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Knowledge of the beneficial effects of perinatal derivatives (PnD) in wound healing goes back to the early 1900s when the human fetal amniotic membrane served as a biological dressing to treat burns and skin ulcerations. Since the twenty-first century, isolated cells from perinatal tissues and their secretomes have gained increasing scientific interest, as they can be obtained non-invasively, have anti-inflammatory, anti-cancer, and anti-fibrotic characteristics, and are immunologically tolerated in vivo. Many studies that apply PnD in pre-clinical cutaneous wound healing models show large variations in the choice of the animal species (e.g., large animals, rodents), the choice of diabetic or non-diabetic animals, the type of injury (full-thickness wounds, burns, radiation-induced wounds, skin flaps), the source and type of PnD (placenta, umbilical cord, fetal membranes, cells, secretomes, tissue extracts), the method of administration (topical application, intradermal/subcutaneous injection, intravenous or intraperitoneal injection, subcutaneous implantation), and the type of delivery systems (e.g., hydrogels, synthetic or natural biomaterials as carriers for transplanted cells, extracts or secretomes). This review provides a comprehensive and integrative overview of the application of PnD in wound healing to assess its efficacy in preclinical animal models. We highlight the advantages and limitations of the most commonly used animal models and evaluate the impact of the type of PnD, the route of administration, and the dose of cells/secretome application in correlation with the wound healing outcome. This review is a collaborative effort from the COST SPRINT Action (CA17116), which broadly aims at approaching consensus for different aspects of PnD research, such as providing inputs for future standards for the preclinical application of PnD in wound healing.

Goals-of-care conversations led by the oncologist are key to advancing the prognostic awareness of the patient and family, but too frequently do not occur or are ineffective in leading to advance care planning and appropriate planning for end-of-life care. At our institution, a phase 3 trial of palliative care added to usual care of phase 1 clinical trial patients gave us the opportunity to develop an electronic medical record–based goals-of-care template for discussions. We can complete all or parts of the form with patients, use it to ensure full coverage of important tasks such as planning for transition to hospice and legacy work, and make sure all the providers are “on the same page” about treatment plans. We have this within our EMR as a SmartPhrase that can be brought up for completion, and have found that it helps to clarify patient understanding. The form can also be used to document advance care planning for both clinical care and billing. Although this tool has not been formally tested, we have found that it is effective in day-to-day practice as well as in research, and we share it here.

Perinatal brain injury (PBI) in preterm birth is associated with substantial injury and dysmaturation of white and gray matter, and can lead to severe neurodevelopmental deficits. Mesenchymal stromal cells (MSC) have been suggested to have neuroprotective effects in perinatal brain injury, in part through the release of extracellular vesicles like exosomes. We aimed to evaluate the neuroprotective effects of intranasally administered MSC-derived exosomes and their potential to improve neurodevelopmental outcome after PBI. Exosomes were isolated from human Wharton&rsquo, s jelly MSC supernatant using ultracentrifugation. Two days old Wistar rat pups were subjected to PBI by a combination of inflammation and hypoxia-ischemia. Exosomes were intranasally administered after the induction of inflammation and prior to ischemia, which was followed by hypoxia. Infrared-labeled exosomes were intranasally administered to track their distribution with a LI-COR scanner. Acute oligodendrocyte- and neuron-specific cell death was analyzed 24 h after injury in animals with or without MSC exosome application using terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay and immunohistochemical counterstaining. Myelination, mature oligodendroglial and neuronal cell counts were assessed on postnatal day 11 using immunohistochemistry, Western blot or RT-PCR. Morris water maze assay was used to evaluate the effect of MSC exosomes on long-term neurodevelopmental outcome 4 weeks after injury. We found that intranasally administered exosomes reached the frontal part of the brain within 30 min after administration and distributed throughout the whole brain after 3 h. While PBI was not associated with oligodendrocyte-specific cell death, it induced significant neuron-specific cell death which was substantially reduced upon MSC exosome application prior to ischemia. MSC exosomes rescued normal myelination, mature oligodendroglial and neuronal cell counts which were impaired after PBI. Finally, the application of MSC exosomes significantly improved learning ability in animals with PBI. In conclusion, MSC exosomes represent a novel prevention strategy with substantial clinical potential as they can be administered intranasally, prevent gray and white matter alterations and improve long-term neurodevelopmental outcome after PBI.

