An Antibody-Based Leukocyte-Capture Microarray for the Diagnosis of Systemic Lupus Erythematosus

The diagnosis of Systemic Lupus Erythematosus (SLE) is challenging due to its heterogeneous clinical presentation and the lack of robust biomarkers to distinguish it from other autoimmune diseases. Further, currently used laboratory tests do not readily distinguish active and inactive disease. Several groups have attempted to apply emerging high throughput profiling technologies to diagnose and monitor SLE. Despite showing promise, many are expensive and technically challenging for routine clinical use. The goal of this work is to develop a better diagnostic and monitoring tool for SLE. We report a highly customisable antibody microarray that consists of a duplicate arrangement of 82 antibodies directed against surface antigens on peripheral blood mononuclear cells (PMBCs). This high-throughput array was used to profile SLE patients (n = 60) with varying disease activity, compared to healthy controls (n = 24), patients with rheumatoid arthritis (n = 25), and other autoimmune diseases (n = 28). We used a computational algorithm to calculate a score from the entire microarray profile and correlated it with SLE disease activity. Our results demonstrate that leukocyte-capture microarray profiles can readily distinguish active SLE patients from healthy controls (AUROC = 0.84). When combined with the standard laboratory tests (serum anti-dsDNA, complements C3 and C4), the microarrays provide significantly increased discrimination. The antibody microarrays can be enhanced by the addition of other markers for potential application to the diagnosis and stratification of SLE, paving the way for the customised and accurate diagnosis and monitoring of SLE.     

Development and SAR of functionally selective allosteric modulators of GABAA receptors

Positive modulators at the benzodiazepine site of α2- and α3-containing GABA(A) receptors are believed to be anxiolytic. Through oocyte voltage clamp studies, we have discovered two series of compounds that are positive modulators at α2-/α3-containing GABA(A) receptors and that show no functional activity at α1-containing GABA(A) receptors. We report studies to improve this functional selectivity and ultimately deliver clinical candidates. The functional SAR of cinnolines and quinolines that are positive allosteric modulators of the α2- and α3-containing GABA(A) receptors, while simultaneously neutral antagonists at α1-containing GABA(A) receptors, is described. Such functionally selective modulators of GABA(A) receptors are expected to be useful in the treatment of anxiety and other psychiatric illnesses.     

The Positive Inside Rule Is Stronger When Followed by a Transmembrane Helix

The translocon recognizes transmembrane helices with sufficient level of hydrophobicity and inserts them into the membrane. However, sometimes less hydrophobic helices are also recognized. Positive inside rule, orientational preferences of and specific interactions with neighboring helices have been shown to aid in the recognition of these helices, at least in artificial systems. To better understand how the translocon inserts marginally hydrophobic helices, we studied three naturally occurring marginally hydrophobic helices, which were previously shown to require the subsequent helix for efficient translocon recognition. We find no evidence for specific interactions when we scan all residues in the subsequent helices. Instead, we identify arginines located at the N-terminal part of the subsequent helices that are crucial for the recognition of the marginally hydrophobic transmembrane helices, indicating that the positive inside rule is important. However, in two of the constructs, these arginines do not aid in the recognition without the rest of the subsequent helix; that is, the positive inside rule alone is not sufficient. Instead, the improved recognition of marginally hydrophobic helices can here be explained as follows: the positive inside rule provides an orientational preference of the subsequent helix, which in turn allows the marginally hydrophobic helix to be inserted; that is, the effect of the positive inside rule is stronger if positively charged residues are followed by a transmembrane helix. Such a mechanism obviously cannot aid C-terminal helices, and consequently, we find that the terminal helices in multi-spanning membrane proteins are more hydrophobic than internal helices.     

Feeding and digestive responses to fatty acid intake in two South American passerines with different food habits

Specific fatty acids (FA) such as unsaturated (UFA) and saturated (SFA) fatty acids contained in foods are key factors in the nutritional ecology of birds. By means of a field and experimental approach, we evaluated the effect of diet on the activity of three esterases involved in FA hydrolysis; carboxylesterase (CE: 4-NPA-CE and a-NA-CE) and butyrylcholinesterase, in two South American passerines: the omnivorous rufous-collared sparrow (Zonotrichia capensis) and the granivorous common diuca-finch (Diuca diuca). The activity of the three esterases was measured in the intestines of freshly caught individuals over two distinct seasons and also after a chronic intake of a UFA-rich or SFA-rich diet in the laboratory. In turn, we assessed the feeding responses of the birds choosing amongst diets contrasting in the kind of specific FA (UFA- vs. SFA-treated diets). During summer, field CE activities (4-NPA-CE and a-NA-CE) in the small intestine were higher in the rufous-collared sparrow (25.3 ± 3.3 and 81.4 ± 10.8 µmol min^?1 g tissue^?1, respectively) than in the common diuca-finch (10.0 ± 3.0 and 33.9 ± 13.1 µmol min^?1 g tissue^?1, respectively). Two hour feeding trial test indicated that both species exhibited a clear preference for UFA-treated diets. On average, the rufous-collared sparrow consumed 0.46 g 2 h^?1 of UFA-rich diets and 0.12 g 2 h^?1 of SFA-rich diets. In turn, the consumption pattern of the common diuca-finch averaged 0.73 and 0.16 g 2 h^?1 for UFA-rich and SFA-rich diets, respectively. After a month of dietary acclimation to UFA-rich and SFA-rich diets, both species maintained body mass irrespective of the dietary regime. Additionally, the intestinal 4-NPA-CE activity exhibited by birds fed on a UFA-rich or SFA-rich diet was higher in the rufous-collared sparrow (39.0 ± 5.3 and 44.2 ± 7.3 µmol min^?1 g tissue^?1, respectively) than in the common diuca-finch (13.3 ± 1.9 and 11.2 ± 1.4 µmol min^?1 g tissue^?1, respectively). Finally, the intestinal a-NA-CE activity exhibited by the rufous-collared sparrow was about two times higher when consuming an UFA-rich diet. Our results suggest that the rufus-collared sparrow exhibits a greater capacity for intestinal FA hydrolysis, which would allow it to better deal with fats from different sources.     

