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B-cell leukemia/lymphoma 11A (BCL11A) is a transcription factor that regulates the expression of genes involved in cell division or apoptosis. A link between high BCL11A expression and a worse prognosis has been demonstrated in patients with various cancers. The aim of this study was to investigate the expression pattern of BCL11A in breast cancer (BC) cases and mastopathy samples and to correlate the results with the clinicopathological data. The expression of the BCL11A protein was investigated using immunohistochemistry (IHC) on 200 cases of BC and 13 mastopathy samples. The level of BCL11A mRNA was determined using real-time PCR in 22 cases of BC and 6 mastopathy samples. The expression of BCL11A was also examined at the protein and mRNA levels in BC cell lines. A higher expression level of BCL11A in BC cases was shown compared to mastopathy samples. The expression level of BCL11A in BC cases and in the studied cell lines decreased with the increasing grade of histological malignancy (G). It was also negatively correlated with the primary tumor size. A significantly lower expression of BCL11A was found in BC that did not express estrogen or progesterone receptors and in triple-negative cases. The results of our research suggest that BCL11A may be relevant in the development of BC.

Methotrexate (MTX) is recommended as a first-line treatment for rheumatoid arthritis (RA). There are no strict guidelines regarding monitoring for liver damage in RA patients. This study aimed to evaluate noninvasive diagnostic procedures in assessing liver fibrosis in RA patients. Ninety-six RA patients were recruited for this study. The procollagen III N-terminal peptide (PIIINP) serum level was measured in all patients. The Enhanced Liver Fibrosis score (ELF-1) was calculated for 82 patients. Transient elastography (TE) was performed in 91 patients, those examined were divided into two groups: a study and control group, comprising patients with and without risk factors for liver fibrosis, respectively. The TE result correlated only with the body mass index—BMI (p p = 0.33). The TE result was significantly higher in those with risk factors for liver fibrosis than in those without risk factors (TE result > = 7.1 kPa 28/42 vs 13/41, HR = 2.103, Mann–Whitney U test, approximately 0.02). There was a positive correlation between the PIIINP level and body weight (p = 0.028), cumulative MTX dose (p = 0.007), RA activity (p = 0.028) and diabetes mellitus (DM) (p = 0.001). There was a positive correlation between the ELF-1 score and age (p p = 0.007) and RA activity (p

Atherogenesis leads to the development of atherosclerosis, a progressive chronic disease characterized by subendothelial lipoprotein retention and endothelial impairment in the arterial wall. It develops mainly as a result of inflammation and also many other complex processes, which arise from, among others, oxidation and adhesion. Cornelian cherry (Cornus mas L.) fruits are abundant in iridoids and anthocyanins—compounds with potent antioxidant and anti-inflammatory activity. This study aimed to determine the effect of two different doses (10 mg and 50 mg per kg of body weight, respectively) of iridoid and anthocyanin-rich resin-purified Cornelian cherry extract on the markers that are important in the progress of inflammation, cell proliferation and adhesion, immune system cell infiltration, and atherosclerotic lesion development in a cholesterol-rich diet rabbit model. We used biobank blood and liver samples that were collected during the previous original experiment. We assessed the mRNA expression of MMP-1, MMP-9, IL-6, NOX, and VCAM-1 in the aorta, and the serum levels of VCAM-1, ICAM-1, CRP, PON-1, MCP-1, and PCT. The application of the Cornelian cherry extract at a dose of 50 mg/kg bw resulted in a significant reduction in MMP-1, IL-6, and NOX mRNA expression in the aorta and a decrease in VCAM-1, ICAM-1, PON-1, and PCT serum levels. The administration of a 10 mg/kg bw dose caused a significant decrease in serum ICAM-1, PON-1, and MCP-1. The results indicate the potential usefulness of the Cornelian cherry extract in the prevention or treatment of atherogenesis-related cardiovascular diseases, such as atherosclerosis or metabolic syndrome.

