CLINTEC

To assess the safety, tolerability, pharmacokinetics, and pharmacodynamics of the Fc-inactivated anti-β amyloid (Aβ) monoclonal antibody (mAb) GSK933776 in patients with mild Alzheimer's disease (AD) or mild cognitive impairment (MCI). This was a two-part, single blind, placebo-controlled, first-time-in-human (FTIH) study of single (n = 18) and repeat dose (n = 32) intravenous GSK933776 0.001-6 mg/kg (ClinicalTrials.gov: NCT00459550). Additional safety data from an open-label, uncontrolled, single dose study of intravenous GSK933776 1-6 mg/kg (n = 18) are included (ClinicalTrials.gov: NCT01424436). There were no cases of amyloid-related imaging abnormalities-edema (ARIA-E) or -hemorrhage (ARIA-H) after GSK933776 administration in both studies. Three patients across the two studies developed anti-GSK933776 antibodies. Plasma GSK933776 half-life (t1/2) was 10-15 days after repeat dosing. After each of three administrations of GSK933776, plasma levels of total Aβ42 and Aβ increased...

The concentration-effect auinine-induced hearing Quinine-induced reversible hearing impairment at the relationship of impairment frequencies of 1000 and 2000 Hz was investigated in healthy volunteers to analyze the plasma concentration-effect relationship of the drug. Six subjects were given two identical oral doses of quinine and a constant rate infusion of quinine over 6 hours (15 mg . kg-') on three separate occasions. A simple pharmacodynamic model, E = k . CY (in which E is effect, k is a proportionality constant, C is drug concentration, and the exponent y is a fitting parameter), was found to describe well the relationship between hearing impairment and quinine concentrations in a hypothetical effect compartment. No statistical differences were found in the estimated pararneters when the three dosings were compared, indicating that quinine-induced hearing impairment is independent of route of administration. The first-order rate constant (k,), linking plasma concentrations to the concentrations in the effect compartment, was (mean 2 SD) 0.71 2 0.19 and 0.99 2 0.37 hr-' for 1000 and 2000 Hz, respectively. The corresponding values of k were 0.15 2 0.10 and 0.12 2 0.19 and the values of y were 2.13 2 0.57 and 3.44 2 1.04 for 1000 and 2000 Hz, respectively. Effect was also analyzed by semiparametric pharmacodynamic modeling, which gave results comparable to those obtained with the link model. We conclude that a simple power function is a reliable pharmacodynamic model for describing quinine-induced hearing impairment in healthy subjects. (CLIN PHARMACOL THER 1994;55:317-23.)

A LTHOUGH ketamine has been on the market for 40 odd years, its role as an analgesic is still debated (1). In this issue of Acta Anaesthesiologica Scandinavica, Jensen et al. (2) present a trial of ketamine added to morphine in PCA for pain after uterine artery embolization. The rationale for the

Background: The effects of ketamine on respiration, alone, or in combination with opioids, have not been completely clarified. Both stimulant and depressant effects have been reported, as well as attenuation of opioid-induced hypoventilation at the expense of increased oxygen consumption. These conflicting results might partly be due to dose-dependent mechanisms. We have, therefore, determined the ventilatory effects of ketamine, in combination with alfentanil, using infusions to different pseudo steady-state concentrations. Methods: On two separate days, eight healthy male volunteers were given alfentanil as a continuous computer-controlled infusion, aiming at a plasma concentration of 50 ng ¡ mL ª1. After reaching apparent steady-state for alfentanil, racemic ketamine or placebo was administered in a protocol randomised for the two days. On the ketamine days a computer-controlled infusion, aiming for escalating ketamine plasma concentrations of 50, 100 and 200 ng ¡ mL ª1 , was added to the alfentanil infusion. On the placebo days saline was added. Using a face-mask with an occlusion valve, respiratory parameters were measured during airbreathing and after 6 repetitive 30-s CO 2 challenges. 744 Results: The alfentanil infusion induced hypoventilation by decreasing respiratory rate, while tidal volume and respiratory drive were unaffected. This hypoventilation was antagonised by ketamine in a concentration-dependent manner mainly through an increase in respiratory rate. The CO 2 response was not affected by alfentanil or ketamine. Conclusion: In the dose range of interest for postoperative, intensive-care and pain-clinic settings, ketamine antagonises the resting hypoventilation induced by alfentanil.