BackgroundPreterm newborns are at high risk of developing neurodevelopmental deficits caused by neuroinflammation leading to perinatal brain injury. Human Wharton’s jelly mesenchymal stem cells (hWJ-MSC) derived from the umbilical cord have been suggested to reduce neuroinflammation, in part through the release of extracellular vesicle-like exosomes. Here, we studied whether exosomes derived from hWJ-MSC have anti-inflammatory effects on microglia-mediated neuroinflammation in perinatal brain injury.MethodsUsing ultracentrifugation, we isolated exosomes from hWJ-MSC culture supernatants. In an in vitro model of neuroinflammation, we stimulated immortalized BV-2 microglia and primary mixed glial cells with lipopolysaccharide (LPS) in the presence or absence of exosomes. In vivo, we introduced brain damage in 3-day-old rat pups and treated them intranasally with hWJ-MSC-derived exosomes.ResultshWJ-MSC-derived exosomes dampened the LPS-induced expression of inflammation-related genes by BV-2 microglia and primary mixed glial cells. The secretion of pro-inflammatory cytokines by LPS-stimulated primary mixed glial was inhibited by exosomes as well. Exosomes interfered within the Toll-like receptor 4 signaling of BV-2 microglia, as they prevented the degradation of the NFκB inhibitor IκBα and the phosphorylation of molecules of the mitogen-activated protein kinase family in response to LPS stimulation. Finally, intranasally administered exosomes reached the brain and reduced microglia-mediated neuroinflammation in rats with perinatal brain injury.ConclusionsOur data suggest that the administration of hWJ-MSC-derived exosomes represents a promising therapy to prevent and treat perinatal brain injury.

Feature selection is often necessary when implementing classifiers in practice. Most approaches to feature selection are motivated by the curse of dimensionality, but few seek to mitigate the overall computational cost of feature extraction. In this work, we propose a model-based approach for addressing both objectives. The model is based around a sparse kernel machine with feature scaling parameters controlled by a beta-Bernoulli prior. The hyperparameters are controlled by each feature’s computational cost. Experiments were carried out using publicly-available data sets, and the proposed Cost-Constrained Feature Optimization (CCFO) was compared to related methods in terms of accuracy and computational reduction.

Collaborative writing has become essential in organizations. However, current collaborative writing tools are not accessible and usable, which prevents persons who are blind from independently participating in group work when collaborative writing activities are involved. Based on several usability studies, we present a prototype of an interface that improves the collaborative writing experience for persons who are blind.

Perinatal complications in both term- and preterm-born infants are a leading cause of neonatal morbidities and mortality. Infants face different challenges in the neonatal intensive care unit with long-term morbidities such as perinatal brain injury and bronchopulmonary dysplasia being particularly devastating. While advances in perinatal medicine have improved our understanding of the pathogenesis, effective therapies to prevent and/or reduce the severity of these disorders are still lacking. The potential of mesenchymal stem/stromal cell (MSC) therapy has emerged during the last two decades, and an increasing effort is conducted to address brain- and lung-related morbidities in neonates at risk. Various studies support the notion that MSCs have protective effects. MSCs are an easy source and may be readily available after birth in a clinical setting. MSCs' mechanisms of action are diverse, including migration and homing, release of growth factors and immunomodulation, and the potential to replace injured cells. Here, we review the pathophysiology of perinatally acquired brain and lung injuries and focus on MSCs as potential candidates for therapeutic strategies summarizing preclinical and clinical evidence.

The discovery of mesenchymal stem cells (MSCs) in perinatal sources, such as the amniotic fluid (AF) and the umbilical connective tissue, the so-called Wharton's jelly (WJ), has transformed them into promising stem cell grafts for the application in regenerative medicine. The advantages of AF-MSCs and WJ-MSCs over adult MSCs, such as bone marrow-derived mesenchymal stem cells (BM-MSCs), include their minimally invasive isolation procedure, their more primitive cell character without being tumourigenic, their low immunogenicity and their potential autologous application in congenital disorders and when cryopreserved in adulthood. This chapter gives an overview of the biology of AF-MSCs and WJ-MSCs, and their regenerative potential based on the results of recent preclinical and clinical studies. In the end, open questions concerning the use of WJ-MSCs and AF-MSCs in regenerative medicine will be emphasized.

