Management of patients with a neurovascular conflict involving the optic nerve and a non-diseased intracranial artery: Three cases.

Introduction: Determining whether a neurovascular conflict (NVC) involving the anterior visual pathway (AVP) and a non-diseased intracranial artery is amenable for microvascular decompression is challenging. Moreover, it is unclear whether microvascular decompression of the optic nerve is an effective therapy. Research question: What are the outcomes of different treatment strategies for NVCs involving the AVP and a non-diseased intracranial artery? Material and methods: Data on patients with symptomatic NVCs involving the AVP and a non-diseased intracranial artery was collected and included treatment and outcome parameters. The case series was drafted in accordance with the CARE guidelines. Results: Three patients aged 53,53 and 55 visited our out-patient clinic with a suspected symptomatic NVC between the optic nerve and a non-diseased intracranial artery. A conservative treatment was opted for in the first patient aimed at treating her glaucoma, with temporary improvement of symptoms. Microvascular decompression of the optic nerve was performed in two patients. One operated patient developed post-operative complications resulting in posterior circulation perfusion decline, while the other experienced a worse tunnel vision with a decrease in visual acuity. Discussion and conclusion: The diagnosis of a symptomatic NVC between the AVP and a non-diseased intracranial artery should be considered with caution, i.e. after exclusion of all other causes. Microvascular decompression can be performed but does not necessarily improve symptoms. A better understanding of the pathophysiological mechanisms underlying these NVCs is warranted to determine the benefit of microvascular decompression of the optic nerve.

Food system actor perspectives on future-proofing European food systems through plant breeding.

Crop improvement is a key innovation area in the pursuit of sustainable food systems. However, realising its potential requires integration of the needs and priorities of all agri-food chain stakeholders. In this study, we provide a multi-stakeholder perspective on the role of crop improvement in future-proofing the European food system. We engaged agri-business, farm- and consumer-level stakeholders, and plant scientists through an online survey and focus groups. Four of each group's top five priorities were shared and related to environmental sustainability goals (water, nitrogen and phosphorus efficiency, and heat stress). Consensus was identified on issues including considering existing alternatives to plant breeding (e.g. management strategies), minimising trade-offs, and addressing geographical variation in needs. We conducted a rapid evidence synthesis on the impacts of priority crop improvement options, highlighting the urgent need for further research examining downstream sustainability impacts to identify concrete targets for plant breeding innovation as a food systems solution.

Blood flow restriction training in South Africa - a panel discussion.

Background: Blood flow restriction (BFR) training uses a cuff to partially occlude venous blood flow and improve musculoskeletal training outcomes. Over the past 25 years, numerous studies have demonstrated its relative safety and efficacy. Objectives: Blood flow restriction training is under review by the Health Professions Council of South Africa due to safety and ethical concerns. The objective of this roundtable discussion is to gain better insight into the current use and perception of blood flow restriction training in South Africa. Formation of panel: The expert panel had experience with the use of BFR training and included one representative from each of the following professions, namely, sports medicine, physiotherapy and biokinetics. Discussion: The panellists provided their unique perspectives on BFR training, whilst reaching a relative consensus on its safety, screening, efficacy, and appropriate use. Agreement on appropriate loading and occlusion pressure protocols during different phases of rehabilitation was less clear. Conclusion: Although BFR is a safe and effective modality, the development of evidence-based protocols among different health professionals in South Africa is required to ensure good clinical practice.

The effectiveness of an online intervention in stimulating injury-preventive behaviour in adult novice runners: Results of a randomised controlled trial.

