[Results of efficacy investigation of the toothpastes with natural compounds].

The examination of hard tissues, periodontal and hygiene indices was conducted in 56 patients to determine the efficacy of toothpastes with natural compounds. According to the results, the natural compounds did not alter the color of the hard tissues, but also improved the hygienic status and lessened the periodontal inflammation manifestation in the oral cavity.

The role of N-acetyltransferase 2 polymorphism in the etiopathogenesis of inflammatory bowel disease.

BACKGROUND AND PURPOSE: Inflammatory bowel disease (IBD) consists of ulcerative colitis (UC) and Crohn's disease (CD), which are complex genetic disorders resulting from the interplay between several genetic and environmental risk factors. The arylamine N-acetyltransferase 2 (NAT2) enzyme detoxifies a wide spectrum of naturally occurring xenobiotics including carcinogens and drugs. Acetylation catalyzed by NAT2 is an important process in metabolic activation of arylamines to electrophilic intermediates that initiate carcinogenesis. The aim of our study was to determine whether there is any association between the susceptibility to inflammatory bowel disease among the variations of NAT2 genotypes. METHODS: This study was carried out in 80 patients with IBD. The control group consisted of 100 healthy volunteers. The most common mutations found in the Caucasian population are at the positions 481T, 803G, 590A and 857A on the NAT2 gene. This was determined using the polymerase chain reaction-restriction fragment length polymorphism method with DNA extracted from peripheral blood. RESULTS: Risk of IBD development was 3.86 for the carriers of the NAT2*5/NAT2*7 genotype and 2.53 for the carriers with NAT2*6/NAT2*7, but it was not statistically significant. A statistically significant correlation between the NAT2*7 allele prevalence and the risk for developing IBD was found (OR = 5.8; P = 0.005). CONCLUSIONS: Higher prevalence of the NAT2*7 allele in patients with IBD and the obtained OR values could suggest that this mutation has the effect of increasing IBD development. Future studies are needed to confirm our assumptions on larger group of patients.

Genetic polymorphisms of CYP2D6 oxidation in patients with inflammatory bowel disease.

Inflammatory bowel disease (IBD) consists of ulcerative colitis and Crohn's disease, both of which are associated with increased colorectal cancer risk. The relationship between genetically determined polymorphic metabolism of exogenous substances by oxidation catalyzed by CYP2D6 isoenzyme and susceptibility to cancer has aroused great interest. We determined whether there was an association between susceptibility to inflammatory bowel disease and particularly to CYP2D6 genotypes. The study was carried out in 39 patients with IBD. The control group consisted of 129 healthy volunteers. The CYP2D6 genotypes were analyzed by polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP) method with DNA extracted from peripheral blood. Among 39 patients with inflammatory bowel disease, extensive metabolizer (EM) genotype constituted 97.4%. One patient (2.6%) was poor metabolizer with CYP2D6*4/CYP2D6*4 genotype. Results obtained in the inflammatory bowel disease group did not differ significantly from those of the control group. Although the odds ratio for EM metabolizers was about 3.8-fold greater in the group of patients with inflammatory bowel disease, this association was not statistically significant. This data also showed no overall statistically significant association between alleles and incidence risk of inflammatory bowel disease [odds ratio (OR) of 1.36 for CYP2D6*1 allele, 0.83 for CYP2D6*3 allele, and 0.74 for CYP2D6*4 allele]. The present results suggest that EM genotype may be the risk factor of inflammatory bowel disease. Future studies are needed to confirm our assumptions on larger group of patients.

Lack of association between arylamine N-acetyltransferase 2 (NAT2) polymorphism and systemic sclerosis.

