Post-traumatic osteoarthritis progression is diminished by early mechanical unloading and anti-inflammatory treatment in mice.

OBJECTIVE: Post-traumatic osteoarthritis (PTOA) is a degenerative joint disease initiated by injury. Early phase (0-7 days) treatments often include rest (unloading) and anti-inflammatory medications, but how those early interventions impact PTOA progression is unknown. We hypothesized that early unloading and anti-inflammatory treatment would diminish joint inflammation and slow PTOA progression. DESIGN: Mice were injured with non-invasive ACL rupture followed by hindlimb unloading (HLU) or normal cage activity (ground control: GC) for 7 days, after which all mice were allowed normal cage activity. HLU and GC mice were treated with daily celecoxib (CXB; 10 mg/kg IP) or vehicle. Protease activity was evaluated using in vivo fluorescence imaging, osteophyte formation and epiphyseal trabecular bone were quantified using micro-computed tomography, and synovitis and articular cartilage were evaluated using whole-joint histology at 7, 14, 21, and 28 days post-injury. RESULTS: HLU significantly reduced protease activity (-22-30% compared to GC) and synovitis (-24-50% relative to GC) at day 7 post-injury (during unloading), but these differences were not maintained at later timepoints. Similarly, trabecular bone volume was partially preserved in HLU mice at during unloading (-14-15% BV/TV for HLU mice, -21-22% for GC mice relative to uninjured), but these differences were not maintained during reloading. Osteophyte volume was reduced by both HLU and CXB, but there was not an additive effect of these treatments (HLU: -46%, CXB: -30%, HLU + CXB: -35% relative to vehicle GC at day 28). CONCLUSIONS: These data suggest that early unloading following joint injury can reduce inflammation and potentially slow PTOA progression.

National expenditure on health research in South Africa: How has the landscape changed in the past decade?

BACKGROUND: Over the past 18 years, the South African (SA) Ministry of Health has committed to allocate 2% of the national health budget to research, while the National Health Research Policy (2001) proposed that the health research budget should be 2% of total public sector health expenditure. A review was conducted by the National Health Research Committee (NHRC) in 2014 to determine whether these goals had been met, using available data up to 2009/10. It revealed that public sector health research funding remained below 2% of the national health budget, supporting the perception of reduced public sector health research funding. OBJECTIVES: To provide an update on the previous review to investigate changes in the health research landscape since 2009/10 and whether goals have been met. METHODS: Various publicly available sources of information on public and private expenditure on health research in SA were used to investigate health research funding and expenditure. In addition, questionnaires were sent to 35 major national and international funders of health research in SA to obtain data on the level of funding provided and the fields of research funded. RESULTS: Total health research expenditure in SA was ZAR6.9 billion in 2016/17, or 19.2% of gross expenditure on research and development, with 1.7% of the ZAR38.6 billion National Department of Health budget from National Treasury being spent on health research through the South African Medical Research Council (ZAR658 million), corresponding to 0.4% of the consolidated government expenditure on health. However, although the total government plus science council spend on health research in 2016/17 was ZAR1.45 billion, this represents just 0.033% of the gross domestic product (GDP), thus remaining well below the aspirational target of 0.15% of the GDP set by the NHRC in 2014. Based on feedback from the funders, the estimated baseline health research funding in 2016/17 was in excess of ZAR4.1 billion, which is considerably higher than many researchers may realise. Three-quarters of this funding originated from foreign sources, suggesting both strengths and opportunities for health research in SA, but also highlighting increasing dependence on foreign funding. Notably, the majority of funders approached were not able to readily break down expenditure according to disease area. CONCLUSIONS: Health research funding has changed significantly since our previous review, although the government's own commitments to it remain unmet. Improved mechanisms to track health research expenditure are urgently required for better alignment of funding priorities and increased co-ordination between science councils in health research funding.

Privacy rights of human research participants in South Africa must be taken seriously.

Letter by Thaldar and Townsend, following an article by the same authors (Thaldar D, Townsend B. Genomic research and privacy: A response to Staunton et al. S Afr Med J 2020;110(3):172-174. https://doi.org/10.7196/SAMJ.2020.v110i3.14431) and both commenting on an article by Staunton et al. (Staunton C, Adams R, Botes M, et al. Safeguarding the future of genomic research in South Africa: Broad consent and the Protection of Personal Information Act No. 4 of 2013. S Afr Med J 2019;109(7):468-470. https://doi.org/10.7196/SAMJ.2019.v109i7.14148); and response to article and letter by Staunton et al.

Osteogenic preconditioning in perfusion bioreactors improves vascularization and bone formation by human bone marrow aspirates.

