Objectives Enteric neuropathies are severe gastrointestinal disorders with unsatisfactory outcomes. We aimed to investigate the potential of enteric neural stem cell therapy approaches for such disorders by transplanting mouse enteric neural crest cells (ENCCs) into ganglionic and aganglionic mouse gut in vivo and analysing functional integration and long-term safety. Design Neurospheres generated from yellow fluorescent protein (YFP) expressing ENCCs selected from postnatal Wnt1-cre;R26R-YFP/YFP murine gut were transplanted into ganglionic hindgut of wild-type littermates or aganglionic hindgut of Ednrbtm1Ywa mice (lacking functional endothelin receptor type-B). Intestines were then assessed for ENCC integration and differentiation using immunohistochemistry, cell function using calcium imaging, and long-term safety using PCR to detect off-target YFP expression. Results YFP+ ENCCs engrafted, proliferated and differentiated into enteric neurons and glia within recipient ganglionic gut. Transplanted cells and their projections spread along the endogenous myenteric plexus to form branching networks. Electrical point stimulation of endogenous nerve fibres resulted in calcium transients (F/F0 = 1.16±0.01;43 cells, n = 6) in YFP+ transplanted ENCCs (abolished with TTX). Long-term follow-up (24 months) showed transplanted ENCCs did not give rise to tumours or spread to other organs (PCR negative in extraintestinal sites). In aganglionic gut ENCCs similarly spread and differentiated to form neuronal and glial networks with projections closely associated with endogenous neural networks of the transition zone. Conclusions Transplanted ENCCs successfully engrafted into recipient ganglionic and aganglionic gut showing appropriate spread, localisation and, importantly, functional integration without any long-term safety issues. This study provides key support for the development and use of enteric neural stem cell therapies.
Abstract Background Further investigation of confirmed UTI in children aims to prevent renal scarring and future complications. Methods We conducted a systematic review to determine the most effective approach to the further investigation of confirmed urinary tract infection (UTI) in children under five years of age. Results 73 studies were included. Many studies had methodological limitations or were poorly reported. Effectiveness of further investigations: One study found that routine imaging did not lead to a reduction in recurrent UTIs or renal scarring. Diagnostic accuracy: The studies do not support the use of less invasive tests such as ultrasound as an alternative to renal scintigraphy, either to rule out infection of the upper urinary tract (LR- = 0.57, 95%CI: 0.47, 0.68) and thus to exclude patients from further investigation or to detect renal scarring (LR+ = 3.5, 95% CI: 2.5, 4.8). None of the tests investigated can accurately predict the development of renal scarring. The available evidence supports the consideration of contrast-enhanced ultrasound techniques for detecting vesico-ureteric reflux (VUR), as an alternative to micturating cystourethrography (MCUG) (LR+ = 14.1, 95% CI: 9.5, 20.8; LR- = 0.20, 95%CI: 0.13, 0.29); these techniques have the advantage of not requiring exposure to ionising radiation. Conclusion There is no evidence to support the clinical effectiveness of routine investigation of children with confirmed UTI. Primary research on the effectiveness, in terms of improved patient outcome, of testing at all stages in the investigation of confirmed urinary tract infection is urgently required.
