A case study: Glycosaminoglycan profiles of autologous chondrocyte implantation (ACI) tissue improves as the tissue matures

Background Autologous chondrocyte implantation (ACI) has been used to treat cartilage defects in thousands of patients worldwide with good clinical effectiveness 10–20 years after implantation. Information concerning the quality of the repair cartilage is still limited because biopsies are small and rare. Glycosaminoglycan structure influences physiological function and is likely to be important in the long term stability of the repair tissue. The aim of this study was to assess glycosaminoglycans in ACI tissue over a two year period. Methods Biopsies were taken from one patient (25 years old) at 12 months and 20 months post-ACI-treatment and from three normal cadavers (21, 22 and 25 years old). Fluorophore-assisted carbohydrate electrophoresis (FACE) was used to quantitatively assess the individual glycosaminoglycans. Results At 12 months the ACI biopsy had 40% less hyaluronan than the age-matched cadaveric biopsies but by 20 months the ACI biopsy had the same amount of hyaluronan as the controls. Both the 12 and 20 month ACI biopsies had less chondroitin sulphate disaccharides and shorter chondroitin sulphate chains than the age-matched cadaveric biopsies. However, chondroitin sulphate chain length doubled as the ACI repair tissue matured at 12 months (3913 Da ± 464) and 20 months (6923 Da ± 711) and there was less keratan sulphate as compared to the controls. Conclusions Although the glycosaminoglycan composition of the repair tissue is not identical to mature articular cartilage its quality continues to improve with time

Uplift histories of Africa and Australia from linear inverse modeling of drainage inventories

We describe and apply a linear inverse model which calculates spatial and temporal patterns of uplift rate by minimizing the misfit between inventories of observed and predicted longitudinal river profiles. Our approach builds upon a more general, non-linear, optimization model, which suggests that shapes of river profiles are dominantly controlled by upstream advec- tion of kinematic waves of incision produced by spatial and temporal changes in regional uplift rate. Here, we use the method of characteristics to solve a version of this problem. A damped, non-negative, least squares approach is developed that permits river profiles to be inverted as a function of up- lift rate. An important benefit of a linearized treatment is low computational cost. We have tested our algorithm by inverting 957 river profiles from both Africa and Australia. For each continent, the drainage network was constructed from a digital elevation model. The fidelity of river profiles extracted from this network was carefully checked using satellite imagery. River profiles were inverted many times to systematically investigate the trade-off between model misfit and smoothness. Spatial and temporal patterns of both uplift rate and cumulative uplift were calibrated using independent geologic and geophys- ical observations. Uplift patterns suggest that the topography of Africa and Australia grew in Cenozoic times. Inverse modeling of large inventories of river profiles demonstrates that drainage networks contain coherent signals that record the regional growth of elevation.This is the final version. It first appeared at http://onlinelibrary.wiley.com/wol1/doi/10.1002/2014JF003297/abstract

Can auditory objects be subitized?

In vision, humans have the ability to mentally 'tag' approximately four objects, allowing us to monitor, attend, and interact with them. As a consequence, we can rapidly and accurately enumerate up to four objects – a process known as subitizing. Here, we investigate whether a similar ability exists for tagging auditory stimuli and find that only two or three auditory stimuli can be enumerated with high accuracy. We assess whether this high accuracy indicates the existence of an auditory subitizing mechanism, and if it is influenced by factors known to influence visual subitizing. Based on accuracy, Experiments 1 and 2 reveal a potential auditory subitizing mechanism only when stimuli are spatially separated, as is the case for visual subitizing. Experiment 3 failed to show any evidence of auditory subitizing when objects were separated in time, rather than space. All three experiments provide only limited evidence for an age-related decline in auditory enumeration of small numbers of objects. This suggests that poor auditory tagging does not contribute significantly to older adults’ difficulties in multi-talker conversations. We hypothesize that although auditory subitizing might occur, it is restricted to approximately two spatially-separated objects due to the difficulty of parsing the auditory scene into its constituent parts

