The effect of supplementing sow with bioactive substances on neonatal small intestinal epithelium.

Development of the small intestinal epithelium in early postnatal period has a significant influence on pig's survival rate and further productivity. The aim of this research was to verify whether the diet supplementation of pregnant and lactating sow with a blend of bioactive substances (flax seed, rapeseed, linden inflorescence, taurine, L-carnitine and tocopherol acetate) had an effect on the development of intestinal epithelium in their offspring. The doses of bioactive substances were calculated to meet the demands for optimal supply of the pig fetuses and newborns. Pig neonates from two groups of sows, control and supplemented, were sacrificed at the day 1, 2, 4, 7 and 14 of life. The samples taken from mid-jejunum were evaluated for mitosis (Ki67), apoptosis (active caspase 3), autophagy (MAP I LC3), and DNA damage (p53). Increase of mitotic index was noticed at day 1, 4 and 7 for supplemented group when compared to the control. Reduction of apoptotic index was observed at day 2 as compared to control. A tendency toward elevated autophagy was observed during the first 2-4 postnatal days in both groups. p53 expression was significantly lower in supplemented group as compared to control. Overall, the mitosis to programmed cell death ratio was increased and the maturation of epithelial cells quickened. We suppose that the supplementation of pregnant and lactating sow diet with bioactive substances enhanced maturation of the small intestinal epithelium in their offspring during the early postnatal period.

Muon decay: measurement of the transverse polarization of the decay positrons and its implications for the fermi coupling constant and time reversal invariance.

The two transverse polarization components P(T1) and P(T2) of the e(+) from the decay of polarized mu(+) have been measured as a function of the e(+) energy. Their energy averaged values are P(T1)=(6.3+/-7.7+/-3.4) x 10(-3) and P(T2)=(-3.7+/-7.7+/-3.4) x 10(-3). From the energy dependence of P(T1) and P(T2) the decay parameters eta,eta('') and alpha(')/A,beta(')/A are derived, respectively. Assuming only one additional coupling besides the dominant V-A interaction one gets improved limits on eta, beta(')/A, and the scalar coupling constant g(S)(RR): eta=(-2.1+/-7.0+/-1.0) x 10(-3), beta(')/A=(-1.3+/-3.5+/-0.6) x 10(-3), Re{g(S)(RR)}=(-4.2+/-14.0+/-2.0) x 10(-3), and Im{g(S)(RR)}=(5.2+/-14.0+/-2.4) x 10(-3).

[Physiology and mechanisms of blood circulation in vascular bed]. / Fizjologia i mechanizmy krazenia krwi w lozysku naczyniowym.

The Authors have presented the essential physiological aspects of the blood flow vital to all physicians considering use of the autohaemotransfusion, extracorporeal circulation, extrapulmonary oxygenation and controlled haemodilution.

Calcification of the aortic wall in hypercalcemic rabbits.

The mineralization process was investigated in the aortic wall of hypercalcemic rabbits. The elevated calcium level in serum was induced by intramuscular injection of vitamin D3. The animals were killed at different times of the experiment (max. 246 d). The freeze-dried tissue homogenates were used for elemental composition studies by means of proton induced X-ray emission (PIXE) and atomic absorption spectroscopy. The structural information was obtained from infrared (IR) and X-ray diffraction (XRD) spectra. Moreover, the ascending part of the aortic arch was separated and used for micro-PIXE (PIXE in combination with proton microprobe) and histochemical examinations. It was found that hypercalcemia (blood serum Ca content elevated by about 20%) induced calcification of the aortic wall. The mineral phase within the aortic wall consisted of Ca-P salts. The Ca/P ratio continuously increased during the experiment and approached 2 after 246 d of the vitamin D3 treatment. The IR and XRD studies made possible the identification of the complex phase composition of the samples. The hydroxyapatite crystals were detected after 196 days, however, in earlier phases of the experiment, amorphous calcium phosphate, dicalcium phosphate dihydrate and octacalcium phosphate were also observed. On the basis of the data obtained, the mechanism of the precipitation and growth of inorganic deposits in the tunica media of the aortic wall was discussed.