Background: The online intervention Runfitcheck was developed to stimulate injury-preventive behaviour among adult novice runners. Objectives: This study evaluated the effectiveness of Runfitcheck on injury-preventive behaviour among adult novice runners. Methods: A randomised controlled trial was conducted among adult novice runners. The intervention group had access to the Runfitcheck intervention, the control group performed their running activities as usual. One, three, and five months after enrolment, participants reported retrospectively what they had done regarding injury-preventive behaviour (operationalised as (i) using a (personalised) training schedule; (ii) performing strength and technique exercises; and (iii) performing a warm-up routine prior to running). Relative Risks (RR) and 95% Confidence Interval (95%CI) were used to analyse behavioural change. Results: The intervention group (n=715) searched more often for information about a warm-up routine (RR 1.211; 95%CI 1.080-1.357), and added more often strength exercises to their warm-up routine (RR 1.228; 95%CI 1.092-1.380). The intervention group performed more often running technique exercises compared to the control group (n=696) (RR 1.134; 95%CI 1.015-1.267), but less often strength exercises (RR 0.865 (95%CI 0.752-0.995). Within the group of runners that did not perform any warm-up routine at enrolment (n=272), the intervention group started to perform a regular warm-up routine more often than the control group (RR 1.461; 95%CI 1.084-1.968). No significant results were found for using a training schedule. Conclusion: The online intervention Runfitcheck was effective in stimulating aspects of injury-preventive behaviour in adult novice runners, mostly related to a warm-up routine. Trial registration: NL6225, Registered April 24th 2007 - Retrospectively registered, https://www.trialregister.nl/trial/6225.

Invited review: Piglet survival: benefits of the immunocompetence.

Piglet mortality has a negative impact on animal welfare and public acceptance. Moreover, the number of weaned piglets per sow mainly determines the profitability of piglet production. Increased litter sizes are associated with lower birth weights and piglet survival. Decreased survival rates and performance of piglets make the control of diseases and infections within pig production even more crucial. Consequently, selection for immunocompetence becomes an important key aspect within modern breeding programmes. However, the phenotypic recording of immune traits is difficult and expensive to realize within farm routines. Even though immune traits show genetic variability, only few examples exist on their respective suitability within a breeding programme and their relationships to economically important production traits. The analysis of immune traits for an evaluation of immunocompetence to gain a generally improved immune response is promising. Generally, in-depth knowledge of the genetic background of the immune system is needed to gain helpful insights about its possible incorporation into breeding programmes. Possible physiological drawbacks for enhanced immunocompetence must be considered with regards to the allocation theory and possible trade-offs between the immune system and performance. This review aims to discuss the relationships between the immunocompetence of the pig, piglet survival as well as the potential of these traits to be included into a breeding strategy for improved robustness.

High prevalence of virological failure and HIV drug mutations in a first-line cohort of Malawian children.

Background: Drug resistance mutations (DRMs) increasingly jeopardize paediatric HIV programmes in sub-Saharan Africa. As individual monitoring of DRMs and viral loads has limited availability, population data on DRMs are essential to determine first-line susceptibility. Paediatric data from sub-Saharan Africa are scarce and unavailable for Malawi. Objectives: To determine the prevalence of virological failure (VF) and DRMs among ART-naive HIV-infected Malawian children during the first year of first-line ART. Methods: In a prospective cohort of HIV-infected Malawian children, on first-line treatment, children were followed monthly; blood was collected for viral load testing (6 and 12 months) and genotypic resistance testing (12 months). VF was defined as at least one viral load >1000 copies/mL or death after 6 months of ART. DRMs were identified and susceptibility to NRTIs and NNRTIs was scored using the Stanford algorithm and by calculating genotypic susceptibility scores (GSSs). Results: VF occurred in 66% (23/35) of the children during 12 months of follow-up. DRMs were detected in 44% (15/34); all had NNRTI resistance and 12% (4/34) had dual-class NNRTI/NRTI resistance. Reduced susceptibility (DRMs and GSS <3) was seen in 41% (14/34) to their current first-line regimen. High-level resistance was most common for nevirapine [26% (9/34)]. Conclusions: In this first report on VF and DRMs in children on first-line ART in Malawi, the rates of VF and DRMs were alarmingly high. Paediatric HIV programmes in sub-Saharan Africa should emphasize programmatic evaluation of VF and include detection of DRMs to adjust and design adequate first- and second-line regimens and prevent widespread resistance in children.

Estimating variance components and breeding values for number of oocytes and number of embryos in dairy cattle using a single-step genomic evaluation.

