Parameters of Oxidative and Inflammatory Status in a Three-Month Observation of Patients with Acute Myocardial Infarction Undergoing Coronary Angioplasty-A Preliminary Study.

Background and Objectives: Patients with acute myocardial infarction (MI) are usually treated with percutaneous transluminal coronary angioplasty (PTCA), which is burdened with a risk of postoperative complications, often accompanied by biochemical disturbances. The aim of our study was to evaluate a set of selected parameters of oxidative and inflammatory status, which could be useful in the management of post-procedural care in MI patients after PTCA. Materials and Methods: In this preliminary study, ischemia modified albumin (IMA), advanced oxidation protein products (AOPP), thiol groups (SH), total antioxidant status (TAS), insulin growth factor-1 (IGF-1), presepsin (PSP), and trimethylamine N-oxide (TMAO) were chosen as candidate biomarkers, and were determined in patients with MI who underwent PTCA at two time points: During cardiac episodes (at admission to the hospital, T0) and 3 months later (T3). Results: Most of the examined parameters were significantly different between patients and control subjects (except for IMA and TAS), but only hsCRP changed significantly during the time of observation (T0 vs. T3). Discriminant analysis created a model composed of AOPP, hsCRP, PSP, and TMAO, which differentiated male subjects into a group with MI and a control (without cardiovascular diseases). Conclusion: This set of parameters seems useful in evaluating inflammatory and oxidative status in MI patients after PTCA.

Association of Serum Proteases and Acute Phase Factors Levels with Survival Outcomes in Patients with Colorectal Cancer.

Colorectal cancer (CRC) represents a substantial burden on global healthcare, contributing to significant morbidity and mortality worldwide. Despite advances in screening methodologies, its incidence remains high, necessitating continued efforts in early detection and treatment. Neoplastic invasion and metastasis are primary determinants of CRC lethality, emphasizing the urgency of understanding underlying mechanisms to develop effective therapeutic strategies. This study aimed to explore the potential of serum biomarkers in predicting survival outcomes in CRC patients, with a focus on cathepsin B (CB), leukocytic elastase (LE), total sialic acid (TSA), lipid-associated sialic acid (LASA), antitrypsin activity (ATA), C-reactive protein (CRP), and cystatin C (CC). We recruited 185 CRC patients and 35 healthy controls, assessing demographic variables, tumor characteristics, and 7 serum biomarker levels, including (1) CB, (2) LE, (3) TSA, (4) LASA, (5) ATA, (6) CRP, and (7) CC. Statistical analyses included ANOVA with Tukey's post hoc tests and MANOVA for continuous variables. Student's t-test was used for dependent samples, while non-parametric tests like Mann-Whitney U and Wilcoxon signed-rank tests were applied for variables deviating from the normal distribution. Categorical variables were assessed using chi-square and Kruskal-Wallis tests. Spearman's rank correlation coefficient was utilized to examine variable correlations. Survival analysis employed the Kaplan-Meier method with a log-rank test for comparing survival times between groups. Significant associations were observed between CB (p = 0.04), LE (p = 0.01), and TSA (p = 0.008) levels and survival outcomes in CRC patients. Dukes' classification stages also showed a significant correlation with survival (p = 0.001). However, no significant associations were found for LASA, ATA, CRP, and CC. Multivariate analysis of LE, TSA, and ATA demonstrated a notable correlation with survival (p = 0.041), notwithstanding ATA's lack of significance in univariate analysis (p = 0.13). CB, LE, and TSA emerged as promising diagnostic markers with prognostic value in CRC, potentially aiding in early diagnosis and treatment planning. Further research is needed to validate these findings and explore additional prognostic indicators.

[The influence of sleep deprivation on the cognitive processes in medical students during exam session]. / Wplyw deprywacji snu na procesy poznawcze studentów medycyny w czasie sesji egzaminacyjnej.

