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Brain-derived neurotrophic factor (BDNF) is involved in the survival and maturation of neurons, and also promotes and controls neurogenesis. Its levels are lowered in many neurodegenerative diseases, including Huntington’s disease (HD). Clinical pictures of HD can be very diverse, which makes it difficult to assess its severity, however, molecular markers may be helpful. The aim of the study was to determine the relationship between HD severity and the plasma BDNF concentration in HD patients. The study recruited 42 patients with diagnosed and genetically confirmed HD and 40 healthy volunteers. BDNF levels were determined in plasma with the enzyme-linked immunosorbent assay (ELISA). Correlations between BDNF levels and clinical profiles and HD severity were also investigated. The BDNF level was significantly lower in HD patients compared to the control. There was no correlation between the BDNF level and motor symptoms and cognitive impairment. In the early disease stages, BDNF levels were associated with a better neurological examination, independence, and functional evaluation, in contrast to later HD stages, where the correlations were inverse. Multidirectional correlations between parameters of saccadic disorders and the BDNF level do not allow for drawing a conclusion, whether or not there is a relationship between the severity of saccadic disorders and the BDNF concentration.

Introduction One of the examples of genes whose expression can be altered by the action of ustekinumab is TGF-β. It is a pleiotropic cytokine whose activity affects psoriatic changes and the state of homeostasis of the whole organism. Aim To evaluate the effect of ustekinumab on the transcriptional activity of TGF-b family genes in patients with psoriatic arthritis and to check whether the results obtained can be helpful in monitoring the progress of treatment. Material and methods From total PBMCs obtained from peripheral blood of 14 patients with psoriatic arthritis, total RNA was isolated. The expression level of the TGF-β1, TGF-β2 and TGF-β3 genes was determined by RT-qPCR in real time. Results In all the analysed samples, the presence of mRNA of three TGF-β isoforms was quantitated in each week of therapy. TGF-β3 and the smallest TGF-β2 showed the highest expression. Statistically significant correlations were observed in the amount of TGF-β1 and TGF-β3/µg mRNA RNA, TGF-β2 and TGF-β2/µg RNA and TGF-β3 and TGF-β3/µg RNA. Conclusions Ustekinumab influences the transcriptional activity of TGF-β genes, and the changes caused have a bearing on the patient’s health.

Huntington’s disease (HD) is an autosomal dominant genetic disease that can be divided into preclinical and symptomatic stages. Due to the diverse HD phenotype, there is an urgent need to identify markers that would independently assess its severity. The aim of this study was to evaluate the use of plasma levels of TGF-β1 in the assessment of HD severity. One hundred HD patients and 40 healthy volunteers were included in the study. All HD patients underwent neurological and cognitive function assessment. TGF-β1 levels were determined in the plasma of all patients. The correlations between TGF-β1 levels and clinical profile and HD severity were also investigated. In symptomatic patients, cognitive decline was demonstrated, while in preclinical patients, no symptoms were found. Plasma levels of TGF-β1 in HD patients did not differ significantly from the control group and did not change with the progression of the disease. In addition, TGF-β1 levels also did not correlate with the severity of motor dysfunction. Positive correlations between plasma TGF-β1 concentration and intensity of cognitive impairment were found, but only in the early disease stage. There was no clear benefit in assessing plasma TGF-β1 levels in HD patients as a marker of disease severity.

Background Endoglin is a marker of active, proliferating endothelial cells of blood vessels. In many cancers, it is present in both peripheral vessels and vessels located inside the tumor. Endoglin is more specific and sensitive compared to other tumor angiogenesis markers. It is suggested that endoglin can be considered a reliable marker of disease outcome. Objective The aim of the study was to assess the expression of endoglin and to determine its potential usefulness as a complementary molecular marker of endometrial cancer. Method The study included 60 women who underwent hysterectomy: 45 with endometrioid endometrial cancer (study group) and 15 without neoplastic changes (control group). The study group was further divided according to the degree of histological differentiation: G1, 17; G2, 15; and G3, 13. The expression of endoglin was determined immunohistochemically with mouse anti-Endoglin monoclonal antibody. The obtained reactions were evaluated using light microscopy. Results Analysis of endoglin expression in endothelium showed that it reached 145% of the control. In G2, we observed that the endoglin level decreased and was similar to the control, while in G3 it increased and was even higher than in G1. In cancer cells, endoglin expression increased with the grade of endometrial cancer. Conclusion Endoglin can be considered a valuable complementary molecular marker, allowing to visualize the advancement of the cancer process, including endometrial cancer.

Background:Many experimental studies have demonstrated the importance of COX-2 in the tumor angiogenesis. Inducible iNOS is responsible for a high and stable level of nitric oxide and is expressed in response to pro-inflammatory factors.Objective:The aim of this study was to evaluate the expression of COX-2 and iNOS at the protein level and to assess their potential prognostic significance in patients with endometrial cancer.Methods:The study group consisted of 45 women with endometrial cancer divided according to the degree of histological differentiation i.e. G1, 17; G2, 15; G3, 13. The control group consisted of 15 women without neoplastic changes. The expression of studied proteins was determined immunohistochemically with specific polyclonal antibodies.Results:Analysis of the COX-2 expression showed that the optical density of the reaction product in G1 reached 186% in the control group, while the values in G2 and G3 reached 243% and 293%, respectively. In the case of iNOS, the optical density of the reaction product reached the following percentages in the control group: 147% in G1, 243% in G2, and 241% in G3.Conclusions:Our findings suggest that changes in the expression of COX-2 and iNOS may be potentially useful in predicting the progression of endometrial cancer and treatment effectiveness.

