Metallothionein as a Scavenger of Free Radicals - New Cardioprotective Therapeutic Agent or Initiator of Tumor Chemoresistance?

Cardiotoxicity is a serious complication of anticancer therapy by anthracycline antibiotics. Except for intercalation into DNA/RNA structure, inhibition of DNA-topoisomerase and histone eviction from chromatin, the main mechanism of their action is iron-mediated formation of various forms of free radicals, which leads to irreversible damage to cancer cells. The most serious adverse effect of anthracyclines is, thus, cardiomyopathy leading to congestive heart failure, which is caused by the same mechanisms. Here, we briefly summarize the basic types of free radicals formed by anthracyclines and the main processes how to scavenge them. From these, the main attention is paid to metallothioneins. These low-molecular cysteine-rich proteins are introduced and their functions and properties are reviewed. Further, their role in detoxification of metals and drugs is discussed. Based on these beneficial roles, their use as a new therapeutic agent against oxidative stress and for cardioprotection is critically evaluated with respect to their ability to increase chemoresistance against some types of commonly used cytostatics.

Evaluation of the Impact of Different Types of Health Education on the Adoption and Preservation of Prohealth Attitudes in Preventing Cancer in Juveniles Younger than 18 Years.

Reduction in the incidence of cancer can be achieved through appropriate health behaviors. We hypothesized that education would improve knowledge of cancer prevention, and this, in turn, will affect and individual's readiness to modify lifestyle. The aim of this study was to assess the impact of cancer prevention education on adopting and preserving prohealth attitudes among high school students in Poland. Research participants were 307 high school students varying by gender, place of residence, parents' education, and type of school education. Participants were divided into five groups, of which four were educated using different methods according to classification methods based on the concept of multilateral learning. The fifth (control) group was not educated. The effects of education were assessed 1 month and 1 year after education. General knowledge about cancer and healthy lifestyle level before education was low. After education, both increased compared with the control group. There was a clear relationship between level of knowledge and readiness to adopt and healthy attitudes and behavior. The most effective method of education was a discussion and a lecture by means of teaching complex. Education significantly improved generally low knowledge about cancer and healthy lifestyle in high school students. This indicates the urgent need to implement such educational programs.

The application of capillary electrophoresis in the determination of glutathione in healthy women's blood.

The purpose of this work was to compare glutathione status in the blood of women taking oral contraceptives, female smokers and pregnant women using both the capillary electrophoresis (CE) and alloxan methods. The studies were performed on the whole blood prepared by deproteinization by 25% metaphosphoric acid. The reduced glutathione (GSH) was measured by alloxan and CE methods. Oxidized glutathione (GSSG) was measured by CE and the GSH/GSSG ratio was calculated. In pregnant women (Group A), women taking oral contraceptives (Group B) and female smokers (Group C), lower concentrations of GSH were observed compared with the control group (Group D) as measured by CE and alloxan methods, suggesting the presence of oxidative stress. The level of GSSG in Group C was higher than in Group D, indicating glutathione oxidation. In Groups A and B, reduced levels of GSSG were observed, which indicates that other processes besides oxidation affect glutathione status. In Groups A and C, a lower GSH/GSSG ratio was observed than in Group D, while in Group B no statistical change was observed. In conclusion, the advantage of CE is the possibility of measuring GSSG, which could allow for a more accurate interpretation of the status of GSH in the human body. Oxidation of glutathione in female smokers was indicated, while in pregnant women and women taking oral contraceptives processes other than oxidation can be associated with a decrease in glutathione levels.

The influence of biological and environmental factors on metallothionein concentration in the blood.