BACKGROUND: Along with socio-economic pression increase in developed countries, the progressive shortening of night sleep has been observed. Sleep plays a vital role in human organism regeneration, and its deprivation leads to a series of adverse psychosomatic effects, including intellectual performance limitation or reducing body immunity, which increases susceptibility to diseases. Chronic sleep deprivation, quite often affecting medical students, significantly contributes to hypersomnia and leads to chronic fatigue. MATERIAL AND METHODS: The results of 60 students were analysed; students were divided into 2 subgroups, depending on the average sleep duration during previous 2-3 nights: IA (2.3±0.8 h on average) and IB (4.9±0.5 h on average). The control group consisted of 50 students, whose night sleep duration in that period was 7.5±0.62 h. In all subjects under analysis visual and auditory evoked potentials were registered, which is a non-invasive method of cognitive performance tests. RESULTS: The obtained results showed that people with average night sleep duration 2.3±0.8 h (subgroup IA) had worse rate and precision of stimulus response, and thereby significantly worse (p < 0.001) sensorimotor performance, then those from subgroup IB. The study of evoked potentials showed significant (from p < 0.04 to p < 0.001) elongation of all latency p-waves connected with the stimulus perception (N75) and attention span (P100, N135). Moreover, significant elongation of visual latency wave P100 and auditory wave V was related to psychophysical fatigue occurring in sleep deprivation. CONCLUSIONS: The analysis of this study results, obtained in medical students showed that sleep deprivation occurring during exam session is closely related to cognitive abilities, which in turn adversely affects the academic achievement. These results indicate that night sleep duration is a differentiating factor for cognitive abilities quality. Also, psychosomatic fatigue adversely affects cognitive processes. Med Pr. 2023;74(1):27-40.

EEG phase synchronization during absence seizures.

Absence seizures-generalized rhythmic spike-and-wave discharges (SWDs) are the defining property of childhood (CAE) and juvenile (JAE) absence epilepsies. Such seizures are the most compelling examples of pathological neuronal hypersynchrony. All the absence detection algorithms proposed so far have been derived from the properties of individual SWDs. In this work, we investigate EEG phase synchronization in patients with CAE/JAE and healthy subjects to explore the possibility of using the wavelet phase synchronization index to detect seizures and quantify their disorganization (fragmentation). The overlap of the ictal and interictal probability density functions was high enough to preclude effective seizure detection based solely on changes in EEG synchronization. We used a machine learning classifier with the phase synchronization index (calculated for 1 s data segments with 0.5 s overlap) and the normalized amplitude as features to detect generalized SWDs. Using 19 channels (10-20 setup), we identified 99.2% of absences. However, the overlap of the segments classified as ictal with seizures was only 83%. The analysis showed that seizures were disorganized in approximately half of the 65 subjects. On average, generalized SWDs lasted about 80% of the duration of abnormal EEG activity. The disruption of the ictal rhythm can manifest itself as the disappearance of epileptic spikes (with high-amplitude delta waves persisting), transient cessation of epileptic discharges, or loss of global synchronization. The detector can analyze a real-time data stream. Its performance is good for a six-channel setup (Fp1, Fp2, F7, F8, O1, O2), which can be implemented as an unobtrusive EEG headband. False detections are rare for controls and young adults (0.03% and 0.02%, respectively). In patients, they are more frequent (0.5%), but in approximately 82% cases, classification errors are caused by short epileptiform discharges. Most importantly, the proposed detector can be applied to parts of EEG with abnormal EEG activity to quantitatively determine seizure fragmentation. This property is important because a previous study reported that the probability of disorganized discharges is eight times higher in JAE than in CAE. Future research must establish whether seizure properties (frequency, length, fragmentation, etc.) and clinical characteristics can help distinguish CAE and JAE.

Diagnostic usefulness of selected proteases and acute phase factors in patients with colorectal adenocarcinoma.