Introduction: The pathogenesis of nasal polyps is still not fully understood. Aim : To analyze the topography and intensity of interleukin 1β (IL-1β), tumor necrosis factor α (TNF-α), cyclooxygenase 2 (COX-2), nitric oxide synthase 2 (NOS-2), and nuclear factor-κB (NF-κB) expressions in eosinophilic and neutrophilic polyps and in normal nasal mucosa. Material and methods : The study included specimens from 20 patients with eosinophilic polyps (more than 10% of eosinophils in inflammatory infiltrate), 20 individuals with neutrophilic polyps (predominance of neutrophils and less than 10% of eosinophils), and samples of normal nasal mucosa from 10 controls. The expressions of studied proteins in vascular endothelial cells, epithelial, stromal and glandular cells were determined immunohistochemically with specific monoclonal antibodies. Results : Irrespective of the cellular type, the intensity of expressions in eosinophilic and neutrophilic polyps was significantly higher than in the normal mucosa. Eosinophilic polyps were characterized by stronger expressions of TNF-α (in all cellular types), IL-1β (in endothelial, glandular and epithelial cells), NF-κB (in stromal and epithelial cells), COX-2 (in glandular and stromal cells), and NOS-2 (in endothelial and stromal cells). In contrast, neutrophilic polyps showed significantly stronger expressions of COX-2 (in epithelial and endothelial cells) and NOS-2 (in glandular and epithelial cells). In both phenotypes, the strongest expressions of all studied markers were documented in vascular endothelial cells. Conclusions : Inflammatory markers are involved in pathogenesis of both eosinophilic and neutrophilic polyps. Endothelial defects can play an important role in the development of nasal polyps.

Background Brain-derived neurotrophic factor (BDNF) is a widespread cytokine, which may be found both in central nervous system and peripheral tissues. It plays multiple roles and there are some literature suggestion that it may be involved in HD pathophysiology. Aims To asses BDNF plasma level utility as HD biomarker. Methods The blood was collected from 42 HD patients vs 39 healthy control subject matching for age and sex. Plasma level of BDNF was evaluated in both groups. All HD subjects went UHDRS evaluation. Study group was divided into different severity group on the basis of motor UHDRS score, which resulted in: 13 early stage subjects, 21 moderate, 3 advanced and 3 preclinical. Results A total of 42 HD patients (24 female) were enrolled in the study. Mean age was 49,95±12,38, CAG level 44,18±4,89, motor UHDRS 43,78±21,31. The mean level of BDNF was 1,64±1,62 vs 3,71±2,53 in controls, which was a statistically significant difference (p=0,00003). We found several correlation with plasma BDNF level – in early stage disease: with motor UHDRS score (p=0,05, R=– 0,55), bradykinesia (p=0,005, R=– 0,49), lower limbs (p=0,02, R=0,62) and trunk dystonia (p=0,05, R=0,51), gait disorders (p=0,05 R=– 0,50); severe HD: motor UHDRS score(p=0,05 R=– 0,85), eye movements and speech disorders (p=0,04 R=0,89) and upper limbs dystonia (p=0,04, R=– 0,97). Cognitive evaluation results and plasma BDNF correlation were noted too. Conclusions Plasma BDNF correlates with HD severity, mostly in fully symptomatic stages of HD.

Objectives: The aim of the study was to determine the expression of VEGF (vascular endothelial growth factor) isoforms and their receptors in uterine myomas. Material and methods: The study included 40 women with myomas of reproductive age and 40 perimenopausal women (the study group). Myometrial samples (the control group) were taken from 10 women undergoing hysterectomy for ovarian tumors and 10 older women undergoing hysterectomy for uterine prolapse. Results: A significantly increased expression of VEGF-A has been found in myomas, both small and large, in the younger women, which may by a sign of increased angiogenesis and intensive tumor growth. In perimenopausal women, the increase of VEGF expression was observed only in the endothelium and vascular smooth muscle. Conclusion: An important conclusion of this study is that angiogenesis is independent of myoma size, which may suggest intensive tumor growth and the related increased angiogenesis. High expression of VEGF-A and VEGF-R1 receptors in large myomas can probably cause malignant transformation and more extensive growth, regardless of patient age.

Objectives Introduction: Survivin is a member of the inhibitor of apoptosis protein (IAP) family, which are selectively overexpressed in human neoplasms, and its expression has been shown to be connected with cell proliferation. We analyzed survivin expression in ovarian epithelial neoplasms to evaluate its role in the development of ovarian tumors. Material and methods: Immunohistochemistry assays were conducted in 137 cases (48 ovarian carcinoma, 43 borderline ovarian carcinoma, 46 benign ovarian tumor, and 20 samples of normal ovarian tissue of ovarian epithelial neoplasms. Histological types included serous (n=68) and mucinous (n=69) tumors. All tumors were reviewed histopathologically and classified according to the WHO criteria. Results: Survivin expression in the group of serous neoplasms was detected in 24.0% (6 of 25) of benign cases, in 60.0% (12 of 20) of borderline tumors, and 91.0% (24 of 47) of ovarian carcinomas. In the group of mucinous tumors, survivin expression was found in 33.5% (7 of 21) of benign cases, 43.5% (10 of 23) of borderline tumors, and 80.0% (20 of 25) of malignant tumors. Conclusions: Our results demonstrate that survivin overexpression may play a crucial role in the development of epithelial ovarian neoplasms and be an important prognostic factor for the influence of survivin expression on epithelial ovarian cancers.

Epithelial ovarian cancer represents one of the most deadly gynaecological neoplasms in developed countries and is a highly heterogeneous disease. Epidemiological studies show that anti-inflammatory drugs reduce the incidence and mortality of several types of cancer, indicating the potential role of pro-inflammatory factors in carcinogenesis. The expression of pro-inflammatory factors in various cancer types, including ovarian cancer, was assessed in many studies, yielding in consistent results, often due to the histological heterogeneity of various cancers. The aim of the study was to investigate the expression of IL-1, IL-6, TGF-β, TNF-α, COX-2, iNOS, and NF-kB in serous and mucinous ovarian cancers. Ninety cases of ovarian tumors classified into mucous and serous type (45 patients in each group) were selected. Each group was classified into subgroups according to the three stages of tumor differentiation, i.e. into (i) benign, (ii) borderline and (iii) malignant tumors. The presence of proteins of interest in paraffin sections was analysed by immunohistochemistry. The expression of most of the studied factors depended on the histological tumor subtype and the degree of malignancy. Expression of NF-κB appears to be related to the level of the neoplastic differentiation only in the group of serous tumors, while the presence of IL-6 in the mucinous tumor subtype was observed only in the case of benign lesions. Expression of IL-1, TNF-α and COX-2 increased with the stage of the disease in both serous and mucinous tumors. The highest level of TGF-β expression was observed in serous borderline tumors. The different levels of iNOS immunoreactivity between the groups of serous and mucinous tumors were observed only in borderline tumors. The results of our study may be helpful in designing therapeutic strategies depending on the type of ovarian cancer.

