Cellular and humoral immune responses associated with protection in sheep vaccinated against Teladorsagia circumcincta.

Due to increased anthelmintic resistance, complementary methods to drugs are necessary to control gastrointestinal nematodes (GIN). Vaccines are an environmentally-friendly and promising option. In a previous study, a Teladorsagia circumcincta recombinant sub-unit vaccine was administered to two sheep breeds with different levels of resistance against GIN. In the susceptible Canaria Sheep (CS) breed, vaccinates harboured smaller worms with fewer eggs in utero than the control group. Here, we extend this work, by investigating the cellular and humoral immune responses of these two sheep breeds following vaccination and experimental infection with T. circumcincta. In the vaccinated CS group, negative associations between antigen-specific IgA, IgG2 and Globule Leukocytes (GLs) with several parasitological parameters were established as well as a higher CD4+/CD8+ ratio than in control CS animals, suggesting a key role in the protection induced by the vaccine. In the more resistant Canaria Hair Breed (CHB) sheep the vaccine did not significantly impact on the parasitological parameters studied and none of these humoral associations were observed in vaccinated CHB lambs, although CHB had higher proportions of CD4+ and CD8+ T cells within the abomasal lymph nodes, suggesting higher mucosal T cell activation. Each of the component proteins in the vaccine induced an increase in immunoglobulin levels in vaccinated groups of each breed. However, levels of immunoglobulins to only three of the antigens (Tci-MEP-1, Tci-SAA-1, Tci-ASP-1) were negatively correlated with parasitological parameters in the CS breed and they may be, at least partially, responsible for the protective effect of the vaccine in this breed. These data could be useful for improving the current vaccine prototype.

Impacts of breed type and vaccination on Teladorsagia circumcincta infection in native sheep in Gran Canaria.

Vaccines and genetic resistance offer potential future alternatives to the exclusive use of anthelmintics to control gastrointestinal nematodes (GIN). Here, a Teladorsagia circumcincta prototype vaccine was administered to two sheep breeds which differ in their relative levels of resistance to infection with GIN. Vaccination of the more susceptible Canaria Sheep (CS) breed induced significant reductions in worm length and numbers of worm eggs in utero (EIU) when compared to control CS sheep. In the more resistant Canaria Hair Breed (CHB), although vaccination induced a reduction in all parasitological parameters analysed, differences between vaccinated and control sheep were not statistically significant. Such interactions between sheep breed and vaccination may allow better integrated control of GIN in future.

Changes in distribution, morphology and ultrastructure of chloride cell in Atlantic salmon during an AGD infection.

Amoebic gill disease (AGD) is emerging as one of the most significant health challenges affecting farmed Atlantic salmon in the marine environment. It is caused by the amphizoic amoeba Neoparamoeba perurans, with infestation of gills causing severe hyperplastic lesions, compromising overall gill integrity and function. This study used histology, transmission electron microscopy (TEM), immunohistochemistry and transcript expression to relate AGD-associated pathological changes to changes in the morphology and distribution of chloride cells (CCs) in the gills of Atlantic salmon (Salmo salar L.) showing the progression of an AGD infection. A marked reduction in numbers of immunolabelled CCs was detected, and a changing pattern in distribution and morphology was closely linked with the level of basal epithelial hyperplasia in the gill. In addition, acute degenerative ultrastructural changes to CCs at the lesion site were observed with TEM. These findings were supported by the early-onset downregulation of Na+ /K+ -ATPase transcript expression. This study provides supportive evidence that histological AGD lesion assessment was a good qualitative tool for AGD scoring and corresponded well with qPCR genomic Paramoeba perurans quantification. Ultrastructural changes induced in salmon CCs as a result of AGD are reported here for the first time.

Gene silencing by RNA interference in the ectoparasitic mite, Psoroptes ovis.

