Use of a water-soluble busulfan formulation--pharmacokinetic studies in a canine model

In a canine model we investigated the toxicity and pharmacokinetics of a water soluble busulfan preparation. Busulfan was dissolved in dimethylsulfoxide (DMSO) and administered either orally or intravenously in a single dose of 1 mg/kg. The application in either preparation was well tolerated. In seven dogs, peak levels in the range of 730 ng/mL to 1,000 ng/mL were measured after intravenous injection with an area under curve (AUC) of 75 ng.h/kg.mL to 146 ng.h/kg.mL. It was of note that even the oral administration of the same busulfan preparation resulted in AUC values in the same range as observed after parenteral application. The absorption rate of busulfan tablets in our model was as unpredictable as documented in clinical trials. On the basis of the present study, clinical trials using busulfan dissolved in DMSO given either intravenously or orally appear warranted. This approach should lead to predictable blood levels, reduced toxicity, and increased efficacy of busulfan-containing regimens.     

Expression and function of CD40 on Hodgkin and Reed-Sternberg cells and the possible relevance for Hodgkin's disease

CD40 was originally described as a B-cell-restricted antigen and was subsequently found to be a member of the tumor necrosis factor (TNF) receptor superfamily. CD40 is also expressed on dendritic cells, thymic epithelium, monocytes, and some carcinoma cell lines, and plays a critical role in cell contact-dependent activation. Primary and cultured Hodgkin and Reed-Sternberg (H-RS) cells, the presumed malignant cells of Hodgkin's disease (HD); were found to express high levels of cell surface CD40. We found that recombinant CD40 ligand (CD40L) induced interleukin-8 (IL-8) secretion and enhanced IL-6, TNF, and lymphotoxin-alpha (LT-alpha/TNF-beta) release from cultured H-RS cells. These cytokines play a significant role in the clinical presentation and pathology of HD, a tumor of cytokine-producing cells. CD40L had no mitogenic activity for HD-derived cell lines. In contrast, CD40L enhanced expression of costimulatory molecules intracellular adhesion molecule-T and B7-1 on cultured H-RS cells, both of which are overexpressed on primary H-RS cells. In addition, CD40L induced a 40% to 60% reduction of the expression of the HD-associated CD30 antigen, another member of the TNF receptor superfamily. Primary and cultured H- RS cells express not only CD30, but also CD40. CD40L has pleiotropic biologic activities on H-RS cells, and the CD40-CD40L interaction might be a critical element in the deregulated cytokine network and cell contact-dependent activation cascade typical for HD.     

Whiplash injuries in Finland: a prospective 1-year follow-up study

OBJECTIVES: The aim of the study was to define how many whiplash injuries occur in Finland in traffic accidents and the degree of severity of these injuries using the whiplash-associated disorders (WAD) classification presented by the Quebec Task Force, and to define possible long-term health effects caused by whiplash injury as well as the duration of whiplash-associated sick-leaves. METHODS: This was a prospective one-year-follow-up study. Fourteen insurance companies paving compensations for traffic accidents in Finland sent the accident reports and medical certificates of all neck injuries attributable to traffic accidents to the research team. The material was collected from neck injuries that had occurred in traffic accidents during the year 1998. RESULTS: The majority of those suffering a whiplash injury were women. On the basis of the WAD classification, most whiplash injuries were mild, belonging to grades WAD I and II. At one year from the accident nearly 10% considered that their health had been impaired significantly as a result of their neck injury. Over 10% of those questioned had been on sick-leave for over a month but only 1.5% had been on sick-leave associated with the injury for more than 6 months. The most common symptom after one year was neck pain or neck pain combined with headache and symptoms in the upper extremities. No major changes related to the seasons of the year were found. CONCLUSIONS: The number of reported neck injuries in proportion to all traffic accidents involving physical injuries is small, even in proportion to rear-end collisions. In a considerable proportion of collision patients, whiplash injury does result in significant impairment which can last as long as a year after the accident. The WAD classification predicts the duration of work disability and the long-term health damage caused by the injury. Since the appearance of symptoms and the individual need for rehabilitation due to impaired functional capacity do not depend solely on the tissue damage and biomechanical forces involved in the collision, in the future it will be important to determine which factors are responsible for the differences in coping after a collision.     