Mesenchymal stem cells (MSC) have multiorgan differentiation capacity, providing the potential for prenatal treatment of genetic disorders. We address the question if in utero transplantation of MSC results in short-term organ-specific engraftment in the fetal sheep.Sheep fetal liver-derived MSC selected by adherence culture (passage 1) were transplantated into the fetal peritoneal cavity with ultrasound-guidance (mean gestational age, 59 days). After 14 days recipient fetuses were analyzed by fluorescence-activated cell sorting (FACS), real-time polymerase chain reaction (PCR), and immunohistochemistry.Fetuses (n = 11) were transplanted with 7.7 x 10(6) MSCs (mean). All surviving fetuses (n = 5) showed engraftment with mean levels of 3.2% (lung), 0.8% (spleen), 0.6% (liver, brain), 0.4% (bone marrow), 0.1% (blood, thymus), and0.1% (kidneys) by flow cytometry. Immunohistochemistry showed organ-specific distribution.In utero transplantation of allogeneic MSC results in low level, multiorgan engraftment at 14 days post transplant. This supports the potential of in utero MSC transplantation for the treatment of nonhematopoietic genetic disorders of the fetus.

Objective: We investigated the influence of treatment with simvastatin and vitamins C and E on serum levels of soluble cell adhesion molecules (sCAM) as markers of atherosclerosis and endothelial activation. Methods: In 11 hypercholesterolemic patients, serum levels of sCAM were measured by enzyme immunoassay before and after 6 weeks of treatment with either simvastatin (20 mg) or vitamin C (1 g) plus vitamin E (600 mg) and after an additional 6 weeks of combined treatment. In 8 of the patients angiography was performed at study entry. Results: Baseline levels of soluble intercellular adhesion molecule-1 (sICAM-1) showed an age-dependent increase (coefficient of correlation 0.71, p = 0.014) and were higher in patients with established atherosclerotic lesions than in patients with no such lesions (369.5 ± 61.8 vs. 238.4 ± 56.4 ng/ml, p = 0.023). Whereas either treatment alone had no influence on the serum levels of sCAM, combined treatment with simvastatin and vitamins C and E reduced serum levels of soluble E-selectin from 43.2 ± 18.9 ng/ml at baseline to 39.7 ± 16.5 ng/ml (p = 0.027) at study end. Although not significant, levels of soluble P-selectin were also reduced from a mean of 125.9 ± 64.1 ng/ml at baseline to 115 ± 64.9 ng/ml (p = 0.1). Serum levels of sICAM-1 and soluble vascular cell adhesion molecule-1 (sVCAM-1) remained unchanged by this treatment. Conclusion: Combined treatment with simvastatin and vitamins C and E reduced serum levels of soluble E-selectin but had no influence on sICAM-1 and sVCAM-1 levels.

Objective The purpose of this study was to assess the feasibility of autologous stem cell transplantation in fetal sheep and to compare short-term engraftment of allogeneic and autologous fetal liver stem cells in an immunocompetent large animal model. Study design Fetal liver stem cells were collected from preimmune sheep fetuses with an open or ultrasound-guided technique. After being labeled with PKH26, the cells were transplanted intraperitoneally into allogeneic and autologous fetal recipients at 48 to 64 days of gestation. Engraftment was determined by flow cytometry and real-time polymerase chain reaction 1 to 2 weeks after transplantation. Results Fetal loss rate was 29% (allogeneic transplantation) and 73% (autologous transplantation). Engraftment of donor cells was found in all fetuses, with a level of ≤4.7% in fetal liver, spleen, bone marrow, blood and thymus. Overall, there was no difference between allogeneic and autologous grafts. Conclusion Autologous in utero transplantation of fetal liver stem cells in fetal sheep is feasible, but yields a high loss rate. Differences in the major histocompatibility complex between donor and recipient seems not to have a major impact on stem cell engraftment early in gestation; major histocompatibility complex–independent donor/host competition might be responsible for low engraftment in immunocompetent recipients.