The presence of components of the RNA interference (RNAi) pathway in Psoroptes ovis, an ectoparasitic mite responsible for psoroptic mange, was investigated through interrogation of the P. ovis genome. Homologues of transcripts representing critical elements for achieving effective RNAi in the mite, Tetranychus urticae and the model organisms Caenorhabditis elegans and Drosophila melanogaster were identified and, following the development of a non-invasive immersion method of double stranded RNA delivery, gene silencing by RNAi was successfully demonstrated in P. ovis. Significant reductions in transcript levels were achieved for three target genes which encode the Group 2 allergen (Pso o 2), mu-class glutathione S-transferase (PoGST-mu1) and beta-tubulin (Poßtub). This is the first demonstration of RNAi in P. ovis and provides a mechanism for mining transcriptomic and genomic datasets for novel control targets against this economically important ectoparasite.

Facial nerve repair after operative injury: Impact of timing on hypoglossal-facial nerve graft outcomes.

PURPOSE: Reanimation of facial paralysis is a complex problem with multiple treatment options. One option is hypoglossal-facial nerve grafting, which can be performed in the immediate postoperative period after nerve transection, or in a delayed setting after skull base surgery when the nerve is anatomically intact but function is poor. The purpose of this study is to investigate the effect of timing of hypoglossal-facial grafting on functional outcome. MATERIALS AND METHODS: A retrospective case series from a single tertiary otologic referral center was performed identifying 60 patients with facial nerve injury following cerebellopontine angle tumor extirpation. Patients underwent hypoglossal-facial nerve anastomosis following facial nerve injury. Facial nerve function was measured using the House-Brackmann facial nerve grading system at a median follow-up interval of 18months. Multivariate logistic regression analysis was used determine how time to hypoglossal-facial nerve grafting affected odds of achieving House-Brackmann grade of ≤3. RESULTS: Patients who underwent acute hypoglossal-facial anastomotic repair (0-14days from injury) were more likely to achieve House-Brackmann grade ≤3 compared to those that had delayed repair (OR 4.97, 95% CI 1.5-16.9, p=0.01). CONCLUSIONS: Early hypoglossal-facial anastomotic repair after acute facial nerve injury is associated with better long-term facial function outcomes and should be considered in the management algorithm.

Mapping autism risk loci using genetic linkage and chromosomal rearrangements.

Autism spectrum disorders (ASDs) are common, heritable neurodevelopmental conditions. The genetic architecture of ASDs is complex, requiring large samples to overcome heterogeneity. Here we broaden coverage and sample size relative to other studies of ASDs by using Affymetrix 10K SNP arrays and 1,181 [corrected] families with at least two affected individuals, performing the largest linkage scan to date while also analyzing copy number variation in these families. Linkage and copy number variation analyses implicate chromosome 11p12-p13 and neurexins, respectively, among other candidate loci. Neurexins team with previously implicated neuroligins for glutamatergic synaptogenesis, highlighting glutamate-related genes as promising candidates for contributing to ASDs.

Evaluation of copy number variations reveals novel candidate genes in autism spectrum disorder-associated pathways.

Autism spectrum disorders (ASDs) are highly heritable, yet relatively few associated genetic loci have been replicated. Copy number variations (CNVs) have been implicated in autism; however, the majority of loci contribute to <1% of the disease population. Therefore, independent studies are important to refine associated CNV regions and discover novel susceptibility genes. In this study, a genome-wide SNP array was utilized for CNV detection by two distinct algorithms in a European ancestry case-control data set. We identify a significantly higher burden in the number and size of deletions, and disrupting more genes in ASD cases. Moreover, 18 deletions larger than 1 Mb were detected exclusively in cases, implicating novel regions at 2q22.1, 3p26.3, 4q12 and 14q23. Case-specific CNVs provided further evidence for pathways previously implicated in ASDs, revealing new candidate genes within the GABAergic signaling and neural development pathways. These include DBI, an allosteric binder of GABA receptors, GABARAPL1, the GABA receptor-associated protein, and SLC6A11, a postsynaptic GABA transporter. We also identified CNVs in COBL, deletions of which cause defects in neuronal cytoskeleton morphogenesis in model vertebrates, and DNER, a neuron-specific Notch ligand required for cerebellar development. Moreover, we found evidence of genetic overlap between ASDs and other neurodevelopmental and neuropsychiatric diseases. These genes include glutamate receptors (GRID1, GRIK2 and GRIK4), synaptic regulators (NRXN3, SLC6A8 and SYN3), transcription factor (ZNF804A) and RNA-binding protein FMR1. Taken together, these CNVs may be a few of the missing pieces of ASD heritability and lead to discovering novel etiological mechanisms.

