Neonatal umbilical cord blood transplantation halts skeletal disease progression in the murine model of MPS-I

Umbilical cord blood (UCB) is a promising source of stem cells to use in early haematopoietic stem cell transplantation (HSCT) approaches for several genetic diseases that can be diagnosed at birth. Mucopolysaccharidosis type I (MPS-I) is a progressive multi-system disorder caused by deficiency of lysosomal enzyme α-L-iduronidase, and patients treated with allogeneic HSCT at the onset have improved outcome, suggesting to administer such therapy as early as possible. Given that the best characterized MPS-I murine model is an immunocompetent mouse, we here developed a transplantation system based on murine UCB. With the final aim of testing the therapeutic efficacy of UCB in MPS-I mice transplanted at birth, we first defined the features of murine UCB cells and demonstrated that they are capable of multi-lineage haematopoietic repopulation of myeloablated adult mice similarly to bone marrow cells. We then assessed the effectiveness of murine UCB cells transplantation in busulfan-conditioned newborn MPS-I mice. Twenty weeks after treatment, iduronidase activity was increased in visceral organs of MPS-I animals, glycosaminoglycans storage was reduced, and skeletal phenotype was ameliorated. This study explores a potential therapy for MPS-I at a very early stage in life and represents a novel model to test UCB-based transplantation approaches for various diseases

INTELLIGENT TECHNIQUES FOR HANDLING UNCERTAINTY IN THE ASSESSMENT OF NEONATAL OUTCOME

Objective assessment of the neonatal outcome of labour is important, but it is a difficult and challenging problem. It is an invaluable source of information which can be used to provide feedback to clinicians, to audit a unit's overall performance, and can guide subsequent neonatal care. Current methods are inadequate as they fail to distinguish damage that occurred during labour from damage that occurred before or after labour. Analysis of the chemical acid-base status of blood taken from the umbilical cord of an infant immediately after delivery provides information on any damage suffered by the infant due to lack of oxygen during labour. However, this process is complex and error prone, and requires expertise which is not always available on labour wards. A model of clinical expertise required for the accurate interpretation of umbilical acid-base status was developed, and encapsulated in a rule-based expert system. This expert system checks results to ensure their consistency, identifies whether the results come from arterial or venous vessels, and then produces an interpretation of their meaning. This 'crisp' expert system was validated, verified and commercially released, and has since been installed at twenty two hospitals all around the United Kingdom. The assessment of umbilical acid-base status is characterised by uncertainty in both the basic data and the knowledge required for its interpretation. Fuzzy logic provides a technique for representing both these forms of uncertainty in a single framework. A 'preliminary' fuzzy-logic based expert system to interpret error-free results was developed, based on the knowledge embedded in the crisp expert system. Its performance was compared against clinicians in a validation test, but initially its performance was found to be poor in comparison with the clinicians and inferior to the crisp expert system. An automatic tuning algorithm was developed to modify the behaviour of the fuzzy model utilised in the expert system. Sub-normal membership functions were used to weight terms in the fuzzy expert system in a novel manner. This resulted in an improvement in the performance of the fuzzy expert system to a level comparable to the clinicians, and superior to the crisp expert system. Experimental work was carried out to evaluate the imprecision in umbilical cord acid-base parameters. This information, in conjunction with fresh knowledge elicitation sessions, allowed the creation of a more comprehensive fuzzy expert system, to validate and interpret all acid-base data. This 'integrated' fuzzy expert system was tuned using the comparison data obtained previously, and incorporated vessel identification rules and interpretation rules, with numeric and linguistic outputs for each. The performance of each of the outputs was evaluated in a rigorous validation study. This demonstrated excellent agreement with the experts for the numeric outputs, and agreement on a par with the experts for the linguistic outputs. The numeric interpretation produced by the fuzzy expert system is a novel single dimensional measure that accurately represents the severity of acid-base results. The development of the crisp and fuzzy expert systems represents a major achievement and constitutes a significant contribution to the assessment of neonatal outcome.Plymouth Postgraduate Medical Schoo

Standardization as situation-specific achievement: regulatory diversity and the production of value in intercontinental collaborations in stem cell medicine