Cell-derived extracellular matrix (ECM) provides a niche to promote osteogenic differentiation, cell adhesion, survival, and trophic factor secretion. To determine whether osteogenic preconditioning would improve the bone-forming potential of unfractionated bone marrow aspirate (BMA), we perfused cells on ECM-coated scaffolds to generate naïve and preconditioned constructs, respectively. The composition of cells selected from BMA was distinct on each scaffold. Naïve constructs exhibited robust proangiogenic potential in vitro, while preconditioned scaffolds contained more mesenchymal stem/stromal cells (MSCs) and endothelial cells (ECs) and exhibited an osteogenic phenotype. Upon implantation into an orthotopic calvarial defect, BMA-derived ECs were present in vessels in preconditioned implants, resulting in robust perfusion and greater vessel density over the first 14 days compared to naïve implants. After 10 weeks, human ECs and differentiated MSCs were detected in de novo tissues derived from naïve and preconditioned scaffolds. These results demonstrate that bioreactor-based preconditioning augments the bone-forming potential of BMA.

Safeguarding the future of genomic research in South Africa: Broad consent and the Protection of Personal Information Act No. 4 of 2013.

Genomic research has been identified in South Africa (SA) as important in developing a strong bio-economy that has the potential to improve human health, drive job creation and offer potential solutions to the disease burden harboured by low- and middle-income countries. Central to the success of genomic research is the wide sharing of biological samples and data, but the true value of data can only be unlocked if there are laws and policies in place that foster the legal and ethical sharing of genomic data. The introduction and entry into force of SA's Protection of Personal Information Act (POPIA) No. 4 of 2013 is to be welcomed, but the wording of POPIA as it pertains to consent for the processing of personal information for research purposes has sparked a debate about the legal status of broad consent. We argue that a purposive interpretation of the legislation would permit broad consent for the processing of personal information for research. Although there are ongoing debates surrounding the ethical use of broad consent in Africa, the objective of this article is not to engage with the ethics of broad consent itself, but rather to focus on the legal status of broad consent for genomic data sharing under POPIA.

Cutting the cost of South African antiretroviral therapy using newer, safer drugs.

Antiretrovirals are a significant cost driver for HIV programmes. Current first-line regimens have performed well in real-life programmes, but have a low barrier to virological resistance and still carry toxicity that limits adherence. New drug developments may mean that we have access to safer, more robust and cheaper regimens, but only if the appropriate clinical trials are conducted. We briefly discuss these trials, and demonstrate the large cost savings to the South African HIV programme if these are successful.

Key issues in the clinical development and implementation of TB vaccines in South Africa.

Significant progress has been made in advancing the development pipeline for a new and more effective TB vaccine with some candidate vaccines now in late stage clinical evaluation. However, progress has been hampered by an incomplete understanding of the components of a protective immune response and limited animal models, rendering the field unable to reliably predict vaccine efficacy earlier in preclinical development, including by evaluation in animal models, and limiting the predictive utility of comparing immunogenic effects across vaccine candidates in phase I/II studies. Consequently, new candidate vaccines have to be evaluated for efficacy in large-scale phase II/III trials using clinical endpoints. Apart from the technical challenges of characterising TB incidence in target populations at high risk of acquiring TB disease and standardising case definitions in order to improve both the sensitivity and more importantly the specificity of trial endpoints, there is an urgency in expanding and supporting the considerable trial infrastructure that will be required to evaluate and ultimately license a new TB vaccine. In the longer term, implementation strategies are dependent on what policy makers most value. Economic analyses will be essential to guide policy and implementation. This paper outlines the gaps and challenges and identifies solutions for effectively developing and efficiently introducing a new TB vaccine.

Parents' reactions to the diagnosis of cerebral palsy: associations between resolution, age and severity of disability.

BACKGROUND: For parents, receiving a diagnosis, typically in early childhood, that their child has cerebral palsy may conjure up high distress and anxiety. Resolution of these initial reactions may help parents to focus on the challenges and needs of their children. AIMS: of the study were to test whether parents of older children displayed resolution more often than parents of younger children, and whether parents of children with less severe cerebral palsy also showed more resolution. METHOD: Resolution of reactions to diagnosis was assessed with the Reaction to Diagnosis Interview, in a clinic-based sample of 255 parents of children with cerebral palsy aged between 1.4 and 17.3 years. Physicians rated motor ability using the Gross Motor Function Classification System. RESULTS: Overall, the responses of 81.6% of the parents were predominantly indicative of resolution. Unresolved reactions were significantly more often found among parents of younger children and parents of children with more severe motor disabilities. Among parents of teenage children, resolution was more often apparent from a focus on action to better the lives of their children, whereas in parents of younger children, it was more apparent from their focus on constructive thoughts and information seeking. CONCLUSIONS: Given time, the large majority of parents may resolve their reactions to the diagnosis that their child has cerebral palsy. Parents of the most severely affected children may need specific support which, given the age trends, might be aimed at different resolution processes for parents of younger and older children.

