In a previous study from this institution, patients at high risk for preterm labour were screened for the presence of bacterial vaginosis (BV). When BV was present, they were randomised to receive either treatment (metronidazole) or placebo (vitamin C). There were significantly more patients with preterm labour in the metronidazole group. The aim of this double-blind randomised placebo-controlled trial study was to determine whether vitamin C could indeed reduce the recurrence risk of preterm labour. Patients with a history of preterm labour in a preceding pregnancy were randomised to receive 250 mg vitamin C or a matching placebo twice daily until 34 weeks' gestation. They attended a dedicated premature labour clinic. Significantly more women delivered before term in the group that received vitamin C, but there was no difference in the outcome of the babies between the two groups. Supplementation with vitamin C did not prevent premature labour.

Cell suspension cultures of Vitis vinifera cv. Dauphine berries were used to study the response to the vascular pathogen, Eutypa lata, in comparison with a biological control agent, Trichoderma atroviride, that was previously shown to be effective in pruning wound protection. The expression of genes coding for enzymes of the phenylpropanoid pathway and pathogenesis-related (PR) proteins was profiled over a 48-h period using quantitative reverse transcriptase PCR. The cell cultures responded to elicitors of both fungi with a hypersensitive-like response that lead to a decrease in cell viability. Similar genes were triggered by both the pathogen and biocontrol agent, but the timing patterns and magnitude of expression was dependent on the specific fungal elicitor. Culture filtrates of both fungi caused upregulation of phenylalanine ammonia-lyase (PAL), 4-coumaroyl Co-A ligase (CCo-A) and stilbene synthase (STS), and a downregulation of chalcone synthase (CHS) genes. The pathogen filtrate caused a biphasic pattern in the upregulation of PAL and STS genes which was not observed in cells treated with filtrates of the biocontrol agent. Analytical assays showed significantly higher total phenolic content and chitinolytic enzyme activity in the cell cultures treated with the T. atroviride filtrate compared to the pathogen filtrate. These results corresponded well to the higher expression of PAL and chitinase class IV genes. The response of the cell cultures to T. atroviride filtrate provides support for the notion that the wound protection by the biocontrol agent at least partially relies on the induction of grapevine resistance mechanisms.

Background and Aims:  Carotenoids and chlorophylls perform a number of essential roles in plants making their accurate quantification important to a variety of studies. We aimed to develop an extraction protocol to accurately determine the photosynthetic pigments in grapevine leaf and berry tissue, specifically focusing on limiting the degradation of these pigments. Methods and Results:  An extraction protocol for grapevine leaf and berry tissue was systematically optimised by identifying a number of critical parameters. Extracted pigments were analysed using Reversed Phase‐High Performance Liquid Chromatography (RP‐HPLC). Specific parameters that were optimised included avoiding freeze‐drying the material; the volume of acetone and the time required to extract all the pigments from the tissue; the addition of 0.1% (v/v) N‐ethyldiisopropylamine to berry extracts to minimise pigment degradation during the extraction procedure; and avoiding concentration of the extracts that otherwise resulted in differential degradation of pigments. Additionally, the method of extraction and normalisation with an internal standard was adapted and improved for accuracy. The optimised protocol was validated using authentic standards and its utility shown by analysing the pigment content of berries and leaves at different growth stages. Conclusions:  A method has been developed that is able to extract and accurately quantify, by means of HPLC profiling, the levels of photosynthetic pigments from grape berries and leaves. The method avoided any degradation of the pigments during the extraction and was applicable to both berries and leaves in different stages of growth and development, indicating its general usefulness to vegetative and reproductive organs, even if their metabolic states are very different. Significance of Study:  The divergence of methods used for photosynthetic pigment analysis in plants, each with specific advantages and disadvantages were considered and used to optimise a number of parameters in a single method that proved to be applicable to plant organs in different developmental stages. The method is fast, applicable to vegetative and reproductive grapevine tissues, avoids degradation of pigments and ensures maximum accuracy when quantifying these important pigments.

BackgroundThis work describes a proteomics profiling method, optimized and applied to berry cell suspensions to evaluate organ-specific cultures as a platform to study grape berry ripening. Variations in berry ripening within a cluster(s) on a vine and in a vineyard are a major impediment towards complete understanding of the functional processes that control ripening, specifically when a characterized and homogenous sample is required. Berry cell suspensions could overcome some of these problems, but their suitability as a model system for berry development and ripening needs to be established first.Methodology/Principal FindingsIn this study we report on the proteomic evaluation of the cytosolic proteins obtained from synchronized cell suspension cultures that were established from callus lines originating from green, véraison and ripe Vitis vinifera berry explants. The proteins were separated using liquid phase IEF in a Microrotofor cell and SDS PAGE. This method proved superior to gel-based 2DE. Principal component analysis confirmed that biological and technical repeats grouped tightly and importantly, showed that the proteomes of berry cultures originating from the different growth/ripening stages were distinct. A total of twenty six common bands were selected after band matching between different growth stages and twenty two of these bands were positively identified. Thirty two % of the identified proteins are currently annotated as hypothetical. The differential expression profile of the identified proteins, when compared with published literature on grape berry ripening, suggested common trends in terms of relative abundance in the different developmental stages between real berries and cell suspensions.ConclusionsThe advantages of having suspension cultures that accurately mimic specific developmental stages are profound and could significantly contribute to the study of the intricate regulatory and signaling networks responsible for berry development and ripening.