BACKGROUND: Uncontrolled growth and loss of control over basic metabolic functions, leading to invasive proliferation and metastases, are the salient traits of malignant tumors in general and colorectal cancer in particular. Invasion and metastases hinder effective tumor treatment. While surgical techniques and radiotherapy can be used to remove tumor focus, only chemotherapy can eliminate dispersed neoplastic cells. However, the efficacy of the latter method is limited in the advanced stages of the disease. Therefore, recognition of the mechanisms involved in neoplastic cell spreading is indispensable for developing effective therapies. AIM: To use a number of biomarkers involved in cancer progression and identify a panel that could be used for effective early diagnosis. METHODS: We recruited 185 patients with colorectal adenocarcinoma (98 men, 87 women with median age 63). Thirty-five healthy controls were sex and age-matched. Dukes' staging was as follows: A = 22, B = 52, C = 72, D = 39. We analyzed patients' blood serum before surgery. We determined: (1) Cathepsin B (CB) with Barrett's method (fluorogenic substrate); (2) Leukocytic elastase (LE) in a complex with alpha 1 trypsin inhibitor (AAT) using the immunoenzymatic MERCK test; (3) Total sialic acid (TSA) with the colorimetric periodate-resorcinol method; (4) Lipid-bound sialic acid (LASA) with the colorimetric Taut's method; and (5) The antitrypsin activity (ATA) employing the colorimetric test. RESULTS: In patients, the values of the five biochemical parameters were as follows: CB = 16.1 ± 8.8 mU/L, LE = 875 ± 598 µg/L, TSA = 99 ± 31 mg%, LASA = 0.68 ± 0.33 mg%, and ATA = 3211 ± 1504 U/mL. Except for LASA, they were significantly greater than those of controls: CB = 11.4 ± 6.5 mU/L, LE = 379 ± 187 µg/L, TSA = 71.4 ± 15.1 mg%, LASA = 0.69 ± 0.28 mg%, and ATA = 2016 ± 690 U/mL. For CB and LASA, the differences between the four Dukes' stages and controls were not statistically significant. The inter-stage differences for CB and LASA were also absent. The receiver operating characteristic (ROC) analysis revealed the potential diagnostic value of CB, TSA, and ATA. The area under ROC, sensitivity, and specificity for these three parameters were: 0.85, 72%, 90%; 0.75, 66%, 77%; and 0.77, 63%, 84%, respectively. The sensitivity and specificity for the three-parameter panel CB-TSA-ATA were equal to 88.2% and 100%, respectively. CONCLUSION: The increased value of CB, TSA, and ATA parameters are associated with tumor biology, invasion, and metastasis of colorectal cancer. The presented evidence suggests the potential value of the CB-TSA-ATA biochemical marker panel in early diagnostics.

Absence Seizure Detection Algorithm for Portable EEG Devices.

Absence seizures are generalized nonmotor epileptic seizures with abrupt onset and termination. Transient impairment of consciousness and spike-slow wave discharges (SWDs) in EEG are their characteristic manifestations. This type of seizure is severe in two common pediatric syndromes: childhood (CAE) and juvenile (JAE) absence epilepsy. The appearance of low-cost, portable EEG devices has paved the way for long-term, remote monitoring of CAE and JAE patients. The potential benefits of this kind of monitoring include facilitating diagnosis, personalized drug titration, and determining the duration of pharmacotherapy. Herein, we present a novel absence detection algorithm based on the properties of the complex Morlet continuous wavelet transform of SWDs. We used a dataset containing EEGs from 64 patients (37 h of recordings with almost 400 seizures) and 30 age and sex-matched controls (9 h of recordings) for development and testing. For seizures lasting longer than 2 s, the detector, which analyzed two bipolar EEG channels (Fp1-T3 and Fp2-T4), achieved a sensitivity of 97.6% with 0.7/h detection rate. In the patients, all false detections were associated with epileptiform discharges, which did not yield clinical manifestations. When the duration threshold was raised to 3 s, the false detection rate fell to 0.5/h. The overlap of automatically detected seizures with the actual seizures was equal to ~96%. For EEG recordings sampled at 250 Hz, the one-channel processing speed for midrange smartphones running Android 10 (about 0.2 s per 1 min of EEG) was high enough for real-time seizure detection.

Interhemispheric cerebral asymmetry detected by VEPS in diabetic patients with recognized depression.

OBJECTIVES: The present study was undertaken in order to verify the hypothesis stating that patients with depression exhibit some abnormalities concerning a cerebral symmetry. For this purpose, an analysis of the relationship between the VEPs (Visual Evoked Potentials) results and the depressive symptoms intensification, as well interrelation between depressive and diabetic symptoms were performed. MATERIAL AND METHODS: VEPs recordings were obtained from the two study groups (both aged 20-45 years), 20 healthy subjects and 32 diabetic patients with clinically documented depression. The VEPs examination was carried out using a computer system called STELLA (Stimulated Electroencephalogram on Line Analyzer). RESULTS: VEPs examinations revealed a cerebral symmetry in the all control subjects and in 10 out of 32 diabetic patients with depressive disorders. It is noteworthy that 22 of 32 diabetic patients (68.8%) showed a hemispheric asymmetry in the VEPs recordings. Of 12 patients with a moderate depression, 10 showed the left cerebral laterality, and 2--the right laterality. In turn, all 10 patients with major depressive disorder (MDD) demonstrated in the VEPs recordings a significant cerebral laterality with the right hemisphere dominance. CONCLUSIONS: The obtained results suggest that interhemispheric cerebral asymmetry might be considered a specific feature of depression, and, if this is a case, it could justify a conclusion that right hemisphere dominance could be a neurophysiological marker of MDD. It should be emphasized that intensification of the depressive symptoms has an unfavorable influence on course of diabetes mellitus, its self-control and severity of following complications.