In this study the expression of GnRH, FSH, LH, ER-α, ER-β, and PR receptors was examined in uterine myomas of women in reproductive and perimenopausal age. In cases of GnRH and tropic hormones a membranous and cytoplasmic immunohistochemical reaction was detected, in cases of ER-αand PR the reaction was located in cell nucleus, and in the case of ER-βit manifested also a cytoplasmic location. In some of the examined cases the expression was detected in endometrium, myocytes, and endothelium of blood vessels, in uterine glands and myoma cells. In myometrium the level of GnRH and LH receptors increases with age, whereas the level of progesterone and both estrogen receptors decreases. In myomas of women in reproductive age, independently of their size, expression of GnRH, FSH, and LH receptors was more pronounced than in myometrium. In women of perimenopausal age, independently of myoma size, expression of LH and estrogenαreceptors was higher while expression of GnRH receptors was lower than in myometrium. FSH receptor expression was not observed. Expression of estrogen receptorβwas not affected by age of the woman or size of myoma. Analysis of obtained results indicates on existing in small myomas local feedback axis between GnRH-LH-progesterone.

Uterine myomas represent one of the most frequently manifested benign tumors in women. They originate from smooth muscle cells of myometrium or its blood vessels. Many studies suggest that inflammation and pro-inflammatory factors may play a role in the carcinogenesis with an involvement of the transcription factor NF-kappaB which activity can be controlled by various environmental factors, including many cytokines. The aim of the study was to investigate the expression of NF-B, interleukin-1β (IL-1β), tumor necrosis factor a (TNF-α), cyclooxygenase 2 (COX-2) and inducible nitric oxide synthase (iNOS) in myometrium and uterine myomas of women of various age. The expression of NF-kappaB, selected cytokines and enzymes was estimated in women of reproductive or perimenopausal age by semiquantitative immunohistochemistry. The expression of the examined proteins was higher in myomas than in control myometrium and was dependent on the size of myomas and the age of women. However, the expression of the cytoplasmic NF-kappaB observed in uterine myomas was independent on the size of myomas and no significant differences were observed in the number of stained nuclei between control and myoma groups. Thus, the expression of proinflammatory factors in myomas was not accompanied by the nuclear activation of NF-kappaB p65. The results of our study indicate that the examined factors may be involved in the pathogenesis of benign tumors and not only malignant diseases.

Introduction Parkinson disease (PD) is the common neurodegenerative disease. α-Synuclein (ASN), main aggregating protein in neural cells of CNS in PD, was found in peripheral fluids. Testing ASN in plasma is potential test for diagnose PD, but previous studies are controversial. The aim of this study was to investigate if plasma ASN level may be a valuable biomarker, is the level of plasma ASN concentration different in various motor subtypes of diseases, is there a relation between the level of plasma ASN and the severity of motor symptoms. Methods Patients with PD hospitalized in Neurology Department, Medical College were performed sequencing the 8th and 9th exon of GBA gene. Next plasma ASN level was tested in 58 patients with sequenced GBA gene and in 38 healthy volunteers (HV), matched by the age (respectively 68.43 vs. 64.57 years of age) and sex (female %, respectively: 43.10 vs.44.74). Patients were assessed with the scales: UPDRS (II, III, IV), Hoehn–Yahr (HY) and qualified to PIGD or TD subtype. For homogeneity of the group patients with GBA mutation were excluded from the analysis. Results The ASN level did not differ between patients and HV (respectively: 4.53 vs. 3.73 ng/ml) and between patients with different subtypes. There was inverse correlation between ASN and HY in PIGD subtype. Conclusions Plasma ASN level is not valuable marker of the disease. It does not differ in subtypes of the disease. There is relation between plasma ASN level and the severity of the disease in PIGD subtype.

The publication of the human genome sequence provided direction in the search for novel diagnostic and therapeutic methods for the treatment of human diseases. The aim of the present study was to investigate the hypothesis that the expression profile of genes involved in the regulation of angiogenesis may be a marker in endometrial cancer that facilitates the diagnosis and prognosis of patients, as well as the identification of novel therapeutic targets. The current study included 36 patients with grade (G) 1 to 3 endometrial cancer, and a control group of patients consisting of females that qualified for the removal of the uterus. Out of these, 28 samples (control, 3; G1, 7; G2, 12; and G3, 6) were selected for microarray analysis. Molecular analysis of the endometrial samples involved the extraction of total RNA, purification of the obtained extracts and subsequent analysis of the gene expression profiles using an oligonucleotide microarray technique (GeneChip® Human Genome U133A plates). The results indicated that the mRNA expression profile of genes involved in the regulation of angiogenesis varies depending on the degree of histological differentiation of endometrial adenocarcinoma. Similar results were obtained from descriptive statistics characterizing the expression profile of 691 mRNAs associated with the regulation of angiogenesis in the groups of patients with endometrial adenocarcinoma. In addition, the results of the present study indicated that neuropilin2 (NRP2) may serve an important role in the activity of endothelial cells, and may affect vascular endothelial growth factor, and potentially plexins and integrins via regulation of their functions. An understanding of how these proteins interact remains to be determined; however, elucidating these interactions may provide an explanation for the mechanisms underlying angiogenesis. In conclusion, the results of the present study suggest that NRP2 may be a valuable target for investigation in future pharmacological studies involving angiogenesis in endometrial cancer.