The article examines the role and challenges of scientific self-governance and standardization in inter-continental clinical research partnerships in stem cell medicine. The paper shows that – due to a high level of regulatory diversity – the enactment of internationally recognized standards in multi-country stem cell trials is a complex and highly situation-specific achievement. Standardization is imposed on a background of regulatory, institutional and epistemic-cultural heterogeneity, and implemented exclusively in the context of select clinical projects. Based on ethnographic data from the first trans-continental clinical trial infrastructure in stem cell medicine between China and the USA, the article demonstrates that locally evolved and international forms of experimental clinical research practices often co-exist in the same medical institutions. Researchers switch back and forth between these schemas, depending on the purposes of their research, the partners they work with, the geographic scale of research projects, and the contrasting demands for regulatory review, that result from these differences. Drawing on Birch’s analysis of the role of standardization in international forms of capital production in the biosciences, the article argues that the integration of local knowledge institutions into the global bioeconomy does not necessarily result in the shutting down of localized forms of value production. In emerging fields of medical research, that are regulated in highly divergent ways across geographical regions, the coexistence of distinct modes of clinical translation allows also for the production of multiple forms of economic value, at varying spatial scales. This is especially so in countries with lenient regulations. As this paper shows, the long-standing absence of a regulatory framework for clinical stem cell applications in China, permits the situation-specific adoption of internationally recognized standards in some contexts, while enabling the continuation of localized forms of value production in others

Correlation of Ultrasonographic and Pathophysiologic Measurements of Umbilical Vessels in Gestational Diabetes

Aim: The resistance of placental blood vessels might be increased in diabetic pregnancies. This increased resistance can affect uteroplacental blood flow and influence the oxygen and nutrient supply of the fetus and fetal growth. Our aim was to compare the ultrasonographic, pathomorphologic data and vasoreactivity of umbilical and placental vessels of gestational diabetic newborns with that of normal pregnancy newborns. Methods: In this case-control study the placental vascularization of pregnant women was determined by 3D power Doppler ultrasound technique. We calculated the vascularization index (VI), flow index (FI) and vascularization flow index (VFI). We performed a tissue bath experiment (treatment with oxytocin and desmopressin) on umbilical vessels and collected pathomorphologic data according to the Royal College of Pathologists’ 2011 protocol. Results: The placental vascularization index and the umbilical artery S/D were significantly lower in the case group. The mean VI was 6.21% (±2.69 SD) in control versus 3.5% (±2.97 SD) (p<0.05) in GDM. The mean value of the umbilical artery S/D was 2.27 (±0.22 SD) and 2.18 (±0.45 SD) (p<0.05) respectively. In an isolated tissue bath experiment, oxytocin and desmopressin did not elicit significant contraction on umbilical cord vessels. Conclusion: Our results suggest that umbilical vessels might have a different receptor pathway regulation that can compensate for the rheological changes in the pregnant woman’s blood flow and gives opportunity for selective therapy to fetuses more vulnerable to hypoxia

Metabolomic and Proteomic Analysis of the Mesenchymal Stem Cells’ Secretome

Mesenchymal stem cells (MSCs) are multipotent stromal cells with a strong potential in human regenerative medicine due to their ability to renew themselves and differentiate into various specialized cell types under certain physiological or experimental conditions. MSCs secrete a broad spectrum of autocrine and paracrine factors (MSCs’ secretome) that could exert significant effects on cells in their vicinity. MSCs have been clinically tested and have displayed a great potential in the treatment of bone/cartilage fractures and disorders, diabetes, cardiovascular diseases and immune, neurodegenerative and inflammatory diseases. The therapeutic efficacy of MSCs was initially attributed to their multipotent character and ability to engraft and differentiate at the site of injury. However, in recent years, it has been revealed that either undifferentiated or differentiated MSCs’ secretome plays an important role in the therapeutic potential of MSCs. The deciphering of the composition of MSCs’ secretome through proteomic and metabolic analyses and implementation of certain advanced analytical (nuclear magnetic resonance (NMR) spectroscopy, mass spectrometry (MS), chromatography, etc.) and immunological methods could contribute to the understanding of the mechanisms underlying the therapeutic effects of MSCs

