Optimization of a solid phase extraction procedure for prostaglandin E2, F2 alpha and their tissue metabolites

The primary prostaglandins PGE(2) and PGF(2 alpha) are metabolized in tissues by a series of enzymatic and non-enzymatic reactions. To measure metabolic rates and individual reaction rates it is necessary to extract the parent prostaglandins and metabolites before the separation and quantification of each compound is achieved. Here we have established and optimized a solid phase extraction (SPE) procedure to recover PGE(2), PGF(2 alpha) and their six enzymatic and non-enzymatic tissue metabolites from aqueous solutions including urine, plasma and tissue homogenate. We have used octadecyl-bonded silica gel as the stationary phase and methanol-water mixtures as binary mobile phases. The volumes and concentrations of the washing and elution solutions were optimized individually for each PG. Recoveries of all PG standards were quantitative except for PGEM, which was recovered at 80% efficiency. Biological matrix components interfered with the extraction in a PG- and matrix-specific fashion. Inclusion of 1% formic acid in the loading mixture raised recoveries from urine, plasma and tissue homogenate to >or=90%. This SPE method is the first that has been optimized by systematic elution studies for PGE(2), PGF(2 alpha) and the complement of their tissue metabolites. The procedure is simple, robust and can serve as an effective pre-purification step before downstream separation and quantification of each tissue metabolite of PGE(2) and PGF(2 alpha) from complex biological matrices.     

Acute phase reactants add little to composite disease activity indices for rheumatoid arthritis: validation of a clinical activity score

Introduction  

Frequent assessments of rheumatoid arthritis (RA) disease activity allow timely adaptation of therapy, which is essential in preventing disease progression. However, values of acute phase reactants (APRs) are needed to calculate current composite activity indices, such as the Disease Activity Score (DAS)28, the DAS28-CRP (i.e. the DAS28 using C-reactive protein instead of erythrocyte sedimentation rate) and the Simplified Disease Activity Index (SDAI). We hypothesized that APRs make limited contribution to the SDAI, and that an SDAI-modification eliminating APRs – termed the Clinical Disease Activity Index (CDAI; i.e. the sum of tender and swollen joint counts [28 joints] and patient and physician global assessments [in cm]) – would have comparable validity in clinical cohorts.     

Results of studies on the effect of radiologic contamination after the Czernobyl catastrophe and prophylactic iodine on thyroid morphology and function of inhabitants of North-East Poland]

The results of the investigations of radioactive contamination after the Chernobyl catastrophe and subsequent iodine prophylaxis on the thyroid gland function and morphology in Northeast Poland. The aim of the study was to determine whether kalium iodine in one dose during radioactive contamination in Poland limited the radioactive dose in the thyroid gland and if significant disadvantageous side-effects in the intrathyroid and extrathyroid occurred. Additionally during the studies we tried to determine if radioactive iodine contamination which occurred in the region of the Medical Academy in Bia?ystok caused an increase in thyroid disease. It is interesting to note the different results obtained after radioactive contamination with the results from the investigations in this same territory in 1983-1985. In 1983-1985, before the Chernobyl catastrophe, 6,921 persons in Northeast Poland were investigated. In 1986-1988, immediately after the disaster 4,010 persons were investigated. The main study according to grant No MZ-XVII was carried out in three provinces: Bia?ystok, Suwa?ki and Olsztyn. In this investigation 10,011 persons born before April 26, 1986 and after January 1, 1936 participated, 5,789 townspeople and 4,222 villagers, 3,987 children up to 16 years of age it the time of the disaster 1,973 boys and 2,009 girls; 6,024 adults 2,509 men and 3,516 women were drawn from a register. Committed doses to the thyroid in the investigated region were one of the highest in Poland and depended on age group and were depended on time of prophylaxis non proportional. Iodine prophylaxis was provided mainly with one dose of Lugol solution about 90%, 95% children and 30% adults took iodine. The majority of the population (53.3%-74%) were given iodine in April. From May 1st to 5th 23.0-43.4% received iodine, but after May 5th very few persons. Iodine was well tolerated, but Lugol Solution was better tolerated than other kinds of iodine. Only 241 (4.4%) cases had side effects, mainly vomiting (143), symptoms such as stomach ache, diarrhea, dyspnoe, skinrash etc. in lesser numbers. 12% (29 persons) were seen by a physician. In the investigated population were 200 pregnant women aged 19-40 years of which the majority (177) delivered full term healthy babies. Only 1 interrupted pregnancy and 7 had spontaneous abortion. Changes in the thyroid were noticed by 187 persons (2.3%-11.7%) most of which were enlargement of the thyroid, but only a few were confirmed by a physician. In the studied population from 1989 to 1990 over 30% of the population had struma.(ABSTRACT TRUNCATED AT 400 WORDS)     

Histochemische Untersuchungen über den Involutionsmechanismus der Milchdrüse

Zusammenfassung Es wurden mit histochemischen Methoden Milchdrüsen von Meerschweinchen in physiologischer und durch Kastration während der Laktationsperiode hervorgerufener Involution untersucht. Nachgewiesen wurden alkalische und saure Phosphatase (APh und SPh), unspezifische Esterase, Sukzinodehydrase, SH-Gruppen, Nukleinsäuren und Lipide. — Die Aktivität der SPh nimmt während der Involution zu. Die Bedeutung dieses Enzyms für den Rückbildungsprozeß des Drüsengewebes wird diskutiert. — Das Vorkommen der APh-Aktivität in Drüsenendstücken im Involutionsstadium kann auf die Beteiligung dieses Enzyms an der Rückresorption der mit der Milch ausgeschiedenen Substanzen hinweisen. — Zwischen der Aktivität der unspezifischen Esterase und dem Gehalt sowie der Lokalisation von Lipiden besteht eine Abhängigkeit. — Es konnten keine Unterschiede in der Sukzinodehydraseaktivität und dem DNS-Gehalt aufgezeigt werden. — Die Verminderung von SH-Gruppen und RNS hängt mit dem Aufhören der Sekretionsproduktion durch die Zellen der Drüsenendstücke der Milchdrüsen zusammen.

---

Histochemical studies on the involution mechanism of the mammary gland

Summary Histochemical methods were used to study mammary glands of guinea pigs in the course of physiologic involution and that induced by castration during lactation. Alkaline and acid phosphatases, unspecific esterase, succinic dehydrogenase, SH groups, nucleic acids and lipids were determined. Acid phosphatase activity was found to be increased in mammary glands, subject to involution. The participation of the enzyme in the involutionary process of the gland tissue is discussed. The distribution of alkaline phosphatase in the secretory sections of the gland during involution would suggest the participation of the enzyme in the reabsorption of the substance secreted with milk. A correlation existing between the activity of unspecific esterase, the level and distribution of lipids in the mammary gland could be established. No differences were detected in the activity of succinic dehydrogenase and DNA level. A decrease in SH groups and RNA content is related to cessation of milk secretion.