Background: In case of overdose of paracetamol the ability of hepatic biotransformation is saturated and accumulation of toxic metabolite – NAPQI takes place. Main CYP isoforms considered to be responsible for bioactivation of APAP and promoting the same liver intoxication are CYP2E1, CYP1A2, CYP3A4 and in animals 2B1/2 isoforms additionally. Purpose of this work was examination of paracetamol influence and/or trichloroethylene on the composition of hepatic cytochrome P450 isoforms. Materials and method: Tests were carried out on rats which were treated with trichloroethylene, paracetamol and/or N-acetylcysteine. In the microsomal fraction content of three isoforms of cytochrome P450 i.e. CYP2E1, CYP2B1/2 and CYP1A2 were determined. Results: Paracetamol slightly stimulated CYP2B1/2 lowering simultaneously level of CYP1A2. Trichloroethylene stimulated CYP2B1/2. N-acetylcysteine stimulated all tested P450 isoforms. N-acetylcysteine given together with examinated xenobiotics induced studied P450 isoforms. Conclusions: N-acetylcysteine demonstrated a protective effect on studied CYP isoforms especially when was given upon termination of xenobiotics exposure

Background: Feature of modern existing hazards both environmental and occupational is cumulative exposure often leading to unexpected response of the organism resulting, among other things, in interactions with cytochrome P450 system involved in biotransformation of trichloroethylene and paracetamol. Hepatotoxity of paracetamol is closely connected with hepatic glutathione level. „In therapy of acute paracetamol poisoning application of N-acetylcysteine as a factor, which protects GSH level in cells, is recommended.” Materials and method: Tests were performed on rats which were treated with trichloroethylene, paracetamol and/or N-acetylcysteine. In rat liver total level of glutathione was determined i.e. reduced and oxidized form. Results: Paracetamol just after completion of the exposure affected the glutathione level. Trichloroethylene throughout the period of observation stimulated growth of glutathione level in liver. N-acetylcysteine didn’t have any influence on the level of investigated tripeptyde. Conclusions: N-acetylcysteine removes negative effect of paracetamol especially when it’s applied with 2-hour delay. After exposure for trichloethylene immediate application of N-acetylcysteine caused noticeable lowering of glutathione level. Cumulative exposure for three xenobiotics had positive influence for glutathione level in rat liver.

Background: There is a number of factors which potentially affect occurrence of toxic change in liver after overdosing of paracetamol. Hepatic metabolism of trichloroethylene has primary impact on hepatotoxic effect of this solvent. This means that the combined exposure to these xenobiotics can be particularly harmful for human. The influence of N-acetylcysteine (NAC) as a protective factor after paracetamol intoxication was studies. Materials and method: Tests were carried out on rats which were treated with trichloroethylene, paracetamol and/or N-acetylcysteine. In the hepatic microsomal fraction activity of the components of cytochrome P450- dependent monooxygenases was determined Results: Paracetamol slightly stimulated cytochrome P450 having no effect on reductase activity cooperating with it. Cytochrome b5 and its reductase were inhibited by this compound. Trichloroethylene was the inhibitor of compounds of II microsomal electron transport chain. N-acetylcysteine inhibited activity of reductase of NADH-cytochrome b5. Conclusions: Tested doses of the xenobiotics influenced on II microsomal electron transport chain. Protective influence of N-acetylcysteine was better if this compound was applied 2 hours after exposure on xenobiotics

We studied uterine myomas originating from females of reproductive age and from females of perimenopausal age. Uterine myomas represent benign tumors of the myometrium, and they develop frequently in women of reproductive age. The frequency of uterine myomas increases with age until women reach the menopause. The study included patients with a myomatous uterus, in the reproductive age or peri-menopausal age, independently evaluating small and large myomas. Myometrial alterations in their direct vicinity were evaluated independently of the myomas. The study included evaluation of immunolocalization of two index proteins which participate in myoma cells growth control: Ki-67 nuclear antigen and caspase 3. In women of reproductive age, both in small and large myomas, elevated immunostaining of Ki-67 was noted in parallel to low levels of caspase 3 staining, which indicated the ongoing process of proliferation. In women of peri-menopausal age with small or large myomas, no Ki-67 immunostaining was detected, while staining of caspase 3 manifested low levels. Proliferation in reproductive age women myomas is higher than in the peri-menopausal age. (Folia Histochemica et Cytobiologica 2011; Vol. 49, No. 3, pp. 398–404)

Uterine myomas represent one of the most common female pathologies. Uterine smooth muscle myomas or fibromas are benign tumours which respond to hormones and their etiology induces wide interest. The myomas were found to contain aromatase and, in addition, cells of the myomas were found to synthesize estrogen. This study was conducted on patients with the myomas, in either generative age or in the perimenopausal period. Expression of aromatase was detected in patients of various age, with large or small uterine myomas, using an immunohistochemical technique. In addition expression of the enzyme was examined at the periphery of every myoma.

Summary Nucleolar organizer regions (NORs) were evaluated in 11 women with adenomyosis. The material, sampled during operations, was silver-stained using the technique of Ploton et al. and verified histopathologically. A hundred cell nuclei per slide were assessed. The examined variables comprised the number of argyrophylic nucleolar organizer regions (AgNORs) in the nucleus, the surface area of a single AgNOR and the position of the AgNORs in the cell nucleus, as well as the AgNORs’ coefficient, which were all estimated under a microscope. The parameters were quantitatively assessed using computer image analysis software Multi-Scan Base V.8. Adenomyosis was found to be associated with a significant decrease (up to 60%) in the nuclear area, as well as with a marked reduction (up to 50% as compared with normal endometrium) in the number of AgNOR granules. The total area of AgNORs in the cell nucleus also decreased from 3.55 to 1.57 μm2. There were no significant differences in the number of granules per nucleus either in the control group or in the adenomyosis group. The AgNOR coefficient was found to be lower in adenomyosis compared to normal endometrium.

