The effect of LPS injections on non-specific immune response in affected pigeons.

The aim of the study was to investigate the effect of LPS injections on non-specific mechanisms of immunity in pigeons. On the first day of observation the experimental birds (n=18) were intravenously injected with Escherichia coli LPS (10 microg/kg b.w.), while the control animals (n=6) received in the same way apyrogenic physiological saline. On the second and the third day of the experiment LPS in the same doses was injected again. Four and a half hours after the saline and each pyrogen administration blood samples were collected from the control and experimental pigeons. The total protein, gamma globulin, lysozyme, acute phase protein (Cp, CRP, Tf, ferritin, Alb) and selected trace element (Fe, Cu, Zn) concentrations were investigated. The obtained results showed the increase in the concentration of total protein, Cp, CRP and Tf in endotoxin fever resulting from LPS injection in pigeons. In contrast, the concentration of gamma globulins, ferritin and A1lb were decreased in response to the first LPS injection. However, the consecutive injections of LPS caused a decrease in the concentration of total protein, CRP and Tf. In opposition to those results, a significant rise in the lysozyme and ferritin concentrations was observed. On the other hand, the first LPS injection caused a decline in the iron and zinc concentrations which remaining lower than the control values following repeated administration of LPS. On the contrary, the copper concentration increased successively in response to the next LPS injections.

Alpha-ketoglutarate protects the liver of piglets exposed during prenatal life to chronic excess of dexamethasone from metabolic and structural changes.

Glucocorticoids play a role in the origin of the features of the metabolic diseases. Alpha-ketoglutarate (AKG) is defined as glutamine homologue and derivative, conditionally an essential amino acid. In the liver, glutamine serves as a precursor for ureagenesis, gluconeogenesis and acute phase protein synthesis The aim of the study was to determine the effect of AKG administered to piglets prenatally exposed to dexamethasone, on the structure of the liver and its metabolic function. Sows were administered with dexamethasone (3 mg/sow/48 h) from day 70 of pregnancy to the parturition, and then after the birth, the piglets were divided into the group administered with AKG (0.4 g/kg body weight) or physiological saline. Biochemical markers, lysozyme and ceruloplasmin serum activities, concentrations of selected free amino acids, macro- and microelements and histomorphometry of the liver tissue were determined. The total cholesterol concentrations in the sows and their newborns from the Dex groups were higher by 72% and 64%, respectively, compared with the control groups. Triacylglycerol concentration was higher by 50% in sows from the Dex group and 55% in the new-born piglets. Alpha-ketoglutarate administered to the piglets after prenatal influence of dexamethasone lowered the total cholesterol concentration by 40%, and enhanced aspartate by 41%, serine by 76%, glutamate by 105%, glutamine by 36%, glycine by 53% and arginine by 105%, as well as methionine and cystathionine, but increased the sulphur concentration compared with the control (p < 0.01). Intracellular space D decreased after AKG administration in comparison with the piglets from Dex/Control group not treated with AKG. Postnatal administration of AKG had a protective effect on liver structure, and lowered the total cholesterol concentration in piglets prenatally exposed to dexamethasone, and also influenced selected macro- and microelement serum concentrations and amino acids plasma concentration.

The role of the biomedical physicist in the education of the healthcare professions: an EFOMP project.

The role of the biomedical physicist in the education of the healthcare professions has not yet been studied in a systematic manner. This article presents the first results of an EFOMP project aimed at researching and developing this important component of the role of the biomedical physicist. A background to the study expands on the reasons that led to the need for the project. This is followed by an extensive review of the published literature regarding the role. This focuses mainly on the teaching contributions within programmes for physicians, diagnostic radiographers, radiation therapists, and the postgraduate medical specializations of radiology, radiotherapy, interventional radiology and cardiology. Finally a summary list of the specific research objectives that need to be immediately addressed is presented. These are the carrying out of a Europe-wide position audit for the role, the construction of a strategic role development model and the design of a curriculum development model suitable for modern healthcare professional education.