Physicochemical characterization of the inorganic phases in the aortic wall of young individuals.

The autopsy samples of human aortic wall were investigated to determine the structure of the inorganic deposits formed at the onset of the mineralization process. The studies were concentrated on the material for which histochemical staining of randomly selected sections did not reveal the presence of minerals. The highly mineralized human media samples as well as broad spectrum of model compounds were investigated for comparative purposes. It was found that the inorganic phase at the onset of tunica media mineralization is composed of a few compounds including octacalcium phosphate as the predominant Ca-P compound, and a Mg-compound, probably magnesium carbonate or double/triple salts containing magnesium carbonate.

Calcification of aortic wall in cholesterol-fed rabbits.

Development of the mineralization process in the course of atherogenesis was studied using the cholesterol-fed rabbit model. The aorta samples were investigated by means of proton and electron microprobes, infrared spectroscopy and X-ray diffraction as well as selected histochemical staining. Blood serum was analysed every 2 weeks to determine the content of cholesterol, triglycerides, inorganic phosphorus, ionized calcium, elemental composition as well as activity of alkaline phosphatase. It was found that the administered diet did not disturb the calcium and phosphorus homeostasis. Histochemical findings confirmed the formation of lipid-rich lesions blocking the lumen of the vessel. The dystrophic calcification was observed only in the atheroma, while in the tunica media a slight mineralization similar to that found in controls was observed after 210 days of the diet. In the atheroma the only phase detected was a defective hydroxyapatite. The perfection of the crystals, as well as the diameter of the deposits, increased during the course of the diet reaching about 2 microns after 210 days. The crystals were not contaminated with carbonate groups regardless of the duration of the diet.

Intramembranaceous ossification analyses by a proton microprobe.

The proton induced X-ray emission method in combination with a proton microprobe was applied to study the intramembranaceous ossification. As material sections of mouse embryo skulls from the 17th and 19th day of gestation were used. The morphology of the sample was examined by routine histochemical procedure performed on the sections adjacent to that irradiated by the proton microprobe. The measurements were made in line scan and raster scan mode. The concentrations of P, S, Cl, K, Ca, Fe and Zn were determined at each irradiated point. The average element concentrations were calculated for four parts of each section (bone, cartilage, mesenchymal tissue close to the bone and mesenchymal tissue in other places). The distributions of Ca and P (less markedly than Ca) concentrations almost exclusively correlate with localization of the bone while S, Cl and K concentrations show preference to the cartilage. The amount of inorganic material in flat bones of the 17-day embryo amounts to 14% of the dry mass. The material is characterized by a Ca/P ratio of about 1.6. In the embryo 2 days older the amount of the inorganic phase is practically the same (15%) while the Ca/P ratio approaches 2. This suggests the presence of the precursor phase in the flat bone calcification. It is possible that octacalcium phosphate (Ca/P ratio equals to 1.72) is formed at the onset of the flat bone mineralization which transforms rapidly (in 2 days) to a more stable mineral (defective hydroxyapatite).

Artery wall calcification: correlation of atherosclerosis with mineralization.

Popliteal arteries from 14 individuals (17-85 y old) were investigated. The concentrations and localizations of P, S, Cl, K, Ca, Fe, Cu, Zn and Br were measured by means of PIXE and micro-PIXE methods. The presence of PO4(3-) and CO3(2-) groups was assessed using the IR technique. The amount of P and Ca increased with age approaching at places 9% and 20% and mineral deposits were detected in tunica media. At the same time an increase in the Ca/P ratio and in the crystallinity of deposits was observed. The samples from old individuals also contained more CO3(2-) groups. The concentrations and localization of Zn and Br showed artery wall layer-dependent changes. In some places of the artery wall, minerals were also found in young persons. They were not correlated with places of blood vessel branching.

The analysis of mineral deposits and proteoglycans content in the cartilage of mouse trachea using PIXE in combination with proton microprobe.