Evaluating mitochondrial DNA variation in autism spectrum disorders.

Despite the increasing speculation that oxidative stress and abnormal energy metabolism may play a role in Autism Spectrum Disorders (ASD), and the observation that patients with mitochondrial defects have symptoms consistent with ASD, there are no comprehensive published studies examining the role of mitochondrial variation in autism. Therefore, we have sought to comprehensively examine the role of mitochondrial DNA (mtDNA) variation with regard to ASD risk, employing a multi-phase approach. In phase 1 of our experiment, we examined 132 mtDNA single-nucleotide polymorphisms (SNPs) genotyped as part of our genome-wide association studies of ASD. In phase 2 we genotyped the major European mitochondrial haplogroup-defining variants within an expanded set of autism probands and controls. Finally in phase 3, we resequenced the entire mtDNA in a subset of our Caucasian samples (∼400 proband-father pairs). In each phase we tested whether mitochondrial variation showed evidence of association to ASD. Despite a thorough interrogation of mtDNA variation, we found no evidence to suggest a major role for mtDNA variation in ASD susceptibility. Accordingly, while there may be attractive biological hints suggesting the role of mitochondria in ASD our data indicate that mtDNA variation is not a major contributing factor to the development of ASD.

Carbon Dioxide Laser Rejuvenation of the Facial Skin.

This article reviews the evaluation and techniques for facial skin rejuvenation using the fractionated carbon dioxide laser. It includes a detailed overview of laser skin rejuvenation and discusses the potential complications associated with this procedure. A review of clinical outcomes in the literature is also included.

Free and open-source software for object detection, size, and colour determination for use in plant phenotyping.

BACKGROUND: Object detection, size determination, and colour detection of images are tools commonly used in plant science. Key examples of this include identification of ripening stages of fruit such as tomatoes and the determination of chlorophyll content as an indicator of plant health. While methods exist for determining these important phenotypes, they often require proprietary software or require coding knowledge to adapt existing code. RESULTS: We provide a set of free and open-source Python scripts that, without any adaptation, are able to perform background correction and colour correction on images using a ColourChecker chart. Further scripts identify objects, use an object of known size to calibrate for size, and extract the average colour of objects in RGB, Lab, and YUV colour spaces. We use two examples to demonstrate the use of these scripts. We show the consistency of these scripts by imaging in four different lighting conditions, and then we use two examples to show how the scripts can be used. In the first example, we estimate the lycopene content in tomatoes (Solanum lycopersicum) var. Tiny Tim using fruit images and an exponential model to predict lycopene content. We demonstrate that three different cameras (a DSLR camera and two separate mobile phones) are all able to model lycopene content. The models that predict lycopene or chlorophyll need to be adjusted depending on the camera used. In the second example, we estimate the chlorophyll content of basil (Ocimum basilicum) using leaf images and an exponential model to predict chlorophyll content. CONCLUSION: A fast, cheap, non-destructive, and inexpensive method is provided for the determination of the size and colour of plant materials using a rig consisting of a lightbox, camera, and colour checker card and using free and open-source scripts that run in Python 3.8. This method accurately predicted the lycopene content in tomato fruit and the chlorophyll content in basil leaves.