Reproductive technologies such as multiple ovulation and embryo transfer (MOET) and ovum pick-up (OPU) accelerate genetic improvement in dairy breeding schemes. To enhance the efficiency of embryo production, breeding values for traits such as number of oocytes (NoO) and number of MOET embryos (NoM) can help in selection of donors with high MOET or OPU efficiency. The aim of this study was therefore to estimate variance components and (genomic) breeding values for NoO and NoM based on Dutch Holstein data. Furthermore, a 10-fold cross-validation was carried out to assess the accuracy of pedigree and genomic breeding values for NoO and NoM. For NoO, 40,734 OPU sessions between 1993 and 2015 were analyzed. These OPU sessions originated from 2,543 donors, from which 1,144 were genotyped. For NoM, 35,695 sessions between 1994 and 2015 were analyzed. These MOET sessions originated from 13,868 donors, from which 3,716 were genotyped. Analyses were done using only pedigree information and using a single-step genomic BLUP (ssGBLUP) approach combining genomic information and pedigree information. Heritabilities were very similar based on pedigree information or based on ssGBLUP [i.e., 0.32 (standard error = 0.03) for NoO and 0.21 (standard error = 0.01) for NoM with pedigree, 0.31 (standard error = 0.03) for NoO, and 0.22 (standard error = 0.01) for NoM with ssGBLUP]. For animals without their own information as mimicked in the cross-validation, the accuracy of pedigree-based breeding values was 0.46 for NoO and NoM. The accuracies of genomic breeding values from ssGBLUP were 0.54 for NoO and 0.52 for NoM. These results show that including genomic information increases the accuracies. These moderate accuracies in combination with a large genetic variance show good opportunities for selection of potential bull dams.

Gill infection model for columnaris disease in common carp and rainbow trout.

Challenge models generating gill lesions typical for columnaris disease were developed for the fry of both Common Carp Cyprinus carpio and Rainbow Trout Oncorhynchus mykiss by means of an immersion challenge and Flavobacterium columnare field isolates were characterized regarding virulence. Carp inoculated with highly virulent isolates revealed diffuse, whitish discoloration of the gills affecting all arches, while in trout mostly unilateral focal lesions, which were restricted to the first two gill arches, occurred. Light microscopic examination of the gills of carp exposed to highly virulent isolates revealed a diffuse loss of branchial structures and desquamation and necrosis of gill epithelium with fusion of filaments and lamellae. In severe cases, large parts of the filaments were replaced with necrotic debris entangled with massive clusters of F. columnare bacterial cells enwrapped in an eosinophilic matrix. In trout, histopathologic lesions were similar but less extensive and much more focal, and well delineated from apparently healthy tissue. Scanning and transmission electron microscopic observations of the affected gills showed long, slender bacterial cells contained in an extracellular matrix and in close contact with the destructed gill tissue. This is the first study to reveal gill lesions typical for columnaris disease at macroscopic, light microscopic, and ultrastructural levels in both Common Carp and Rainbow Trout following a challenge with F. columnare. The results provide a basis for research opportunities to examine pathogen-gill interactions.

Assessing the 'system' in safe systems-based road designs: using cognitive work analysis to evaluate intersection designs.

While a safe systems approach has long been acknowledged as the underlying philosophy of contemporary road safety strategies, systemic applications are sparse. This article argues that systems-based methods from the discipline of Ergonomics have a key role to play in road transport design and evaluation. To demonstrate, the Cognitive Work Analysis framework was used to evaluate two road designs - a traditional Melbourne intersection and a cut-through design for future intersections based on road safety safe systems principles. The results demonstrate that, although the cut-through intersection appears different in layout from the traditional intersection, system constraints are not markedly different. Furthermore, the analyses demonstrated that redistribution of constraints in the cut-through intersection resulted in emergent behaviour, which was not anticipated and could prove problematic. Further, based on the lack of understanding of emergent behaviour, similar design induced problems are apparent across both intersections. Specifically, incompatibilities between infrastructure, vehicles and different road users were not dealt with by the proposed design changes. The importance of applying systems methods in the design and evaluation of road transport systems is discussed.

Purification of white spot syndrome virus by iodixanol density gradient centrifugation.