OBJECTIVES: It has been shown that exposure to some environmental toxins may induce scleroderma-like illness in predisposed individuals, but the etiopathogenesis of the idiopathic form of systemic sclerosis (SSc) remains obscure. The genetic background of this illness has been confirmed in multiple studies. We investigated whether patients with SSc differ from healthy subjects with regard to the enzymatic activity of polymorphic N-acetyltransferase 2 (NAT2). METHODS: The study was carried out in 39 patients with SSc; 15 fulfilled the criteria of diffuse SSc (dSSc) and 24 of limited SSc (lSSc); an ethnically matched control group consisted of 100 healthy volunteers. Acetylation phenotype was estimated using the isoniazid as a model drug. The most common mutations in the Caucasian population at positions 481T, 803G, 590A and 857A on the NAT2 gene were determined using the polymerase chain reaction-restriction fragment length polymorphism method with deoxyribonucleic acid (DNA) extracted from peripheral blood. RESULTS: In the group of patients with SSc, the frequency of fast acetylator genotypes was 38.5% (95% CI 23.4-55.4), while that for the genotypes coding slow acetylator status was 51.3% (95% CI 34.8-67.6). There was a strong correlation between NAT2 phenotype and NAT2 genotype with a concordance of 97%. We did not observe a preponderance of slow acetylators among patients with SSc and in two subsets of SSc. With the sample size analyzed in the present study, there is a 90% probability of detecting significant differences in distribution of slow, fast, and intermediate phenotypes between patients with SSc and controls, there is a difference of at least 30.3, 28.7 and 21.9% in the distribution of these phenotypes in the general population, respectively. CONCLUSION: Acetylator status does not seem to be the significant factor in the development of SSc in patients with both subsets of this autoimmune disease, but further studies are required to confirm this conclusion.

Presenilin I interaction with cytoskeleton and association with actin filaments.

Presenilin I (PSI) has been shown to interact with microfilament-associated proteins of the filamin family. Here, we investigated a possible association of PSI with the cytoskeleton. Immunoblotting of detergent-insoluble fractions of rat brain homogenate revealed enrichment of neuron-specific 36 and 14 kDa proteolytic fragments of PSI, whereas 30 and 20 kDa fragments were found in the detergent-soluble fraction. Specific severing of microfilaments with gelsolin in the detergent-insoluble pellet and subsequent centrifugation led to the detection of both actin and PSI fragments in the supernatant. In addition, in vitro translated PSI cosedimented with actin filaments. Our findings provide biochemical evidence for the association of PSI fragments with actin filaments.

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).

[Specific anesthetic procedures in acute poisoning]. / Specyfika postepowania anestezjologicznego w ostrych zatruciach.

The decline of mortality rate in acute poisoning is connected with a completely altered method of treatment, namely the method of intensive treatment and extracorporeal elimination of the poisons. The anaesthetist plays a specific role in the treatment of acute poisoning since he cooperates with toxicologist and takes active part both in reanimation and intensive treatment of the poisoned. Anaesthetic procedure in toxicology is used as causal therapy and the part it plays is specific especially in extracorporeal elimination of the poison from the body. The authors of these studies discussed the methods of anaesthetic treatment in acute poisoning and presented its dynamic use in the period from 1966 to 1937 in the Toxicological Clinic of the Medical Academy in Kraków.

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.

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.

Elemental composition of the human atherosclerotic artery wall.

The elemental composition of the human atherosclerotic popliteal artery was examined using the proton-induced X-ray-emission (PIXE) method. The application of a narrow proton beam (3 X 10 micron 2) enabled us to determine not only the concentrations of Cl, K, Ca, Fe, Cu, Zn, Br and Pb, but also their localization in different artery-wall regions. The highest mean concentrations of Cl, K, Zn and Br were found in the tunica media. In the investigated sections the distribution of Ca and Fe varied: sometimes, these elements were prevalent in the tunica intima, whereas in other cases, the highest concentrations were observed in the tunica media or tunica adventitia. The concentration profiles of each element were characterized by many sharp, narrow peaks. The highest concentrations of Ca and Fe showed such high levels that only one explanation is possible, i.e. the presence of crystals. The correlation of Ca peaks with those of Zn and Fe is discussed. The usefulness of the micro-PIXE method for the investigation of biomedical materials is also considered.