Space controlled environment agriculture offers pathways to improve the sustainability of controlled environmental agriculture on Earth.

Terrestrial controlled environment agriculture (CEA) will have an increasingly important role in food production. However, present CEA systems are energy- and resource-hungry and rarely profitable, requiring a step change in design and optimization. Here we argue that the unique nature of space controlled environment agriculture (SpaCEA), which needs to be both highly resource efficient and circular in design, presents an opportunity to develop intrinsically circular CEA systems. Life-cycle analysis tools should be used to optimize the provision and use of natural or electrical light, power, nutrients and infrastructure in CEA and/or SpaCEA systems, and to guide research and development into subsystems that bring strong environmental advantages. We suggest that SpaCEA public outreach can also be used to improve the perception of terrestrial CEA on Earth by using space as a gateway for exhibiting CEA food growing technologies. A substantial focus on SpaCEA development should be viewed as an efficient contribution to addressing major current CEA challenges.

Inactivation of recombinant bacteriophage lambda by use of chemical agents and UV radiation.

Several approaches for the inactivation of bacteriophage lambda, including UV germicidal irradiation (UVGI) and the chemical agents Virkon-S, Chloros, Decon-90, and sodium hydroxide (NaOH), were compared. Virkon, NaOH, and UVGI caused a ≥7-log(10) reduction in phage titers. This study successfully describes several methods with potential for bacteriophage inactivation in industrial settings.

Assessment of cathepsin D and L-like proteinases of poultry red mite, Dermanyssus gallinae (De Geer), as potential vaccine antigens.

Vaccination is a feasible strategy for controlling the haematophagous poultry red mite Dermanyssus gallinae. A cDNA library enriched for genes upregulated after feeding was created to identify potential vaccine antigens. From this library, a gene (Dg-CatD-1) encoding a 383 amino acid protein (Dg-CatD-1) with homology to cathepsin D lysosomal aspartyl proteinases was identified as a potential vaccine candidate. A second gene (Dg-CatL-1) encoding a 341 amino acid protein (Dg-CatL-1) with homology to cathepsin L cysteine proteinases was also selected for further study. IgY obtained from naturally infested hens failed to detect Dg-CatD-1 suggesting that it is a concealed antigen. Conversely, Dg-CatL-1 was detected by IgY derived from natural-infestation, indicating that infested hens are exposed to Dg-CatL-1. Mortality rates 120 h after mites had been fed anti-Dg-CatD-1 were significantly higher than those fed control IgY (PF<0·01). In a survival analysis, fitting a proportional hazards model to the time of death of mites, anti-Dg-CatD-1 and anti-Dg-CatL-1 IgY had 4·42 and 2·13 times higher risks of dying compared with controls (PF<0·05). Dg-CatD-1 and L-1 both have potential as vaccine antigens as part of a multi-component vaccine and have the potential to be improved as vaccine antigens using alternative expression systems.

Carbon Dioxide Laser Rejuvenation of the Facial Skin.

This article reviews the evaluation and techniques for facial skin rejuvenation using the fractionated carbon dioxide laser. It includes a detailed overview of laser skin rejuvenation and discusses the potential complications associated with this procedure. A review of clinical outcomes in the literature is also included.

FoamPi: An open-source raspberry Pi based apparatus for monitoring polyurethane foam reactions.

Adiabatic temperature rise is an important method for determining isocyanate conversion in polyurethane foam reactions as well as many other exothermic chemical reactions. Adiabatic temperature rise can be used in conjunction with change in height and mass measurements to gain understanding into the blowing and gelling reactions that occur during polyurethane foaming as well as give important information on cell morphology. FoamPi is an open-source Raspberry Pi device for monitoring polyurethane foaming reactions. The device effectively monitors temperature rise, change in foam height as well as changes in the mass during the reaction. Three Python scripts are also presented. The first logs raw data during the reaction. The second corrects temperature data such that it can be used in adiabatic temperature rise reactions for calculating isocyanate conversion; additionally this script reduces noise in all the data and removes erroneous readings. The final script extracts important information from the corrected data such as maximum temperature change and maximum height change as well as the time to reach these points. Commercial examples of such equipment exist however the price (>£10000) of these equipment make these systems inaccessible for many research laboratories. The FoamPi build presented is inexpensive (£350) and test examples are shown here to indicate the reproducibility of results as well as precision of the FoamPi.