Effects of chitosan and bioactive glass modifications of knitted and rolled polylactide‐based 96/4 L/D scaffolds on chondrogenic differentiation of adipose stem cells

The performance of biodegradable knitted and rolled 3‐dimensional (3D) polylactide‐based 96/4 scaffolds modified with bioactive glass (BaG) 13‐93, chitosan and both was compared with regard to the viability, proliferation and chondrogenic differentiation of rabbit adipose stem cells (ASCs). Scaffold porosities were determined by micro‐computed tomography (μCT). Water absorption and degradation of scaffolds were studied during 28‐day hydrolysis in Tris‐buffer. Viability, number and differentiation of ASCs in PLA96/4 scaffolds were examined in vitro. The dimensions of the scaffolds were maintained during hydrolysis and mass loss was detected only in the BaG13‐93 containing scaffolds. ASCs adhered and proliferated on each scaffold type. Cell aggregation and expression of chondral matrix components improved in all scaffold types in chondrogenic medium. Signs of hypertrophy were detected in the modified scaffolds but not in the plain PLA96/4 scaffold. Chondrogenic differentiation was most enhanced in the presence of chitosan. These findings indicate that the plain P scaffold provided a good 3D‐matrix for ASC proliferation whereas the addition of chitosan to the PLA96/4 scaffold induced chondrogenic differentiation independent of the medium. Accordingly, a PLA96/4 scaffold modified by chitosan could provide a functional and bioactive basis for tissue‐engineered chondral implants. Copyright © 2012 John Wiley & Sons, Ltd.     

Hepatic copper in primary biliary cirrhosis: biliary excretion and response to penicillamine treatment.

Excessive hepatic copper accumulation occurs in long-lasting cholestatic liver disorders especially in primary biliary cirrhosis. As in Wilson's disease, penicillamine has recently been introduced for the treatment of primary biliary cirrhosis. In Wilson's disease there is decreased biliary excretion of copper. The present study shows that as compared with controls the biliary excretion of copper is not decreased in primary biliary cirrhosis; instead it may be increased in some patients. However, when compared with high hepatic copper concentration biliary copper excretion was low. In contrast with copper, biliary secretion of bile acids was decreased in eight of the 17 patients. Treatment with oral penicillamine (600 mg/day) for one year resulted in a significant decrease of hepatic copper concentration, but had no consistent effect on the biliary excretion of copper or on the amount of histologically stainable orcein-positive copper-binding protein. The results suggest that excessive hepatic copper accumulation in primary biliary cirrhosis may not be primarily caused by a decreased biliary excretion, or that a new equilibrium is achieved in advanced primary biliary cirrhosis. D-penicillamine appears to improve significantly the biliary excretion of bile acids.     

Comparison of Chondroitin Sulfate and Hyaluronic Acid Doped Conductive Polypyrrole Films for Adipose Stem Cells

Polypyrrole (PPy) is a conductive polymer that has aroused interest due to its biocompatibility with several cell types and high tailorability as an electroconductive scaffold coating. This study compares the effect of hyaluronic acid (HA) and chondroitin sulfate (CS) doped PPy films on human adipose stem cells (hASCs) under electrical stimulation. The PPy films were synthetized electrochemically. The surface morphology of PPy–HA and PPy–CS was characterized by an atomic force microscope. A pulsed biphasic electric current (BEC) was applied via PPy films non-stimulated samples acting as controls. Viability, attachment, proliferation and osteogenic differentiation of hASCs were evaluated by live/dead staining, DNA content, Alkaline phosphatase activity and mineralization assays. Human ASCs grew as a homogenous cell sheet on PPy–CS surfaces, whereas on PPy–HA cells clustered into small spherical structures. PPy–CS supported hASC proliferation significantly better than PPy–HA at the 7 day time point. Both substrates equally triggered early osteogenic differentiation of hASCs, although mineralization was significantly induced on PPy–CS compared to PPy–HA under BEC. These differences may be due to different surface morphologies originating from the CS and HA dopants. Our results suggest that PPy–CS in particular is a potential osteogenic scaffold coating for bone tissue engineering.     

Physiological responses to emotional excitement in healthy subjects and patients with coronary artery disease

Emotional excitement may trigger cardiovascular (CV) events, particularly in patients with coronary artery disease (CAD). Our aim was to compare changes in various biomarkers in CAD patients and age-matched healthy male subjects during “real-life” emotional excitement. Enthusiastic male ice hockey spectators (CAD n = 18, healthy subjects n = 16) attended Finnish national ice hockey play-off matches. Heart rate variability, plasma catecholamines, endothelin-1 (ET-1) and interleukin-6 (IL-6) were determined at the baseline and during the match. A significantly more marked increase in both ET-1 and IL-6 was observed in CAD patients compared with healthy subjects during the match (time × group interaction p = 0.009 and p = 0.018 for ET-1 and IL-6, respectively). The high-frequency power of R–R intervals decreased in CAD patients (p < 0.001) but did not change in healthy subjects (p = ns, time × group interaction p < 0.001). Changes in adrenaline and noradrenaline did not differ between the groups. Emotional excitement causes more marked increases of markers of vasoconstriction and acute inflammation and withdrawal of cardiac vagal regulation in patients with CAD.