Tissue plasminogen activator mRNA expression in granule neurons coincides with their migration in the developing cerebellum

Tissue plasminogen activator activity in the developing cerebellum, as quantified by zymography of cerebellar homogenates from embryonic day (E) 17 to adult mice, shows a peak of activity at postnatal day (P) 7, followed by a steady 75% decrease into adulthood. Northern blot analysis reveals a similar pattern for tissue plasminogen activator mRNA levels, which are low at E17 but increase dramatically, reaching their highest levels of specific mRNA/micrograms RNA in P1-P7 mice and declining about threefold in the adult mouse. In situ hybridization of whole mouse brain sections with a tissue plasminogen activator antisense cRNA probe shows pronounce reactivity in the cerebellum. Although some binding is associated with the cerebellar meninges, the external granule layer is devoid of tissue plasminogen activator mRNA at all ages. However, highly labeled elongated cells, which also bind antibody to neuronal nuclear antigen and are adjacent to Bergmann glial fibers (i.e., migrating granule neurons), are readily visible throughout the molecular and Purkinje layers at P7 and P14. In the adult mouse cerebellum, tissue plasminogen activator mRNA labeling is restricted to cells in the Purkinje/internal granule layers. Thus, tissue plasminogen activator gene expression is induced as granule neurons leave the external granule layer and begin their inward migration.     

Overview of a quality assurance/quality control compliance program consistent with FDA regulations and policies for somatic cell and gene therapies: a four year experience

Somatic cell and gene therapy involve the application of biological technologies to an individual patient through the use of living cells which provide a therapeutic benefit (Aliski, 1991). Various forms of cellular and gene therapies are being developed and evaluated in an increasing number of clinical trials for congenital and acquired disorders. The potential and progress of these therapeutic applications have resulted in an increasing effort by the Food and Drug Administration (FDA) to develop the regulatory framework under which these therapeutic approaches would insure safety and efficacy, the primary mandate of the FDA. Over five years ago Cellcor began to define the parameters, specifications, and conditions relevant to a Quality Assurance/Quality Control (QA/QC) program that has evolved to insure safety and maximize the efficacy of applications of the company's ex vivo technology, autolymphocyte therapy. Autolymphocyte therapy is an outpatient form of somatic cell immunotherapy based upon the infusion of T cells that have been activated ex vivo using a combination of previously generated autologous cytokines and an anti-CD3 monoclonal antibody. We have been able to demonstrate the feasibility for the safe, controlled, and consistent preparation and delivery of a cellular therapy by application of relevant GMP regulations. This presentation reviews aspects of this program and chronicles our experience which at present amounts to over 4400 in fusions for over 700 patients. This program provides a high degree of assurance that a cellular therapy program can be carried out in a multisite mode involving hundreds of patients through the strict adherence to cGMP as set forth in existing regulations.(ABSTRACT TRUNCATED AT 250 WORDS)     

Comparison of technetium 99m nanocolloid and indium 111 leucocytes in the diagnosis of orthopaedic infections

It has been suggested that technetium 99m nanocolloid is as effective an inflammatory radiopharmaceutical as indium 111 leucocytes. This study compares the efficacy of 99Tcm nanocolloid and 111In leucocytes in the detection of orthopaedic infection in 19 patients with a high clinical suspicion of infection. The two scintigrams were performed within 24 h of each other. A scintigram was considered positive where there was an increase in tracer uptake at the site of interest. Concordance rate of 73% was achieved. The numbers of false positives with 111In leucocytes and 99Tcm nanocolloid were three and six respectively. The single false negative in both was of a patient with tuberculous spondylodiscitis. Sensitivity was 75% in both. Specificities were 79% and 60% for 111In leucocytes and 99Tcm nanocolloid respectively. Positive Predictive Value was only 33% with 99Tcm nanocolloid and 50% with 111In leucocytes. 99Tcm nanocolloid also proved less reliable in accurately detecting infected prostheses. We conclude that 99Tcm nanocolloid cannot replace 111In leucocytes in the diagnosis of orthopaedic infections.     

Clinical significance of thyrotropin measurement by immunoradiometric assay and thyrotropin-releasing hormone test

The levels of serum TSH were measured by a sensitive immunoradiometric assay (IRMA) method and the responses of TSH to TRH stimulation were observed in three groups of hyperthyroidism, primary hypothyroidism and secondary hypothyroidism. The levels of serum TSH were found to be undetected in 98% (1/51) of the patients with hyperthyroidism, very high in 100% (35/35) of the patients with primary hypothyroidism and normal in 91% (30/33) of the patients with secondary hypothyroidism. TRH test showed no responses of TSH in all patients with hyperthyroidism, high responses in all patients with primary hypothyroidism and blunt responses in 69% (11/16) of patients with secondary hypothyroidism. The results indicate that the measurement of serum TSH by IRMA is a sensitive index in the diagnosis of hyperthyroidism and primary hypothyroidism, but can not be differentiated the secondary hypothyroidism from normal conditions. TRH test may be helpful in the differential diagnosis between secondary hypothyroidism and normal conditions.