The concentration of metallothionein (MT), a low-molecular-weight protein, is regulated by many factors, primarily metals (zinc, cadmium, copper), cytokines, glucocorticoides and free radicals. These factors are determined by such aspects of human biology as gender, pregnancy and age, as well as by environmental factors including the use of oral contraceptives and cigarette smoking, all which may affect MT levels in the body. The aim of this study was to investigate the influence of these biological and environmental factors on MT concentrations in erythrocyte lysate and in plasma. MT concentrations were determined by a two-step direct enzyme-linked immunosorbent assay. Evaluation of exposure to cigarette smoking was performed by checking cotinine levels in the plasma of subjects. The studies showed higher MT concentrations in both the erythrocyte lysate and plasma of women when compared to men. Furthermore, pregnancy causes an increase of MT concentration in plasma, while oral contraceptives cause an elevated concentration of MT in erythrocyte lysate. Age impacts plasma MT concentrations in men, whereas it does not affect concentrations of MT in erythrocyte lysate.

Comparison of positron emission tomography/computed tomography with classical contrast-enhanced computed tomography in the initial staging of Hodgkin lymphoma.

We compared initial computed tomography (CT) and positron emission tomography (PET)/CT in 96 patients with Hodgkin lymphoma (HL), assessing the role of baseline PET/CT in stage migration and treatment selection. The number of patients with stage I, II, III and IV disease based on CT versus PET/CT was: 5 vs. 7, 49 vs. 37, 28 vs. 22 and 14 vs. 30, respectively. In 33 (34%) patients, PET/CT changed HL stage: 27 (28%) were upstaged and six (6.3%) downstaged. Upstaging was caused by detection of new extranodal involvements (47 sites in 26 patients): bone marrow (10 patients), spleen (five patients) and lung (two patients). In nine patients≥2 further coexisting locations were detected. Downstaging resulted from the absence of fluorodeoxyglucose (18F-FDG) uptake in enlarged nodes (>15 mm) in the abdomen and pelvis. PET/CT modified HL stage in 34% of patients leading to treatment modification in the majority. Our results indicate that PET/CT should be mandatory in the initial staging of HL.

G protein-coupled receptors: abnormalities in signal transmission, disease states and pharmacotherapy.

The aim of this review is to present the research results and draw new conclusions about the impact of alterations in the signal transmission through the G protein-coupled receptors (GPCRs) on the formation of diseases and drug therapy. GPCR family is the largest and the most diverse group of membrane receptors. They transmit signals into the cell by interaction with different ligands, which include, inter alia, hormones, neurotransmitters, and photons. GPCRs are responsible for the proper conduction of many physiological processes such as vision, intercellular communication, the neuronal transmission, hormonal signaling and are involved in many pathological processes. They are also point on the binding pathway of multiple drugs. They are targets of nearly one third of the drugs at the current pharmaceutical market. The genes encoding GPCRs represent about 4% of the human genome. Mutations that occur in them are associated with a broad spectrum of diseases of diverse etiology. As a mutations result, there is a change in receptor activity (GPCR become inactive, overactive, or constitutively active), in the process of ligand binding and signal transduction. Changes in the GPCRs functioning can cause diseases such as retinitis pigmentosa (rhodopsin mutations), nephrogenic diabetes insipidus (vasopressin receptor mutations), obesity (melanocortin receptor mutations). Many mutational changes in genes encoding GPCR can change drug therapy of already existed diseases: heart failure (adrenergic receptors), asthma (cysteinyl leukotriene receptors). Studies concerning the structure and function of genetically modified GPCRs allow to get know a variety of mechanisms of its action, which in turn can contribute to broaden the knowledge on the etiology and pharmacotherapy of many currently incurable diseases.

The influence of the occupational exposure to heavy metals and tobacco smoke on the selected oxidative stress markers in smelters.