The aim of the study was to evaluate the localization and intensity of RNA-binding motif single-stranded-interacting protein 3 (RBMS3) expression in clinical material using immunohistochemical (IHC) reactions in cases of ductal breast cancer (in vivo), and to determine the level of RBMS3 expression at both the protein and mRNA levels in breast cancer cell lines (in vitro). Moreover, the data obtained in the in vivo and in vitro studies were correlated with the clinicopathological profiles of the patients. Material for the IHC studies comprised 490 invasive ductal carcinoma (IDC) cases and 26 mastopathy tissues. Western blot and RT-qPCR were performed on four breast cancer cell lines (MCF-7, BT-474, SK-BR-3 and MDA-MB-231) and the HME1-hTERT (Me16C) normal immortalized breast epithelial cell line (control). The Kaplan–Meier plotter tool was employed to analyze the predictive value of overall survival of RBMS3 expression at the mRNA level. Cytoplasmatic RBMS3 IHC expression was observed in breast cancer cells and stromal cells. The statistical analysis revealed a significantly decreased RBMS3 expression in the cancer specimens when compared with the mastopathy tissues (p < 0.001). An increased expression of RBMS3 was corelated with HER2(+) cancer specimens (p < 0.05) and ER(−) cancer specimens (p < 0.05). In addition, a statistically significant higher expression of RBMS3 was observed in cancer stromal cells in comparison to the control and cancer cells (p < 0.0001). The statistical analysis demonstrated a significantly higher expression of RBMS3 mRNA in the SK-BR-3 cell line compared with all other cell lines (p < 0.05). A positive correlation was revealed between the expression of RBMS3, at both the mRNA and protein levels, and longer overall survival. The differences in the expression of RBMS3 in cancer cells (both in vivo and in vitro) and the stroma of breast cancer with regard to the molecular status of the tumor may indicate that RBMS3 could be a potential novel target for the development of personalized methods of treatment. RBMS3 can be an indicator of longer overall survival for potential use in breast cancer diagnostic process.

Rackoff and Simon proved that a variant of Chaum's protocol for anonymous communication, later developed as the Onion Routing Protocol, is unlinkable against a passive adversary that controls all communication links and most of the nodes in a communication system. A major drawback of their analysis is that the protocol is secure only if (almost) all nodes participate at all times. That is, even if only n?N nodes wish to send messages, allN nodes have to participate in the protocol at all times. This suggests necessity of sending dummy messages and a high message overhead. Our first contribution is showing that this is unnecessary. We relax the adversary model and assume that the adversary only controls a certain fraction of the communication links in the communication network. We think this is a realistic adversary model. For this adversary model we show that a low message overhead variant of Chaum's protocol is provably secure. Furthermore, all previous security proofs assumed the a priori distribution on the messages is uniform. We feel this assumption is unrealistic. The analysis we give holds for any a priori information on the communication distribution. We achieve that by combining Markov chain techniques together with information theory tools in a simple and elegant way.

In ozone-treated erythrocyte membrane suspension a slow decrease occurs in the EPR signal of 2,2,6,6-tetramethyl-piperidine-1-oxyl (TEMPO). Because of the absence of such a phenomenon in control membranes and ozonized buffer, this effect must be caused by reaction of nitroxide radicals with products of ozone reactions with membrane components. To find out which components are responsible for the decrease in EPR signal we studied this effect in simple model systems. The same phenomenon was observed both in lipid and protein systems treated by ozone. For unsaturated fatty acids, the correlation between the rate of decrease in EPR signal and the number of double bonds in the lipid molecule was very strong. This suggests that the observed decrease in the nitroxide radical TEMPO EPR signal in ozone-treated erythrocyte membranes is a complex process, but probably the most important reaction is recombination of nitroxide radicals with organic free radicals produced both in the process of lipid peroxidation and ozonolysis of double bonds.

Doxyl stearate spin probes which differed in the attachment of the nitroxide free radical to the fatty acid have been used to study membrane fluidity in ozone-treated bovine erythrocytes and liposomes. Analysis of EPR spectra of spin labels incorporated into lipid bilayer of the erythrocyte membranes indicates an increase in the mobility and decrease in the order of membrane lipids. In isolated erythrocyte membranes (ghosts) the most significant changes were observed for 16-doxylstearic acid. In intact erythrocytes statistically significant were differences for 5-doxylstearic acid. The effect of ozone on liposomes prepared from a lipid extract of erythrocyte lipids was marked in the membrane microenvironment sampled by all spin probes. Ozone apparently leads to alterations of membrane dynamics and structure but does not cause increased rigidity of the membrane.

