Isolated pancreatic zymogen granules have been reported to lyse in common electrolyte solutions such as NaCl or KCl or at pH values above 5.5. A new method, based on an isosmotic Percoll density gradient, was developed for the isolation of zymogen granules and applied to rat pancreas. The granules are highly purified as judged by electron microscopic appearance and specific amylase activity. These granules exhibit a high degree of stability at physiological pH and in isotonic NaCl or KCl. Zymogen granule diameters, determined with a Coulter Counter, were 1.0 +/- 0.2 micron in either isotonic NaCl and KCl. These size values, obtained in physiological solutions, are comparable with granule sizes determined in intact cells by microscopy. Amylase activity averaged 0.66 microU per granule and protein content averaged 0.31 pg per granule; these values were not significantly influenced by different conditions of pH between 5.5 and 7.0 and ionic strength from near 0 to 0.15. The granule density estimated from the protein content was 1.13 g/ml, which agrees well with the behavior of granules in a density gradient. The properties of zymogen granules from the new preparation rectify the apparent discrepancy between their role as a storage organelle and their previously reported in vitro instability.

Mg-ATP-dependent 45Ca2+ uptake and Ca2+-ATPase activity have been examined in isolated microsomes obtained by differential centrifugation and in purified subcellular fractions obtained by Ficoll-sucrose density centrifugation in the presence of mitochondrial inhibitors. Mg-ATP-dependent 45Ca2+ uptake increased with increasing EGTA-buffered free [Ca2+], reaching a maximum of 2 nmol 45Ca2+ X 15 min-1 X mg prot-1 at 2 mumol/1 [Ca2+] in the incubation medium. Half-maximal 45Ca2+ uptake was at approximately 0.2 mumol/1 [Ca2+]. Maximal Ca2+ -Mg2+ -ATPase activity was 130 nmol X 15 min-1 X mg prot-1 at 2 mumol/l [Ca2+], with an apparent Km of approximately 0.3 mumol/l [Ca2+]. The Ca2+ ionophore A23187 (10(-6) mol/l), the mercurial compounds mersalyl (10(-5) mol/l) and CH3ClHg (10(-3) mol/l), as well as La3+ (10(-4) mol/l), vanadate (10(-4) mol/l), and saponin (50 micrograms/mg prot), abolished Mg-ATP-promoted 45Ca2+ uptake. In the absence of Mg2+, ATP did not provoke 45Ca2+ uptake. Using the purified smooth membrane fraction (F1) from the Ficoll-sucrose density gradient (enrichment of Na+-K+-ATPase specific activity by ninefold and of NADH-cytochrome c reductase by threefold as compared with total tissue homogenate), Mg-ATP-dependent 45Ca2+ uptake correlated better with Na+-K+-ATPase (r = 0.97) than with the smooth endoplasmic marker NADH-cytochrome c reductase (r = 0.52). No correlation was found with RNA, the marker for rough endoplasmic reticulum. We conclude that pancreatic plasma membranes contain a Ca2+-Mg2+-ATPase that represents the Ca2+ extrusion system from acinar cells. It is also possible that vesicular membrane structures associated with the plasma membrane, or endocytotic plasma membrane vesicles, take up Ca2+ and represent an intracellular Ca2+ pool.

Even when patients with nonsmall cell lung cancer undergo surgical resection at an early stage, recurrent disease often impairs the clinical outcome. There are numerous causes potentially responsible for a relapse of the disease, one of them being extensive angiogenesis. The balance of at least two systems, VEGF VEGFR and Ang Tie, regulates vessel formation. The aim of this study was to determine the impact of surgery on the plasma levels of the main angiogenic factors during the first month after surgery in nonsmall cell lung cancer patients. The study group consisted of 37 patients with stage I nonsmall cell lung cancer. Plasma concentrations of Ang1, Ang2, sTie2, VEGF, and sVEGF R1 were evaluated by ELISA three times: before surgical resection and on postoperative days 7 and 30. The median of Ang2 and VEGF concentrations increased on postoperative day 7 and decreased on day 30. On the other hand, the concentration of sTie2 decreased on the 7th day after resection and did not change statistically later on. The concentrations of Ang1 and sVEGF R1 did not change after the surgery. Lung cancer resection results in proangiogenic plasma protein changes that may stimulate tumor recurrences and metastases after early resection.