Vasa previa: prenatal diagnosis and management

PURPOSE AND VIEW: Vasa previa is a rare disorder of placentation associated with a high rate of perinatal morbidity and mortality when undetected before delivery. We have evaluated the recent evidence for prenatal diagnosis and management of vasa previa. RECENT FINDINGS: Around 85% of cases of vasa previa have one or more identifiable risk factors including in-vitro fertilization, multiple gestations, bilobed, succenturiate or low-lying placentas, and velamentous cord insertion. The development of standardized prenatal targeted scanning protocols may improve perinatal outcomes. There is no clear consensus on the optimal surveillance strategy including the need for hospitalization, timing of corticosteroids administration and the value of transvaginal cervical length measurements. Outpatient management is possible if there is no evidence of cervical shortening on ultrasound and there are no symptoms of bleeding or uterine contractions. Recent national guidelines and expert reviews have recommended scheduled cesarean section of all asymptomatic women presenting with vasa previa between 34 and 36 weeks’ gestation. SUMMARY: Prenatal diagnosis of vasa previa is pivotal to prevent intrapartum fetal death. Although there is insufficient evidence to support the universal mid-gestation ultrasound screening for vasa previa, recent evidence indicates the need for standardized prenatal targeted screening protocols of pregnancies at high-risk of vasa previa

Preeclampsia and intrauterine growth restriction: role of human umbilical cord mesenchymal stem cells-trophoblast cross-talk

Background: Oxidative stress is involved in the pathogenesis and maintenance of pregnancy-related disorders, such as intrauterine growth restriction (IUGR) and preeclampsia (PE). Human umbilical cord mesenchymal stem cells (hUMSCs) have been suggested as a possible therapeutic tool for the treatment of pregnancy-related disorders in view of their paracrine actions on trophoblast cells. Objectives: To quantify the plasma markers of peroxidation in patients affected by PE and IUGR and to examine the role of oxidative stress in the pathophysiology of PE and IUGR in vitro by using hUMSCs from physiological and pathological pregnancies and a trophoblast cell line (HTR-8/SVneo). Study design: In pathological and physiological pregnancies the plasma markers of oxidative stress, arterial blood pressure, serum uric acid, 24h proteinuria, weight gain and body mass index (BMI) were examined. Furthermore, the pulsatility index (PI) of uterine and umbilical arteries, and of fetal middle cerebral artery was measured. In vitro, the different responses of hUMSCs, taken from physiological and pathological pregnancies, and of HTR-8/SVneo to pregnancy-related hormones in terms of viability and nitric oxide (NO) release were investigated. In some experiments, the above measurements were performed on co-cultures between HTR-8/SVneo and hUMSCs. Results: The results obtained have shown that in pathological pregnancies, body mass index, serum acid uric, pulsatility index in uterine and umbilical arteries and markers of oxidative stress were higher than those found in physiological ones. Moreover, in PE and IUGR, a relation was observed between laboratory and clinical findings and the increased levels of oxidative stress. HTR-8/SVneo and hUMSCs showed reduced viability and increased NO production when stressed with H2O2. Finally, HTR-8/SVneo cultured in cross-talk with hUMSCs from pathological pregnancies showed a deterioration of cell viability and NO release when treated with pregnancy-related hormones. Conclusion: Our findings support that hUMSCs taken from patients affected by PE and IUGR have significant features in comparison with those from physiologic pregnancies. Moreover, the cross-talk between hUMSCs and trophoblast cells might be involved in the etiopathology of IUGR and PE secondary to oxidative stress

The role of mucosal immunity in the pathogenesis of necrotizing enterocolitis

Necrotizing enterocolitis (NEC) is the most devastating gastrointestinal disease of prematurity. Although the precise cause is not well understood, the main risk factors thought to contribute to NEC include prematurity, formula feeding, and bacterial colonization. Recent evidence suggests that NEC develops as a consequence of intestinal hyper-responsiveness to microbial ligands upon bacterial colonization in the preterm infant, initiating a cascade of aberrant signaling events, and a robust pro-inflammatory mucosal immune response. We now have a greater understanding of important mechanisms of disease pathogenesis, such as the role of cytokines, immunoglobulins, and immune cells in NEC. In this review, we will provide an overview of the mucosal immunity of the intestine and the relationship between components of the mucosal immune system involved in the pathogenesis of NEC, while highlighting recent advances in the field that have promise as potential therapeutic targets. First, we will describe the cellular components of the intestinal epithelium and mucosal immune system and their relationship to NEC. We will then discuss the relationship between the gut microbiota and cell signaling that underpins disease pathogenesis. We will conclude our discussion by highlighting notable therapeutic advancements in NEC that target the intestinal mucosal immunity