High mortality rate in Amanita phalloides (death cap) intoxications is a result of the acute liver failure following hepatocyte damage due to hepatocellular uptake of amatoxins. α-Amanitin (α-AMA), the major amatoxin, blocks a RNA polymerase II, which results in inhibition of transcription of DNA and protein synthesis processes and leads to hepatocyte death. α-AMA is also a strong apoptosis inductor and may play a significant role in pathogenesis of hepatic damage in course of amanitin intoxication. The aim of this study was to examine mechanisms of α-AMA-induced apoptosis in human hepatocytes, as well as in determining if commonly clinically used antidotes benzylpenicillin (BPCN) and N-acetylcysteine (ACC) are able to protect human hepatocytes against α-AMA-induced apoptosis. The experiment was performed on cultured human hepatocytes. Viability of cultured hepatocytes was assessed using the MTT assay, whereas apoptosis processes were evaluated by the electron microscopy, detection of DNA laddering, determination of caspase-3 activity, and measuring annexin V, p53 and Bcl-2 protein concentration. Cytotoxicity and apoptosis evaluation were performed after 24 h of exposure to α-AMA and/or tested antidotes.Both ACC and BPCN were well tolerated by human hepatocyte cultures, and exposure to those substances did not reduce cell viability nor induce apoptosis. Exposure of hepatocytes to α-AMA at concentration 2 μM resulted in derangement of cell cultures, apoptosis and significant reduction in cell viability. α-AMA-induced apoptosis in human heptocyte cultures is p53- and caspase-3-dependent. Human hepatocyte cultures are exposed simultaneously to α-AMA and tested antidotes (BPCN or ACC) showed significantly higher cell viability and significantly lower values of apoptosis markers compared to the cultures exposed to α-AMA only.

We concern security of shrinking generator against fault attacks. While this pseudorandom bitstream generator is cryptographically strong and well suited for hardware implementations, especially for cheap artefacts, we show that using it for the devices that are not fault resistant is risky. That is, even if a device concerned is tamper-proof, generating random faults and analyzing the results may reveal secret keys stored inside the device. For the attack we flip a random bit and observe propagation of errors. The attack uses peculiar properties of the shrinking generator and presents a new kind of threats for designs based on combining weaker generators. In particular, it indicates that potentially all designs based on combining LFSR generators might be practically weak due to slow propagation of errors in a single LFSR.

The order and the mobility of the lipids in the membrane were measured by the ESR method for the erythrocytes with both normal and modified intracellular concentration of ATP. The lipid order did not depend on the ATP level, but the lipid mobility was affected by the intracellular ATP concentration. The lipid mobility was higher in the cells with a larger concentration of ATP.

Among dipterans of medical and veterinary importance, the mosquitoes (Culicidae) play the most important role both as vectors and nuisance insects. Mass occurrence of floodwater mosquito species in the flooded area in Wrocław district (Lower Silesia, Poland) in 1997 has enhanced the already existed problem of mosquito control. A successful model of control strategy for the city based on the currently recommended integrated methodologies with special emphasis on preventive treatment of aquatic larvae with microbial insecticides has been conducted. Bacillus thuringiensis israelensis-based formulations replaced chemicals to control Culex pipiens and Culiseta annulata--two dominant species out of eight noticed during field study in Wrocław area in the years 1998-2000. Use of Bti larvicide or other biocontrol agent, conserving biodiversity, requires collecting entomological data, mapping and treating all breeding sources as well as the designing appropriate strategies every year.

We present two efficient and simple fault attacks on the shrinking generator. In a first case if the attacker can stop control generator for some small number of steps and observe the output, then with high probability he can deduce the full control sequence, and so the other input bitstream. The second method assumes that the attacker can disturb the control sequence (in an unpredictable and random way) and observe many samples of such experiments. Then he can reconstruct a certain sequence that agrees with the input sequence of the generator on a large fraction of bits.