Up to now, only a few brief procedures for purifying white spot syndrome virus (WSSV) have been described. They were mainly based on sucrose, NaBr and CsCl density gradient centrifugation. This work describes for the first time the purification of WSSV through iodixanol density gradients, using virus isolated from infected tissues and haemolymph of Penaeus vannamei (Boone). The purification from tissues included a concentration step by centrifugation (2.5 h at 60,000 g) onto a 50% iodixanol cushion and a purification step by centrifugation (3 h at 80,000 g) through a discontinuous iodixanol gradient (phosphate-buffered saline, 5%, 10%, 15% and 20%). The purification from infected haemolymph enclosed a dialysis step with a membrane of 1,000 kDa (18 h) and a purification step through the earlier iodixanol gradient. The gradients were collected in fractions and analysed. The number of particles, infectivity titre (in vivo), total protein and viral protein content were evaluated. The purification from infected tissues gave WSSV suspensions with a very high infectivity and an acceptable purity, while virus purified from haemolymph had a high infectivity and a very high purity. Additionally, it was observed that WSSV has an unusually low buoyant density and that it is very sensitive to high external pressures.

Luminal uptake of Vibrio (Listonella) anguillarum by shed enterocytes--a novel early defence strategy in larval fish.

As adhesion and translocation through fish gut enterocytes of the pathogen Vibrio (Listonella) anguillarum are not well investigated, the effective cause of disease and mortality outbreaks in larval sea bass, Dicentrarchus labrax, suffering from vibriosis is unknown. We detected V. anguillarum within the gut of experimentally infected gnotobiotic sea bass larvae using transmission electron microscopy and immunogold labelling. Intact bacteria were observed in close contact with the apical brush border in the gut lumen. Enterocytes contained lysosomes positive for protein A-gold particles suggesting intracellular elimination of bacterial fragments. Shed intestinal cells were regularly visualized in the gut lumen in late stages of exposure. Some of the luminal cells showed invagination and putative engulfment of bacterial structures by pseudopod-like formations. The engulfed structures were positive for protein A-colloidal gold indicating that these structures were V. anguillarum. Immunogold positive thread-like structures secreted by V. anguillarum suggested the presence of outer membrane vesicles (MVs) hypothesizing that MVs are potent transporters of active virulence factors to sea bass gut cells suggestive for a substantial role in biofilm formation and pathogenesis. We put forward the hypothesis that MVs are important in the pathogenesis of V. anguillarum in sea bass larvae.

Evaluation of the behavior of murine and human embryonic stem cells in in vitro migration and invasion assays.

Cell migration and invasion are essential processes in a variety of physiological events in the body, but also in several patho-physiological events. In this paper, the behavior of murine and human embryonic stem cells is examined in in vitro migration and invasion models. mESC and hESC were applied as spheroids, also known as patches, and as single cells, to mimic possible cell therapy application strategies. Two known in vitro migration assays, the ECM (extracellular matrix) assay and the Boyden chamber migration assay were selected. These assays revealed that mESC are statistically significantly more infiltrative than hESC. Application as spheroid results in a slightly higher infiltrative capacity compared single cells. The PHF (precultured chick heart fragment) assay was selected as an invasion assay. In the PHF assay a more 3D examination of the infiltrative nature of the ESC can be observed. The mESC showed infiltrative behavior, as spheroids and as single cells. The hESC were infiltrative as single cells but not as spheroids. The results of these assays are mostly complementary and prove the applicability of these assays, which were originally applied in tumor biology, in migratory behavior studies regarding stem cells and their progeny in basic and other conditions.

Platelet gel supernatant as a potential tool to repopulate acellular heart valves.

OBJECTIVE: The aim of this study was to repopulate decellularized heart valve matrices with ovine mesenchymal stem cells (oMSCs) by the use of platelet gel (PG) supernatant, a storage vehicle for growth factors. METHODS: oMSCs were exposed to different concentrations of PG-released supernatant and cell proliferation was evaluated using the MTS assay. oMSC motility and invasiveness were assayed using a Boyden chamber. A quantitative sandwich enzyme immunoassay was used to examine amounts of bFGF and TGF-ß1 in the PG supernatant. Repopulation of acellular heart valve matrices was stimulated by seeding matrices with oMSCs supplemented with the PG supernatant. RESULTS: The most significant increase in proliferation induced by PG supernatant appeared at 1 × 10(5) plts/ml concentration. Higher concentrations evoked reduction of the stimulatory process. oMSC motility was most significantly stimulated at 1 × 10(6) plts/ml. Stimulating invasiveness of oMSCs needed the much higher concentration of 2 × 10(6) plts/ml. Immunoassays revealed that sheep PG supernatant contains 184.8 pg/ml bFGF and 60.5 ng/ml TGF-ß1. Moreover, repopulation of acellular heart valve matrices was significantly enhanced by PG supernatant addition and resulted in upregulation of the myofibroblast marker alpha-smooth muscle actin. CONCLUSIONS: Growth factors released from platelets had the potential to induce cell repopulation in a heart valve tissue engineering procedure, through stimulation of mesenchymal stem-cell migration and invasion.