Chronic pain is one of the most common reasons for therapeutic cannabis use. To describe therapeutic cannabis use among patients with chronic pain. Patients with chronic pain who voluntarily indicated that they used cannabis therapeutically completed a questionnaire about the type of cannabis used, the mode of administration, the amount used and the frequency of use, and their perception of the effectiveness of cannabis on a set of pain-associated symptoms and side effects. The study was approved by the McGill University Health Centre Research Ethics Board. Fifteen patients (10 male) were interviewed (median age 49.5 years, range 24 to 68 years). All patients smoked herbal cannabis for therapeutic reasons (median duration of use six years, range two weeks to 37 years). Seven patients only smoked at night-time (median dose eight puffs, range two to eight puffs), and eight patients used cannabis mainly during the day (median dose three puffs, range two to eight puffs); the median freq...

Purpose Transdermal buprenorphine patches provide comparable pain relief to that of low-potency opioids in elderly individuals. However, specific data on their use in elderly individuals is limited. This study investigated and compared the PK of buprenorphine transdermal patches in elderly (≥75 years) versus younger (50-60 years) individuals. Methods This was a multiple-dose, open-label, parallelgroup study in healthy volunteers split into two age groups (younger, 50-60 years; elderly, ≥75 years) with 37 individuals in each. Study participants received two consecutive 7day buprenorphine 5 μg/h transdermal patch applications, and blood samples were collected on the week of the second patch application [day 7 (predose), days 8, 9, 10, 12, and 14] to determine PK at steady state. Pharmacokinetic parameters were determined for buprenorphine and norbuprenorphine. Safety was assessed by analyzing adverse events, hematology, clinical chemistry, urine analysis, vital signs, electrocardiogram (ECG), and physical examinations. Results The area under the plasma concentration-time curve at steady state (AUC tau), measured over one dosing interval, was similar for elderly [mean ± standard deviation (SD) 9,940 pg/h/ml (4,827 pg/h/ml] and younger [mean ± SD 11,309 (3,670 pg/h/ml] individuals. Bioequivalence was not demonstrated between groups, which may be attributable to the relatively high level of variability in individual plasma profiles. More adverse events were reported by younger (216) than elderly (164) study participants. Conclusions No dosage alterations are necessary for PK reasons when treating elderly people with buprenorphine transdermal patches.

Imagine the profits in quality of life that can be made by treating inherited diseases early in life, maybe even before birth! Immense cost savings can also be made by treating diseases promptly. Hence, prenatal stem cell therapy holds great promise for developing new and early-stage treatment strategies for several diseases. Successful prenatal stem cell therapy would represent a major step forward in the management of patients with hematological, metabolic, or immunological disorders. However, treatment before birth has several limitations, including ethical issues. In this review, we summarize the past, the present, and the future of prenatal stem cell therapy, which includes an overview of different stem cell types, preclinical studies, and clinical attempts treating various diseases. We also discuss the current challenges and future strategies for prenatal stem cell therapy and also new approaches, which may lead to advancement in the management of patients with severe incurable diseases.