The aim of this study was to evaluate the effects of chronic exposure to cadmium (Cd) on the renal cytochrome P450-dependent monooxygenase system. For this purpose, male Wistar rats were intoxicated with Cd administered in drinking water at a concentration of 5 or 50 mg Cd/l for 6, 12 and 24 weeks. Concentrations of cytochrome P450 and cytochrome b(5) as well as activities of NADPH-cytochrome P450 reductase and NADH-cytochrome b(5) reductase were determined in the kidney microsomal fraction. Protein content of CYP1A1, CYP2E1 and CYP3A1 cytochrome P450 isoforms was evaluated as well. In the rats exposed to 5 mg Cd/l, the concentration of cytochrome P450 decreased (by 41%) after 24 weeks of the experiment. The activity of NADPH-cytochrome P450 reductase decreased (by 24%) after 6 and 12 weeks, whereas after 24 weeks it remained unchanged, compared with the control group. Moreover, a decrease in the concentration of cytochrome b(5) (by 25, 15 and 26% at 6, 12 and 24 weeks, respectively) and the activity of its NADH reductase (by 26 and 31% at 6 and 24 weeks, respectively) was noted in these animals. At the exposure to 50 mg Cd/l, the concentrations of cytochrome P450 and cytochrome b(5) and the activities of their corresponding reductases were decreased at each time-point. Western blot analysis revealed that all isoforms of cytochrome P450 studied were affected by Cd and the effect was dependent on the level and the duration of exposure. The results of this study indicate that chronic exposure to Cd in a dose- and time-dependent manner affects the kidney cytochrome P450-dependent monooxygenase system by decreasing the concentrations of cytochrome P450 and cytochrome b(5) and inhibiting the activities of their corresponding reductases. The effect of Cd on the cytochrome P450 content is associated with its ability to stimulate or inhibit of various P450 isoforms. A very important finding of this study is that Cd affects the kidney cytochrome P450-dependent monooxygenase system at relatively low exposure and low kidney Cd accumulation (2.40+/-0.15 microg/g). As the experimental model used reflects human exposure to Cd, we conclude that Cd can affect the kidney cytochrome P450-dependent monooxygenase system in environmentally exposed humans. Previously we have reported disorders in the system in the liver of rats at the same levels of exposure as in this study. Thus, we hypothesize that the metabolism and detoxification of many substances, including xenobiotics, may be seriously affected in Cd-exposed subjects.

The distribution of tissue kallikrein (TK) and its plasma inhibitor, kallistatin in plasma and intestinal tissue, was studied in patients with active ulcerative colitis (UC) and Crohn's disease (CD). TK was localized to goblet cells and kallistatin to epithelial cells of normal human intestine. Both proteins are visualized in macrophages inside granulomas in CD as well as in plasmocytes in both CD and UC. Intestinal tissue kallikrein (ITK) and kallistatin are significantly decreased in inflamed intestine compared to noninflammatory controls. TK mRNA is significantly decreased in intestinal biopsy samples from active UC patients compared with inactive patients or controls. Immunoreactive TK is present in plasma in very low concentrations in patients and did not differ in normal subjects. Plasma kallistatin was significantly decreased in patients with active disease compared to normal controls. Our data suggest that release of TK during inflammation plays a role in inflammatory bowel disease.

Background: It has been proved that nuclear factor-kappa B (NF-κB) is activated in all cells, promotes proliferation of cells, regulates the immunological and inflammatory response, and contribute to the pathogenesis of many conditions, including cancer. Many studies pointed to constitutive activation of NF-κB in cells of certain malignant tumors. Objective: The aim of the study was to analyze the role of nuclear growth factor κB as colon cancer marker and prognostic factor. Materials and Methods: The study included 59 primary colorectal tumor patients and 15 patients in control group. The tumor samples were taken during partial colectomy and colonoscopy in control group. Tissues samples were fixed and embedded in paraffin blocks and cut. Sections were used for schedule immunohistochemical staining with the application of specific antibody for NF-κB epitope. The marker expression was compared with well-known prognostic factors in colon tumors such as tumor type, stage, and grade to establish if it might be a potential prognostic factor. Results: The results showed statistically significant difference between control group and cancer group. Conclusions: The expression NF-κB did not depend on the stage and grade of colon tumors.

Objective: The saphenous vein subjected to arterial pressure stretches to its elastic limits and constitutes intimal hyperplasia. Sheathing of the vein graft with pressure-resistant tubing might prolong vein graft patency. Methods: Twenty-one sheep received radial vein grafts or hybrid grafts composed of radial vein, collagen fibrin glue and highly flexible torlen/dacron mesh tubing transplanted into the carotid artery position. Veins were examined with the use of light and electron microscopy. Proliferating cell antigen (Ki-67) stains served as markers of proliferation. Results: The mean wall thickness of both intimal and medial layers was evaluated. The mean intimal wall thickness was 19 ^ 11 mm in hybrid grafts vs. 24 ^ 7 mm in unsheathed grafts (P , 0:001); 22 ^ 6 vs. 26 ^ 10 m m( P , 0:001); 23 ^ 8 vs. 52 ^ 15 mm (P , 0:001); 37 ^ 21 vs. 90 ^ 31 m m( P , 0:001); 57 ^ 31 vs. 104 ^ 28 m m( P , 0:001); 58 ^ 21 vs. 133 ^ 32 m m( P , 0:001); and 72 ^ 22 vs. 244 ^ 100 m m( P , 0:001) after respectively 5 days, 9 days, 4 weeks, 6 weeks, 8 weeks, 10 weeks and 12 weeks from implantation. Electronic microscope examination of hybrid grafts revealed a smooth endothelial layer with intact nuclei and an intima composed of layers of collagen and muscle fibers. In unsheathed grafts endothelial edema and nuclear destruction were observed. Conclusions: The external vein graft support with mesh tubing reduces intimal and medial layer thickening and cell proliferation in composite vein grafts transplanted in the arterial position. q 2002 Elsevier Science B.V. All rights reserved.

The study aimed to evaluate the effect of age on the activity of the cytochrome P450-dependent monooxygenase system and on cellular respiration processes in Wistar rats aged 0.5, 1, 2, 4, 8, 12, 20, and 28 months. The following parameters were determined: cytochrome P450 content, cytochrome b5 content, NADPH-cytochrome P450 reductase activity, and NADH-cytochrome b5 reductase activity, succinate dehydrogenase (SDH) activity, and lactate dehydrogenase (LDH) activity. In the study, cytochrome P450 content increased in the first month of life, which was accompanied by increases in SDH and LDH activities. In the subsequent months, SDH activity decreased, whereas LDH activity increased to reach the maximum in month eight and then decreased. Cytochrome b5 content showed a decreasing tendency throughout the experiment. NADH-cytochrome b5 reductase activity showed only slight deviations in individual age groups.