Double protein-coated poly-ε-caprolactone scaffolds: successful 2D to 3D transfer.

In the past decade, tissue engineering has evolved from a promising technology to an established scientific field. Large attention has focussed on developing scaffolds from both biodegradable and nondegradable polymers to be cultivated with cells, to replace human body defects. The major drawback of most polymers is however their limited cell-interactive properties. An additional complication when developing a surface modification protocol for those materials is the transferability of protocols from 2D substrates to 3D scaffolds. In the present work, we therefore report on possible biological effects originating from the transfer of a double protein coating protocol, involving gelatin type B and fibronectin, from 2D poly-ε-caprolactone (PCL) films to 3D PCL scaffolds produced by rapid prototyping. A variety of techniques including scanning electron microscopy, X-ray photoelectron spectroscopy and confocal fluorescence microscopy confirmed a successful and homogeneous protein-coating on both 2D and 3D substrates. Interestingly, the biological performance of the double protein-coated PCL substrates, reflected by the initial cell adhesion, proliferation, and colonization was superior compared to the other surface modification steps, independent of the material dimension.

Bacterial host interaction of GFP-labelled Vibrio anguillarum HI-610 with gnotobiotic sea bass, Dicentrarchus labrax (L.), larvae.

The location and cell damage caused by Vibrio anguillarum, the causative agent of classical vibriosis, within the developing gut of the newly hatched sea bass, Dicentrarchus labrax (L.), is unknown. A gnotobiotic sea bass model was used to investigate the early interactions of V. anguillarum with sea bass larvae. In the present study, germ-free sea bass larvae were orally exposed to a V. anguillarum HI-610 pathogen labelled with the green fluorescent protein (GFP-HI-610) and sampled at regular intervals. Pathogenic colonization of gut enterocytes was observed 2 h post-exposure (p.e.) and onwards, whereas bacteria within the swim bladder were visualized 48 h p.e and onwards. Ultrastructural findings demonstrated direct bacterial contact with the host cell in the oesophageal mucosa and putative attachment to microvilli of mid- and hindgut enterocytes. The present findings form a starting point for studies assessing the impact of potential candidates (probiotics, prebiotics, antimicrobial peptides) to mitigate bacterial virulence.

Persistent subretinal fluid after surgery for rhegmatogenous retinal detachment: hypothesis and review.

BACKGROUND: Persistent subretinal fluid after rhegmatogenous retinal detachment (RRD) surgery is responsible for delayed recovery, and may affect the final visual outcome. Cause, consequences, and treatment remain elusive. DESIGN: Literature review and case series. METHODS: We reviewed the pathophysiological principles and therapeutic options from the literature, and we report the results from a subretinal fluid cytology study. Nine eyes from nine patients with macula-involving RRD underwent surgical repair. The cellular content of subretinal fluid (SRF) was studied by electron microscopy and anti-rhodopsin immunostaining. All eyes were assessed postoperatively with optical coherence tomography for the detection of persistent submacular fluid (PSF) (Ethics Committee Ghent University Hospital, registration number B6702006169). RESULTS: Certain patient characteristics as well as surgical methods were implicated. PSF appears to occur more frequently in patients with longstanding detachments treated with buckling surgery. Several therapeutic options have been suggested but safety and efficacy remain unclear. We found PSF in three eyes on postoperative OCT scans, which corresponded to the three cell-rich subretinal samples. CONCLUSIONS: PSF after successful RRD repair seems to be related to fluid composition. We hypothesize, in the absence of an effective treatment, that a modified surgical drainage, including a washout of the subretinal space, could evacuate the subretinal fluid more completely, and may prevent this complication.

Expansion of human embryonic stem cells: a comparative study.

