Highly sensitive high-performance liquid chromatographic assay for methotrexate in the presence and absence of anti-methotrexate antibody fragments in rat and mouse plasma

Recently, Balthasar and Fung have proposed that anti-methotrexate antibody fragments may be employed to enhance the selectivity of intraperitoneal methotrexate (MTX) therapy. This current work presents a sensitive high-performance liquid chromatographic method for measuring plasma concentrations of total (i.e., bound and unbound) MTX and free (unbound) MTX in rat and mouse plasma, in the presence or absence of therapeutic anti-MTX antibody fragments. The assay involves pre-column derivatization of MTX by sodium hydrosulfite to 2,4-diamino-6-methylpteridine. The limit of quantitation for MTX by this assay was 1.25 ng in rat plasma, mouse plasma and mouse plasma ultrafiltrate, which corresponds to a concentration of 25 ng/ml for a 50 microl sample. The limit of quantitation was found to be 2.5 ng in rat plasma ultrafiltrate (i.e., 50 ng/ml in 50 microl rat plasma ultrafiltrate). The method was shown to be quite accurate, as the mean assayed concentration of quality control samples was within 10% of theoretical values. We have applied the method to the investigation of MTX pharmacokinetics in mice and rats, following the administration of MTX alone or following simultaneous administration of MTX and anti-MTX Fab fragments. The method has been shown to be suitable for the assay of total and free methotrexate in the plasma of these species and will enable the testing of pharmacokinetic hypotheses regarding the influence of anti-MTX Fab fragments on the disposition of MTX.     

Novel imaging techniques for diabetic macular edema

Retinal edema should be defined as any increase of water of the retinal tissue resulting in an increase in its volume. It may be of cytotoxic or vasogenic origin. Development of vasogenic macular edema is dependent on a series of factors such as blood pressure, blood-retinal barrier permeability, retinal cell damage, retinal tissue osmotic pressure and retinal tissue compliance. Objective measurements of retinal thickness are now possible using the Retinal Thickness Analyser. Localised measurements of blood-retinal barrier permeability may also be obtained using the Retinal Leakage Analyser, a modified confocal scanning laser fluorometer, while obtaining simultaneously angiographic images of the choroid and retina. These new imaging techniques show that cytotoxic and vasogenic retinal edema may occur independently in the early stages of diabetic retinopathy. These findings offer new perpectives for designing novel therapeutic strategies. This revised version was published online in July 2006 with corrections to the Cover Date.     

Perspectives in transplantation immunology 1991

Summary The clinical success of organ transplantation depends to a large degree on the immunological acceptance of the grafted organ. This paper summarizes from an immunological point of view the recent progress that has been made to improve graft acceptance, and discusses some future aspects in the field. Over the last few years, major emphasis has been put on the development of new immunosuppressive drugs, including FK 506, rapamycin, and Deoxyspergualin. Together with monoclonal antibodies against defined T-cell surface antigens, there are now new and effective means available to prevent or treat rejection episodes. Progress has also been made in the field of HLA typing, where the introduction of molecular biology-based methods significantly increased the accuracy of HLA class II typing. The ultimate goal of transplantation immunology is the induction of (donor-) specific tolerance. While some protocols are effective in inducing peripheral tolerance in experimental animals, these regimens are at present not yet applicable in the clinical situation. To overcome the shortage of donor organs, alternative strategies are currently being considered. Among these, xenotransplantation may eventually prove successful, despite the massive immunological problems such as, e.g., the presence of preformed xenoreactive antibodies.Abbreviations CTL cytolytic T lymphocyte - HLA human leukocyte antigen - MHC major histocompatibility complex - PCR polymerase chain reaction - mAB monoclonal antibody - RFLP restriction fragment length polymorphism - TCR T-cell receptor Preprint of a lecture to be read at the 22nd Congress of the Gesellschaft für Nephrologie, Heidelberg, September 15–18, 1991 (Editor: Prof. Dr. E. Ritz, Heidelberg)     

Bilateral pneumothorax with pneumomediastinum under anaesthesia in a healthy female

A case of bilateral pneumothorax and Pneumomediastinum developing under general anaesthesia without apparent cause is reported. Possible aetiologic factors and mechanisms of the condition are outlined. Clinical signs and the immediate management of pneumothorax and Pneumomediastinum are discussed.     

Formal retrospective case review and sudden infant death

A review of 24 consecutive sudden infant deaths was undertaken to evaluate the importance of the various stages in the postmortem assessment of such cases. Death in three cases was caused by obvious trauma. Of the remainder, 16 were attributed to sudden infant death syndrome (SIDS), 4 to accidental asphyxia (identified by death scene examination and/or formal case review) and 1 to a lingual thyroglossal duct cyst. Three (14%) of 21 deaths thought to be SIDS after postmortem examination were attributed to asphyxia following subsequent formal case review.     

Chronic illness in adolescents: Transfer or transition to adult services?

It is widely believed that the improved survival of young people with chronic diseases will be associated with the development of appropriate services within the adult healthcare domain. There is, however, little evidence to suggest that this is happening at a rate commensurate with clinical requirements. This paper highlights the multiplicity of barriers that impede the development of transition services to facilitate the transfer of medical care from the paediatric to the adult domain. Different models of transition care are described, and the terms 'transfer' and 'transition' are differentiated. The clinical demand for service development is highlighted, as well as the need for specific research in this area of healthcare delivery.     

Randomised controlled trial of zinc supplementation in malnourished Bangladeshi children with acute diarrhoea

OBJECTIVE: To evaluate the impact of zinc supplementation on the clinical course, stool weight, duration of diarrhoea, changes in serum zinc, and body weight gain of children with acute diarrhoea. DESIGN: Randomised double blind controlled trial. Children were assigned to receive zinc (20 mg elemental zinc per day) containing multivitamins or control group (zinc-free multivitamins) daily in three divided doses for two weeks. SETTING: A diarrhoeal disease hospital in Dhaka, Bangladesh. PATIENTS: 111 children, 3 to 24 months old, below 76% median weight for age of the National Center for Health Statistics standard with acute diarrhoea. Children with severe infection and/or oedema were excluded. MAIN OUTCOME MEASURES: Total diarrhoeal stool output, duration of diarrhoea, rate of weight gain, and changes in serum zinc levels after supplementation. RESULTS: Stool output was 28% less and duration 14% shorter in the zinc supplemented group than placebo (p = 0.06). There were reductions in median total diarrhoeal stool output among zinc supplemented subjects who were shorter (less than 95% height for age), 239 v 326 g/kg (p < 0.04), and who had a lower initial serum zinc (< 14 mmol/l), 279 v 329 g/kg (p < 0.05); a shortening of mean time to recovery occurred (4.7 v 6.2 days, p < 0.04) in those with lower serum zinc. There was an increase in mean serum zinc in the zinc supplemented group (+2.4 v -0.3 mumol/l, p < 0.001) during two weeks of supplementation, and better mean weight gain (120 v 30 g, p < 0.03) at the time of discharge from hospital. CONCLUSIONS: Zinc supplementation is a simple, acceptable, and affordable strategy which should be considered in the management of acute diarrhoea and in prevention of growth faltering in children specially those who are malnourished.