Intensive physical exercise disturbs the entire homeostasis in the body and leads to changes in haemodynamic and metabolic alterations not only in skeletal muscles but also in many distant organs. In response to acute physical exercise, a decrease of the glomerular filtration may occur, followed by stimulation of the renin-angiotensin system (RAS). Recent studies have shown that both AT1 and AT2 angiotensin receptors may play a role in mediating the apoptotic process in the kidney. Our previous studies have demonstrated an occurrence of apoptosis in rat renal tubular cells after an excessive exercise. The aim of the present study was to determine the possible mechanism of exercise-induced apoptosis in rat kidney. The analysis was performed on kidneys of rats, subjected to treadmill running until exhaustion. Apoptosis was detected in paraffin sections by the TUNEL technique. The expression of AT1 and AT2 receptors in renal tubular cells was examined by immunohistochemistry and Western blot. Our results confirmed that apoptosis after physical exercise is present in renal distal tubular cells. Moreover, there was an increased expression of AT1 and AT2 receptors in distal tubular cells. These studies suggest that physical exercise may induce apoptosis by a mechanism, involving the activation of angiotensin AT1 and AT2 receptors.

Encapsulating messages in onions is one of the major techniques providing anonymous communication in computer networks. To some extent, it provides security against traffic analysis by a passive adversary. However, it can be highly vulnerable to attacks by an active adversary. For instance, the adversary may perform a simple so–called repetitive attack: a malicious server sends the same massage twice, then the adversary traces places where the same message appears twice – revealing the route of the original message. A repetitive attack was examined for mix–networks. However, none of the countermeasures designed is suitable for onion–routing. In this paper we propose an “onion-like” encoding design based on universal re-encryption. The onions constructed in this way can be used in a protocol that achieves the same goals as the classical onions, however, at the same time we achieve immunity against a repetitive attack. Even if an adversary disturbs communication and prevents processing a message somewhere on the onion path, it is easy to identify the malicious server performing the attack and provide an evidence of its illegal behavior.

We consider unlinkability of communication problem: given n users, each sending a message to some destination, encode and route the messages so that an adversary analyzing the traffic in the communication network cannot link the senders with the recipients. A solution should have a small communication overhead, that is, the number of additional messages should be kept low.

Recently, David Chaum proposed an electronic voting scheme that combines visual cryptography and digital processing. It was designed to meet not only mathematical security standards, but also to be accepted by voters that do not trust electronic devices.

It is believed that masking is an effective countermeasure against power analysis attacks: before a certain operation involving a key is performed in a cryptographic chip, the input to this operation is combined with a random value. This has to prevent leaking information since the input to the operation is random.We show that this belief might be wrong. We present a Hamming weight attack on an addition operation. It works with random inputs to the addition circuit, hence masking even helps in the case when we cannot control the plaintext. It can be applied to any round of the encryption. Even with moderate accuracy of measuring power consumption it determines explicitly subkey bits. The attack combines the classical power analysis (over Hamming weight) with the strategy of the saturation attack performed using a random sample.We conclude that implementing addition in cryptographic devices must be done very carefully as it might leak secret keys used for encryption. In particular, the simple key schedule of certain algorithms (such as IDEA and Twofish) combined with the usage of addition might be a serious danger.

Purified red blood cells, exposed to an ozone atmosphere, show an increased rate of hemolysis on sudden osmotic stress. To determine this effect of ozone in the presence of natural antioxidants, bovine red blood cells, used as models, were suspended in blood plasma, or in physiological saline with one of the following antioxidants: albumin, glutathione, uric acid, glucose and a vitamin E analog (trolox). After exposure of the suspensions to oxygen and oxygen/ozone atmospheres the rates of osmotic hemolysis were measured, using a stop-flow technique, and compared with rates measured in air-exposed controls. Blood plasma, containing all natural antioxidants, caused a decreased rate of osmotic hemolysis of cells exposed to oxygen and also decreased the effect of ozone. Trolox cancelled the oxygen effects only. Albumin, glutathione and uric acid tended to protect the cells from the hemolytic effects of ozone. The antihemolytic effect of glucose, seen only in some samples, may depend on uncontrolled factors. The alteration of the rates correlates with an increased fluidity of red cell membranes exposed to ozone.

The sulfhydryl group specific spin labels have been used to study the membrane proteins in the bovine and human erythrocyte membranes treated with ozone. The ratio hw/hs, determined from the respective peak amplitudes of the resulting EPR spectra of 3-maleimido-1-oxyl-2,2,5,5-tetramethylpyrrolidine bound to the ozone treated membranes, was shown to be elevated following an increased ozone concentration. The ratio of the central to high-field peak amplitude h0 / h-1, a measure of the rotational correlation time, determined from EPR spectra of ozonized membranes covalently labeled with 4-(iodoacetamido)-l-oxyl-2,2,5,5-tetramethylpyrrolidine was shown to be diminished. The changes in the erythrocyte membranes treated with ozone, observed for both spin labels, indicate a dose dependent increase in the mobility of membrane proteins.