[Evaluation of ambulatory cardiac rehabilitation influence on the intensity of erectile dysfunction in patients with ischemic heart disease]. / Ocena wplywu ambulatoryjnej rehabilitacji kardiologicznej na natezenie zaburzen erekcji u chorych na chorobe niedokrwienna serca.

UNLABELLED: Considering a progressive course of cardiovascular disease, often leading to premature death, and difficulty in obtaining long-lasting stabilization of clinic state, it is deeply justified to take preventive interventions completing repairing actions and pharmacotherapy. Addressing various preventive programs to the patients with CVD, we put a special emphasis to simple and low-cost modification of physical activity, which has a beneficial influence on the circulatory system. As it has been proved, protective action of physical activity on the vessels concerns the whole arterial system including the vessels responsible for erectile dysfunction (ED) creation. THE AIM OF THE STUDY: The analysis of the influence of physical activity modification, taking place within frames of a six-month supervised cardiac rehabilitation, on ED intensification in the population with ischemic heart disease (IHD). MATERIAL AND METHODS: The analysis has been conducted on 129 patients with IHD, whose preliminary test IIEF-5 (International Index of Erectile Function-5) showed < or =21 scores, which justified ED diagnosis. The analyzed group consisted of 98 patients with IHD at the mean age of 62.35 +/- 8.88 years, who were subjected to the six-month cardiologic rehabilitation. The testing group comprises 31 patients with IHD at the mean age of 61.71 +/- 7.35, who were not rehabilitated for objective reasons. The patients of both groups filled in an IIEF-5 questionnaire twice, at the interval of six months. RESULTS: As a result of cardiologic rehabilitation in the analyzed group, a statistically significant increase in scores occurred in the IIEF5 test, from 11.88 +/- 6.2 to 13.69 +/- 7.07, which was not observed in the control group. Moreover, a division of the analyzed group into ED intensity categories (severe, medium-severe, moderate, light) confirmed the occurrence of significant changes of ED intensity for subsequent ED severity categories. For the subsequent ED intensity categories, from the greatest to the lightest, the statistically significant increase of the scores from a sheet IIEF-5: 4.66 +/- 0.98 vs. 5.34 +/- 1.41 (p < 0.01); 9.5 +/- 1.2 vs. 10.9 +/- 1.58 (p < 0.01); 14.67 +/- 1.22 vs. 17.7 +/- 1.80 (p < 0.01) and 19.62 +/- 1.11 vs. 21.85 +/- 1.23 (p < 0.01) has been found. Bearing in mind the dependence of results on the credibility of data from the sheet IIEF-5, the last element was the analysis of 'truthfulness test', which has not shown any statistically significant differences obtained for first and next filling in the questionnaire. CONCLUSIONS: The performed analyses allowed drawing the conclusions. A six-month cardiac rehabilitation cycle led to a significant positive modification of erectile dysfunction intensity. The greatest positive ED modification occurred at the groups of the patients, in whom the erectile dysfunction intensity was the smallest.

Assessment of brainstem auditory evoked potentials (BAEPs) in patients with acromegaly.

OBJECTIVES: Acromegaly is associated by various systemic complications, involving also the nervous system. Other studies revealed peripheral but not central nervous system impairment with somatosensory evoked potentials examinations in acromegaly. Aim of the present study was to assess whether brainstem transmission in acromegaly is disturbed. MATERIAL AND METHODS: The study was carried out in 37 patients. The control group consisted of 47 healthy persons. In all of the subjects, peripheral transmission, reflected by peak I latency, and brainstem transmission, tested by interpeak latency I-V (IPL I-V), were examined. RESULTS: Peak I latency was delayed in 6 out of 37 patients (1 - bilaterally, 2 - right side, 3 - left side). The group-mean latency of peak I was 1.53 msec and 1.56 msec, for the right and left side, respectively. There were found no statistically significant differences between the right and left side, likewise in comparison with control group. In turn, as compared with the controls, IPL I-V was disturbed in 25 out of 37 patients: in most of the cases IPL I-V prolongation was observed (8 - bilateral prolongation, 11 - right side, 6 - left side). Moreover, the statistically significant difference between the brainstem sides (4.27 vs. 4.11 msec; p<0.05) was observed. CONCLUSION: In the examined patients with acromegaly, there was no peripheral disturbance in transmission, as examined by BAEPs registrations. Conversely, in nearly half of the patients with acromegaly, brainstem transmission was found to be delayed, and significant difference between responses from the both sides of the brainstem were noted.