The aim of the study was to verify if there is any association between exposure to Cu, Zn, Cd, Pb, As and the formation of malondialdehyde (MDA), 8-hydroxydeoxyguanosine (8-OHdG), advanced oxidation protein products (AOPP), and whether in this process cigarette smoking plays a role. The investigations were performed in the 352 smelters occupationally exposed to heavy metals and 73 persons of control group. Metals concentration was determined by atomic absorption spectrometry. MDA and AOPP concentrations were determined by spectrophotometric methods. The concentration of 8-OHdG was determined by ELISA method. It was demonstrated an increased Cu concentration in smoking smelters compared to non-smoking control group. It was noted no differences in Zn and Mg concentrations between the examined groups. Pb concentration was more than sixfold higher in the group of smoking smelters and about fivefold higher in the group of non-smoking smelters compared to the control groups (smokers and non-smokers). It was shown that Cd concentration in the blood was nearly fivefold higher in the smoking control group compared to the non-smoking control group and more than threefold higher in the group of smoking smelters compared to non-smoking. It was shown an increased As concentration (more than fourfold) and decreased Ca concentration in both groups of smelters compared to control groups. In groups of smelters (smokers and non-smokers), twofold higher MDA and AOPP concentrations, and AOPP/albumin index compared to control groups (smokers and non-smokers) were shown. Tobacco smoke is the major source of Cd in the blood of smelters. Occupational exposure causes accumulation of Pb in the blood. Occupational exposure to heavy metals causes raise of MDA concentration and causes greater increase in AOPP concentration than tobacco smoke.

The role of metallothionein interactions with other proteins.

Metallothionein (MT) is a protein involved in numerous key processes, and the most important include zinc ion homeostasis, detoxification of heavy metals, and protection against oxidative stress. MT by interaction with other proteins fulfills its function, resulting in different effects in the body. Interaction of MT with ferritin, which causes a redox reaction, resulting in the reduction of Fe(3+) stored in ferritin and a release of harmful Fe(2+) , was observed. Referring to the redox function of MT, it has been shown that the pair of GSH/GSSG modulates transfer of Zn between MT and Zn-binding proteins. Furthermore, it was shown that GSSG, in the presence of GSH, interacts directly with MT. Apothionein-MT can retrieve Zn from the transcription factors or Zn-containing enzymes. Apothionein-MT by taking Zn can deactivate metal-dependent enzymes while Zn-MT has the opposite effect. As the effect of MT interaction with low-density lipoprotein receptors-megalin and lipoprotein receptor related protein 1, the uptake of Cd-MT occurs and results in the disruption of many functions of proximal tubules. MT is involved in numerous processes and many of them are regulated by protein-protein interactions. Possibly in the future MT will become a therapeutic agent, which will result in a breakthrough in the field of pharmacy and medicine.

The prevalence of pulp stones in historical populations from the middle Euphrates valley (Syria).

There are many reports in the literature concerning pulp stones in contemporary living populations, but there are no descriptions of cases of pulp stones and their prevalence in populations of the past. Here we present a study of pulp stones in a series of archaeologically derived samples from the Middle Euphrates Valley (Syria) obtained from two sites: Terqa and Tell Masaikh. The specimens were assigned to five periods: Early Bronze (2650-2350 BC); Middle Bronze (2200-1700 BC); late Roman (AD 200-400); Islamic (AD 600-1200); and Modern Islamic (AD 1850-1950). A total of 529 teeth representing 117 adult individuals of both sexes were examined. Pulp stones were identified by X-ray and 10 selected specimens were sectioned for histological study. Pulp stones were found in 99 of 117 individuals (85%) and in 271 of 529 (51%) teeth. Pulp stone prevalence was found to increase with age, for individuals of older age classes have more pulp stones than younger individuals. Intriguingly, the prevalence of single pulp stones was higher among older individuals (36-45, >46), while younger individuals (17-25, 26-35) more often possessed multiple stones. Individuals with moderate to highly advanced dental wear have pulp stones significantly more often than individuals whose tooth wear is limited to invisible or very small facets. Though there is no statistical significance in the prevalence of pulp stones across chronological periods, it appears that a high level of calcium in the diet is accompanied by a greater prevalence of pulp stones.