Objective: Studies showed that the expression of heat shock protein 70 (HSP70) by whole-body hyperthermia or warming of the heart is associated with protection against ischemia/reperfusion injury. The aim of this study is to determine a time-related response of HSP70 expression through topical cardiac warming with correlation to cytokine production. Methods: 30 rats were divided into three groups: no heat shock, heat shocked, and controls. Heat shock was performed with 42°C saline solution applied to the heart for 5, 30, and 60 min. HSP70 and cytokines were measured. Results: Heat shock treated animals showed a 1.2-fold increase after 5 min (NS) in HSP70 expression, a 2.0-fold increase (p < 0.02) after 30 min, and a 2.3-fold increase (p < 0.012) after 60 min compared to controls. The IL-1β levels decreased from 14.3 pg/ml (normal controls) to 7.1 pg/ml after 5 min, to 1.6 pg/ml after 30 min (p < 0.002), and to 1.4 pg/ ml after 60 min of heat shock treatment (p < 0.002). The TNF-α levels also decreased, but not significantly. Conclusions: Upregulation of HSP70 through this novel method is instant and detectable within hours. The amount of HSP70 expression induced is time dependent, showing an indirect correlation with cytokine levels. These results suggest that the protective effect of HSP70 is immediate and might be explained by reduced cytokine levels. No prior recovery period is needed.

Objective: Cardiac surgery with cardiopulmonary bypass (CPB) results in vascular injury and tissue damage which involves leukocyteendothelial interactions mediated by cytokines and adhesion molecules. This study was designed to demonstrate the effect of normothermic and hypothermic CPB to cytokine and soluble adhesion molecule levels in adults and to determine whether these levels correlate to the patients postoperative course. Design and patients: In 25 patients after normothermic and in 25 patients after hypothermic coronary artery bypass grafting with cardiopulmonary bypass (CPB), blood samples for cytokine and soluble adhesion molecule analysis were taken preoperatively, 24, 36, 48 h, and 6 days postoperatively. Soluble adhesion molecules (sE-selectin, sICAM-1) were measured by ELISA and cytokines (TNF-a, IL-6, IL-8) by chemilumenscent-immunoassay. Clinical data were collected prospectively. Results: Postoperatively, adhesion molecule and cytokine levels were significantly elevated after CPB. Mean plasma levels of sICAM-1 was 2.4-fold higher after 6 days. Mean plasma concentration of sE-selectin peaked after 48 h with a 2-fold increase compared to normothermic conditions. In the hypothermia group sICAM-1, sE-selectin, IL-6, and IL-8 showed significantly higher levels (P , 0:0057, P , 0:0012, P , 0:0419, P , 0:0145) after 24 h compared to the normothermia group. No clinical differences were seen. Conclusion: Adhesion molecules and cytokines are elevated after CPB. Patients after hypothermic CPB show significant higher sICAM-1, sE-selectin, IL-6, and IL-8 levels after 24 h compared to normothermic conditions. These results are mainly due to longer CPB and crossclamp times but do not alter the patient’s postoperative course. q 2000 Elsevier Science B.V. All rights reserved.

This article explores connections between the mental health and educational systems by profiling the Redefining Actions and Decisions (RAD) curriculum, a theory-based, developmentally informed HIV education program for middle and high schools. RAD was developed by Deborah R. Schoeberlein and evaluated by Dr. John Brett. RAD's public health approach, which has gained considerable attention at the national level, argues that HIV prevention, either through risk-abstinence or risk-reduction techniques, is an appropriate strategy for safeguarding community health.

During spring 1998 RAD Educational Programs, in cooperation with the National Middle School Association, conducted a needs assessment of middle level HIV/STD prevention programs from the perspective of representatives from state education agencies (SEA) and local education agencies (LEA). Data were collected from 84% of states (SEAs) and 88% of cities (LEAs) funded by CDC to conduct HIV/STD prevention activities. A range of activities are occurring in school-based HIV/STD prevention for early adolescents in grades 5-8. However, a prevailing characteristic among states was the high degree to which local control and local decision making affects programming in schools. Needs of SEAs and LEAs are similar as reflected by the need for continued improvement of middle level HIV/STD prevention education. A concrete result was the multitude of requests for effective HIV/STD curricula for middle level students. In addition, administrators shared similar needs for an increase in time allotment for health instruction. Most SEAs (65%) and LEAs (77%) believed that time for health education in the classroom would increase if a nation-wide mandate existed resulting in appropriate standards and assessments for all middle level and high school students.