The current interest in CYP expression in the colon results from its uniqueness as a target organ for cancer. To date, the CYP expression profiles in the colon have not yet been subject of comprehensive research. In this study, we investigated 40 patients with Crohn's disease, 40 with ulcerative colitis, and 40 healthy subjects as a control group. Colon tissues were fixed, dehydrated, cleared in xylene and embedded in paraffin. Sections were prepared from paraffin blocks for immunohistochemical staining with specific antibodies. We used antibodies to the human CYP1A1, CYP2B6, CYP2C9, CYP2E1 and CYP3A4 isoforms, as well as antibodies to the human glycoprotein P, glutathione-S transferase and antibody to the UDP-glucuronosyltransferase. The sections were stained immunohistochemically and examined using light microscopy. Cellular localization was determined, and computer image analysis was used. In all cases with Crohn's disease, the proteins studied showed at least a twofold expression. Ulcerative colitis showed a much weaker influence regarding the expression of the proteins studied but in case of CYP2C9 and UDP-glucuronosyltransferase, a decrease of expression was observed.

This study was undertaken to investigate the effects of age and some inducers (BNF-40 mg/kg/day, DEX − 20 mg/kg/day for 3 days each) on hepatic mixed function oxidase. Wistar male rats of very young (0.5 and 1 months), older (2, 4, 8,12 months) and very old (20 and 28 months) ages were used. The cytochrome P-450 content and the activity of NADH-cytochrome b5 reductase were highest in one-month-old animals, while the cytochrome b5 content was highest in the youngest but NADPH-cytochrome P-450 activity in the oldest rats. β-Naphthoflavone (BNF) treatment resulted in an enhanced content of cytochrome P-450 in all age groups but the activity NADPH-cytochrome P-450 reductase was increased only in the oldest rats. Dexamethasone (DEX) treatment led to a decrease in the activity of NADPH-cytochrome P-450 reductase in the oldest animals only. The cytochrome P-450 content was decreased in young animals (0.5-, 1-, 2-month-old), too. BNF and DEX decreased the cytochrome b 5 content and the activity of NADH-cytochrome b5 in all age groups.

Till the end of 1980s transmission of the message carried by the hormone was thought to be mediated by a single estrogen receptor only. In the mid 1990s studies were published which described the discovery of a new estrogen receptor in both animals and humans [4-7]. The new estrogen receptor, ER-proved to be highly homologous to the classical estrogen receptor, ER- Functional differences at the molecular level between ER- and ER- are of principal importance for the body. Just as importantly, in the case of estrogen receptors the same ligand may play role of either an agonist or antagonist, depending on whether it binds to ER- or ER-. Therefore, following binding of the ligand ER- and ER- may induce distinct biological effects, linked to conformational changes of the receptor [8].

delta-Aminolevulinic acid synthase and heme oxygenase are closely functionally connected with the hepatic microsomal mixed function oxydase system. There is a wide range of changes in microsomal heme oxygenase activity, cytochrome P-450 level and mitochondrial delta-aminolevulinic acid synthase activity in rat livers at various stages of life. We tried to investigate the spontaneous phases of these enzymatic protein activities and the levels of cytochrome P-450 as a function of age. We noted that each of these proteins change their activity and level very characteristically during the rats' lifetime. However, correlations between heme oxygenase activity, delta-aminolevulinic acid synthase activity and cytochrome P-450 levels still need to be investigated to elucidate the mechanism of these connections.

An age‐associated alterations in daily and circadian changes of cytochrome P‐450‐linked monooxygenases system activity were studied using 1‐, 6‐ and 12‐months old male Wistar rats, in winter and in spring season. Cytochrome P‐450 and NADPH‐cytochrome P‐450 reductase showed 12h rhythm in all investigated age groups of animals. Cytochrome b5 and NADH‐cytochrome b5 reductase were characterized by a 12h daily rhythm in 1‐month old rats, but in older ones 24h circadian rhythm was found. There was not significant changes of the rhythm pattern in the activity of investigated MFO system ingredients in rats of different age, between spring and winter.

Peptic ulcer disease develops if the balance between aggressive and defensive factors is destroyed. Among the identified risk factors are Helicobacter pylori infection (Hp), stress, cigarette smoking and taking nonsteroidal anti-inflammatory drugs. However, the exact ulcerogenic mechanism of these factors remains not fully elucidated; yet habitual smoking is the most controversial. The aim of this study was to investigate the influence of cigarette smoking and/or Hp infection on gastric mucosa expression of COX-2, iNOS and proliferation and apoptosis processes (Ki-67 and active caspase 3 expressions, respectively). In smokers without Hp all studied parameters were higher than in nonsmoker without Hp group. Hp infection increases iNOS , COX-2 and active caspase 3 expression. Highest values for iNOS, COX-2 and Ki-67 was observed in smoker with Hp infection group. Hp infection in smokers significantly decrease active caspase 3 expression.

Uterine myomas represent one of the most common female diseases. Uterine myomas or fibromas are benign, hormone-responding tumours of, respectively, smooth muscles and fibroblasts and their aetiology induces a significant interest. In myomas the presence of aromatase was detected and, in addition, oestrogen was found to be synthesized in myoma cells. The studies were performed on myoma patients of generative age and those in peri-menopausal age. Expression of TRAF2 and TRAF6 proteins was examined using immunohistochemistry and Western blot approach in small and large uterine myomas isolated from women of various age. In addition, the evaluation was conducted at the periphery of every myoma. We indicated that the level of both tested proteins in myomas is higher than in control. TRAF2 level in myometrium was lower than in myomas but higher than in control. In the case of TRAF6 those changes were ambiguous. Age didn't have influence the level of expression in both tested TRAF in studied structures.

