The oligomeric assembly of galectin-11 is critical for anti-parasitic activity in sheep (Ovis aries)

Galectins are a family of glycan-binding molecules with a characteristic affinity for ß-D-glycosides that mediate a variety of important cellular functions, including immune and inflammatory responses. Galectin-11 (LGALS-11) has been recently identified as a mediator induced specifically in animals against gastrointestinal nematodes and can interfere with parasite growth and development. Here, we report that at least two natural genetic variants of LGALS-11 exist in sheep, and demonstrate fundamental differences in anti-parasitic activity, correlated with their ability to dimerise. This study improves our understanding of the role of galectins in the host immune and inflammatory responses against parasitic nematodes and provides a basis for genetic studies toward selective breeding of animals for resistance to parasites. © 2020, The Author(s). **Please note that there are multiple authors for this article therefore only the name of the first 5 including Federation University Australia affiliate “Dhanasekaran Sakthivel, Sarah Preston, Robin Gasser, Els Meeusen, David Piedrafita” is provided in this record*

Deep Learning for Enhanced Fault Diagnosis of Monoblock Centrifugal Pumps: Spectrogram-Based Analysis

Abstract The reliable operation of monoblock centrifugal pumps (MCP) is crucial in various industrial applications. Achieving optimal performance and minimizing costly downtime requires effectively detecting and diagnosing faults in critical pump components. This study proposes an innovative approach that leverages deep transfer learning techniques. An accelerometer was adopted to capture vibration signals emitted by the pump. These signals are then converted into spectrogram images which serve as the input for a sophisticated classification system based on deep learning. This enables the accurate identification and diagnosis of pump faults. To evaluate the effectiveness of the proposed methodology, 15 pre-trained networks including ResNet-50, InceptionV3, GoogLeNet, DenseNet-201, ShuffleNet, VGG-19, MobileNet-v2, InceptionResNetV2, VGG-16, NasNetmobile, EfficientNetb0, AlexNet, ResNet-18, Xception, ResNet101 and ResNet-18 were employed. The experimental results demonstrate the efficacy of the proposed approach with AlexNet exhibiting the highest level of accuracy among the pre-trained networks. Additionally, a meticulous evaluation of the execution time of the classification process was performed. AlexNet achieved 100.00% accuracy with an impressive execution (training) time of 17 s. This research provides invaluable insights into applying deep transfer learning for fault detection and diagnosis in MCP. Using pre-trained networks offers an efficient and precise solution for this task. The findings of this study have the potential to significantly enhance the reliability and maintenance practices of MCP in various industrial settings

Cloning, expression, purification and crystallographic studies of galectin-11 from domestic sheep (Ovis aries)

Galectins are an evolutionarily conserved family of proteins that translate glycan recognition into cellular effects. Galectin-11 is a unique member of the galectin family that is only expressed in ruminants such as sheep, goat and cattle and that plays a critical role in several important biological processes, such as reproduction and parasite-mediated innate immune responses. Currently, these two areas are of major importance for the sustainability of ruminant livestock production. Despite the emerging biological significance of galectin-11, no structural information is available. It is expected that structural studies will unravel the functional mechanisms of galectin-11 activity. Here, the expression, purification and crystallization of the ruminant-specific galectin-11 from domestic sheep and the collection of X-ray data to 2.0 Å resolution are reported. © 2015

Proteomic identification of galectin-11 and 14 ligands from Haemonchus contortus

Haemonchus contortus is the most pathogenic nematode of small ruminants. Infection in sheep and goats results in anaemia that decreases animal productivity and can ultimately cause death. The involvement of ruminant-specific galectin-11 (LGALS-11) and galectin-14 (LGALS-14) has been postulated to play important roles in protective immune responses against parasitic infection; however, their ligands are unknown. In the current study, LGALS-11 and LGALS-14 ligands in H. contortus were identified from larval (L4) and adult parasitic stages extracts using immobilised LGALS-11 and LGALS-14 affinity column chromatography and mass spectrometry. Both LGALS-11 and LGALS-14 bound more putative protein targets in the adult stage of H. contortus (43 proteins) when compared to the larval stage (two proteins). Of the 43 proteins identified in the adult stage, 34 and 35 proteins were bound by LGALS-11 and LGALS-14, respectively, with 26 proteins binding to both galectins. Interestingly, hematophagous stage-specific sperm-coating protein and zinc metalloprotease (M13), which are known vaccine candidates, were identified as putative ligands of both LGALS-11 and LGALS- 14. The identification of glycoproteins of H. contortus by LGALS-11 and LGALS-14 provide new insights into host-parasite interactions and the potential for developing new interventions

