Co over a surface area of 60 km2. More than 90% of deposited sediments are concentrated in a 10 km2 area of agricultural soils, and we estimated that 18% were plowed into the soil. Nevertheless, the level of activity measured in the vegetable crops and milk was not significantly different from the level measured in similar samples from regions that were not affected by the December 2003 floods.

Comparative evolution history of SINEs in Arabidopsis thaliana and Brassica oleracea: evidence for a high rate of SINE loss.

Brassica oleracea and Arabidopsis thaliana belong to the Brassicaceae(Cruciferae) family and diverged 16 to 19 million years ago. Although the genome size of B. oleracea (approximately 600 million base pairs) is more than four times that of A. thaliana (approximately 130 million base pairs), their gene content is believed to be very similar with more than 85% sequence identity in the coding region. Therefore, this important difference in genome size is likely to reflect a different rate of non-coding DNA accumulation. Transposable elements (TEs) constitute a major fraction of non-coding DNA in plant species. A different rate in TE accumulation between two closely related species can result in significant genome size variations in a short evolutionary period. Short interspersed elements (SINEs) are non-autonomous retroposons that have invaded the genome of most eukaryote species. Several SINE families are present in B. oleracea and A. thaliana and we found that two of them (called RathE1 and RathE2) are present in both species. In this study, the tempo of evolution of RathE1 and RathE2 SINE families in both species was compared. We observed that most B. oleracea RathE2 SINEs are "young" (close to the consensus sequence) and abundant while elements from this family are more degenerated and much less abundant in A. thaliana. However, the situation is different for the RathE1 SINE family for which the youngest elements are found in A. thaliana. Surprisingly, no SINE was found to occupy the same (orthologous) genomic locus in both species suggesting that either these SINE families were not amplified at a significant rate in the common ancestor of the two species or that older elements were lost and only the recent (lineage-specific) insertions remain. To test this latter hypothesis, loci containing a recently inserted SINE in the A. thaliana col-0 ecotype were selected and characterized in several other A. thaliana ecotypes. In addition to the expected SINE containing allele and the pre-integrative allele (i.e. the "empty" allele), we observed in the different ecotypes, alleles with truncated portions of the SINE (up to the complete loss of the element) and of the immediate genomic flanking sequences. The absence of SINEs in orthologous positions between B. oleracea and A. thaliana and the presence in recently diverged A. thaliana ecotypes of alleles containing severely truncated SINEs suggest a very high rate of SINE loss in these species.

Processing of 20S pre-rRNA to 18S ribosomal RNA in yeast requires Rrp10p, an essential non-ribosomal cytoplasmic protein.

Numerous non-ribosomal trans-acting factors involved in pre-ribosomal RNA processing have been characterized, but none of them is specifically required for the last cytoplasmic steps of 18S rRNA maturation. Here we demonstrate that Rio1p/Rrp10p is such a factor. Previous studies showed that the RIO1 gene is essential for cell viability and conserved from archaebacteria to man. We isolated a RIO1 mutant in a screen for mutations synthetically lethal with a mutant allele of GAR1, an essential gene required for 18S rRNA production and rRNA pseudouridylation. We show that RIO1 encodes a cytoplasmic non-ribosomal protein, and that depletion of Rio1p blocks 18S rRNA production leading to 20S pre-rRNA accumulation. In situ hybridization reveals that, in Rio1p depleted cells, 20S pre-rRNA localizes in the cytoplasm, demonstrating that its accumulation is not due to an export defect. This strongly suggests that Rio1p is involved in the cytoplasmic cleavage of 20S pre-rRNA at site D, producing mature 18S rRNA. Thus, Rio1p has been renamed Rrp10p (ribosomal RNA processing #10). Rio1p/Rrp10p is the first non-ribosomal factor characterized specifically required for 20S pre-rRNA processing.

Identification of a regulated pathway for nuclear pre-mRNA turnover.

We have identified a nuclear pathway that rapidly degrades unspliced pre-mRNAs in yeast. This involves 3'-->5' degradation by the exosome complex and 5'-->3' degradation by the exonuclease Rat1p. 3'-->5' degradation is normally the major pathway and is regulated in response to carbon source. Inhibition of pre-mRNA degradation resulted in increased levels of pre-mRNAs and spliced mRNAs. When splicing was inhibited by mutation of a splicing factor, inhibition of turnover resulted in 20- to 50-fold accumulation of pre-mRNAs, accompanied by increased mRNA production. Splicing of a reporter construct with a 3' splice site mutation was also increased on inhibition of turnover, showing competition between degradation and splicing. We propose that nuclear pre-mRNA turnover represents a novel step in the regulation of gene expression.