Wavelet analysis of scaling properties of gastric electrical activity.

We present a novel approach to the analysis of fluctuations in human myoelectrical gastric activity measured noninvasively from the surface of the abdomen. The time intervals between successive maxima of the wavelet transformed quasi-periodic electrogastrographic waveform define the gastric rate variability (GRV) time series. By using the method of average wavelet coefficients, the statistical fluctuations in the GRV signal in healthy individuals are determined to scale in time. Such scaling was previously found in a variety of physiological phenomena, all of which support the hypothesis that physiological dynamics utilize fractal time series. We determine the scaling index in a cohort of 17 healthy individuals to be 0.80 +/- 0.14, which compared with a set of surrogate data is found to be significant at the level P < 0.01. We also determined that the dynamical pattern, so evident in the spectrum of average wavelet coefficients of the GRV time series of healthy individuals, is significantly reduced in a cohort of systemic sclerosis patients having a scaling index 0.64 +/- 0.17. These results imply that the long-term memory in GRV time series is significantly reduced from healthy individuals to those with systemic sclerosis. Consequently, this disease degrades the complexity of the underlying gastrointestinal control system and this degradation is manifest in the loss of scaling in the GRV time series.

Total and lipid-bound plasma sialic acid as diagnostic markers in colorectal cancer patients: correlation with cathepsin B expression in progression to Dukes stage.

PURPOSE: Serum cathepsin B (CB), Total Sialic acid (TSA), total sialic acid (TSA) and lipid bound sialic acid (TSA) concentrations more useful than the other markers investigated for detecting different malignancies. Our aim was to investigate the possible correlation between serum CB with TSA, LSA in colorectal carcinoma with pathological stages progressed of the disease. METHODS: The study was performed on 177 patients (109 patients with colon and 68 patients with rectal) and 50 healthy individuals comprised the control group. Serum CB activity was determined using fluorogenic substrate. Serum TSA and LSA Concentrations were measured according to the method described by Katopodis. RESULTS: Plasma CB and TSA levels in the tumor group were significantly increased in comparison with the controls group (P < or = 0.0001). No significant differences were observed in LSA level between the tumor group and the controls group. T/N ratios for CB, TSA elevated 2.3-fold, 2.5-fold respectively). LSA 1.8-fold. Serum CB activity, TSA concentrations values in plasma samples of patients were increased significantly with pathological stages progressed (P < or = 0.0001). CB is seen to correlate more strongly with TSA in tumor group (P < or = 0.0001, r= 0.7277) in comparison with controls group. These correlations became more significant as the stage of the disease progressed. CONCLUSION: The present investigations indicate that CB activity, serum TSA, concentrations are sensitive markers for detecting and earliest diagnosis of colorectal cancer. These markers with other clinical and biochemical criteria may play important metabolic roles in cancer progression.

Cathepsin D expression in human colorectal cancer: relationship with tumour type and tissue differentiation grade.

OBJECTIVE: Cathepsin D (CD) is one of the main proteolytic enzymes contributing to the development of cancer. The aim of this study was to CD activity assay in the homogenates of tissues from the centre of the tumour (0) and tumour free area 2 cm, and 5 cm from the tumour border in human colorectal cancer. Activity in the centre of the tumour was compared with immunohistochemical expression CD. METHODS: CD activity was measured using acid denatured Hb as a substrate. For immunohistochemical staining peroxidase method was used. RESULTS: Activity of CD was significantly higher (15-fold) in tumour tissue homogenates in comparison to normal mucosa adjacent (control) (p < or = 0.0001) and raised parallel to the stage of tumour tissue differentiation grade. CD activity decreased significantly (p < or = 0.0001) with the distance from the tumour border 2 cm (12.7 fold) and 5 cm (5.7 fold) in comparison to the centre of the tumour. In immunohistochemical examinations CD was detected as diffuse cytoplasmic as well as fine granular staining of the cytoplasm, with occasional coarse cytoplasmic granules staining in the same cases that were positive for both. Positive staining was observed in 2 of 3 in well-differentiated (66%), 4 of 10 in moderately-differentiated (40%) and 4 of 5 in poorly-differentiated (80%), tubular adencarcinomas represented: 3 of 7 (42%) and 9 of 13 in invasive adencarcinoma (69%). CONCLUSION: We have observed a wide range of cathepsin D and their antigen expressions patterns in colorectal tumours with the development the disease stage, this finding may be used as a daignostic tumor marker in colorectal cancer.