The influence of vessel wall elasticity and peripheral resistance on the carotid artery flow wave form: a CFD model compared to in vivo ultrasound measurements.

The Doppler flow wave form and its derived measures such as the pulsatility index provide clinically important tools for the investigation of arterial disease. The typical shape of Doppler flow wave forms is physiologically known to be largely determined by both peripheral resistance and elastic properties of the arterial wall. In the present study we systematically investigate the influence of both vessel wall elasticity and peripheral resistance on the flow wave form obtained from a CFD-simulation of blood flow in the carotid bifurcation. Numerical results are compared to in vivo ultrasound measurements. The in vivo measurement provides a realistic geometry, local elasticities and an input flow wave form for the numerical experiment. Numerical and experimental results are compared at three different sites in the carotid branches. Peripheral resistance has a profoundly decreasing effect on velocities in the external carotid artery. If elasticity is taken into account, the computed peak systolic velocities are considerably lower and a more realistic smoothing of the flow wave form is found. Together, the results indicate that only if both vessel wall elasticity and positive peripheral resistance are taken into account, experimentally obtained Doppler flow wave forms can be reproduced numerically.

Deletion of a coordinate regulator of type 2 cytokine expression in mice.

Mechanisms that underlie the patterning of cytokine expression in T helper (T(H)) cell subsets remain incompletely defined. An evolutionarily conserved approximately 400-bp noncoding sequence in the intergenic region between the genes Il4 and Il13, designated conserved noncoding sequence 1 (CNS-1), was deleted in mice. The capacity to develop T(H)2 cells was compromised in vitro and in vivo in the absence of CNS-1. Despite the profound effect in T cells, mast cells from CNS-1(-/-) mice maintained their capacity to produce interleukin 4. A T cell-specific element critical for the optimal expression of type 2 cytokines may represent the evolution of a regulatory sequence exploited by adaptive immunity.

Active conservation of noncoding sequences revealed by three-way species comparisons.

Human and mouse genomic sequence comparisons are being increasingly used to search for evolutionarily conserved gene regulatory elements. Large-scale human-mouse DNA comparison studies have discovered numerous conserved noncoding sequences of which only a fraction has been functionally investigated A question therefore remains as to whether most of these noncoding sequences are conserved because of functional constraints or are the result of a lack of divergence time.

Experiences of siblings of children with physical disabilities: an empirical investigation.

PURPOSE: This study aimed to investigate the experiences of Dutch siblings of children with physical disabilities (n=43). METHOD: Interviews were conducted to investigate experiences of siblings in 3 areas: the sibling relationship, the relationship with parents, and the relationship with others. The siblings also completed a coping response inventory. RESULTS: The siblings reported difficulties in undertaking activities and in communicating with their brother/sister with a disability. Most siblings worried about the future and the health of their brother/sister with a disability. The siblings reported open communication and trust as the main characteristics of the relationship with their parents. They acknowledged their parents' attempts to treat all children equally, although parents were not always successful in doing so. Having a sibling with a disability did not cause problems in the relationship with friends. The awkward reactions of strangers caused the siblings much annoyance and distress. Siblings in this sample used more approach coping strategies than avoidance strategies, t(34) = 2.37, p < 0.05. CONCLUSION: The brothers and sisters generally did not experience many significant problems, however, a minority of the children did experience problems for which they would like help. In addition, they reported joys as well as problems.

Identification of a coordinate regulator of interleukins 4, 13, and 5 by cross-species sequence comparisons.

Long-range regulatory elements are difficult to discover experimentally; however, they tend to be conserved among mammals, suggesting that cross-species sequence comparisons should identify them. To search for regulatory sequences, we examined about 1 megabase of orthologous human and mouse sequences for conserved noncoding elements with greater than or equal to 70% identity over at least 100 base pairs. Ninety noncoding sequences meeting these criteria were discovered, and the analysis of 15 of these elements found that about 70% were conserved across mammals. Characterization of the largest element in yeast artificial chromosome transgenic mice revealed it to be a coordinate regulator of three genes, interleukin-4, interleukin-13, and interleukin-5, spread over 120 kilobases.

Electron microscopic study of human sperm membrane isolation.