Changes in U937 cell viability induced by stress factors - possible role of calmodulin.

Current studies were aimed to elucidate influence of magnetic field (MF) stimulation on cell viability and its effect on expression of calmodulin (CaM) and Hsp70 protein which plays a role of cell stress indicator and is a Ca2+-dependent CaM-binding protein. For the experimental model we have chosen U937 cell line exposed to chemical- and/or physical stress factors. Puromycin (PMC) was used as a chemical apoptosis inducer. Alternating (AC) (6.5rms mT, 35 Hz) magnetic field combined with 6 mT static (DC) component, or pulsed electromagnetic field (45 ± 5)mT, 50 Hz (PEMF) acted as physical stressors. Cell viability was assessed by flow cytometry, and the Western blot analysis was carried out for CaM and Hsp70 levels in cytosolic extracts of U937 cells. Cell viability in samples exposed to MF alone did not differ from sham sample, for both types of MF exposure systems. Simultaneous action of MF and PMC influenced cell viability in type of MF stimulation-dependent manner. In contrast to PEMF + PMC stimulated samples, combination of ACDCMF with PMC enhanced cell death compared to PMC control. The observed changes in cell viability were correlated with changes in level of CaM and Hsp70 proteins. Immunoblots have shown, that cytosolic content of both CaM and Hsp70 proteins was enhanced in PMC-treated sample, and further elevated for ACDCMF + PMC. For PEMF + PMC stimulated samples, level of CaM was reduced compared to PMC-treated sample. The results suggest that the changes in expression of CaM and CaM-dependent proteins might modulate effectiveness of cell death under stimulation with MF and/or cytotoxic agents.

Stochastic simulations of remodeling applied to a two-dimensional trabecular bone structure.

In this investigation the stochastic dynamics of trabecular bone remodeling (which is a key concept in a range of remodeling models) is verified. For this purpose, two-dimensional binary images of horizontal histological sections of lumbar vertebral bone (L-3) were collected from young and old subjects. Then the stochastic algorithm of remodeling was applied to the images of young trabecular bone. The resulting images of "simulated" old structures and those of "real" old structures were analyzed to evaluate the following architectural parameters: bone volume/trabecular volume; mean area of marrow cavities; mean trabeculae length; marrow space star volume; Euler number density; trabecular thickness; trabecular number; trabecular separation; mean two-point distance along the skeleton of trabecular structure; probability of disconnection; the ratio of marrow space star volume to geometrical volume and dimensionless ratio of mean area of marrow cavities; and mean trabecular length squared (form factor). Using the parametric t-test, the groups of simulated and real old bone images were compared. It is found that the p value of the t-test is never less than 0.20. For eight parameters the p value is >0.45. It is concluded that, as long as the horizontal sections of lumbar vertebrae are considered (sections perpendicular to the direction of the main spine load), the stochastic algorithm of bone remodeling will properly reproduce the architectural properties of trabecular bone.

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.

Characteristics of CD14 shedding from human monocytes. Evidence for the competition of soluble CD14 (sCD14) with CD14 receptors for lipopolysaccharide (LPS) binding.

The accumulation of sCD14 shed from human monocytes in vivo might correlate with other inflammatory parameters and could be of importance in overcoming a sepsis situation. The development of the sCD14 titer in the supernatant of monocyte-enriched MNC cultures isolated from healthy volunteers was studied utilizing a commercially available sCD14 ELISA. These culture experiments revealed the prolonged liberation of sCD14 into the supernatant during a period of several days. A medium-exchange schedule of 2-3 days was found to be superior to a longer incubation period with respect to the sCD14 yield. PMA initially enhanced the CD14 shedding slightly, but after a few hours it strongly repressed the process. Such a reduction was also achieved by protein synthesis inhibitors (cycloheximide, actinomycin D). Additionally, we monitored the concentration of sCD14, CRP, IL-6 and IL-8 in human sera from healthy persons or patients suffering from severe burn injuries with or without sepsis. Our results indicate that sCD14 is strongly correlated with IL-6, but not with IL-8. sCD14 titers were higher in the group of patients with both burn injuries and sepsis. From experiments with monocyte-enriched MNC cultures isolated from healthy volunteers and medium supplemented with sera containing sCD14 as well as radiolabeled LPS, we conclude that the enhanced shedding of CD14 in vivo during sepsis is probably not able to reduce the binding of LPS to monocytes.