Neointimal hyperplasia (NIH) in vein grafts implanted into the arterial system develops after re-endothelialization and is considered a significant risk factor of occlusion. Evidence suggests that VEGF-A expression with VEGFR-2 activation and/or VEGFR-1 down-regulation might be involved in inhibiting NIH formation. The aim was to assess whether a stented vein graft (SV) has an impact on VEGF-A and VEGFR-1 expression compared with non-stented vein grafts.Twelve sheep received a radial vein with an outside stent (SV) and a radial vein (RV) transplanted into their carotid arteries. The covering of the luminal surface of the SV and RV grafts by endothelium was 98.3% and 96.3%, respectively, at 6 weeks. From the 6th to 12th weeks after transplantation, the time course of total VEGF-A expression and VEGFR-1 expression were evaluated separately for the intima and media.VEGF-A and VEGFR-1 expression were significantly lower in the SV than in the RV group in the intima. In the media the SV grafts were associated with higher VEGF-A and VEGFR-1 expression at 6 and 8 weeks, but lower values were observed at weeks 10 and 12 compared with the RV grafts. Comparing the time courses of VEGF-A and VEGFR-1 expression in the intima and media with intimal/medial thickening in the SV and RV groups, negative correlations for the SV grafts were found.These findings indicate that outside stenting of the vein graft decreases VEGF-A expression and induces significant down-regulation of VEGFR-1 in the intima and media after the re-endothelialization.

We have previously showed that the extravascular dacron mesh stent wrapped around a vein graft and implanted into the arterial system prevented the hypertrophy of the graft's wall, impeded the overgrowth of the intima and decreased the proliferation rate of venous graft cellular elements.To determine the role of cellular proliferation and apoptosis in the process of remodelling in the stent in an animal model in a 12-week period.Male sheep (n = 21) received by transplantation the hybrid graft (group 1) or carotid artery radial vein grafts (group 2). A hybrid graft was composed of a radial vein, collagen fibrin glue and highly flexible torlen/dacron mesh tubing. Grafts were retrieved on day 5, 9 and then week 4, 6, 8, 10, 12, respectively. A proliferation process was assessed using a Ki-67 antigen kit. The presence of apoptosis was detected using a TUNEL kit, strictly according to the manufacturer's manual.The number of proliferating cells has presented a decreasing trend in both groups, whereas the mean quantity of apoptotic cells increased over a 12-week period (p0.001) in both groups. Proliferation was more prominent during the first 5 weeks in both groups. The trend had a tendency to reverse during the last 7 weeks of observation. The ratio of proliferating to apoptotic cells differed between groups (1.6 vs. 1.9 on day 5 and 0.2 vs. 0.6 in week 12, in group 1 and group 2, respectively). No linear correlation between proliferation and apoptosis was observed (p0.05).Different kinetics in the trico hybrid graft group in comparison with the radial vein graft group was observed, with a more prominent cellular turnover in the trico hybrid graft. Apoptosis in an unprotected vein wall was overcomed by the proliferation process. In trico hybrid vein grafts, beneficial remodelling of the intimal layer was predominantly dependent on inhibition of intimal proliferation rather than the effect of changes of the apoptosis ratio. There was no linear correlation between proliferation and apoptosis in the investigated grafts.

Telomerase activity could be a potential marker for the neoplastic process, because it is absent in normal cells and present in tumor cells. Immunohistochemical studies were conducted using samples obtained from 32 uterine myomas, each sample having a size of 3-4 cm and obtained from women between 35 and 45 years of age. These studies also concentrated on fragments of macroscopically unaltered myometrium, collected 3-4 cm from a uterine tumor. Immunohistochemistry was performed using antibody to the catalytic unit of telomerase (hTERT; clone 44F12, NCL-L-hTERT, Novocastra Laboratories, UK). This study aimed at detecting a possible presence of potentially neoplastic cells in the margins of healthy tissue, which was removed together with the primary tumor. The results were classified according to the number of telomerase-positive cells. Tumors of the first group had up to 50% telomerase-positive cells, while their content in the second group exceeded 50%. Our study demonstrated an almost two-fold increase in the number of telomerase-positive tumor cells compared with myometrial cells 3-4 cm from the tumor. Hopefully, investigating the presence of telomerase in both uterine myometrium and myoma could facilitate the diagnosis of the neoplastic process.

Mitochondria and lysosomes were evaluated by assessment of changes in activity of selected enzymes: lactate dehydrogenase (LDH), succinate dehydrogenase (SDH), adenosinetriphosphatase (ATPase), acid phosphatase (AcPase) and beta-glucuronidase (BG) in rats under profound hypoxia induced by endotoxemic shock. The study was conducted on adult male Wistar rats. The animals formed the following four groups of 15 rats each: control animals (C);-rats receiving intraperitonally O(2)/O(3) (CO), rats receiving of Escherichia coli toxin (LPS) (CL); rats receiving LPS plus oxygen-ozone mixture (OL). Histoenzymatic examinations of liver, kidney, lungs, and heart muscle were performed. Lipopolysaccharide suppressed activities of all the enzymes except for LDH, the activity of which as high as a fourfold increase. The results demonstrated potent, stabilizing and regenerative effects of ozone therapy on body enzymatic processes in course of induced endotoxemic shock in rats, which might prove to be of clinical significance.

The confirmed advantageous effects of oxygen/ozone therapy in several clinical conditions stimulated experimental studies on effects of the therapy in rats with an induced septic shock. The studies were conducted on adult male rats of Wistar strain. Four groups of the animals, each of 15 rats, included: I--control group, (C); II--animals intraperitoneally administered with O(2)/O(3) (CO), III--rats given of Escherichia coli endotoxin (lipopolysaccharide-LPS) (CL), IV--rats administered with the lipopolysaccharide plus administered with the oxygen/ozone mixture (OL). Activities of catalase and superoxide dismutase and of free radical reactions were estimated. The exposure to LPS augmented activities of SOD and of catalase in liver, lungs and heart. In all the examined organs LPS induced significant changes in levels of free radicals. Except of the lungs, parallel administration of the rats with LPS and ozone/oxygen revoked development of the alterations. The obtained results point to a strong, stabilizing and regenerative effect of ozonotherapy.