The relationship between erythrocyte shape and the critical cell volume was investigated. Agents able to increase the critical cell volume induced three main stable shapes of erythrocytes: discocytic, stomatocytic, and echinocytic. The absence of correlation between shape and critical cell volume under isoosmotic conditions suggests that relative differences between the surface areas of the inner and the outer leaflet of the cell membrane do not influence the critical volume of a cell.

Arsenate, used as an inhibitor of glycolysis, decreases the critical cell volume of erythrocytes, whereas orthovanadate, used as an accelerator of dephosphorylation of phosphatidyl-inositol-4,5-biphosphate, exerts an opposite effect. The ATP-dependent changes of the critical cell volume do not depend on the phosphorylation state of phosphatidylinositol. An interaction of the membrane skeleton with the lipid-protein matrix has been proposed as a regulation mechanism of the critical cell volume of erythrocytes.

The effect of the intracellular level of ATP and of the state of spectrin on the critical cell volume of bovine erythrocyte was studied. The state of spectrin was changed by thermal denaturation, which for the bovine red cell took place at similar temperature as for the human erythrocyte. The increase of the ATP level and the spectrin denaturation increased the critical cell volume, while metabolic starvation decreased it. The changes of the ATP level did not influence the critical volume after the denaturation of spectrin. The results suggest that the ATP-dependent effect on the critical cell volume was caused by an alteration of the membrane extensibility due to the change of the membrane skeleton-lipid bilayer interaction(s).

The effect of DIDS on osmotic properties of bovine erythrocytes was studied. The isoosmotic volume, the amount of intracellular solutes, and the osmotically non-active volume were not influenced by DIDS. An increase of osmotic fragility of erythrocytes upon DIDS treatment was evident and identical both in NaCl and in NaCl + KCl hypotonic solutions. These results suggest that the critical cell volume decreases. The link between the effect of DIDS on the membrane skeleton extractability and on the osmotic fragility was postulated.

The influence of fructose-1,6-bisphosphate aldolase, as a membrane peripheral protein, on some electrical and transport properties of spherical lipid membranes was investigated. It was found that the association of the enzyme with the membrane did not effect markedly the electrical conductance or capacity of the membrane but decreased the water filtration coefficient and the cationic transferance number. The enzyme association also modifies temperature characteristics of the membrane parameters.

The osmotic properties of bovine erythrocytes aged in vivo were studied by the modified microhematocrit method. The osmotic fragility of older red cells decreases due to their larger relative osmotically non-active volume. Relative critical cell volume of bovine erythrocytes does not alter significantly with cell age. The age dependent change in the osmotic fragility of human red blood cells, the reverse of that found for bovine erythrocytes, is due to a different alteration of the critical cell volume during intravascular erythrocyte aging.

Water transport through spherical lipid bilayers consisting of bovine red cell lipids has been studied. A new experimental method for the determination of volume flow induced by concentration gradient is described. Changes of the filtration coefficient indicate phase transition in membrane lipid taking place at 33-36 degrees C.

The phase transitions of spherical lipid bilayers built of bovine red cell lipids have been studied. The electrical and ESR measurements revealed some conformational changes in lipid bilayers taking place at 36-38 degrees C.

A nitroxide spin label probe technique was applied to study the interaction between glutaraldehyde or osmium tetroxide (OsO4) amd the membranes of horse erythrocytes, ghosts and liposomes prepared from erythrocyte lipids. Two major conclusions have been established: (1) Reaction of the fixation reagents with the membrane is selective. OsO4 reacts predominantly with lipids and glutaraldehyde with membrane proteins. (2) The lipid-protein interactions change after pretreatment by OsO4 or glutaraldehyde.