Genetic complementation of the urease-negative Helicobacter pylori mutant N6ureB::TnKm.

Helicobacter pylori produces urease composed of the structural subunits UreA and UreB. Isogenic mutants produced by shuttle mutagenesis from the wild-type strain N6 are widely used in the literature. We describe the genetic complementation of the mutant N6ureB::TnKm by stable transformation with the vector pHel2 containing the cloned genes ureA and ureB and their specific promoter sequence. The orientation of the cloned insert was found to be crucial for urease expression. The majority of complemented clones functionally expressed urease at higher levels than did N6. Homologous recombination between chromosomal and cloned genes occurred at a frequency of 5%.

Comparison of trabecular bone architecture in young and old bones.

In this study, different parameters currently applied for the description of trabecular bone architecture in young and old subjects are compared. Moreover, new parameters are proposed and assessed. For the investigations, the two-dimensional images of vertebral body sections are acquired with the use of a low-magnification digital camera. The parameters were calculated both for the skeleton and for the bone/marrow interface distinguished in the images of the trabecular network. The following methods of the descriptions of the trabecular bone architecture were considered: histomorphometric analysis (BV/TV, asymmetry, mean trabeculae length), fractal geometry technique, Euler characteristic, star volume of the marrow cavity, the mean distance between two points of the trabecular network, and the probability of disconnection (is straightforwardly connected with the number of separated parts of the network). Moreover, bone mineral density (BMD) was determined for comparative purposes. The quantities directly extracted form the images are also combined to produce new indexes, which better differentiate young and old bones. It was found that the BMD, the BV/TV, the star volume, the Euler number, and the probability of disconnection might be used as indicators of the age-related changes of trabecular bone. The parameters could be measured with the precision comparable to that of the BMD. The only exception is the probability of disconnection. Highly significant correlations were demonstrated between bone density (BMD, BV/TV) and trabecular architecture (SV/GV, probability of disconnection).

Optimizing the assessment of age-related changes in trabecular bone.

The goal of this study was to develop an optimal procedure to determine age-related changes in trabecular bone. The investigations were based on two-dimensional images of the human vertebral trabecular bone specimens. The following indices of trabecular structure were considered: bone volume/total volume, star volume of the marrow cavity, Euler number and the probability of disconnection (straightforwardly connected with the number of separated parts of the network). To follow precisely the changes in the trabecular structure with age, a computer simulation model was used. Up to 35 years of physiological remodelling were simulated. The validation of the model calculations was based on a quantitative comparison with the data measured for older individuals. The simulations confirmed that the description of the age-related changes in the trabecular bone by means of the architectural parameter (star volume) constitutes a promising tool for subjects older than approximately 50 years. For individuals younger than approximately 50 years bone mineral density (bone volume/total volume) seems to be the best suited descriptor. The results suggest that the optimal diagnostic procedure is age-dependent and should not be limited to the bone mineral density measurement. The clinical usefulness of the procedure has been validated by examination of the CT images.

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.

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.

Transport of traffic-related aerosols in urban areas.