Objective: Currently used valve substitutes for valve replacement have certain disadvantages that limit their long-term benefits such as poor durability, risks of infection, thromboebolism or rejection. A tissue engineered autologous valve composed of living tissue is expected to overcome these shortcomings with natural existing biological mechanisms for growth, repair, remodeling and development. The aim of the study was to improve cell seeding methods for developing tissue-engineered valve tissue. Methods: Human aortic myofibroblasts were seeded on polyglycolic acid (PGA) meshes. Cell attachment and growth of myofibroblasts on the PGA scaffolds with different seeding intervals were compared to determine an optimal seeding interval. In addition, scanning electron microscopy study of the seeded meshes was also performed to document tissue development. Results: There was a direct correlation between cell numbers assessed by direct counting and MTT(3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltertra-zolium bromide) assay. Both attach rate and cell growth seeded on meshes with long intervals (24 and 36 h) were significantly higher than those seeded with short intervals (2 and 12 h) (P , 0.01), there was no significant difference between 24- and 36-h seeding interval. Scanning electron microscopy also documented more cell attachment with long seeding intervals resulting in a more solid tissue like structure. Conclusion: It is feasible to use human aortic myofibroblasts to develop a new functional tissue in vitro. Twenty-four hours is an optimal seeding interval for seeding human aortic myofibroblasts on PGA scaffolds and MTT test is a rapid and reliable quantitative method to assess the optimal human cell seeding on polymeric meshes. © 1999 Elsevier Science B.V. All rights reserved.

OBJECTIVE: Survivors of premature birth suffer from long term disabilities. Synthetic PreImplantation Factor (sPIF*) modulates inflammatory responses and reverses neuroinflammation. Proteinkinase A (PKA) and protein kinase C (PKC) are crucial signaling molecules. PKA up-regulates IL-10 and brain-derived neurotrophic factor (BDNF) expression, which exert neuroprotective effects. Anti-apoptotic phosphorylation of Bad is mediated by PKA. PKC phosphorylates GAP-43, a marker for neuronal plasticity and structural recovery. We explored sPIF protective role in neuronal (N2a) cells and in a rat model of encephalopathy of prematurity. *proprietary. STUDY DESIGN: Cells were subjected to LPS and treated with sPIF or scrambled sPIF. Neonatal rats (postnatal day 3: P3) were subjected to LPS, ligation of carotid artery, and hypoxia (8% O2, 65min; n¼ 30). sPIF (0.75mg/kg twice daily) was injected (P6-13) and brains harvested at P13. sPIF’s potential and mechanisms were evaluated using immunohistochemistry, ELISA, Western Blot, and qRT-PCR. Data were analyzed using two-tailed Student’s t-test. P

Collaborative writing applications are widely utilized in the work place to co-author documents and to exchange ideas. Unfortunately, persons who are blind have difficulty in identifying the changes and their related context, which prevents them from actively participating in collaborative writing. In order to expand on the knowledge gained from previous research, additional usability studies were carried out with participants who are blind utilizing Microsoft Word with the focus on track change and comment features. Based on results from all the previous usability studies, a Microsoft Word Add-In was proposed to solve the issues identified. This paper briefly presents the results of the usability studies, the prototype design of the Microsoft Word Add-In and a pilot review of the Microsoft Word Add-In for making the track change and comment features accessible and usable.

Collaborative writing applications are widely utilized in organizations to co-author documents and jointly exchange ideas. Unfortunately, for persons who are blind, collaborative writing applications are often difficult to access and use. Therefore, this paper presents the results from several usability studies that examined how visually able persons and persons who are blind interact with collaborative writing applications, and the accessibility and usability issues they encounter.

Objective The aim of this study is to assess early homing of placenta-derived stem cells after perinatal intracerebral transplantation in rats. Study Design Neonatal Wistar rats (2-4 days old) were anesthetized, and 250,000 human placenta–derived mesenchymal stem cells (MSC) injected into the lateral ventricle or the paraventricular white matter using a stereotactic frame. Donor MSC were detected by immunohistochemistry using an antihuman HLA-ABC antibody. Results In all, 84% of the animals survived the transplantation. Donor cells were detected in the brain ventricle 1-2 hours posttransplantation. After 4 hours, donor cells migrated throughout the ventricular system. At 1-4 weeks after transplantation, some cells had migrated into the periventricular white matter. Conclusion Human placenta–derived MSC were successfully transplanted into the lateral ventricles of neonatal rats. Donor cells survived, homed, and migrated in the recipient brains. Proliferation and differentiation analysis and functional tests will assess the therapeutic effects of stem cell transplantation.

