Vibi A

Initiation of respiratory support in the delivery room increases the risk and severity of brain injury in preterm neonates through two major pathways: an inflammatory pathway and a haemodynamic pathway. The relative contribution of each pathway on preterm brain injury is not known. We aimed to assess the role of the inflammatory and haemodynamic pathway on ventilation-induced brain injury (VIBI) in the preterm lamb. Fetal lambs (125 ± 1 day gestation) were exteriorised, instrumented and ventilated with a high tidal-volume (VT) injurious strategy for 15 min either with placental circulation intact to induce the inflammatory pathway only (INJINF; n = 7) or umbilical cord occluded to induce both the inflammatory and haemodynamic pathways (INJINF+HAE; n = 7). Sham controls were exteriorised but not ventilated (SHAM; n = 5) while unoperated controls (UNOP; n = 7) did not undergo fetal instrumentation. Fetuses were returned in utero following intervention and the ewe allowed to recover. Arterial blood gases and plasma were sampled periodically. Twenty-four hours following intervention, lambs were delivered and maintained on non-injurious ventilation for ~40 min then brains were collected post-mortem for immunohistochemistry and RT-qPCR to assess inflammation, vascular pathology and cell death within white matter regions. Compared to INJINF lambs, INJINF+HAE lambs achieved a consistently higher VT during injurious ventilation and carotid blood flow was significantly lower than baseline by the end of ventilation. Throughout the 24 h recovery period, systemic arterial IL-6 levels of INJINF+HAE lambs were significantly higher than SHAM while there was no difference between INJINF and SHAM animals. At 24 h, mRNA expression levels of pro-inflammatory cytokines, tight junction proteins, markers of cell death, and histological injury indices of gliosis, blood vessel protein extravasation, oligodendrocyte injury and cell death were not different between groups. Injurious ventilation, irrespective of strategy, did not increase brain inflammation or injury 24 h later when compared to control animals. However, the haemodynamic pathway did influence carotid blood flow adaptations during injurious ventilation and increased systemic arterial IL-6 that may underlie long-term pathology. Future studies are required to further characterise the pathways and their long-term effects on VIBI.

Recent approaches to wetland assessment have advocated a multilevel approach which incorporates assessments based on landscape (remote sensing) data, on-site but "rapid" methods, and intensive methods where quantitative data is collected. Brown and Vivas (2004) recently pro- posed an assessment method that uses remote sensing information (Landscape Development Index or LDI) and propose that it may also be usable as a quantified human disturbance gradient. The LDI was evaluated using a large reference wetland data set from Ohio using land use percentages within a 1 km radius circle of the wetlands. The LDI had interpretable and significant relationships with another human disturbance gradient (the Ohio Rapid Assessment Method for Wetlands or ORAM) and with most metrics and scores from the Vegetation Index of Biotic Integrity (VIBI) developed for use in the State of Ohio. Metrics from emergent wetlands had the most significant correlations with the LDI (10 of 10 metrics), followed by forested wetlands (8 of 10 metrics) and shrub wetlands (4 of 10). Poor correlation for VIBI scores and metrics of shrub wetlands was due to differences in attainable LDI scores based on ecoregion and natural buffers shielding the wetland from otherwise intensive land uses. The ORAM and VIBI were developed for use in wetlands in Ohio completely independent of the LDI. It is an important test of the LDI concept that so many interpretable and significant relationships occurred between the VIBI scores, VIBI metric values, and the ORAM scores. For the purposes of VIBI development, the LDI is an independent, quantified disturbance gradient that has provided an additional test of the VIBI. Given its theoretical underpinnings and the fact that it uses quantified land use percentages, the LDI has many advantages over more qualitative human disturbance gradients. Using land use percentages from increasingly smaller distances from the wetland edge (100-200 m) may improve the resolution of the LDI to detect on-site disturbances to a wetland which degrade its ecological condition. The LDI should be evaluated with other large reference data sets in other regions to evaluate its validity and usefulness as an assessment tool.

The cyclotides are currently the largest known family of head-to-tail cyclic proteins. The complex structure of these small plant proteins, which consist of approximately 30 amino acid residues, contains both a circular peptide backbone and a cystine knot, the combination of which produces the cyclic cystine knot motif. To date, cyclotides have been found in plants from the Rubiaceae, Violaceace and Cucurbitaceae families, and are believed to be part of the host defence system. In addition to their insecticidal effect, cyclotides have also been shown to be cytotoxic, anti-HIV, antimicrobial and haemolytic agents. In this study, we show that the alpine violet Viola biflora (Violaceae) is a rich source of cyclotides. The sequences of 11 cyclotides, vibi A-K, were determined by isolation and MS/MS sequencing of proteins and screening of a cDNA library of V. biflora in parallel. For the cDNA screening, a degenerate primer against a conserved (AAFALPA) motif in the cyclotide precursor ER signal sequence yielded a series of predicted cyclotide sequences that were correlated to those of the isolated proteins. There was an apparent discrepancy between the results of the two strategies as only one of the isolated proteins could be identified as a cDNA clone. Finally, to correlate amino acid sequence to cytotoxic potency, vibi D, E, G and H were analysed using a fluorometric microculture cytotoxicity assay using a lymphoma cell line. The IC(50)-values of the bracelet cyclotides vibi E, G and H ranged between 0.96 and 5.0 microM while the Möbius cyclotide vibi D was not cytotoxic at 30 microM.