Sandwich ELISA for t-PA antigen employing a monoclonal antibody

A sandwich enzyme-linked immunosorbent assay (ELISA) for determination of tissue-type plasminogen activator (t-PA) was developed. 96-well flat-bottom polystyrene plates were coated with polyclonal (goat) anti t-PA IgG (10 micrograms/ml). After addition of samples monoclonal (murine) anti t-PA IgG (1.5 micrograms/ml) was added. Finally, peroxidase labelled anti-mouse IgG (goat) was used to quantify the bound second antibody. The assay can be used for determination of t-PA antigen in purified systems, in cell culture supernatants, and in human plasma, provided that EDTA (0.005 M) is present in the sample. In 78 healthy volunteers, t-PA antigen levels at rest were 0.4 - 15.2 ng/ml (4.3 +/- 2.7, means +/- S.D.); A significant positive correlation between t-PA antigen and age could be demonstrated.     

Disease expression and molecular genotype in congenital adrenal hyperplasia due to 21-hydroxylase deficiency.

Genotyping for 10 mutations in the CYP21 gene was performed in 88 families with congenital adrenal hyperplasia due to 21-hydroxylase deficiency. Southern blot analysis was used to detect CYP21 deletions or large gene conversions, and allele-specific hybridizations were performed with DNA amplified by the polymerase chain reaction to detect smaller mutations. Mutations were detected on 95% of chromosomes examined. The most common mutations were an A----G change in the second intron affecting pre-mRNA splicing (26%), large deletions (21%), Ile-172----Asn (16%), and Val-281----Leu (11%). Patients were classified into three mutation groups based on degree of predicted enzymatic compromise. Mutation groups were correlated with clinical diagnosis and specific measures of in vivo 21-hydroxylase activity, such as 17-hydroxyprogesterone, aldosterone, and sodium balance. Mutation group A (no enzymatic activity) consisted principally of salt-wasting (severely affected) patients, group B (2% activity) of simple virilizing patients, and group C (10-20% activity) of nonclassic (mildly affected) patients, but each group contained patients with phenotypes either more or less severe than predicted. These data suggest that most but not all of the phenotypic variability in 21-hydroxylase deficiency results from allelic variation in CYP21. Accurate prenatal diagnosis should be possible in most cases using the described strategy.     

Enhanced fibrinolysis caused by tissue plasminogen activator release in hemodialysis

The effect of hemodialysis on components of the fibrinolytic system was measured in 22 patients. Plasma levels of t-PA antigen, t-PA activity, u-PA antigen and plasminogen activator inhibitor were determined by means of immunological and functional assays. During hemodialysis, 11 patients exhibited an increase in t-PA antigen within the first hour to about three times the starting values (P less than 0.05), followed by a decrease to about double of the initial values until the end of the treatment. Eleven patients showed a continuous increase up to 200% of the starting values until the end of hemodialysis. u-PA levels did not change significantly during the time of investigation (P greater than 0.5).     

A solid colloidal drug delivery system for the eye: encapsulation of pilocarpin in nanoparticles

The present study was undertaken in order to encapsulate pilocarpin into nanoparticles. Two principally different methods for manufacturing these particles were investigated. Firstly, pilocarpin was dissolved in an aqueous medium in which the polymerization was carried out, and secondly, the polymerizing monomer was kept saturated with the drug solution under acidic conditions resulting in an incorporation into the nanoparticles in an aqueous environment. The amount of pilocarpin that could be incorporated into the nanoparticles was found to be largely influenced by the temperature at which the nanoparticles were produced and by the stabilizers used. At low temperatures, up to 60 per cent of pilocarpin nitrate could be encapsulated into butylcyanoacrylate nanoparticles using emulsion polymerization techniques. Larger amounts of pilocarpin could not be incorporated because of the hydrophilicity of the salts of this drug. The physico-chemical characteristics of the nanoparticles are reported: the particle size and morphology were determined by scanning and transmission electron microscopy and photon correlation spectrometry. The average particle size was about 100 nm. The results obtained in this study show that photon correlation spectrometry is a suitable method for the sizing of nanoparticles.     

Course of choroid infarction in a case of temporal arteritis (Horton's disease)

Follow-up of a choroidal infarction in a case of temporal arteritis by choroidal angiography with Indocyanin-green. After one month there was an almost total restitution of the choroidal blood flow in the area originally infarcted.     

Die Kompressionsarthrodese des oberen Sprunggelenks

Zusammenfassung Es wird zuerst die Wirkungsweise, dann das notwendige Instrumentarium und die Operationstechnik der Kompressionsarthrodese des oberen Sprunggelenks besprochen. Hierauf folgen die Ergebnisse der Nachuntersuchung von 25 isolierten Druckarthrodesen des oberen Sprunggelenks und 23 kombinierten des oberen und unteren Sprunggelenks (18 davon Triplearthrodesen).Die Hauptindikationen zur Operation waren bei den isolierten Arthrodesen die posttraumatischen Arthrosen, bei den kombinierten die Fußdeformierungen nach Poliomyelitis. Die Resultate der komplikationslos verlaufenen Fälle sind sich bei beiden Gruppen ungefähr gleich: Entfernung der Schraubenspanner und Steinmann-Nägel 4–5 Wochen nach der Operation. Unterschenkelgehgips für 9–10 Wochen. Gesamtdauer der Gipsfixation ca. 14 Wochen. Diese 14 Wochen genügen in der Regel, um klinische Festigkeit der Arthrodese und einen im Röntgenbild gut sichtbaren Knochendurchbau zu erreichen. Bei den kombinierten Arthrodesen trat früher und in mehr Fällen Beschwerdefreiheit ein.Als Vorteile der Kompressionsarthrodese ergeben sich: kurze Fixationsdauer im Gipsverband, große Wahrscheinlichkeit der knöchernen Versteifung und Erhaltenbleiben der primär erreichten Fußstellung.Mit 3 Textabbildungen     

Blood transfusion and cross matching

To avoid hemolytic transfusion reactions different methods of cross match techniques are described and the relation between the recipient's and the donor's blood quantities are discussed in this regard.     

Sink conditions in the flow-through cell during dissolution

An equation is derived which relates the effective surface area of a powder sample dissolving in a flow-through or column type in vitro cell to the average saturation degree of the flowing solvent due to the dissolution process. Three parameters are needed to define the experimental conditions: the volumetric solvent flow rate, the diffusion layer thickness, and the effective surface area of the sample. The equation indicates that sink conditions in a flow-through cell are independent of the solubility of the dissolving species. As a consequence of the equation there are some theoretical limitations for the range of applicability of the column in vitro dissolution method. Dissolution rates of fine and coarse paracetamol powders in doses of 50 mg and 500 mg demonstrate the validity of the equation.