Capillary electrophoresis application in the analysis of the anti-cancer drugs impurities.

Due to the low therapeutic index of anti-cancer drugs, they should be closely monitored for evidence of potential contamination that may be of high toxicity and not to have the desired therapeutic effect. Therefore, analytical methods to detect drugs related substances at low concentrations are necessary. Capillary electrophoresis allows for fast and clear separation of drug derivatives. A multitude of submethods make selection of suitable environment for various types of chemicals possible. Publications concerning separation of drugs such as cisplatin, carboplatin, lobaplatin, methotrexate, tamoxifen, paclitaxel from their derivatives, which are their potential contaminations, show that capillary electrophoresis provides the appropriate tools to analyze the impurities of these anti-cancer drugs and is able to partially displace such technique as thin layer chromatography and high performance liquid chromatography, which still play a major role in this field.

Changes in pro/antioxidant balance in smoking and non-smoking pregnant women with intrauterine growth restriction.

The aim of this study was to investigate the influence of cigarette smoking on the pro/antioxidant balance in pregnant women with intrauterine growth restriction (IUGR). The studies have shown a 2-fold increase of Cd concentration in blood of women with IUGR in labour and a 10-fold increase in smoking pregnant women with IUGR. The increase of malondialdehyde concentration in plasma and 8-hydroxydeoxyguanosine in serum and Cu/Zn superoxide dismutase activity in erythrocyte lysate of pregnants with IUGR, reinforced by smoking, was revealed. We observed a decrease in the concentration of glutathione in blood and glutathione peroxidase activity in plasma and in erythrocyte lysate. A 4-fold higher metallothionein concentration in the plasma of women with IUGR in labour suggests that metallothionein may be one of the IUGR markers. Metallothionein concentration was intensified by smoking up to 7-fold in comparison to the controls. The pro/antioxidant balance during pregnancy is significantly affected by smoking.

Soluble TNF-α receptor I encoded on plasmid vector and its application in experimental gene therapy of radiation-induced lung fibrosis.

Post-radiation inflammatory reaction leads to an irreversible pulmonary fibrosis which may cause lethal respiratory insufficiency. Pathological inflammatory and fibrotic changes might be attenuated by inhibiting tumour necrosis factor (TNF)-α activity using TNF-α soluble receptors. Thus, an experimental antifibrotic gene therapy with the plasmid vector encoding a mouse soluble receptor I for TNF-α (psTNFR-I) was assessed. Soluble TNFR-I encoding gene was cloned into pcDNA3.1 plasmid. The ability of psTNFR-I expressing vector to transfect cells, and its biological activity in vitro and in vivo were examined by PCR, RT-PCR, MTT assay and ELISA. The C57Bl/6J mice received single intramuscular injection of psTNFR-I, conjugated with polyetylenimine (PEI) 25 kDa, equally divided to both hind legs, 3 days before irradiation (20 Gy, Co60), and either a single injection or ten injections once a week after irradiation. The data proved the effectiveness of psTNFR-I product to neutralise TNF-α activity in vitro. The in vivo plasmid incorporation and maintenance was confirmed. Measurements of plasma soluble TNFR-I levels showed that the in vivo gene transfer was effective. PEI was found to enhance transfection efficiency in vivo. The psTNFR-I/PEI complexes caused no toxicity in the transfected mice. C57Bl/6J mice that received prolonged psTNFR-I/PEI injections developed lethal fibrotic syndrome and died 8 weeks later than the mice treated with a double plasmid injection and the control mice treated with a control plasmid. Sequential administration of soluble TNFR-I by a nonviral, intramuscular gene transduction in the early and late post-radiation inflammatory phase prolonged survival of irradiated mice and attenuated the symptoms of lung fibrosis. The psTNFR-I gene transduction may provide a safe and simple method to partially neutralise TNF-α activity and prevent radiation-induced lung injury.