This study was undertaken to assess the influence of traffic on particulate air pollution in an urban area, and to characterise the short-range transport of the aerosols generated by traffic. The study was conducted in Kraków, a city located in southern Poland with a population of approximately 800,000. Aerosol samples were collected using automatic sampling equipment at five sites located at different distances from the main road in Kraków, ranging from 5 to 1500 m. The sampling set-up allowed standardisation of the results due to continuous determination of the meteorological parameters (temperature, atmospheric pressure, wind speed and direction, rainfall and humidity). Aerosol particles were separated according to aerodynamic diameter into two size fractions: > 1.9 microm (coarse fraction); and 1.9-72 microm (fine fraction). The concentrations of 27 elements were measured in both size fractions (Na, Mg, Al, Si, P, S, Cl, K, Ca, Ti, V, Cr, Mn, Fe, Ni, Cu, Zn, Ga, Ge, As, Se, Br, Rb, Sr, Zr, Ba, Pb). The multielement analyses were performed by Particle-Induced X-ray Emission (PIXE) spectrometry. Traffic contribution to particulate air pollution was determined on the basis of 13 elements which were present above the detection limit in all samples (Mg, Al, Si, P, S, K, Ca, Ti, Mn, Fe, Cu, Zn, Pb). It was found that the traffic contribution in the coarse size fraction was approximately 80% up to 150 m from the road; it dropped abruptly by a factor of 2 over a distance of 150-200 m and declined further to 20% at 1500 m from the road. Traffic contribution for the fine particle concentrations of individual elements was 50-70% in the close vicinity of the road (5 m); then there was a decrease, followed by an increase at a greater distance from the road. Possible explanations for this behaviour of the fine particles are given.

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.

Application of a proton microprobe to biomedical research : Limits and potentialities.

The capability of PIXE analysis to simultaneously detect trace elements with Z≥14, with a high power of detection, can be exploited in biomedical research if the diameter of the proton beam is reduced to micrometer dimensions. In this case, trace analyses of small particles or small parts of a larger specimen are rendered possible without deteriorating the detection limits of PIXE. The measurements yield a completely new type of information on the biological microstructure. In order to fully utilize the abilities of the combined method, however, sample preparation techniques, and irradiation procedures have to be adapted to each analysis problem. Examples of application of the Bochum Proton Microprobe will be used to demonstrate how and to what extent this can be achieved for different types of biomedical problems.

Distribution of elements in rabbit liver lobule.

The elemental composition of rabbit liver was determined by the PIXE and micro-PIXE methods. The mean concentrations of P, S, Cl, K, Fe, Cu, Zn, and Rb measured by both methods were similar. The latter method also allowed for localization of elements within lobule territory. It has been found that some elements are more prevalent in the veins (Cl, Fe) and others in the liver parenchyma (P, Cu, Zn). Moreover, Zn showed the characteristic intralobular distribution. Some methodological aspects of microbeam application to biological materials were also discussed.

[Presence of the trace elements from carbon dioxide absorbent containing lime using a circular apparatus during general anesthesia. ]. / Obecnosc pierwiastków sladowych w wapnie absorbujacym dwutlenek wegla w ukladzie okreznym aparatu do znieczulenia.

The aim of the study was to evaluate release of the trace elements from carbon dioxide absorbent containing soda lime during general anesthesia. We compared two suppliers Polish "Polfa" and German "Dräger". Following trace elements were evaluated: chromium, copper, zinc, cadmium, lead, nickel in soda lime. In blood of the patients we evaluated: copper, zinc, lead, cadmium, bromine, rubidium, iron, mercury. Proton Induced X-ray Emission (PIXE) was used to measure concentrations of the elements. Probes of soda lime were analyzed before anesthesia (Polfa, Dräger), 6 hr after the use (Polfa only) and after 10 weeks (Dräger only). 10 patients were divided in two equal groups, one was anesthetized using soda lime from Polfa and another one from Dräger. Blood samples were taken before anesthesia, immediately after and the next day. Mean values of the concentrations of the elements in soda lime coming from Polfa ranged from 0.20 ppm (nickel) to 7.19 ppm (zinc). In Dräger the measurements were from 0.22 ppm (nickel) to 3.70 ppm (zinc). Mean concentrations of trace elements in blood samples were between 0.20 ppm (lead) and 487 ppm (iron) for the patients anesthetized with Polfa soda lime. In Dräger the measurements ranged from 0.15 ppm (lead) to 485 ppm (iron). Concentrations of cadmium and mercury were below the method's limit. Mean values were almost the same in all time points. Statistical analysis was done using paired t-tests. Values of P < 0.05 were consider significant. We concluded that there were no statistically significant differences between examined groups. Thus, we can say that trace elements were not released from soda lime and concentrations of examined elements in patients' blood were not affected by general anesthesia.