Disrupted organogenesis leads to permanent malformations that may require surgical correction. Autologous tissue grafts may be needed in severe lack of orthotopic tissue but include donor site morbidity. The placenta is commonly discarded after birth and has a therapeutic potential. The aim of this study was to determine if the amnion from placenta or plasma rich of growth factors (PRGF) with mononuclear cells (MNC) from umbilical cord blood (UCB), collected noninvasively, could be used as bio-constructs for autologous transplantation as an easy-accessible no cell culture-required method. Human amnion and PRGF gel were isolated and kept in culture for up to 21 days with or without small intestine submucosa (SIS). The cells in the constructs showed a robust phenotype without induced increased proliferation (Ki67) or apoptosis (caspase 3), but the constructs showed decreased integrity of the amnion-epithelial layer at the end of culture. Amnion-residing cells in the SIS constructs exp...

Background: There are multiple promising treatment strategies for central nervous system trauma and disease. However, to develop clinically potent and safe treatments, models of human-specific conditions are needed to complement in vitro and in vivo animal model-based studies. Methods: We established human brain stem and spinal cord (cross-and longitudinal sections) organotypic cultures (hOCs) from first trimester tissues after informed consent by donor and ethical approval by the Regional Human Ethics Committee, Stockholm (lately referred to as Swedish Ethical Review Authority), and The National Board of Health and Welfare, Sweden. We evaluated the stability of hOCs with a semi-quantitative hOC score, immunohistochemistry, flow cytometry, Ca 2+ signaling, and electrophysiological analysis. We also applied experimental allogeneic human neural cell therapy after injury in the ex vivo spinal cord slices. Results: The spinal cord hOCs presented relatively stable features during 7-21 days in vitro (DIV) (except a slightly increased cell proliferation and activated glial response). After contusion injury performed at 7 DIV, a significant reduction of the hOC score, increase of the activated caspase-3 + cell population, and activated microglial populations at 14 days postinjury compared to sham controls were observed. Such elevation in the activated caspase-3 + population and activated microglial population was not observed after allogeneic human neural cell therapy. Conclusions: We conclude that human spinal cord slice cultures have potential for future structural and functional studies of human spinal cord development, injury, and treatment strategies.

Mesenchymal stem cells (MSCs) have vast potential in cell therapy, and are experimentally used in the clinic. Therefore, it is critical to find a serum- and xeno-free cryopreservation method. The aim of this study was to compare two serum- and xeno-free cryoprotectants for MSCs. Adipose tissue MSCs (Ad-MSCs) and bone marrow MSCs (BM-MSCs) were cryopreserved in two cryoprotectants: the defined serum- and xeno-free STEM-CELLBANKER™ (CB) and 10 % dimethyl sulfoxide (DMSO) in a xeno-free serum replacement cell culture medium and compared to non-cryopreserved MSCs. MSCs cryopreserved in CB or DMSO had similar morphology and surface marker expression compared to their respective non-cryopreserved MSC. Ad-MSCs and BM-MSC in both cryoprotectant media exhibited reduced mean fluorescence intensity (MFI) for CD105, BM-MSCs for CD73 and Ad-MSC increased MFI for HLA class I compared to non-cryopreserved MSCs. Population doubling time of CB cryopreserved and non-cryopreserved Ad-MSCs was similar ...

Disrupted organogenesis leads to permanent malformations that may require surgical correction. Autologous tissue grafts may be needed in severe lack of orthotopic tissue but include donor site morbidity. The placenta is commonly discarded after birth and has a therapeutic potential. The aim of this study was to determine if the amnion from placenta or plasma rich of growth factors (PRGF) with mononuclear cells (MNC) from umbilical cord blood (UCB), collected noninvasively, could be used as bio-constructs for autologous transplantation as an easy-accessible no cell culture-required method. Human amnion and PRGF gel were isolated and kept in culture for up to 21 days with or without small intestine submucosa (SIS). The cells in the constructs showed a robust phenotype without induced increased proliferation (Ki67) or apoptosis (caspase 3), but the constructs showed decreased integrity of the amnion-epithelial layer at the end of culture. Amnion-residing cells in the SIS constructs expressed CD73 or pan-cytokeratin, and cells in the PRGF-SIS constructs expressed CD45 and CD34. This study shows that amnion and UCB are potential sources for production of autologous grafts in the correction of congenital soft tissue defects. The constructs can be made promptly after birth with minimal handling or cell expansion needed.