Antifungal Activity In Vitro and In Vivo of Mesquite Extract (Prosopis glandulosa) Against Phytopathogenic Fungi

Fungi are the primary infectious agents in plant crops and many post-harvest fungal diseases of fruit and vegetables causing significant economic losses worldwide. Here, the antifungal effect of Prosopis glandulosa extract (PgE) against phytopathogenic fungi was evaluated. The effect with PgE (5, 4, 2, 1, 0%) as AI (%) and radial growth rate reduction (K_r %) were determined in vitro in Colletotrichum gloeosporoides, Fusarium oxysporum, Rhizopus oryzae and R. stolonifer (1 × 10⁵ spores/mL). The phytopathogenicity of fungal strains was performed under in vivo conditions (room temperature, 25–30 °C and refrigeration, ~ 4 °C) by fruit surface inoculation method on strawberries, tomatoes and carrots by recording the development of mycelial growth, necrosis, soft rot and dehydration symptoms showed on each fruit at 14 days. The extract (5%) showed the highest AI against C. gloesporioides (~ 96%), and F. oxysporum (~ 79%) and growth rate reduction of 74.92% and 64.82% respectively. Likewise, the extract controls the development of phytopathogenicity symptoms against C. gloesporioides and F. oxysporum in vivo conditions, nevertheless, was less efficiency against both Rhizopus species. The P. glandulosa extract represents an efficient, economical, and eco-friendly alternative to preserve the quality of the agricultural products and to increase their shelf life.     

Affordable and robust phenotyping framework to analyse root system architecture of soil‐grown plants

The phenotypic analysis of root system growth is important to inform efforts to enhance plant resource acquisition from soils; however, root phenotyping remains challenging because of the opacity of soil, requiring systems that facilitate root system visibility and image acquisition. Previously reported systems require costly or bespoke materials not available in most countries, where breeders need tools to select varieties best adapted to local soils and field conditions. Here, we report an affordable soil‐based growth (rhizobox) and imaging system to phenotype root development in glasshouses or shelters. All components of the system are made from locally available commodity components, facilitating the adoption of this affordable technology in low‐income countries. The rhizobox is large enough (approximately 6000 cm² of visible soil) to avoid restricting vertical root system growth for most if not all of the life cycle, yet light enough (approximately 21 kg when filled with soil) for routine handling. Support structures and an imaging station, with five cameras covering the whole soil surface, complement the rhizoboxes. Images are acquired via the Phenotiki sensor interface, collected, stitched and analysed. Root system architecture (RSA) parameters are quantified without intervention. The RSAs of a dicot species (Cicer arietinum, chickpea) and a monocot species (Hordeum vulgare, barley), exhibiting contrasting root systems, were analysed. Insights into root system dynamics during vegetative and reproductive stages of the chickpea life cycle were obtained. This affordable system is relevant for efforts in Ethiopia and other low‐ and middle‐income countries to enhance crop yields and climate resilience sustainably.     

Sublethal effects on seabirds after the Prestige oil-spill are mirrored in sexual signals

It has been suggested that sexual signals may be a useful measure of environmental quality as they represent the sum of environmental pressures on the animal. Accordingly, it has been proposed that carotenoid-based coloration may be especially valuable in monitoring and detecting the sublethal effects of toxic pollutants in the environment. Here, we evaluate whether the carotenoid-based coloration in the bill of adult yellow-legged gulls reflects oil-induced sublethal effects in breeding colonies affected by the Prestige oil spill. In 2004, we took blood samples from 27 adult birds at four insular breeding colonies located in the pathway of the Prestige oil spill. We measured the size of the red bill spot area and analysed plasma biochemical parameters indicative of sublethal effects of oil contamination in gulls, including glucose, total protein, creatinine, inorganic phosphorus, aspartate aminotransferase (AST) and gamma-glutamyl transferase. We showed that the size of their red bill spot area was positively related to body condition, while negatively related with AST levels, an enzyme that is commonly used as an indication of hepatic damage in birds. Hence, the present study provides support for the idea that carotenoid-based colour integuments may be a useful measure of environmental quality.     

Genome-wide analysis of the world's sheep breeds reveals high levels of historic mixture and strong recent selection

Through their domestication and subsequent selection, sheep have been adapted to thrive in a diverse range of environments. To characterise the genetic consequence of both domestication and selection, we genotyped 49,034 SNP in 2,819 animals from a diverse collection of 74 sheep breeds. We find the majority of sheep populations contain high SNP diversity and have retained an effective population size much higher than most cattle or dog breeds, suggesting domestication occurred from a broad genetic base. Extensive haplotype sharing and generally low divergence time between breeds reveal frequent genetic exchange has occurred during the development of modern breeds. A scan of the genome for selection signals revealed 31 regions containing genes for coat pigmentation, skeletal morphology, body size, growth, and reproduction. We demonstrate the strongest selection signal has occurred in response to breeding for the absence of horns. The high density map of genetic variability provides an in-depth view of the genetic history for this important livestock species.