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.

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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...

During the adhesive locomotion of land snails a series of short dark transverse bands, called pedal or foot waves, is visible if a moving snails ventral surface is observed through a sheet of glass. Moreover, the mucus secreted from the pedal glands and some pedal epithelial cells forms a thin layer which acts as a glue augmenting adherence, while also acting as a lubricant under the moving parts of the snails foot. The relationships between velocity and the frequency of pedal waves as well as changes in the volume of small air bubbles under foot waves were analyzed by means of digital recordings made through a glass sheet on which the snails were moving. On the ventral surface of a moving snail foot, the adhering parts of the foot constituted about 80% of the total area, while several moving parts only about 20%. The single moving region of the foot (the pedal wave) amounted to about 3% of snail length. The epithelium in the region of the pedal wave was arched above the substrate and was also more wrinkled than the stationary epithelium, which enabled the forward motion of each specific point of epithelium during the passage of a pedal wave above it. The actual area of epithelium engaged by a pedal wave was at least 30% greater than the area of the epithelium as recorded through a glass sheet. In the region of the pedal wave, the tiny subepithelial muscles acting on the epithelium move it up in the front part of the wave, and then down at the end of the wave, operating vertically in relation to the substrate. In the middle part of the wave, the epithelium only moves forward. In summary, during the adhesive locomotion of snails, the horizontal movement of the ventral surface epithelium proceeds as temporally separate phases of upward, forward and downward movement.

An important electrophysiological variable-the transepithelial potential difference reflects the electrogenic transepithelial ion currents, which are produced and modified by ion transport processes in polarized cells of epithelium. These processes result from coordinated function of transporters in apical and basolateral cell membranes and have been observed in all epithelial tissues studied so far. The experiments were performed on isolated specimens of snail foot. In the experiments, the baseline transepithelial electrical potential difference-PD, changes of transepithelial difference during mechanical stimulation-dPD and the transepithelial resistance were measured with an Ussing apparatus. A total of 60 samples of foot ventral surface of 28 snails were studied. The transepithelial electrical potential difference of isolated foot ranged from-6.0 to 10.0 mV under different experimental conditions. Mechanical stimulation of foot ventral surface caused changes of electrogenic ion transport, observed as transient hyperpolarization (electrical potential difference became more positive). When the transepithelial electrical potential difference decreased during stimulation, the reaction was described as depolarization. When amiloride and bumetanide were added to the stimulating fluid so that the sodium and chloride ion transport pathways were inhibited, prolonged depolarization occurred. Under the influence of different stimuli: mechanical (gentle rinsing), chemical (changes of ion concentrations) and pharmacological (application of ion inhibitors), transient changes of potential difference (dPD) were evoked, ranging from about-0.7 to almost 2.0 mV. Changes in transepithelial potential difference of the pedal surface of the snail's foot related to these physiological stimuli are probably involved in the locomotion of the animal and are under control of the part of the nervous system in which tachykinin related peptides (TRP) act as transmitters.

The immediate reaction of the isolated rabbit tracheal wall after exposure to TDI depends on the duration of exposure and on the physiological condition of the tissue in respect of sodium and chloride ion transport.