Nucleolar organizer regions (NORs) were assessed in 27 women affected by uterine leiomyoma. Tissue samples obtained during surgery were silver-stained according to the method of Ploton et al. The assessed parameters were as follows: the number of argyrophylic nucleolar organizer regions (AgNORs) per nucleus, single AgNOR area, AgNOR intranuclear distribution, and AgNOR coefficient. The parameters were assessed quantitatively. It was found that the AgNOR coefficient was higher in uterine leiomyoma compared to the normal smooth muscle cells. The development of leiomyoma is associated with a marked decrease in myocyte nucleolar area, which accounts for 30% of the nucleus in the normal myometrium. With the comparable AgNOR number in the single nucleus, there were no differences in the single AgNOR granule area (1.21mm 2 70.047 and 1.11mm 2 70.025 in normal myometrium and in leiomyoma, respectively). In the normal myometrium, there was a positive correlation between nuclear area and the single AgNOR granule area, as well as between the AgNOR coefficient and the single AgNOR granule area. There was also a negative correlation between the number of granules per nucleus and their central and peripheral intranuclear distribution. The development of leiomyoma was associated with loss of all correlations observed in the control group. r 2005 Elsevier GmbH. All rights reserved.

The study aimed to evaluate the effect of L-carnitine on hepatic cytochrome P450-dependent monooxygenases exposed to methanol. Male Spraque-Dawley rats were given methanol (1/4 LD50 and 1/2 LD50) together with L-carnitine (1g/kg body weight). The parameters of microsome electron transport chains I and II and the levels of CYP2E1, CYP2B1/2 and CYP1A2 were measured 8, 12, 24, 48, 72 and 96 h after exposure. L-carnitine did not affect cytochrome P450 but it significantly increased at 72 and 96 h NADPH-cytochrome P450 reductase. It stimulated cytochrome b5 at 48 and 96 h and NADH-cytochrome b5 reductase activity at 12, 72 and 96 h. Methanol, especially the lower dose, inhibited cytochrome P450 after 48 h, but the higher methanol dose inhibited NADH-cytochrome b5 reductase activity in this time. L-carnitine, combined with the lower dose of methanol, stimulated NADPH-cytochrome P450 reductase after 48 h and cytochrome b5 and NADH-cytochrome b5 reductase over the whole period of observation. L-carnitine stimulated CYP2B1/2 but not CYP2E1 and CYP1A2. Methanol stimulated CYP2E1 at 24 h, but CYP1A2 at 96 h in the studied doses. CYP2B1/2 was induced by the lower dose of methanol at 24 h but by the higher one at 96 h. When given together, L-carnitine and methanol (1/2 LD50) significantly stimulated CYP2E1 up to 170% at 24 h and 145% at 96 h.

Glucose-6-phosphatase catalyzes the final reactions in both gluconeogenesis and glycolysis. It occurs mainly in glycogenic tissues, such as the liver, where it plays an important role in the synthesis of glucose, a carbohydrate essential for tissue functioning. The effect of age on liver glucose-6-phosphatase activity was evaluated in male Wistar rats treated with mixed function oxidase system (MFO) inducers. The rats were divided into the following age groups: 0.5, 1, 2, 4, 8, 12, 20 and 28 months of age. Glucose-6-phosphatase activity was evaluated biochemically and histochemically. Biochemical glucose-6-phosphatase activity increased up to the 20th month of rat life and then decreased rapidly. A similar tendency was observed in inducer-treated groups, though only dexamethasone stimulated this enzyme activity in all age groups studied. Histochemical glucose-6-phosphatase activity was strongest in the periportal zones. Glucose-6-phosphatase activity decreased significantly at month 8 and then it increased significantly until month 20. In the oldest age group, glucose-6-phosphatase activity decreased again. On histochemical analysis, the inducers used variably affected glucose-6-phosphatase activity.

UDP-glucuronyltransferase (UDP-GT) activity was examined in male Wistar rats aged 0.5, 1, 2, 4, 8, 12, 20, and 28 months. The rats were treated with phenobarbital (75 mg/kg, 72 and 48 h before death), β-naphthoflavone or dexamethasone (40 mg/kg and 20 mg/kg, respectively, for three days before death). Prior to decapitation the rats were fasted for 12 h. Hepatic microsomes were prepared according to the method of Dallner. UDP-GT activity was determined by the method of Burchell and Weatherill. p-Nitrophenol was used as an aglucone. UDP-GT activity decreased rapidly in the control rats aged from two weeks to four months. In the older control rats the activity tended to increase. Two-week-old rats treated with phenobarbital showed a slightly increased UDP-GT activity. In the older animals (up to one year) UDP-GT activity increased to 150% of the control value and stayed at this level in the remaining age groups. β-Naphthoflavone was a more potent inducer of UDP-GT than phenobarbital. The activity of β-naphthoflavone-induced UDP-GT was low in the youngest rats. It was about 180% in two-month-old rats and reached 260% of the control value in eight-month-old rats. Although the activity decreased in the older rats, it still exceeded 200%. Dexamethasone did not affect UDP-GT activity. Only in two-week-old and two-month-old rats did we observe a slight increase in the activity of UDP-GT.

The influence of the cholesterol and protein diet on the mixed-function oxidases system activity and desaturation of fatty acids were examined. The phenobarbital innducibility of these parameters was analyzed too. Investigations were carried out on Wistar adult male rats. The animals were on the cholesterol (0.25%) and protein diet (32%) during 45 days. The cytochrome P-450 and cytochrome b5 contents, the NADPH-cytochrome P-450 reductase and NADH-cytochrome b5 reductase activities, aniline hydroxylase and 4-aminopyrine N-demethylase activities were measured in the hepatic microsomal fraction. In both sexes, the cholesterol diet decreased all the examined parameters with the exception of NADH-cytochrome b5 reductase activity. The protein diet did not change the examined enzyme activities or levels with the exception of induction of NADH-cytochrome b5 reductase and 4-aminopyrine N-demethylase activities. Simultaneous treatment with phenobarbital had heterogenous effect dependent on the type of diet and examined parameter.

Summary The experiments were carried out on male Wistar rats aged 6, 12 and 24 months, over the four seasons of the year. Analysis of the results obtained in all age groups disclosed that changes in activity of the p.a.S (periodic acid Schiff) reaction and in concentrations of Mg and Ca in the liver showed rhythmic oscillations with a period of 12 hours. The maximal p.a.S reaction activity and of Ca and Mg levels were generally found to coincide throughout all seasons and in all age groups. The rhythms of change in these parameters in 12- and 24-month-old rats showed a phase shift as compared to the 6-month-old animals.