OBJECT: Our purpose was to isolated pure, homogeneous human sperm membranes, free of cellular contaminants. METHODS: Donor semen samples collected after masturbation were stored at -70 degrees C and eventually pooled. Each attempt at sperm membrane isolation required 800 x 10(6) spermatozoa which were sonicated by ultrasound (40% output; Vibra Cell). The effect of sonication time (3 x 5, 3 x 15, and 180 sec) on membrane isolation was investigated. Sonicated samples were centrifuged (500 g, 5 min) and the supernatant was pipetted off. The supernatant of the centrifuged sample was layered on either a sucrose cushion (supernatant on 1.6 M sucrose) or a discontinuous sucrose gradient and centrifuged (100,000 g, 1 hr). Contents of supernatants of sonicated samples and fractions (sucrose interfaces) were then fixed in 1.0% tannic acid and 2.5% buffered glutaraldehyde and examined electron microscopically using standard procedures. RESULTS: (1) The optimal sonification time was found to be 3 x 15 sec. (2) Membrane isolation using a sucrose cushion was found to be inadequate, showing significant cellular contamination. (3) Sperm membrane isolation from the sucrose interface between 0.75 and 1.05 M sucrose was found to be most effective. CONCLUSION: The advantage of this method is its simplicity. The drawback of this method is the large number of spermatozoa required for membrane purification.

Ultrastructure of the cardiac and pyloric glands of the gastric mucosa of the South African hedgehog, Atelerix frontalis.

The cardiac and pyloric glands in the gastric mucosa of the South African hedgehog, Atelerix frontalis, are described. The cardiac area of the stomach contains proper cardiac glands and lacks undifferentiated fundic glands. The cardiac glands are simple tubular, coiled, and lined with columnar cells ultrastructurally similar to those of the gastric surface epithelium. Secretory granules with varying electron densities fill the apical cytoplasm of these cells. In contrast to other mammals, these glands lack mucous neck cells. The neck of the pyloric glands contains only a single cell type, whereas the basal regions of these glands contain "light" and "dark" cells. The secretory granules in the "dark" cells and the pyloric neck cells have a moderate electron density and often contain an electron dense core. An electron-lucent cytoplasm with numerous polysomes is characteristic of the "light" cells. Some "light" cells contain electron-dense granules in the apical cytoplasm. The presence of only neutral mucins in the cardiac gland cells denotes the absence of mucous neck cells. The acidic mucins within the pyloric neck cells seem to indicate that these cells are mucous neck cells, whereas the neutral mucins within the basally located pyloric gland cells show at least a partial functional difference from the pyloric neck cells. © 1993 Wiley-Liss, Inc.

Microcirculation of the rat omentum studied by means of corrosion casts.

A study of the microvasculature of the omentum using corrosion casts was undertaken. The object was to supply information regarding the morphology of the three-dimensional structure of the microvasculature, information which may be valuable in understanding the functions of the omentum. Corrosion casts of rat omenta were prepared and studied. Characteristic glomerular-like capillary beds were identified. Some of these beds were densely and others loosely arranged. Two types of capillaries made up these beds, tortuous and straight capillaries. These beds also showed a planate surface giving the impression that they lie directly under the mesothelium.

Early stages of ciliogenesis in the respiratory epithelium of the nasal cavity of rabbit embryos.

Previous studies have shown that ciliogenesis in the epithelial cells of various species exhibits similarities as well as differences. In an attempt to establish whether this process is identical in epithelial cells of a single species, early stages of centriole formation not previously described were encountered. Ciliogenesis was investigated in the respiratory epithelium of the nasal cavity of 18 to 23-day-old rabbit embryos. The appearance of groups of deuterosomes and fibrous granules is followed by the radial formation of procentrioles around the deuterosomes and parent centrioles. The majority of the procentrioles, forming acentriolarly, occur in pairs, with their distal ends facing each other, between the deuterosomes. Subsequent growth of these procentrioles between deuterosomes in a group results in separating the deuterosomes from one another. The deuterosomes, however, still remain interconnected by means of the growing procentrioles. Accordingly, long chains and networks consisting of the above-mentioned structures are formed. After the procentrioles have attained a certain length, the chains and networks split into separate deuterosome-procentriole complexes. During these earlier stages of ciliogenesis fibrous granules are present, however, their function is yet to be determined.

[A multidisciplinary approach to the management of deaf infants]. / Een multidisciplinaire aanpak in het begeleiden van dove baby's.

An important epistemological change characterizes the education of hearing impaired and profoundly deaf children. The integration into ordinary schools and early home guidance are manifestations of this change. Early home guidance has been recognized as a promising new training approach for hearing impaired children. One breaks through the usual confinement in a specialized institution and strict individually based guidance. The family of the hearing impaired baby is directly involved in its early training. The authors have followed this new approach for several years now and explain their daily experience. An integrated training model is outlined in this article.