The epithelial-mesenchymal transition (EMT) has been observed in progression of in situ breast cancer to the invasive form and might be initiated by snail family zinc finger 2 (SLUG) and twist family bHLH transcription factor 1 (TWIST) protein overexpression. During this phenomenon, cells lose their epithelial phenotype and acquire mesenchymal features. The aim of the study was to examine the association of EMT markers SLUG and TWIST with clinicopathological data and the possibility of using these proteins as prognostic markers of breast cancer.Immunohistochemical analysis (IHC) of SLUG and TWIST expression was performed on archival paraffin samples of 19 cases with fibrocystic breast changes (control group), 148 cases of invasive ductal breast cancer (IDC) and 26 of invasive lobular breast cancer (ILC). Laser capture microdissection for isolation of cells from 17 frozen samples of IDC was employed and subsequently SLUG and TWIST mRNA expression in cancer and stromal cells was detected separately by real-time polymerase chain reaction.SLUG and TWIST expression in IDC was significant higher in stromal cells regardless of the method of quantification used (p0.001 for SLUG mRNA, and p0.0001 for SLUG IHC, TWIST IHC and TWIST mRNA expression). Positive correlation of SLUG and TWIST protein and mRNA expression was observed in stromal cells of IDC (r=0.347; p0.0001 and r=0.704; p0.01, respectively). Expression of TWIST protein in IDC was higher in cancer cells of cases with shorter event-free survival period, as well as in stromal cells of cases with shorter overall survival period (p0.05 for both).Stromal cells could play a role in the regulation of EMT in breast cancer.

Metallothioneins (MTs) are low-molecular-weight and cysteine-rich proteins that bind heavy metal ions and oxygen-free radicals. MTs are commonly expressed in various tissues of mammals and are involved in regulation of cell proliferation and differentiation, and may be engaged in angiogenesis. Expression of MTs has been studied in many cancer types, especially breast cancer. The research results indicate that MTs may play important, although not yet fully known, roles in cancer angiogenesis. The aim of this study was to analyze the level of gene expression of selected MT isoforms induced with zinc ions in correlation with vascular endothelial growth factor (VEGF) isoforms in in vitro models of breast cancer.The studies were carried out in three breast cancer cell lines (MCF-7, SK-BR-3, MDA-MB-231). An epithelial cell line derived from normal breast tissue (Me16c) was used as a control. The levels of expression of selected MT isoforms and selected genes involved in angiogenesis were studied with real-time PCR.Expression of different MT isoforms was induced by zinc ions to differing degrees in individual breast cancer cell lines. An increase in the expression of some MT isoforms was associated with a slight increase in the level of expression of VEGFA.The research results may indicate certain correlation between an increased expression of selected MT isoforms and a pro-angiogenic factor VEGF in specific types of breast cancer cells.

A5/1 pseudo-random bit generator, known from GSM networks, potentially might be used for different purposes, such as secret hiding during cryptographic hardware testing, stream encryption in piconets and others. The main advantages of A5/1 are low cost and a fixed output ratio. We show that a hardware implementation of A5/1 and similar constructions must be quite careful. It faces a danger of a new kind of attack, which significantly reduces possible keyspace, allowing full recovery of A5/1 internal registers' content. We use “fault analysis” strategy: we disturb the A5/1 encrypting device (namely, clocking of the LFSR registers) so it produces an incorrect keystream, and through error analysis we deduce the state of the internal registers. If a secret material is used to initialize the generator, like in GSM, this may enable recovering the secret. The attack is based on unique properties of the clocking scheme used by A5/1, which is the basic security component of this construction. The computations that have to be performed in our attack are about 100 times faster than in the cases of the previous fault-less cryptanalysis methods.

Many studies are dedicated to exploring the molecular mechanisms of chemotherapy-resistance in breast cancer (BC). Some of them are focused on searching for candidate genes responsible for this process. The aim of this study was typing the candidate genes associated with the response to standard chemotherapy in the case of invasive ductal carcinoma. Frozen material from 28 biopsies obtained from IDC patients with different responses to chemotherapy were examined using gene expression microarray, Real-Time PCR (RT-PCR) and Western blot (WB). Based on the microarray results, further analysis of candidate gene expression was evaluated in 120 IDC cases by RT-PCR and in 224 IDC cases by immunohistochemistry (IHC). The results were correlated with clinical outcome and molecular subtype of the BC. Gene expression microarray revealed Prolactin-Induced Peptide (PIP) as a single gene differentially expressed in BC therapy responder or non-responder patients (p