Actin content and actin polymerization in hepatoma Morris 5123 tumor bearing rats after treatment with cysteine protease inhibitor and vitamin E.

AIM: Actin microfilaments are components of cytoskeleton, the structure involved in many cellular functions, including cell motility, and morphology, its necessary for tumor progression and metastasis. We investigated the effects of vitamin E and human placetal cysteine protease inhibitor (CPI) on actin content and polymerization after implantation the hepatoma Morris 5123 tumor in Buffalo rat. METHODS: We measured the size and survival animals treated with human placental cysteine protease inhibitor (CPI) plus vitamin E and none treated rats. Also measured the actin content and polymerization in the tumor and liver tissues by the inhibition of DNase I from bovine pancreas under standard assay conditions RESULTS: We observed that the combination 10 mg of vitamin E plus 200 microg CPI obtained the best results than authors. In those cases the animals survived for longer than 4 weeks. In numerous cases the 70 % of tumors disappeared following CPI and vitamin E application. The number of complete tumor responses was higher after combination 10 mg of vitamin E plus CPI 200 microg i.e. 5/7 rats than others group. We showed statistically significant decrease of monomeric (G), filamentous (F) and total actin as well as the F:G ratio level in tumor tissues after the rats was treated with CPI plus vitamin E in comparison with control animals. CONCLUSIONS: The new therapy may be the way for therapeutic intervention, aimed at stopping and possibly reversing the process of metastatic growth, with the use of drugs affecting actin polymerization.

Expression of cystatin C in clinical human colorectal cancer tissues.

We studied the relation between the antipapain activity of cysteine proteinase inhibitors (CPI) and immunohistochemical staining for cystatin C, using anti-chicken cystatin antibodies, in the colorectal cancer tissues. In primary tumour tissues immuno-peroxidase reactivity was present in the cytoplasm and on the cell surface membranes. Sections of non malignant tissues showed no staining. The percentages of positive staining were greater for adenocarcinoma than carcinoma,100% and 77% respectively. Antipapain activity which was increased in malignant tissues in comparison to control, rose successively from well differentiated carcinomas through moderately to poor differentiated. Invasive adenocarcinomas had higher antipapain activity than noninvasive ones. The results indicated that immunohistochemical detection of cystatin using anti-chicken cystatin antibodies could be useful in studying the prognostic significance of cystatin C expression in colorectal cancer.

Effect of vitamin E and human placenta cysteine peptidase inhibitor on expression of cathepsins B and L in implanted hepatoma Morris 5123 tumor model in Wistar rats.

AIM: To examine the effectiveness of human placental inhibitors, by injecting vitamin E to rats with transplanted Morris-5123 hepatoma, on the expression of cathepsins B and L in tumor, liver, lung and blood sera after transplantation of Morris 5123 hepatoma. METHODS: Animals were divided into 10 groups receiving three different concentrations of vitamin E and inhibitors along or in combination and compared with negative control (healthy rats) and positive control (tumor rats). Effectiveness of treatment was evaluated with regard to survival time, tumor response and determination of the activities of proteolytic enzymes and their inhibitors using flurogenic substrates. RESULTS: Cathepsins B and L activities were elevated by 16-fold in comparison with negative control tissues, and their endogenous inhibitor activity decreased by 1.2-fold before treatment. In several cases, tumors completely disappeared following vitamin E plus human placental cyteine protease inhibitor (CPI) compared with controls. The number of complete tumor responses was higher when 20 m/kg vitamin E plus 400 microg of CPI was used, i.e. 7/10 rats survived more than two mo. Cathepsins B and L were expressed significantly in tumor, liver, lung tissues and sera in parallel to the increasing of the endogenous inhibitor activity compared with the controls after treatment (P<0.0001). CONCLUSION: The data indicate formation of metastasis significantly reduced in treated rats, which might provide a therapeutic basis for anti-cancer therapy.

Effects of combined in vivo treatment of transplantable solid mammary carcinoma in wistar rats using vitamin E and cysteine peptidase inhibitors from human placenta.