Evaluation of the role of galectins in parasite immunity

Galectin-11 (LGALS-11) and galectin-14 (LGALS-14) are ruminant specific galectins, first reported in sheep. Although their roles in parasite immunity are still being elucidated, it appears that they influence protection against parasites. In gastrointestinal infections with the nematode Haemonchus contortus, both galectin-11 and galectin-14 appear to be protective. However, in a chronic infection of liver fluke, Fasciola hepatica, these galectins may aid parasite survival. To unravel the structural, functional, and ligand profile of galectin-11 and galectin-14, recombinant production of these proteins is vital. Here we present the recombinant production of soluble galectin-11 and galectin-14 from domestic sheep for in vitro and structural biology studies. These methods include parasite cultivation and infection, galectin staining of host and parasite tissue, surface staining of parasites with recombinant galectins, pull-down assays to identify endogenous galectin binding proteins, and in vitro assays to monitor the effect of galectins on parasite development. © 2022, Springer Science+Business Media, LLC, part of Springer Nature

Strategies That Facilitate Extraction-Free SARS-CoV-2 Nucleic Acid Amplification Tests

The COVID-19 pandemic has resulted in an unprecedented global demand for in vitro diagnostic reagents. Supply shortages and hoarding have impacted testing capacity which has led to inefficient COVID-19 case identification and transmission control, predominantly in developing countries. Traditionally, RNA extraction is a prerequisite for conducting SARS-CoV-2 nucleic acid amplification tests (NAAT); however, simplified methods of sample processing have been successful at bypassing typical nucleic acid extraction steps, enabling extraction-free SARS-CoV-2 NAAT workflows. These methods involve chemical and physical approaches that are inexpensive and easily accessible alternatives to overcome extraction kit supply shortages, while offering acceptable test performance. Here we provide an overview of three main sample preparation strategies that have been shown to facilitate extraction-free SARS-CoV-2 NAATs

TaqMan primer/Probe sequences for assessing basal expression levels of TLR3 and TLR7 mRNA.

<p>TaqMan primer/Probe sequences for assessing basal expression levels of TLR3 and TLR7 mRNA.</p

Virus replication in PBMCs stimulated with TLR3 and TLR7 agonists.

<p>a) Reduction in TCID₅₀ values of PPRV in goat PBMC on imiquimod treatment. Reduction in PPRV H gene expression levels in goat PBMC on treatment with b) poly I:C and c) imiquimod. Bars with the same superscript do not differ significantly. Significance is indicated when p<0.05. Values represent mean ± SD of 40-corrected CT in 5 individual animals per goat breed. Significantly higher PPRV viral loads observed in PBMC of Barbari and Tellicherry breeds as compared to Kanni and Salem Black. Significant reduction in PPRV levels on poly I:C and imiquimod treatment in all breeds.</p

Cytokine levels in imiquimod, poly I:C or PPRV treated PBMCs.

<p>a) <b>TNFα,</b> b) IFNα, and c) IFNγ from supernatants of PPRV infected PBMC of different goat breeds. Bars with the same superscript do not differ significantly. Significance is indicated when p<0.05. PPRV infected PBMC from Kanni and Salem black breeds show higher production of TNFα, IFNα and IFNγ than Barbari. Poly I:C and imiquimod treated PBMC, similarly, show higher production of TNFα and IFNα in Kanni and Salem black breeds than in Barbari. TNFα and IFNγ levels are expressed as the corrected mean ± SD of optical density [OD] of treatment groups from which the OD of mock infected supernatants is subtracted. IFNα concentrations in the experimental samples are expressed as pg/ml.</p