Rational Design of a Polyurethane Foam.

Polyurethane (PU) foams are exceptionally versatile due to the nature of PU bond formation and the large variety of polymeric backbones and formulation components such as catalysts and surfactants. This versatility introduces a challenge, namely a near unlimited number of variables for formulating foams. In addition to this, PU foam development requires expert knowledge, not only in polyurethane chemistry but also in the art of evaluating the resulting foams. In this work, we demonstrate that a rational experimental design framework in conjunction with a design of experiments (DoE) approach reduces both the number of experiments required to understand the formulation space and reduces the need for tacit knowledge from a PU expert. We focus on an in-depth example where a catalyst and two surfactants of a known formulation are set as factors and foam physical properties are set as responses. An iterative DoE approach is used to generate a set of foams with substantially different cell morphology and hydrodynamic behaviour. We demonstrate that with 23 screening formulations and 16 final formulations, foam physical properties can be modelled from catalyst and surfactant loadings. This approach also allows for the exploration of relationships between the cell morphology of PU foam and its hydrodynamic behaviour.

Differences in the protection elicited by a recombinant Teladorsagia circumcincta vaccine in weaned lambs of two Canarian sheep breeds.

Gastrointestinal nematode (GIN) infections are a serious drawback on small ruminant production. Since anthelmintic resistance has extended, optimisation of alternative non-chemical control strategies has attracted interest. Recently, a prototype recombinant vaccine protected immunologically mature sheep from Texel-cross and Canaria Sheep breeds against Teladorsagia circumcincta. The level of protective immunity stimulated by the vaccine varied between individuals and with age. Previous studies suggest that Canaria Hair Breed (CHB) sheep is naturally resistant to GIN infection, with some evidence suggesting that this protection is present in young lambs. Here, we sought to enhance this resistance by immunising three-month-old CHB lambs with a T. circumcincta prototype recombinant vaccine. Following vaccination and a larval challenge period, levels of protection against T. circumcincta infection were compared in CHB lambs with Canaria Sheep (CS) lambs (a breed considered less resistant to GIN). Lambs from the resistant CHB breed appeared to respond more favourably to vaccination, shedding 63% fewer eggs over the sampling period than unvaccinated CHB lambs. No protection was evident in CS vaccinated lambs. At post-mortem, CHB vaccine recipients had a 68% reduction in mean total worm burden, and female worms were significantly shorter and contained fewer eggs in utero compared to unvaccinated CHB lambs. A higher anti-parasite IgG2 level was detected in immunised CHB lambs compared to unvaccinated control CHB animals, with data suggesting that IgA, globular leucocytes, CD45RA+, CD4+ and CD8+ T cells are implicated in this protective response. The development of effective immunity in vaccinated CHB lambs did not reduce lamb growth rate as immunised CHB lambs had a significantly higher average daily weight gain after challenge than their unvaccinated counterparts. Therefore, the protection of CHB lambs was enhanced by immunisation at weaning, suggesting a synergistic effect when combining vaccination with presumed genetic resistance.

Microduplications in an autism multiplex family narrow the region of susceptibility for developmental disorders on 15q24 and implicate 7p21.