Rrp8p is a yeast nucleolar protein functionally linked to Gar1p and involved in pre-rRNA cleavage at site A2.

Chemical modifications and processing of the 18S, 5.8S, and 25S ribosomal RNAs from the 35S pre-ribosomal RNA depend on an important set of small nucleolar ribonucleoprotein particles (snoRNPs). Genetic depletion of yeast Gar1p, an essential common component of H/ACA snoRNPs, leads to inhibition of uridine isomerizations to pseudo-uridines on the 35S pre-rRNA and of the early pre-rRNA cleavages at sites A1 and A2, resulting in a loss of mature 18S rRNA synthesis. To identify Gar1p functional partners, we screened for mutations that are synthetically lethal with a gar1 mutant allele encoding a Gar1p mutant protein lacking its two glycine/arginine-rich (GAR) domains. We identified a previously uncharacterized Saccharomyces cerevisiae open reading frame, YDR083W (now designated RRP8), that encodes a highly conserved protein containing motifs found in methyltransferases. Rrp8p localizes to the nucleolus. A yeast strain lacking this protein is viable at 30 degrees C but displays strong growth impairment at lower temperatures. In this strain, cleavage of the pre-rRNA at site A2 is strongly affected whereas cleavages at sites A0 and A1 are only slightly inhibited or delayed.

Nhp2p and Nop10p are essential for the function of H/ACA snoRNPs.

The small nucleolar ribonucleoprotein particles containing H/ACA-type snoRNAs (H/ACA snoRNPs) are crucial trans-acting factors intervening in eukaryotic ribosome biogenesis. Most of these particles generate the site-specific pseudouridylation of rRNAs while a subset are required for 18S rRNA synthesis. To understand in detail how these particles carry out these functions, all of their protein components have to be characterized. For that purpose, we have affinity-purified complexes containing epitope-tagged Gar1p protein, previously shown to be part of H/ACA snoRNPs. Under the conditions used, three polypeptides of 65, 22 and 10 kDa apparent molecular weight specifically copurify with epitope-tagged Gar1p. The 22 and 10 kDa polypeptides were identified as Nhp2p and a novel protein we termed Nop10p, respectively. Both proteins are conserved, essential and present in the dense fibrillar component of the nucleolus. Nhp2p and Nop10p are specifically associated with all H/ACA snoRNAs and are essential to the function of H/ACA snoRNPs. Cells lacking Nhp2p or Nop10p are impaired in global rRNA pseudouridylation and in the A1 and A2 cleavage steps of the pre-rRNA required for the synthesis of mature 18S rRNA. These phenotypes are probably a direct consequence of the instability of H/ACA snoRNAs and Gar1p observed in cells deprived of Nhp2p or Nop10p. Our results suggest that Nhp2p and Nop10p, together with Cbf5p, constitute the core of H/ACA snoRNPs.

The box H + ACA snoRNAs carry Cbf5p, the putative rRNA pseudouridine synthase.

Many or all of the sites of pseudouridine (Psi) formation in eukaryotic rRNA are selected by site-specific base-pairing with members of the box H + ACA class of small nucleolar RNAs (snoRNAs). Database searches previously identified strong homology between the rat nucleolar protein Nap57p, its yeast homolog Cbf5p, and the Escherichia coli Psi synthase truB/P35. We therefore tested whether Cbf5p is required for synthesis of Psi in the yeast rRNA. After genetic depletion of Cbf5p, formation of Psi in the pre-rRNA is dramatically inhibited, resulting in accumulation of the unmodified rRNA. Protein A-tagged Cbf5p coprecipitates all tested members of the box H + ACA snoRNAs but not box C + D snoRNAs or other RNA species. Genetic depletion of Cbf5p leads to depletion of all box H + ACA snoRNAs. These include snR30, which is required for pre-rRNA processing. Depletion of Cbf5p also results in a pre-rRNA processing defect similar to that seen on depletion of snR30. We conclude that Cbf5p is likely to be the rRNA Psi synthase and is an integral component of the box H + ACA class of snoRNPs, which function to target the enzyme to its site of action.