Markery komórek śródbłonka w chorobie niedokrwiennej sercaM Ma ag gd da al le en na a L La am mp pk ka a 1 1 , , Z Zo of fi ia a G Gr rą ąb bc cz ze ew ws sk ka a 2 2 , , M Ma ar ri ia a K Kr ra aj je ew ws sk ka a 1 1 , , I Ig ga a H Ho oł ły yń ńs sk ka a--I Iw wa an n 1 1 , , J Ja ac ce ek k K Ku ub bi ic ca a 2 2 , , T To om ma as sz z T Ty yr ra ak ko ow ws sk ki i We examined 57 patients with coronary artery disease (CAD): 27 patients with acute myocardial infarction (AMI) and 30 patients with stable angina pectoris (SA). The control group comprised 23 patients without symptoms of CAD. The concentration of von Willebrand factor (vWF), thrombomodulin (sTM), endothelin-1 (ET-1), and adhesion molecules (soluble intercellular adhesion molecule-1 (sICAM-1) and soluble vascular cell adhesion molecule-1 (sVCAM-1)) was analysed in plasma or serum.R Re es su ul lt ts s: : A significant increase in vWF, sICAM-1, sVCAM-1 and ET-1 concentrations was found in AMI compared to the control group. Increased vWF and sICAM-1 concentrations were found in SA compared to the control group. The AMI group was characterized by significantly higher vWF concentration than the SA group. Thrombomodulin concentration did not differ significantly between any patient groups and the control group. There was a positive correlation between vWF concentration and sVCAM-1 and sTM concentrations, and an inverse correlation between ET-1 and sICAM-1 concentrations in AMI. A sICAM-1 correlated positively with sVCAM-1 concentration in SA.C Co on nc cl lu us si io on ns s: : von Willebrand factor is more useful than sTM, endothelin-1, and cell adhesion molecules sICAM-1 and sVCAM-1 to assess endothelium state in patients with CAD. The increase in plasma vWF concentration confirms endothelial injury and/or activation in CAD and indicates a greater severity of these disorders in AMI than in SA.K Ke ey y w wo or rd ds s: : coronary artery disease, endothelial markers S t r e s z c z e n i e W Ws st tę ęp p: : Uszkodzenie, aktywacja lub zaburzenie funkcji śródbłonka naczyniowego odgrywają ważną rolę w progresji zmian miaż-dżycowych i rozwoju chorób układu krążenia.C Ce el l: : Ocena wskaźników biochemicznych określających stan śródbłonka naczyń krwionośnych u osób z chorobą niedokrwienną serca.M Ma at te er ri ia ał ł i i m me et to od dy y: : Badaniami objęto 57 osób z chorobą niedokrwienną serca (coronary artery disease -CAD): 27 pacjentów z ostrym zawałem serca (acute myocardial infarction -AMI) oraz 30 pacjentów ze stabilną chorobą wieńcową (stable angina -SA). Grupę kontrolną stanowiły 23 osoby bez objawów choroby wieńcowej. Stężenia czynnika von Willebranda (von Willebrand factorvWF), trombomoduliny (thrombomodulin -sTM), endoteliny-1 (endothelin-1 -ET-1), cząsteczek adhezyjnych [międzykomórkowej (soluble intercellular adhesion molecule-1 -sICAM-1) i naczyniowej (soluble vascular cell adhesion molecule-1 -sVCAM-1)] oznaczano w osoczu lub surowicy.W Wy yn ni ik ki i: : U pacjentów z AMI obserwowano istotnie zwiększone w stosunku do grupy kontrolnej...