Population diversity and epidemiology of Bremia lactucae the cause of lettuce downy mildew

Bremia lactucae is an obligate biotroph that causes Lettuce Downy Mildew (LDM), a foliar disease of lettuce which negatively affects crop value though reduced yield and quality. Emergence of new strains of B. lactucae that can overcome host resistance or result in reduced sensitivity to fungicide active ingredients is a consistent risk to LDM management and crop production. From the 254 UK samples of B. lactucae collected and genotyped successfully using ten SSR loci, 135 multilocus genotypes were identified. Evidence was found of widespread incidence of heterokaryosis and overwintering clonal lineages. At least one UK isolate was able to overcome each differential line in IBEB differential set-C, of which cv Dandie containing Dm3 was least overcome by isolates tested. Of 15 B. lactucae samples tested with the fungicide active ingredients axoystrobin, mandipropamid and dimethomorph, no fungicide insensitivity was observed. No conclusive evidence of association between genotype to fungicide insensitivity nor virulence was obtained. When trials were inoculated with isolates of various pathogen genotypes, cultivar choice was found to greatly influence the population diversity of B. lactucae, with specific MLLs associated with some of the trial cultivars. The study of aerial dispersal of sporangia using the LAMP assay and aerial samplers found it was predominantly local to infected plants (<5m). DNA of B. lactucae was detected up to 100m from an inoculum source. In commercial environments detection using the LAMP assay was as early as two days before LDM was reported in field, with symptomatic plants observed ~50-80m away from samplers. The UK population of B. lactucae, based on samples collected, is diverse in genotype and phenotype. LDM management should account for the genetic flexibility of B. lactucae heterokaryons, which may contribute to overwintering soil-borne inoculum, and aerial dispersion of sporangia, which transmits the pathogen and can contribute to gene flow

Valorization of spruce needle waste via supercritical extraction of waxes and facile isolation of nonacosan-10-ol

Supercritical carbon dioxide was utilized as a sustainable alternative to solvent extraction of waxes from the waste needles of two spruce species, namely Norwegian and Sitka spruce. These extracts were rich in nonacosan-10-ol, an organic compound with hydrophobic properties that lends its use in the preparation of superhydrophobic coatings. The highest crude yields were 1.7% w/w of needles obtained at 400 bar and 60 °C, while nonacosan-10-ol was selectively extracted at 200 bar and 60 °C (8070 ± 91.1 μg/g of needles). Purification of nonacosan-10-ol from the wax extracts was conducted using a simple rapid green recrystallization technique. This yielded a recovery of 44.6% ± 2% and 48.4% ± 2% of the total nonacosan-10-ol from the original crude Sitka (3600 μg/g of needles) and Norwegian wax (1920 μg/g of needles) respectively. Application of nonacosan-10-ol to a filter paper led to the formation of highly hydrophobic surfaces, with preliminary contact angles of up to 149°. This sustainable production method may develop opportunities to valorize forestry waste within a holistic biorefinery

Placental Mesenchymal Stromal Cells: Preclinical Safety Evaluation for Fetal Myelomeningocele Repair.

BackgroundMyelomeningocele (MMC) is the congenital failure of neural tube closure in utero, for which the standard of care is prenatal surgical repair. We developed clinical-grade placental mesenchymal stromal cells seeded on a dural extracellular matrix (PMSC-ECM), which have been shown to improve motor outcomes in preclinical ovine models. To evaluate the long-term safety of this product prior to use in a clinical trial, we conducted safety testing in a murine model.MethodsClinical grade PMSCs obtained from donor human placentas were seeded onto a 6 mm diameter ECM at a density of 3&nbsp;×&nbsp;105 cells/cm2. Immunodeficient mice were randomized to receive either an ECM only or PMSC-ECM administered into a subcutaneous pocket. Mice were monitored for tumor formation until two study endpoints: 4 wk and 6 mo. Pathology and histology on all tissues was performed to evaluate for tumors. Quantitative polymerase chain reaction (qPCR) was performed to evaluate for the presence of human DNA, which would indicate persistence of PMSCs.ResultsFifty-four mice were included; 13 received ECM only and 14 received PMSC-ECM in both the 4-wk and 6-mo groups. No mice had gross or microscopic evidence of tumor development. A nodular focus of mature fibrous connective tissue was identified at the subcutaneous implantation pocket in the majority of mice with no significant difference between ECM only and PMSC-ECM groups (P&nbsp;=&nbsp;0.32 at 4 wk, P &gt; 0.99 at 6 mo). Additionally, no human DNA was detected by qPCR in any mice at either time point.ConclusionsSubcutaneous implantation of the PMSC-ECM product did not result in tumor formation and we found no evidence that PMSCs persisted. These results support the safety of the PMSC-ECM product for use in a Phase 1/2a human clinical trial evaluating fetal MMC repair augmented with PMSC-ECM

GH deficiency status combined with GH receptor polymorphism affects response to GH in children.