Background Mesenchymal stem cells (MSC) are non-hematopoietic multipotent stromal cells that can differentiate into lineages such as adipocytes, osteocytes and chondrocytes. MSC are immune privileged and also possess immunosuppressive properties, which in combination with their differentiative properties makes them excellent candidates for tissue engineering, an alternative treatment solution for of congenital malformations. This study will investigate mesenchymal stem cells from amniotic fluid and umbilical cords to evaluate which tissue that is superior for tissue engineering. Methods and Results Mesenchymal stem cells were isolated from amniotic fluid (afMSC) obtained at routinely performed amniocenteses and from umbilical cords (ucMSC) collected at births from elective caesarean sections. afMSC were cultured in Dulbecco's modified Eagle medium-low glucose (DMEM) and ucMSC isolated with 3 different protocols were cultured in two different media; modified Eagle medium Alpha (MEMα) and DMEM. After expansion, the cell populations were characterized in regards to their phenotype, immunological properties, proliferative capacity and in vitro differentiation abilities. Mixed lymphocytes culture (MLC) showed that the af and ucMSC were immune privileged and also possessed immunosuppressant properties. Furthermore, the cells cultured in MEMα suppressed the immune response to a greater extent than cells cultured in DMEM. MSC from both sources showed varied differentiation potential towards the osteogenic and adipogenic lineages, but overall with low efficiency. All cells were positive for CD105, CD44, CD73 and HLA I and negative for CD45, CD31, CD14, CD80, HLA-DR and HLA II. MSC from uc were negative for CD34 and positive for CD90, whereas afMSC were intermediately positive for CD34 and CD90. The cells were cultured for 10 passages to investigate their proliferative capacity and their population doubling times were calculated. The average doubling time for afMSC was 49.3 ± 12,7 hours at passages 1 to 5, 42.3 ± 7.5 hours for ucMSC. Conclusions Based on this data we will in the forthcoming studies isolate cells from amitotic fluid using positive selection for various markers and culture the cells in MEMα. With these changes we hope to obtain a potent homogenous cell population that can be used in the treatment of congenital malformations.

Scaffolds of recombinant spider silk protein (spidroin) and hyaluronic acid (HA) hydrogel hold promise in combination with cell therapy for spinal cord injury. However, little is known concerning the human immune response to these biomaterials and grafted human neural stem/progenitor cells (hNPCs). Here, we analyzed short- and long-term in vitro activation of immune cells in human peripheral blood mononuclear cells (hPBMCs) cultured with/without recombinant spidroins, HA hydrogels, and/or allogeneic hNPCs to assess potential host–donor interactions. Viability, proliferation and phenotype of hPBMCs were analyzed using NucleoCounter and flow cytometry. hPBMC viability was confirmed after exposure to the different biomaterials. Short-term (15 h) co-cultures of hPBMCs with spidroins, but not with HA hydrogel, resulted in a significant increase in the proportion of activated CD69+ CD4+ T cells, CD8+ T cells, B cells and NK cells, which likely was caused by residual endotoxins from the Es...

Disrupted organogenesis leads to permanent malformations that may require surgical correction. Affected new-borns without spare tissue for the primary closure are dependent of other solutions. Because of the perinatal tissue shortage, synthetic or biological prosthetic materials are being used, which may be associated with complications. The progress in cellbased therapies makes autologous tissue engineering a rational alternative to current treatments, and infants with prenatally diagnosed soft tissue defects should benefit from having autologous tissue readily available for surgical implantation in the perinatal period. The aim of this thesis was to investigate cell and tissue sources harvested preor postnatally for development of autologous constructs that could be used in the correction of congenital soft tissue defects. We sought a reliable and clinical feasible isolation and culture method, a cell or tissue source with minimal or no donor site morbidity and high yield of viabl...