Cysteine cathepsin B and its endogenous inhibitor play an important role in tumor progression. Increase in cathepsin B expression and reduced levels of its inhibitors were associated with tumor malignancy in breast cancer. The objective of this study was to investigate the effects of a new therapy combining vitamin E and placental inhibitor on the level of endogenous protease inhibitor in sera and tumor tissues with mammary cancer. The inhibitor was used in doses of 100 and 200 micrograms per animal for 8 days. Vitamin E was added after the last treatment with inhibitor and was injected daily in doses of 10 and 20 mg per animal for one mouth. The size and survival time of treated animals as well as cathepsin B and the inhibitor activity in tumor and sera before and after treatment in comparison with the control groups were determined. The activity of cathepsin B significantly decreased both in tumor tissues and in sera (P < or = 0.0001). Cathepsin B activity in tumor tissue homogenates and in sera decreased two-fold and three-fold, respectively, after the animals were treated with vitamin E at a dose of 20 mg, and decreased five-fold and 15-fold, respectively, when treated with vitamin E plus inhibitor in comparison with untreated animals. Endogenous inhibitor activity increased six-fold and 12-fold in the sera and tissue homogenates, respectively, after the animals were treated with 200 micrograms of cysteine protease inhibitor plus 20 mg of vitamin E, in comparison with untreated animals. The total cure responses were higher in eight of 10 rats, as compared with untreated animals. The combination of placental inhibitor and vitamin E resulted in a significant reduction in breast metastasis and might provide a therapeutic basis for anti-metastasis therapy.

Inhibition of cathepsin B activity in human breast cancer tissue by cysteine peptidase inhibitor isolated from human placenta: immunohistochemical and biochemical studies.

Cysteine peptidases and their endogenous inhibitors (CPI) have been shown to be involved in tumor progression and metastasis. Since their activity has been found to be changed in tumor tissue and/or body fluids of cancer patients, the determination of the peptidase/inhibitor levels is considered as a procedure of diagnostic value. Determination of cathepsin B, its precursor and inhibitor activity in homogenates of tumors and control breast tissue samples of patients with invasive ductal and lobular breast carcinoma and with benign breast disease (BBD) was performed using fluorometric assay. Immunohistochemical staining of the breast tissue samples was carried out using polyclonal antibody against cysteine peptidase inhibitor isolated from human placenta. Procathepsin B and cathepsin B were found to be significantly increased and their endogenous inhibitors decreased in homogenates of tumors from patients with breast cancer. A correlation between procathepsin B or cathepsin B activities as well as cysteine peptidase inhibitor activity and the histopathological grading of the tumor was observed. All samples of the tumor tissue showed positive immunostaining with antibody raised against cysteine peptidase inhibitor, while in the control tissue samples the immunostaining was much weaker. Significant difference observed between the activities of cathepsin B and/or its precursor in malignant and benign tumors might serve as a useful clinical indicator in discrimination between benign and invasive tumors.

The role of exercise ventilation in clinical evaluation and risk stratification in patients with chronic heart failure.