Group support applications are widely used in workplace to support group work. Unfortunately, persons who are blind often found it difficult to access group support applications, due to the highly graphical nature of the application; this hinders their ability to contribute to the group. As a result, persons who are blind often face problems in gaining and retaining employment. In order to expand on the knowledge gained from previous research, three additional focus group studies were conducted. The focus group studies show that accessibility and usability issues of group support applications are impeding persons who are blind from productive group work. The implications of the results from this research project are also discussed.

Accessibility has been one of the biggest problems that people with disabilities face in the work place, due to today's rapid change in computer technology. This paper presents the evaluation of several console-based and web-based groupware applications including Outlook, AIM, Google Blog, and Group System's ThinkTank in terms of their accessibility. These applications were evaluated for accessibility based on various characteristics of the applications such as accessible front-end, hierarchy or list structures, input support, output support; screen reader adaptability; and keyboard access. Additionally, web-based groupware applications were evaluated using Web Content Accessibility Guidelines (WCAG) and U.S. Government's Section 508 guidelines. Fujitsu's Web Accessibility Inspector tool was also utilized to help evaluate the web-based applications. It is found that groupware applications have very limited accessibility through the support of keyboard access. Additional audio support and flattened hierarchies should be considered, to enable some persons with disabilities easy access to groupware applications. Future research should include persons with disabilities in evaluating groupware applications, to determine preferences. Since many groupware applications provide Application Program Interfaces (API), custom front-ends should be developed to include audio content and to flatten hierarchies and lists.

The accessibility issues of Groupware applications prevent visually impaired and other persons with disabilities access to these highly graphical interfaces. To address the accessibility issues persons with disabilities have with Groupware, a recent literature review on Groupware accessibility was performed. This paper reviews recent research to identify the accessibility issues; the input devices and output forms utilized to improve accessibility; and, the proposed solutions for Groupware accessibility. The conclusion is that encapsulating or overlaying the Groupware interface with an accessible auditory interface was a common solution to accessibility. Future research is needed in the following areas: guidelines for accessibility; identifying accessibility issues that users with disabilities may face; promoting awareness of the accessibility issues; evaluating groupware accessibility; design of accessible auditory interfaces; system/software support for persons with disabilities working in isolation; support for alternative input and output devices for persons with disabilities.

Objective Recently we could show that mesenchymal stem cells (MSCs) exist in placental tissue. These cells could be differentiated towards both mesodermal and neurogenic lineages. In regard with possible autologous transplantation we evaluated the potential of these cells. Aims: 1.To manufacture an osteogenic composite graft consisting of placental MSCs and chorion scaffold for site-specific perinatal repair of bone defects. 2.To assess the neurogenic potential of these cells in regard with possible neuroregeneration in the premature infant.

OBJECTIVE: New routes for cell transplantation into the brain need to be explored as intracerebral or intrathecal applications have a high risk to cause damage to the central nervous system. It has been hypothesized that transnasally administrated cells bypass the blood-brain barrier and migrate along the olfactory neural route into the brain and cerebrospinal fluid. Our goal is to confirm this hypothesis by transnasally administrating Wharton’s Jelly mesenchymal stem cells (WJ-MSC) and neural progenitor cells (NPC) to perinatal rats in a model of hypoxic-ischemic brain injury. STUDY DESIGN: Four-day-old Wistar rat pups, previously brain-damaged by combined hypoxic-ischemic and inflammatory insult, either received WJ-MSC or green fluorescent protein-expressing NPC: The heads of the rat pups were immobilized and 3 ml drops containing the cells (50’000 cells/ml) were placed on one nostril allowing it to be snorted. This procedure was repeated twice, alternating right to left nostril with an interval of one minute between administrations. The rat pups received a total of 600’000 cells. Animals were sacrificed 24h, 48h or 7 days after the application of the cells. Fixed brains were collected, embedded in paraffin and sectioned. RESULTS: Transplanted cells were found in the layers of the olfactory bulb (OB), the cerebral cortex, thalamus and the hippocampus. The amount of cells was highest in the OB. Animals treated with transnasally delivered stem cells showed significantly decreased gliosis compared to untreated animals. CONCLUSION: Our data show that transnasal delivery of WJ-MSC and NPC to the newborn brain after perinatal brain damage is successful. The cells not only migrate the brain, but also decrease scar formation and improve neurogenesis. Therefore, the non-invasive intranasal delivery of stem cells to the brain may be the preferred method for stem cell treatment of perinatal brain damage and should be preferred in future clinical trials.