[Retrospective analysis of ocular disorders and frequency of cesarean sections for ocular indications in 2000-2008--our own experience]. / Retrospektywna analiza patolog i narz du wzroku i czestosci ciec cesarskich ze wskazan okulistycznych w latach 2000-2008 w materiale wlasnym.

UNLABELLED: 1. Evaluation of frequency of cesarean sections for ocular indications. 2. Analysis of ophthalmological disorders as indications for cesarean section. MATERIAL AND METHODS: 4895 cesarean sections were performed (100 due to ocular indications) in the Department of Obstetrics, Female Pathology and Oncological Gynecology between 2000 and 2008. Medical documentation was analyzed. RESULTS: Among 4895 patients undergoing cesarean sections, 100 (2.04%) presented a written certification from an ophthalmologist suggesting this way of delivery. The frequency of c-sections due to ocular indications continued to increase between 2000-2005 and has been in decline since 2006. The most common ophthalmological disorders included myopia (57%), retinopathy (20%), glaucoma (5%), imminent retinal detachment (4%) and past retinal detachment (3%). In 45% of patients an eye pathology was the only reason for a cesarean section. CONCLUSION: 1. The frequency of cesarean sections due to ocular reasons in our material was 0.7%- 3.44%, average 2.04%. 2. Since 2006 the number of ocular indications for cesarean section has been decreasing. Nevertheless, it remains to be twice as high as in 2000. 3. The most common eye disorders leading to cesarean section were myopia and retinopathy 4. In almost half of the patients the decision to conduct a cesarean section was based solely on ophthalmological indications.

[History of cigarette smoking. The effect of tobacco smoking on women's health]. / Nikotynizm--rys historyczny. Uzaleznienie od nikotyny a zdrowie kobiety.

History of cigarette smoking started on XV century, when Columb imported tobacco to Europe. Popular using of tobacco we are indebted Jaen Nicot Villeman, the name of nicotine originate from his surname. Tobacco first was exploited like a drug, however now it is a very harmful stimulant. Cigarette smoking is still an actual problem and increased risk of many diseases. Very toxic components of smoke get inside all the organs and upsetting their activities and proper running of the life processes. It is common knowledge that smoking badly influences women's health. Nicotine makes the negative influence on function of ovaries metabolism of hormones and state of osseous tissue. Maternal smoking during pregnancy is associated with obstetrical and pediatrician complication with fetus, newborn and child. The aim of the article is to summarize the role of tobacco smoking on women's health.

[The application of immunoperoxidase test for the detection of antibodies various classes (IgA, IgG and IgM) coating bacteria in urine]. / Zastosowanie immunoperoksydazowego testu ACB (IP ACB) do wykrywania przeciwcial róznych klas (IgA, IgM, IgG) oplaszczajacych bakterie w moczu.

For the detection of bacteria coated with immunoglobulins in urine the monoclonal antibodies against human IgA, IgG and IgM conjugated with peroxidase were used. For comparison, the immunofluorescence technique was also employed. The results obtained by two methods revealed that immunofluorescence were less sensitive. It was found that bacteria were predominantly coated with IgA (41,9 +/- 22,4%) and IgG (34,1 +/- 15,3 %) immunoglobulins. The IgM antibodies were found rarely (12,8 +/- 8%).

[Adaptation of immunoenzymatic method for detection of antibody coated bacteria in urine]. / Adaptacja metody immunoenzymatycznej do wykrywania bakterii oplaszczonych przeciwcialami w moczu.

The modified immunoenzymatic method for detection of antibody coated bacteria (IP ACB) was compared with immunofluorescence technique (IF ACB) in the diagnosis of urinary tract infection. For the study 100 patients were employed with significant and insignificant bacteriuria. It was found that 81% of the results obtained by IP ACB and IF ACB were identical, however the immunoenzymatic method was more sensitive than immunofluorescence. Moreover, the IP ACB technique is simpler, less time consuming and may be performed by using the ordinary optic microscope.