Old and New Targets in Antimalarial Drug Discovery

The increasing emergence of resistance to commonly used therapies has placed a huge strain on the prevention and control of malaria; therefore, there is an urgent need to develop novel antimalarial agents. The aim of this research was to design and synthesise a library of potent antimalarial compounds, with desirable pharmacokinetic profiles, in order to identify a drug candidate suitable for preclinical development. This research was divided into two main sections: x The synthesis of compounds deigned to inhibit IspD, a novel target in antimalarial drug discovery x The late stage development of a series of endoperoxide-based antimalarials, which are derived from the structure of artemisinin A library of benzisothiazolinone compounds was generated to target the IspD enzyme. Many of these compounds displayed low micromolar inhibitory activity against both enzymatic and phenotypic assays in vitro and an investigation into structure-activity relationships around the core of these benzisothiazolinones was also conducted. The most potent compound to emerge, a CH2 linked benzisoselenazolone, had an IC50 of 0.17 μM against PfIspD and 5.54 μM against Pf3D7. These compounds represent a novel class of IspD inhibitor, which have the potential for further development as antimalarial agents. A number of 1,2,4,5-tetraoxane analogues were also prepared in order to develop an antimalarial agent suitable for a single-dose cure. The most potent analogue, N205, had an IC50 of 1.3 nM and an average mouse survival of 26.3 days (66% cure rate) following a single dose. A less than optimal stability profile for N205 led to the further development of another potent tetraoxane analogue, E209. Optimisation of the synthetic pathway led to the generation of E209 in a series of five high-yielding steps that are suitable for large-scale production. E209 represents the first 1,2,4,5-tetraoxane that is comparable, in terms of both efficacy and PK/PD profiles, to OZ439, and is a candidate for pre-clinical development

Abnormal loading and functional deficits are present in both limbs before and after unilateral knee arthroplasty

Abstract Unilateral knee replacement is often followed by a contralateral replacement in time and the biomechanics of the other knee before and after knee replacement remains poorly understood. The aim of this paper is to distinguish the features of arthritic gait in the affected and unaffected legs relative to a normal population and to assess the objective recovery of gait function post-operatively, with the aim of defining patients at risk of poor post-operative function. Twenty patients with severe knee OA but no pain or deformity in any other lower limb joint were compared to twenty healthy subjects of the same age. Gait analysis was performed and quadriceps and hamstrings co-contraction was measured. Fifteen subjects returned 1 year following knee arthroplasty. Moments and impulses were calculated, principal component analysis was used to analyse the waveforms and a classification technique (the Cardiff Classifier) was used to select the most discriminant data and define functional performance. Comparing pre-operative function to healthy function, classification accuracies for the affected and unaffected knees were 95% and 92.5% respectively. Post-operatively, the affected limb returned to the normal half of the classifier in 8 patients, and 7 of those patients returned to normal function in the unaffected limb. Recovery of normal gait could be correctly predicted 13 out of 15 times at the affected knee, and 12 out of 15 times at the unaffected knee based on pre-operative gait function. Focused rehabilitation prior to surgery may be beneficial to optimise outcomes and protect the other joints following knee arthroplasty

An In Vitro System to Study the Effect of Subchondral Bone Health on Articular Cartilage Repair in Humans.

Chondrocyte-based cartilage repair strategies, such as articular chondrocyte implantation, are widely used, but few studies addressed the communication between native subchondral bone cells and the transplanted chondrocytes. An indirect co-culture model was developed, representing a chondrocyte/scaffold-construct repair of a cartilage defect adjoining bone, where the bone could have varying degrees of degeneration. Human BM-MSCs were isolated from two areas of subchondral bone in each of five osteochondral tissue specimens from five patients undergoing knee arthroplasty. These two areas underlaid the macroscopically and histologically best and worst cartilage, representing early and late-stage OA, respectively. BM-MSCs were co-cultured with normal chondrocytes suspended in agarose, with the two cell types separated by a porous membrane. After 0, 7, 14 and 21 days, chondrocyte-agarose scaffolds were assessed by gene expression and biochemical analyses, and the abundance of selected proteins in conditioned media was assessed by ELISA. Co-culture with late-OA BM-MSCs resulted in a reduction in GAG deposition and a decreased expression of genes encoding matrix-specific proteins (COL2A1 and ACAN), compared to culturing with early OA BM-MSCs. The concentration of TGF-β1 was significantly higher in the early OA conditioned media. The results of this study have clinical implications for cartilage repair, suggesting that the health of the subchondral bone may influence the outcomes of chondrocyte-based repair strategies

The role of pharmacists in caring for young people with chronic illness

PURPOSE: To explore the perceived and potential roles of pharmacists in the care of young people aged 10–24 years with chronic illness, through the exemplar of juvenile arthritis, from the perspectives of UK community and hospital pharmacists, health service commissioners, rheumatology health professionals, and lay advocates. METHODS: A sequential mixed methods study design comprises the following: focus groups with community and hospital pharmacists; telephone interviews with pharmacy and rheumatology stakeholders and commissioners; and multidisciplinary group discussions to prioritize roles generated by the first two qualitative phases. RESULTS: The high priority roles for pharmacists, identified by pharmacists and rheumatology staff, were developing generic health care skills among young people; transferring information effectively across care interfaces; building trusting relationships with young people; helping young people to find credible online health information; and the need to develop specialist expertise. Participants identified associated challenges for pharmacists in supporting young people with chronic illness. These challenges included parents collecting prescription refills alone, thus reducing opportunities to engage, and pharmacist isolation from the wider health care team. CONCLUSIONS: This study has led to the identification of specific enhancements to pharmacy services for young people, which have received the endorsement of a wide range of stakeholders. These suggestions could inform the next steps in developing the contribution of community and hospital pharmacy to support young people with chronic illness in the optimal use of their medication