The tracheal cartilage of mature mice have been investigated using PIXE (proton induced X-ray emission) in combination with a proton microprobe on snap frozen cryosectioned material. The localization and quantitative measurements of P, S, Cl, K, Ca, Fe, and Zn concentrations as well as direct and indirect assessment of glycosaminoglycans by measurement of S content and measurement of bound colloidal iron at pH = 1.8 has been performed. Adjacent sections were stained with the Hale method in Müller modifications and the v. Koss method for sulphated mucins and inorganic deposits respectively. It has been found that hyaline cartilage in trachea contains mineral deposits and that P + Ca amounts up to 22% of cartilage dry mass. The Ca/P ratio approaches 2 what indicates hydroxyapatite type crystals. The cartilage contains substantial amounts of S reflecting the presence of sulphate groups. It was found that the cartilage binds also colloidal iron at low pH. There is a good correlation between places with high amount of bound colloidal iron assessed by PIXE and places showing strong Prussian Blue staining. The Fe/S ratio was, however, much lower in the cartilage than in other tissues what indicates that the colloidal iron method does not give quantitative results. There were no regions showing substantial decrease in Fe/S ratio which we found previously as typical for degenerating and calcifying growth plate cartilage. This may be connected with a relatively low degree of calcification degree of the tracheal cartilage.

Measurement of colloidal iron binding at low pH in cartilage using the proton microprobe.

Quantitative micro-PIXE analysis was performed on mouse embryo epiphyseal cartilage and on the rib cartilage of mature animals after incubation of sections with colloidal iron at pH 1.8. The iron content as well as that of sulphur and phosphorus and Fe/S, Fe/P ratios were determined. It was found that colloidal iron content was higher in the cartilage than in other tissues. The cartilage also displayed the highest content of sulphur. The Fe/S ratio was however not constant, being highest in the degeneration zone close to the mineralization front, where the binding of iron was strongest while the amount of sulphur decreased. This indicates that factors other than number of sulphate groups influence the binding of positively charged molecules to glycosaminoglycans. This is confirmed by differences in the results obtained for embryonic and mature rib cartilage.

Investigation of inorganic deposits in selected organic matrices.

Inorganic deposits in the wall of human and animal arteries and in experimental tumor (Morris hepatoma 7777) were examined using proton induced X-ray emission (PIXE) and PIXE in combination with proton microprobe (micro-PIXE) techniques. The sections adjacent to the irradiated ones part were submitted to histological investigations and one part of the material was additionally investigated by infrared (IR) spectroscopy. For identification of mineral deposits, the micro-PIXE method appeared the most sensitive. The mineral deposits were detected in the artery samples, even in those without visible morphological changes, as well as in tumor samples. The deposites showed different localization and composition, depending on age and type of vessel. There were also differences between human and animal arteries. IR spectroscopy revealed the presence of carbonate apatite within the artery samples from old individuals. Matching of histological observations with data obtained by micro-PIXE method allows a better correlation of morphological and analytical results.

Measurements of mineralization process in the femur growth plate and rib cartilage of the mouse using pixe in combination with a proton microprobe.

The femoral bone from the 18-day pregnancy embryo and an rib cartilage of mature mice have been investigated using PIXE (proton induced X-ray emission) in combination with a proton microprobe on snap frozen cryosectioned material. The localization and the results of quantitative measurement of P, S, Cl, K, Ca, Fe and Zn have been correlated with the histochemical localization of inorganic deposits. It has been found that in calcifying and degenerating cartilage of the growth plate there is substantial loss of S; this element being indicative for sulphate groups of glycosaminoglycans. This change seems to be an important factor conditioning the process of mineralization. Zn is found in higher concentration in mineralized tissues, both in embryonal and mature cartilage as well as in the bone, and this suggests that Zn is also involved in the mineralization process. The mineralization of rib cartilage exceeds that of embryonal bone, and the Ca/P ratio is higher in the former than in the hydroxyapatite of the latter. The method described is a useful analytical tool especially for such types of studies in which elements are not easily redistributed by freezing, cutting and drying; e.g. in investigations of mineral deposits.