The Johns Hopkins University Applied Physics Laboratory is testing a prototype 600 KHz Broad Band Acoustic Doppler Current Profiler (BB-ADCP) built by RD Instruments. The device uses four acoustic beams in a Janus configuration, similar to existing commercial ADCPs from RDI. However, this new device uses phase coded pulse pairs with coherent correlation processing. Velocity estimates are derived from the phase of the correlation function. Theoretical predictions indicate velocity errors are reduced by roughly an order of magnitude compared with a 600 KHz narrow band ADCP. The BB-ADCP is designed to make current measurements on one meter vertical scales. The device has four primary modes of operation consisting of combinations of different pulse widths (1 and 0.01 m) and different pulse separations ( I and 2 m). The 1 m pulse modes are discussed. In early 1991, the BB-ADCP was deployed on a submarine in the Narragansett Bay off the coast of Rhode Island. A preliminary assessment of instrument performance is presented and compared with theoretical predictions.

Objective The fetal sheep in utero transplantation model has developed into an important tool to study the efficacy of human in utero stem cell transplantation and gene therapy because of similarities in both the scale and development of immunocompetence relative to gestational age. The aim of this study was to determine whether human stem cells can be successfully transplanted to the first-trimester ovine fetus by use of a newly developed minimally invasive technique. Study design Human cord blood–derived, CD34 + -enriched stem cells were injected into the peritoneal cavity of 45- to 60-day-old ovine fetuses by using ultrasound-guided transabdominal percutaneous needle puncture. Engraftment was determined 1 to 3 months after birth by flow cytometry with use of human-specific anti-CD45 antibodies. Results In contrast to previous studies that used surgical techniques, we observed a fetal loss rate of 24%, significantly below previous values and only marginally higher than natural loss. Successful human cell engraftment was achieved in 18% of lambs available for analysis. Engraftment levels of human cells in bone marrow of the recipient were up to 0.8% of total nucleated cells. Conclusion Ultrasound-guided percutaneous transplantation of stem cells to fetal sheep in the first trimester is feasible. Although we were unable to observe a significant improvement in the level of engraftment of human cells in sheep, the decreased fetal loss rate associated with this technique allows greater use for further studies that use this model of in utero transplantation.

Objective: Prenatal ex vivo gene therapy might be an effective and safe strategy with which to treat severe genetic disorders in utero. For this purpose, autologous fetal stem cells must be collected before the second trimester, transfected in vitro, and transplanted back to the fetus. The aim of this study was to determine whether stem cells can be sampled from the first trimester fetal liver in ongoing gestation. Study Design: Fetal liver stem cell sampling was performed in 21 ovine fetuses. Pregnant ewes at 57 ± 2 gestational days were generally anesthesized. A 20-gauge needle was inserted transcutaneously into the fetal liver under ultrasound guidance. Fetal liver cells were sampled by suction. The numbers of nucleated cells and progenitor/stem cells were determined. Results: All 21 fetuses showed normal heart rate 5 minutes after the procedure. A mean (±SEM) of 2.07 ± 0.5 × 10 7 nucleated cells and 172 ± 53 colony-forming units per 10 5 cells (hematopoietic progenitors/stem cells) were collected. Fetal loss rate at term was 7 of 21 fetuses (33%). Conclusion: This study shows that fetal liver cells can be collected in the early fetus with an ultrasound-guided technique. The number of fetal liver cells that are collectable is large enough for autologous transplantation and engraftment of genetically engineered (transfected) stem cells. ( Am J Obstet Gynecol 2002;187:960–3.)

The technical issues of a standoff electro-optic tripwire detector are discussed. Significant advances in short-wave infrared (SWIR) laser diodes and InGaAs detector technologies have made it possible for the demonstration of a passive and active eyesafe (1.5 micron) laser illuminated tripwire (ELIT) detector. The demonstrated system utilizes COTS laser diodes and cameras. The Hough Transform was used for the detection of tripwires in images. System trade-offs are discussed and images are shown.