Novel ulcerative leg lesions in yearling lambs: Clinical features, microbiology and histopathology

Here we report an outbreak of an atypical, ulcerative dermatitis in North Country mule lambs, located in South Gloucestershire, UK. The lesions, which appeared to be contagious, occured between the coronary band and the carpal joint as a focal, well demarcated, circular, ulcerative dermatitis. Histopathological examination of the lesion biopsies revealed areas of ulceration, epidermal hyperplasia, suppurative dermatitis and granulation tissue. Clumped keratohyalin granules and intracellular keratinocyte oedema (ballooning degeneration) were evident within lesion biopsies, consistent with an underlying viral aetiology. A PCR-based microbiological investigation failed to detect bovine digital dermatitis-associated treponeme phylogroups, Dichelobacter nodosus, Staphylococcus aureus, Dermatophilus congolensis or Chordopoxvirinae virus DNA. However, 3 of the 10 (30 %) and 6 of 10 (60 %) lesion samples were positive for Fusobacterium necrophorum and Streptococcus dysgalactiae DNA, respectively. Contralateral limb swabs were negative by all standard PCR assays. To better define the involvement of F. necrophorum in the aetiology of these lesions, a qPCR targeting the rpoB gene was employed and confirmed the presence of F. necrophorum DNA in both the control and lesions swab samples, although the mean F. necrophorum genome copy number detected in the lesion swab samples was ∼19-fold higher than detected in the contralateral control swab samples (245 versus 4752 genome copies/μl, respectively; P < 0.001). Although we have not been able to conclusively define an aetiological agent, the presence of both F. necrophorum and S. dysgalactiae in the majority of lesions assayed supports their role in the aetiopathogenesis of these lesions

Two-week dietary soy supplementation has an estrogenic effect on normal premenopausal breast.

An association has been reported between consumption of a high soy diet and a low incidence of breast cancer within populations of Southeast Asia. Phytoestrogens present in soy act as partial estrogen agonists or antagonists and can inhibit breast cancer cell proliferation in vitro. The effect of 14-day dietary soy supplementation with 60 g (45 mg isoflavones) on the normal breast of 84 premenopausal patients was determined. Serum concentrations of the isoflavanoids, genistein, daidzein, and equol, were raised in patients after soy supplementation (P < or = 0.025). Nipple aspirate (NA) levels of genistein and daidzein were higher than paired serum levels, both before (P < 0.001 and P = 0.001, respectively) and after soy supplementation (P < 0.001 and P = 0.049, respectively); however, there was no significant increase in NA isoflavone levels in response to soy. NA levels of apolipoprotein D were significantly lowered and pS2 levels raised in response to soy supplementation (P < or = 0.002), indicative of an estrogenic stimulus. No effect of soy supplementation on breast epithelial cell proliferation, estrogen and progesterone receptor status, apoptosis, mitosis, or Bcl-2 expression was detected. In conclusion, short term dietary soy has a weak estrogenic response on the breast, as measured by nipple aspirate apolipoprotein D and pS2 expression. No antiestrogenic effect of soy on the breast was detected

Large-Scale Tectonic Forcing of the African Landscape

Abstract: Successful inverse modeling of observed longitudinal river profiles suggests that fluvial landscapes are responsive to continent‐wide tectonic forcing. However, inversion algorithms make simplifying assumptions about landscape erodibility and drainage planform stability that require careful justification. For example, precipitation rate and drainage catchment area are usually assumed to be invariant. Here, we exploit a closed‐loop modeling strategy by inverting drainage networks generated by dynamic landscape simulations in order to investigate the validity of these assumptions. First, we invert 4,018 African river profiles to determine an uplift history that is independently calibrated, and subsequently validated, using separate suites of geologic observations. Second, we use this tectonic forcing to drive landscape simulations that permit divide migration, interfluvial erosion and changes in catchment size. These simulations reproduce large‐scale features of the African landscape, including growth of deltaic deposits. Third, the influence of variable precipitation is investigated by carrying out a series of increasingly severe tests. Inverse modeling of drainage inventories extracted from simulated landscapes can largely recover tectonic forcing. Our closed‐loop modeling strategy suggests that large‐scale tectonic forcing plays the primary role in landscape evolution. One corollary of the integrative solution of the stream‐power equation is that precipitation rate becomes influential only if it varies on time scales longer than ∼1 Ma. We conclude that calibrated inverse modeling of river profiles is a fruitful method for investigating landscape evolution and for testing source‐to‐sink models