Meta-analysis has shown a modest improvement in first-year growth response to recombinant human GH (r-hGH) for carriers of the exon 3-deleted GH receptor (GHRd3) polymorphism but with significant interstudy variability. The associations between GHRd3 and growth response to r-hGH over 3 years in relation to severity of GH deficiency (GHD) were investigated in patients from 14 countries. Treatment-naïve pre-pubertal children with GHD were enrolled from the PREDICT studies (NCT00256126 and NCT00699855), categorized by peak GH level (peak GH) during provocation test: ≤4 μg/l (severe GHD; n=45) and >4 to <10 μg/l mild GHD; n=49) and genotyped for the GHRd3 polymorphism (full length (fl/fl, fl/d3, d3/d3). Gene expression (GE) profiles were characterized at baseline. Changes in growth (height (cm) and SDS) over 3 years were measured. There was a dichotomous influence of GHRd3 polymorphism on response to r-hGH, dependent on peak GH level. GH peak level (higher vs lower) and GHRd3 (fl/fl vs d3 carriers) combined status was associated with height change over 3 years (P<0.05). GHRd3 carriers with lower peak GH had lower growth than subjects with fl/fl (median difference after 3 years -3.3 cm; -0.3 SDS). Conversely, GHRd3 carriers with higher peak GH had better growth (+2.7 cm; +0.2 SDS). Similar patterns were observed for GH-dependent biomarkers. GE profiles were significantly different between the groups, indicating that the interaction between GH status and GHRd3 carriage can be identified at a transcriptomic level. This study demonstrates that responses to r-hGH depend on the interaction between GHD severity and GHRd3 carriage

Long-term safety evaluation of placental mesenchymal stromal cells for in utero repair of myelomeningocele in a novel ovine model.

PurposeAugmentation of in utero myelomeningocele repair with human placental mesenchymal stromal cells seeded onto extracellular matrix (PMSC-ECM) improves motor outcomes in an ovine myelomeningocele model. This study evaluated the safety of PMSC-ECM application directly onto the fetal spinal cord in preparation for a clinical trial.MethodsLaminectomy of L5-L6 with PMSC-ECM placement directly onto the spinal cord was performed in five fetal lambs at gestational age (GA) 100-106 days. Lambs and ewes were monitored for three months following delivery. Lambs underwent magnetic resonance imaging (MRI) of the brain and spine at birth and at three months. All organs from lambs and uteri from ewes underwent histologic evaluation. Lamb spinal cords and brains and ewe placentas were evaluated for persistence of PMSCs by polymerase chain reaction for presence of human DNA.ResultsMRIs demonstrated no evidence of abnormal tissue growth or spinal cord tethering. Histological analysis demonstrated no evidence of abnormal tissue growth or treatment related adverse effects. No human DNA was identified in evaluated tissues.ConclusionThere was no evidence of abnormal tissue growth or PMSC persistence at three months following in utero application of PMSC-ECM to the spinal cord. This supports proceeding with clinical trials of PMSC-ECM for in utero myelomeningocele repair.Level of evidenceN/A TYPE OF STUDY: Basic science

Efficacy of clinical-grade human placental mesenchymal stromal cells in fetal ovine myelomeningocele repair.

BackgroundWhile fetal repair of myelomeningocele (MMC) revolutionized management, many children are still unable to walk independently. Preclinical studies demonstrated that research-grade placental mesenchymal stromal cells (PMSCs) prevent paralysis in fetal ovine MMC, however this had not been replicated with clinical-grade cells that could be used in an upcoming human clinical trial. We tested clinical-grade PMSCs seeded on an extracellular matrix (PMSC-ECM) in the gold standard fetal ovine model of MMC.MethodsThirty-five ovine fetuses underwent MMC defect creation at a median of 76 days gestational age, and defect repair at 101 days gestational age with application of clinical-grade PMSC-ECM (3&nbsp;×&nbsp;105 cells/cm2, n&nbsp;=&nbsp;12 fetuses), research-grade PMSC-ECM (3&nbsp;×&nbsp;105 cells/cm2, three cell lines with n&nbsp;=&nbsp;6 (Group 1), n&nbsp;=&nbsp;6 (Group 2), and n&nbsp;=&nbsp;3 (Group 3) fetuses, respectively) or ECM without PMSCs (n&nbsp;=&nbsp;8 fetuses). Three normal lambs underwent no surgical interventions. The primary outcome was motor function measured by the Sheep Locomotor Rating scale (SLR, range 0: complete paralysis to 15: normal ambulation) at 24&nbsp;h of life. Correlation of lumbar spine large neuron density with SLR was evaluated.ResultsClinical-grade PMSC-ECM lambs had significantly better motor function than ECM-only lambs (SLR 14.5&nbsp;vs. 6.5, p&nbsp;=&nbsp;0.04) and were similar to normal lambs (14.5&nbsp;vs. 15, p&nbsp;=&nbsp;0.2) and research-grade PMSC-ECM lambs (Group 1: 14.5&nbsp;vs. 15, p&nbsp;=&nbsp;0.63; Group 2: 14.5&nbsp;vs. 14.5, p&nbsp;=&nbsp;0.86; Group 3: 14.5&nbsp;vs. 15, p&nbsp;=&nbsp;0.50). Lumbar spine large neuron density was strongly correlated with motor function (r&nbsp;=&nbsp;0.753, p&lt;0.001).ConclusionsClinical-grade placental mesenchymal stromal cells seeded on an extracellular matrix rescued ambulation in a fetal ovine myelomeningocele model. Lumbar spine large neuron density correlated with motor function, suggesting a neuroprotective effect of the PMSC-ECM in prevention of paralysis. A first-in-human clinical trial of PMSCs in human fetal myelomeningocele repair is underway

Randomized, double-blind, placebo-controlled study of interferon-γ 1b in Friedreich Ataxia.