BACKGROUND: Patients with chronic heart failure (CHF) are characterised by an increased ventilatory response to exercise. The role of exercise ventilation in the risk stratification and evaluation of patients with CHF has not yet been established. AIM: To examine the relationship between exercise ventilation indices and clinical parameters of CHF and to assess the prognostic value of the ventilatory response to exercise. METHODS: The study group consisted of 87 patients with CHF (72 males, mean age 58 years) with a mean left ventricular ejection fraction of 32%. Ten patients were in NYHA class I, 38 - in NYHA class II, 34 - in NYHA class III, and 5 - in NYHA class IV. The control group consisted of 20 patients without CHF (13 males, mean age 58 years, mean LVEF - 61%). All studied subjects underwent maximal exercise test with gas-exchange measurement. The following parameters were analysed: peak exercise oxygen consumption [peak VO(2) (ml/kg/min)], VE-VCO(2) index [a coefficient of linear regression analysis depicting an association between ventilation (VE) and carbon dioxide production (VCO(2)) during exercise] and VE/VCO(2) ratio at peak exercise to VE/VCO(2) ratio while at rest (VE/VCO(2 peak/rest)). RESULTS: Ventilatory response indices were significantly higher in patients with CHF compared with controls: VE-VCO(2) - 37.9+/-11.1 vs 27.1+/-4.1; VE-VCO(2 peak/rest) - 0.89+/-0.14 vs 0.75+/-0.10 (p<0.001). In CHF patients a significant positive correlation between ventilatory response parameters and NYHA class (VE-VCO(2) - r=0.52; VE/VCO(2 peak/rest) - r=0.47) and a negative correlation with peak VO(2) (VE-VCO(2) - r=-0.52; VE/VCO(2 peak/rest) - r=-0.49) were noted (p<0.0001 for all correlations). No correlation was found between ventilatory parameters and echocardiographic variables or CHF aetiology. During the follow-up period lasting at least 12 months, 17 (22%) patients died. In the univariate Cox model, NYHA class III-IV, decreased peak VO(2) and increased VE-VCO(2) and VE/VCO(2 peak/rest) values were significantly associated with the risk of death. The multivariate analysis revealed that VE/VCO(2 peak/rest) > or =1.0 was the adverse prognostic factor, independent of peak VO(2) (p=0.02) and NYHA class (p=0.01). The Kaplan-Meier analysis showed that prognosis during the 18-month follow-up period in patients with enhanced exercise ventilation was worse than in the remaining patients (59% survival in patients with VE/VCO(2 peak/rest) > or =1.0 59% vs 91% survival in patients with VE/VCO(2 peak/rest) <1.0, p=0.001). CONCLUSIONS: In patients with stable CHF simple exercise ventilation parameters may provide important clinical and prognostic information.

Ovocystatin affects actin cytoskeleton organization and induces proapoptotic activity.

Ovocystatin is a chicken egg white protein, generally known for its inhibitory activity against cysteine proteases. However, biological activity of ovocystatin does not seem to be well recognized in respect to other possible cellular effects. Our attention has been focused on ovocystatin cytotoxic effects in relation to its influence on actin cytoskeleton organization and apoptosis induction. In vitro studies with human melanoma A375, human cervix HeLa cancer cells and normal human fibroblasts - NHDF were done. Cytotoxic activity of ovocystatin was seen in respect to apoptosis induction - manifested by cell shape changes, phosphatydylserine translocation and actin cytoskeleton reorganization. Normal human fibroblasts have shown lower sensitivity to ovocystatin as compared with human melanoma A375 and human cervix HeLa cancer cells. In conclusion, ovocystatin affects actin cytoskeleton organization and displays proapoptotic activity towards applied cell lines. This implicates its application as a potential anticancer drug. However, its adverse effects on normal cells should be taken into consideration.

Polymorphisms in methyl-group metabolism genes and risk of sporadic colorectal cancer with relation to the CpG island methylator phenotype.

BACKGROUND: The CpG island methylator phenotype (CIMP), together with extensive promoter methylation, is regarded as one of the mechanisms involved in colorectal carcinogenesis. The mechanisms underlying CIMP in sporadic colorectal cancer are poorly understood. Genes involved in methyl-group metabolism are likely to affect DNA methylation and thereby influence an individual's risk of CIMP. The aim of the present study was to evaluate whether polymorphisms in the genes encoding methyl-group metabolism pathway predispose to CIMP+ and/or CIMP- CRC. METHODS: We examined the potential association between the polymorphisms of MTHFR 677C>T, TS 5'UTR 2R/3R, TS 3'UTR 1494del6, DeltaDNMT3B -149C>T and DNMT3B -283T>C in a group of 46 CIMP+ CRC cases, 140 CIMP- CRC cases and 140 healthy controls. The CIMP status of the CRC cases was determined by MS-PCR in tumor tissue by a panel of five markers (CACNA1G, IGF2, NEUROG1, RUNX3 and SOCS1), which was also followed by analyzing hMLH1 methylation and BRAF V600E mutation. RESULTS: The variant allele homozygote genotype for the DeltaDNMT3B -283T>C polymorphism was associated with a decreased risk for CIMP+ CRC (OR: 0.31, 95%CI: 0.09-0.73, p=0.009). Individuals with TS 3R/3R had an increased risk of CIMP- CRC (OR: 2.21, 95%CI: 1.23-4.91, p=0.01). Moreover, the carriers of 3R allele had an increased risk of CIMP- CRC (OR: 1.45, 95%CI: 1.10-2.13, p=0.01). CONCLUSION: This study provides support to the hypothesis that methyl-group metabolism plays a role in the etiology of both CIMP+ and CIMP- colorectal cancers but has a different impact on a distinct molecular subgroups of colorectal cancer.