Objective: In cardiovascular tissue engineering, three-dimensional scaffolds serve as physical supports and templates for cell attachment and tissue development. Currently used scaffolds are still far from ideal, they are potentially immunogenic and they show toxic degradation and inflammatory reactions. The aim of this study is to develop a new method for a three-dimensional completely autologous human tissue without using any scaffold materials. Methods: Human aortic tissue is harvested from the ascending aorta in the operation room and worked up to pure human myofibroblasts cultures. These human aortic myofibroblasts cultures (1.5 £ 10 6 cells, passage 3) were seeded into 15-cm culture dishes. Cells were cultured with Dulbecco’ s modified Eagle’s medium supplemented with 1 mM l-ascorbic acid 2-phosphate for 4 weeks to form myofibroblast sheets. The harvested cell sheets were folded to form four-layer sheets. The folded sheets were then framed up and cultured for another 4 weeks. Tissue development was evaluated by biochemical assay and light and electron microscopy. Results: After 4 weeks of culture in ascorbic acid supplemented medium, myofibroblasts formed thin cell sheets in culture dishes. The cell sheets presented in a multi-layered pattern surrounded by extracellular matrices. Cultured for additional 4 weeks on the frames, the folded sheets further developed into more solid and flexible tissues. Light microscopy documented a structure resembling to a native tissue with confluent extracellular matrix. Under transmission electron microscope, viable cells and confluent bundles of striated mature collagen fibers were observed. Hydroxyproline assays showed significant increase of collagen content after culturing on the frames and were 80.5% of that of natural human pericardium. Conclusions: Improved cell culture technique may render human aortic myofibroblasts to a native tissue-like structure. A three-dimensional completely autologous human tissue may be further developed on the base of this structure with no show toxic degradation or inflammatory reactions. q 2000 Elsevier Science B.V. All rights reserved.

Objective: Cardiac surgery with cardiopulmonary bypass (CPB) results in expression of cytokines and adhesion molecules (AM) with subsequent inflammatory response. The purpose of the study was to evaluate the clinical impact of modified ultrafiltration (MUF) and its efficacy in reducing cytokines and AM following coronary artery bypass grafting (CABG) in adults. Methods: A prospective randomized study of 97 patients undergoing elective CABG was designed. Fifty patients were operated on using normothermic and 47 patients using hypothermic CPB. The normothermic group was subdivided into a group with modified ultrafiltration (na 30) and a group without MUF (na 20). In the hypothermic group 30 patients received MUF compared to 17 patients serving as controls. MUF was instituted after CPB for 15 min through the arterial and venous bypass circuit lines. Cytokines (IL-6, IL-8, TNF-a , IL-2R) and adhesion molecules (sE-selectin, sICAM-1) were measured preoperatively, pre-MUF, in the ultrafiltrate, 24 h, 48 h and 6 days after surgery by chemiluminescent enzyme immunometric assay or enzyme-linked immunosorbent assay (ELISA). Clinical parameters were collected prospectively until discharge. Results: In all patients AM and cytokines were significantly elevated after normothermic and hypothemic CPB. AM and cytokines were significantly higher in hypothermia compared to normothermia. In hypothermic CPB sE-selectin was decreased after 24 h by 37% (P , 0:0063) and by 40% (P , 0:0027) after 48 h postoperatively. ICAM-1 was reduced by 43% (P , 0:0001) after 24 h and by 60% (P , 0:0001) after 6 days. Similar results were seen in cytokines with reduction up to 60% after 24 h. Changes after 48 h were noticeable but not significant. Reduction of AM and cytokines after normothermic CPB was minimal. Neither in normothermia, nor in hypothermia has sIL2R been effectively removed from the circulation. There were no significant differences in the clinical variables between the patients with or without MUF. Conclusion: AM and cytokines are significantly elevated after hypothermic CPB compared to normothermic CPB. MUF led to a significant reduction in cytokine and AM levels after hypothermic CPB, except for IL-2R. MUF showed minimal effect in normothermia. We conclude that MUF is an efficient way to remove cytokines and AM. However, we were unable to demonstrate any significant impact of MUF in outcome of adults after elective CABG. q 2000 Elsevier Science B.V. All rights reserved.