[Physical aspects of selected densitometry methods used in the diagnosis of osteoporosis]. / Aspekty fizyczne wybranych metod densytometrycznych stosowanych w diagnostyce osteoporozy.

Physical background of diagnostic procedures currently applied to assess bone mineral density (single and dual X-ray absorptiometry, quantitative computed tomography and ultrasonography) are described. Special emphasis is given to physical interpretation of diagnostic parameters and to the comparison of the procedures. Systematic errors of the procedures is discussed in detail.

[Calcification of allogenic aortic heart valves]. / Mineralizacja allogennych zastawek aortalnych serca.

The material used for the studies consisted of allogenic aortic valves (AAV) collected from 14 individuals. The necessity of AAV replacement arose from growing circulation insufficiency and AAV dysfunction. The aim the study was the determination of the elemental composition and crystallographic structure of the inorganic deposits in AAV. Moreover, the results of the physicochemical investigations were correlated with clinical data (age of the patient, time between valve replacement surgeries, endomyocarditis, number of infections during last 12 months, arterial hypertension and disturbance of the lipid balance) and with echocardiographic examinations (cusp mineralization and perforation, vegetation, systolic and diastolic dimensions of the left ventricle, maximal and average gradient through allograft valve as well as range of the recoil wave to left ventricle). It was found that mineralization of the AAV cusps was a time-dependent process and took place predominantly at the surface of the cusp. The elemental composition and crystallographic data revealed that the inorganic deposits in AAV were composed of hydroxyapatite crystals. However, the presence of other calcium salts was also found. The development of the mineralization process in AAV does not correlate with endomyocarditis, arterial hypertension and the disturbance of the lipid balance. Probably, endomyocarditis and arterial hypertension induce the pathologic alternations of AAV independently from the mineralization process. The echocardiographic estimations of the pathomorphologic changes of the aortic valve cups are not always consistent with the results of the physicochemical studies.

[Adaptation of computed tomography and roentgenography to bone density measurements]. / Adaptacja tomografii komputerowej i rentgenografii do pomiarów densytometrycznych tkanki kostnej.

The increasing interest in issues connected with osteoporosis has recently caused the development of many new diagnostic methods which allow the measurement of bone density. The DEXA method, performed by specialised densitometers, is one of the most developed and reliable methods. However, the high cost od densitometers and DEXA investigation prevent this method from becoming easily accessible for everyday diagnosis. The adaptation of computerised tomography and rentgenography to densitometric measurements could be one of the methods by which the problem of densitometric diagnosis accessibility could be solved. Both methods are usually applied in the imaging of human tissues, working on the basis of differences in tissue X-ray absorption. X-ray absorption and density are related by linear function in the energy range used in rentgenography and tomography; therefore, quantitative information concerning density should be easily received. The procedure adapting computerised tomography and rentgenography to quantitative measurements of bone density in the lumbar spine is outlined in this work. The quantitative information is obtained from digitalised tomographic and rentgenographic images through use of a personal computer. Both methods were tested using a set of phantoms imitating the lumbar spine and the surrounding tissues. The precision and accuracy of both methods were assessed and compared to the precision and accuracy of the DEXA method. The outlined results confirm the usefulness of the described method in diagnosis.