Copy number variations (CNVs) play a crucial role in the intricate genetics of autism spectrum disorders. A region on chromosome 15q24 vulnerable to both deletions and duplications has been previously implicated in a range of phenotypes including autism, Asperger's syndrome, delayed development, and mild to severe mental retardation. Prior studies have delineated a minimal critical region of approximately 1.33 Mb. In this study, a multiplex autism family was evaluated for CNVs using genotyping data from the Illumina 1 M BeadChip and analyzed with the PennCNV algorithm. Variants were then identified that co-segregate with autism features in this family. Here, we report autistic first cousins who carry two microduplications concordant with disease. Both duplications were inherited maternally and found to be identical by descent. The first is an approximately 10,000 base pair microduplication within the minimal region on 15q24 that falls across a single gene, ubiquitin-like 7. This is the smallest duplication in the region to result in a neuropsychiatric disorder, potentially narrowing the critical region for susceptibility to developmental and autism spectrum disorders. The second is a novel, 352 kb tandem duplication on 7p21 that replicates part of the neurexophilin 1 and islet cell autoantigen 1 genes. The breakpoint junction falls within the intronic regions of these genes and demonstrates a microhomology of four base pairs. Each of these microduplications may contribute to the complex etiology of autism spectrum disorders.

Lessons of the month: A misunderstood teenager with paediatric inflammatory multisystem syndrome - temporarily associated with SARS-CoV-2 admitted under adult medicine.

The emergence of SARS-CoV-2 has proven to be a challenge to healthcare bodies globally. The virus has been associated with a spectrum of clinical features, from anosmia to gastrointestinal upset to multiorgan dysfunction in the most severe cases. Given the range of features observed, it is important to be aware that infectious diseases can present atypically. Furthermore, in many hospitals, including our own, teenagers aged 16 to 18 years old are admitted under the care of adult medical services. Clinicians should be aware of patients presenting with the novel condition of paediatric inflammatory multisystem disorder - temporarily associated with SARS-CoV-2 (PIMS-TS).

Septal Perfoplasty for Management of Symptomatic Nasal Septal Perforation: An Alternative to Surgical Closure.

Importance: Symptomatic septal perforations are often difficult to manage and can have a significant impact on patient quality of life. Available surgical techniques for repair have demonstrated a varying rate of success, presenting a need for reliable interventions targeting symptom control. Objectives: To describe the modified surgical technique here termed septal perfoplasty. To demonstrate that creation of favorable septal perforation characteristics is effective in managing symptoms and improving patient quality of life. Design, Setting, and Participants: A retrospective review of the medical record was performed of patients who underwent the procedure of interest between July 1, 2006 and October 1, 2019 at Vanderbilt University Medical Center. All patients with symptomatic septal perforation who underwent septal perfoplasty within the timeframe reviewed were included. Septal perfoplasty was standardly performed in combination with turbinate reduction in all cases. This was combined with other indicated procedures for chronic sinusitis, repair of vestibular stenosis or nasal deformity. Main Outcomes and Measures: Creation of a well-mucosalized septal perforation, combined with patient-reported acceptable symptom control, was the primary outcome. Secondary outcomes include time to resolution, duration of follow-up, postsurgical complications, and need for further intervention. Results: Twenty patients (70% female; mean [range] age, 45.8 [15-72] years) underwent septal perfoplasty over the course of 13 years. The most common etiology of perforation was trauma (40%), presenting symptom was crusting (95%), and size of perforation repaired was large (60%). Mean follow-up was 37.6 months (range, 1-153 months). Overall, favorable perforation characteristics were created in 95% of cases by the first postoperative appointment. Acceptable symptomatic control was achieved in 18 out of 20 patients (90%), with a median time to improvement of 66 days. Eight patients required additional surgery to address chronic sinusitis or vestibular stenosis. Two patients experienced postoperative infections, treated conservatively with antibiotics. Conclusion and Relevance: Septal perfoplasty is a safe, simple, and effective method for management of symptomatic nasal septal perforation, which provides an alternative to more complicated interventions with comparable rates of symptomatic resolution. This procedure should particularly be considered for patients in which difficult repair is anticipated.