Abstract Background Four-weekly intramuscular (IM) benzathine penicillin G (BPG) injections to prevent acute rheumatic fever (ARF) progression have remained unchanged since 1955. A Phase-I trial in healthy volunteers demonstrated the safety and tolerability of high-dose S ub C utaneous Infusions of B P G (SCIP) which resulted in a much longer effective penicillin exposure, and fewer injections. Here we describe the experiences of young people living with ARF participating in a Phase-II SCIP trial. Methodology Participants (n=20) attended a clinic in Wellington, New Zealand (NZ). After a physical examination, participants received 2% lignocaine followed by 13.8mL (6 vials) to 20.7mL (9 vials) of BPG (Bicillin-LA ® ; determined by weight), into the abdominal subcutaneous tissue. Semi-structured interviews and observations were taken during and after the infusion, as well as on days 28 and 70. All interviews were recorded, transcribed verbatim, and thematically analysed. Principal Findings Low levels of pain were reported on needle insertion, during and following the infusion. Some participants experienced discomfort and bruising on days one and two post dose; however, the pain was reported to be less severe than their usual IM BPG. Participants were ‘relieved’ to only need injections quarterly and the overwhelming majority preferred to continue with SCIP. Conclusions Participants preferred SCIP over their usual regimen, reporting less pain and a preference for the longer time gap between treatments. Recommending SCIP as standard of care for most patients needing long-term ARF/RHD prophylaxis has the potential to transform secondary prophylaxis of ARF/RHD in NZ and globally. Synopsis Acute rheumatic fever (ARF) is a preventable inflammatory disease that occurs as a delayed sequelae to group A streptococcus (GAS) infection. ARF and its complication rheumatic heart disease (RHD) have significant negative effects on health, often resulting in chronic illness and premature death. For 70 years, the only proven way to prevent ARF progression has been benzathine penicillin G (BPG), given as a monthly intramuscular (IM) injection for a minimum of 10 years. The effectiveness of this approach is limited by pain and the frequency of injection which leads to suboptimal adherence. There is an urgent need to improve penicillin formulations for all children living with ARF and RHD. Here we describe the experiences of 20 young people living with ARF participating in a Phase-II trial delivering high-dose S ub C utaneous Infusions of P enicillin (SCIP) in order to provide longer effective penicillin exposure, and therefore fewer injections. Participants in the trial overwhelmingly preferred high-dose SCIP over their usual monthly IM penicillin regimen, reporting less pain and a preference for the longer time gap (28 versus 70 days) between treatments. Reducing injection frequency from 13 to four-or-five per year, may improve adherence and reduce disease progression. Offering widespread SCIP to ARF/RHD patients to evaluate long-term adherence, preferences and disease progression has the potential to transform secondary prophylaxis of ARF/RHD both in New Zealand and globally.
The use of evolutionary methods for the optimisation of engineering systems is reviewed and a number of the most common methods are compared in terms of their philosophical basis and implementation. The use of these approaches, including genetic algorithms, evolutionary programming, evolutionary strategies, simulated annealing and population-based incremental learning, are illustrated using the shape optimisation problem of a steel plate with buckling and stress constraints. The solutions obtained using the various methods are compared. (10 pages)
Care management is the next step from primary nursing, sayJulie CooperandPatrick Mitchell, who describe their outcomes-oriented initiative for developing nurses’ roles
John Goodwill and Anne Chamberlain briefly but successfully cover just about every aspect of care required by a patient undergoing rehabilitation. They cover Doth inpatient and outpatient care and include community chapters written by all members of the team.
The prospect of using neural cell replacement for the treatment of severe enteric neuropathies has seen significant progress in the last decade. The ability to harvest and transplant enteric neural crest cells (ENCCs) that functionally integrate within recipient intestine has recently been confirmed by in vivo murine studies. Although similar cells can be harvested from human fetal and postnatal gut, no studies have as yet verified their functional viability upon in vivo transplantation. We sought to determine whether ENCCs harvested from human fetal bowel are capable of engraftment and functional integration within recipient intestine following in vivo transplantation into postnatal murine colon. Enteric neural crest cells selected and harvested from fetal human gut using the neurotrophin receptor p75NTR were lentivirally labeled with either GFP or calcium-sensitive GCaMP and transplanted into the hindgut of Rag2- /γc- /C5- -immunodeficient mice at postnatal day 21. Transplanted intestines were assessed immunohistochemically for engraftment and differentiation of donor cells. Functional viability and integration with host neuromusculature was assessed using calcium imaging. Transplanted human fetal gut-derived ENCC showed engraftment within the recipient postnatal colon in 8/15 mice (53.3%). At 4 weeks posttransplantation, donor cells had spread from the site of transplantation and extended projections over distances of 1.2 ± 0.6 mm (n = 5), and differentiated into enteric nervous system (ENS) appropriate neurons and glia. These cells formed branching networks located with the myenteric plexus. Calcium transients (change in intensity F/F0 = 1.25 ± 0.03; 15 cells) were recorded in transplanted cells upon stimulation of the recipient endogenous ENS demonstrating their viability and establishment of functional connections.
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