OBJECTIVE: In vitro, in&nbsp;vivo, and open-label studies suggest that interferon gamma (IFN-γ 1b) may improve clinical features in Friedreich Ataxia through an increase in frataxin levels. The present study evaluates the efficacy and safety of IFN-γ 1b in the treatment of Friedreich Ataxia through a double-blind, multicenter, placebo-controlled trial. METHODS: Ninety-two subjects with FRDA between 10 and 25&nbsp;years of age were enrolled. Subjects received either IFN-γ 1b or placebo for 6&nbsp;months. The primary outcome measure was the modified Friedreich Ataxia Rating Scale (mFARS). RESULTS: No difference was noted between the groups after 6&nbsp;months of treatment in the mFARS or secondary outcome measures. No change was noted in buccal cell or whole blood frataxin levels. However, during an open-label extension period, subjects had a more stable course than expected based on natural history data. CONCLUSIONS: This study provides no direct evidence for a beneficial effect of IFN-γ1b in FRDA. The modest stabilization compared to natural history data leaves open the possibility that longer studies may demonstrate benefit

Randomized, double‐blind, placebo‐controlled study of interferon‐ γ

OBJECTIVE: In vitro, in&nbsp;vivo, and open-label studies suggest that interferon gamma (IFN-γ 1b) may improve clinical features in Friedreich Ataxia through an increase in frataxin levels. The present study evaluates the efficacy and safety of IFN-γ 1b in the treatment of Friedreich Ataxia through a double-blind, multicenter, placebo-controlled trial. METHODS: Ninety-two subjects with FRDA between 10 and 25&nbsp;years of age were enrolled. Subjects received either IFN-γ 1b or placebo for 6&nbsp;months. The primary outcome measure was the modified Friedreich Ataxia Rating Scale (mFARS). RESULTS: No difference was noted between the groups after 6&nbsp;months of treatment in the mFARS or secondary outcome measures. No change was noted in buccal cell or whole blood frataxin levels. However, during an open-label extension period, subjects had a more stable course than expected based on natural history data. CONCLUSIONS: This study provides no direct evidence for a beneficial effect of IFN-γ1b in FRDA. The modest stabilization compared to natural history data leaves open the possibility that longer studies may demonstrate benefit

Early investigations into improving bowel and bladder function in fetal ovine myelomeningocele repair.

IntroductionFetal myelomeningocele (MMC) repair improves lower extremity motor function. We have previously demonstrated that augmentation of fetal MMC repair with placental mesenchymal stromal cells (PMSCs) seeded on extracellular matrix (PMSC-ECM) further improves motor function in the ovine model. However, little progress has been made in improving bowel and bladder function, with many patients suffering from neurogenic bowel and bladder. We hypothesized that fetal MMC repair with PMSC-ECM would also improve bowel and bladder function.MethodsMMC defects were surgically created in twelve ovine fetuses at median gestational age (GA) 73 days, followed by defect repair at GA101 with PMSC-ECM. Fetuses were delivered at GA141. Primary bladder function outcomes were voiding posture and void volumes. Primary bowel function outcome was anorectal manometry findings including resting anal pressure and presence of rectoanal inhibitory reflex (RAIR). Secondary outcomes were anorectal and bladder detrusor muscle thickness. PMSC-ECM lambs were compared to normal lambs (n&nbsp;=&nbsp;3).ResultsEighty percent of PMSC-ECM lambs displayed normal voiding posture compared to 100% of normal lambs (p&nbsp;=&nbsp;1). Void volumes were similar (PMSC-ECM 6.1&nbsp;ml/kg vs. normal 8.8&nbsp;ml/kg, p&nbsp;=&nbsp;0.4). Resting mean anal pressures were similar between cohorts (27.0&nbsp;mmHg PMSC-ECM vs. normal 23.5&nbsp;mmHg, p&nbsp;=&nbsp;0.57). RAIR was present in 3/5 PMSC-ECM lambs that underwent anorectal manometry and all normal lambs (p&nbsp;=&nbsp;0.46). Thicknesses of anal sphincter complex, rectal wall muscles, and bladder detrusor muscles were similar between cohorts.ConclusionOvine fetal MMC repair augmented with PMSC-ECM results in near-normal bowel and bladder function. Further work is needed to evaluate these outcomes in human patients