OBJECTIVES: Human embryonic stem cells (hESC) are promising for tissue engineering (TE) purposes due to their unique properties. However, current standard mechanical passaging techniques limit rates of possible TE experiments, as it is difficult to obtain high enough numbers of the cells for experimentation. In this study, several dissociative solutions and application methods are tested for their applicability to, and influence on, hESC culture and expansion. MATERIALS AND METHODS: Expansion of two hESC lines, H1 and VUB01, subjected to different passaging techniques, was evaluated. Four dissociative solutions - TrypLE™ Express, Trypsin-EDTA, Cell Dissociation Solution and Accutase™- were combined with two application protocols. As reference conditions, manual and bead-based passaging techniques were used. RESULTS: Results showed that use of Cell Dissociation Solution in combination with a slow adaptation protocol, generated the best expansion profile for both cell lines. The hESC single cell lines remained pluripotent, had good expansion profiles and were capable of differentiation into representatives of all three germ layers. Reproducibility of the results was confirmed by adaptation for three other hESC lines. CONCLUSION: Use of Cell Dissociation Solution, combined with slow adaptation protocol, allows a fast switch from the mechanical passaging technique to a single-cell split technique, generating stable and robust hESC cell lines, which allow for large scale expansion of hESC for TE purposes.

Enhancing bone healing and regeneration: present and future perspectives in veterinary orthopaedics.

Methods currently used to restore bone defects in human and veterinary orthopaedics are often not satisfactory. This is especially the case in the healing of large, irregular defects which result in the formation of tissues with inferior qualities compared to the original structures. For these reasons, several new approaches are currently being explored to improve bone healing capacities in different situations. This review will examine the different techniques used to enhance bone regeneration, highlighting both experimental and clinically applicable methods with regard to veterinary orthopaedics.

Correlation between cryogenic parameters and physico-chemical properties of porous gelatin cryogels.

In the present work, we have performed an in-depth physico-chemical and bio-physical evaluation of a series of previously described porous gelatin scaffolds (S. VanVlierberghe, V. Cnudde, P. Dubruel, B. Masschaele, A. Cosijns, I. DePaepe, P.J.S. Jacobs, L. VanHoorebeke, J.P. Remon and E. Schacht, Biomacromolecules 8, 331 (2007)). All scaffolds were prepared by a cryogenic treatment and subsequent freeze-drying. Three types of scaffolds were prepared by using different gelatin concentrations and cooling protocols. Type-I hydrogels were composed of cone-like pores with decreasing diameter from top (330 microm) to bottom (20-30 microm). Type-II and type-III scaffolds contained spherical pores with an average diameter of 135 (type II) and 65 microm (type III), respectively. The physico-chemical and bio-physical properties studied include the water uptake capacity and kinetics, the mechanical properties and the enzyme-mediated degradation. We can conclude that the pore geometry affects the water uptake capacity, the mechanical properties and the degradation profile of the hydrogels. Type-I hydrogels possess the highest water uptake, the lowest compression modulus and the fastest enzyme mediated degradation, indicating a clear effect of the pore morphology (elongated channels for type I versus spherical pores for types II and III) on the physico-chemical and bio-physical properties of the materials. In contrast to the effect of the pore geometry (channel-like versus spherical), the pore size does not significantly affect the water uptake, the mechanical properties and the enzyme mediated degradation in the investigated pore size range (65-135 microm). To the best of our knowledge, this is the first report in which the effects of a cryogenic treatment on the hydrogel network properties are investigated in such detail.

Immunohistochemical analysis of low-temperature methylmethacrylate resin-embedded goat tissues.

Goats are frequently used as a suitable animal model for tissue engineering. Immunohistochemistry can be helpful in improving the understanding and evaluation of the in vivo tissue responses at a molecular level. Several commercially available antibodies (KI67, vimentin, CD31, core-binding factor alpha-1, osteocalcin, alkaline phosphatase, MAC387, CD3, CD20, CD20cy, CD79 and CD45) were evaluated on Technovit 9100 New embedded goat tissues. Only vimentin, osteocalcin, MAC387 and CD3 revealed positive staining. These antibodies can be routinely used to evaluate goat tissues at molecular level. The use and development of alternative antibodies might further supplement and complete the possibilities for immunohistochemical analysis of goat tissue samples.