Agnieszka Pedrycz

Immunohistochemical study of proteins linked to apoptosis in rat fetal kidney cells following prepregnancy adriamycin administration in the mother.

Adriamycin is an antibiotic of the anthracycline group. In a previous study, we showed that administration of a single dose of adriamycin (i.p. injection, 5mg/kg body weight) 4 weeks before pregnancy in female Wistar rats induced histological changes in the fetal renal cells typical of apoptosis and also over-expression of heat shock proteins (HSP70). Using a similar experimental model, we have now examined renal cells in fetuses (gestation day 20) to investigate the pathways of the transduction signal of apoptosis in these cells that is induced by prepregnancy maternal administration of adriamycin. Immunolocalization of several proteins - p53, Bax, Apaf-1 and caspase 9 - which take part in the mitochondrial pathway of apoptosis and caspase 12, which takes part in the endoplasmic reticulum pathway of apoptosis, was determined. The results showed that adriamycin administered to the mother rat before pregnancy subsequently induced changes in fetal kidneys involving both the mitochondrial pathway of apoptosis, with increased labeling of the proteins p53, Bax, Apaf-1 and caspase 9, and the endoplasmic reticulum pathway of apoptosis, with increased labeling of caspase 12. Immunolabeling of these proteins was quantified using an image analysis program.

Chromoendoscopy with a Standard-Resolution Colonoscope for Evaluation of Rectal Aberrant Crypt Foci

Colorectal cancer (CRC) is the second most common cause of death worldwide. According to the theory by Vogelstein, colorectal carcinogenesis involves a series of successive changes in the normal colonic mucosa, starting with excessive proliferation and focal disorders of intestinal crypts, followed by adenoma and its subsequent malignant transformation. The first identifiable changes in CRC carcinogenesis are aberrant crypt foci (ACF). ACF are invisible during routine colonoscopy yet are well identifiable in chromoendoscopy using methylene blue or indigo carmine. High-resolution colonoscopes are used for assessment of ACF. The aim of the present study was to evaluate the usefulness of standard-resolution colonoscopy for identification of rectal ACF. The following parameters were evaluated: duration of chromoendoscopy of a given rectal segment, type of ACF, sensitivity and specificity of endoscopy combined with histopathological evaluation. The mean duration of colonoscopy and chromoendoscopy was 26.8 min. In the study population, typical ACF were found in 73 patients (p = 0.489), hyperplastic ACF in 49 (p = 0.328), and dysplastic ACF in 16 patients (p = 0.107). Mixed ACF were observed in 11 individuals (p = 0.073). The sensitivity of the method was found to be 0.96 whereas its specificity 0.99. Identification of rectal ACF using standard-resolution colonoscopy combined with rectal mucosa staining with 0.25% methylene blue is characterised by high sensitivity and specificity.

Influence of L-arginine on expression of HSP70 and p-53 proteins – early biomarkers of cellular danger in renal tubular cells. Immunohistochemical assessment

Introduction The aim of the present study was to investigate expression of HSP70 and p-53 proteins as mechanisms of protection of the renal tubular epithelial cells from l-arginine that induces cellular stress. Material and methods The study material consisted of 16 white Wistar female rats. The rats were divided into 2 equal groups. The rats in the experimental group received L-arginine 40 mg/kg body weight per capita every other day for 2 weeks and were decapitated after 3 weeks of the experiment. After decapitation, specimens from the kidney were collected, fixed in 10% formalin, and then embedded in paraffin blocks. Proteins HSP70 and p-53 on slides were detected using the standard three-step immunohistochemical method. Results The quantitative evaluation of HSP70 and p-53 expression showed that the area occupied with positive HSP70 and p-53 reaction in the rat renal tubular cells of the experimental group (p-53: 2835.44 ±254.72 µm2; HSP70: 24111.42 ±4290.88 µm2) was more statistically significant than the control group (p-53: 1882.05 ±466.43 µm2; HSP70: 11388.63 ±1455.24 µm2). In the present study, the dose of L-arginine was similar to the one that was used in the gestosis treatment of pregnant women. Conclusions The renal epithelial cells responded to L-arginine therapy, increasing expression of HSP70 and p-53 proteins. The study showed that L-arginine as a donor of exogenous nitric oxide has a disruptive effect on the renal tubular cells of rat kidneys. Thus it is going to be a subject of the authors future investigations.

Body Posture, Postural Stability, and Metabolic Age in Patients with Parkinson’s Disease

Background The study aims to analyze the relationship between body posture and composition, as well as postural stability in Parkinsons disease patients. Material and Methods 32 people were evaluated. The study was conducted in the Laboratory of Posturology at Jan Kochanowski University in Kielce (Poland). Body posture was examined using the optoelectronic body posture Formetric Diers Method III 4D. Postural stability was evaluated using the Biodex Balance System platform. Body composition was assessed with the method of bioelectrical impedance analysis using the Tanita MC 780 MA analyzer. Results 11 patients (34.37%) had hyperkyphosis, 10 (31.25%) hyperlordosis, and 3 (9.37%) hyperkyphosis-hyperlordosis posture. Scoliosis (>10°) was observed in 28 (87.5%) subjects, whereas 4 (12.5%) presented scoliotic body posture (1–9°). In the examined population, all parameters of postural stability were within normal limits. Conclusions A significant positive correlation was observed between surface rotation (°), General Stability Index (r = 0.4075, p = 0.0206), and Anteroposterior Stability Index (r = 0.3819, p = 0.0310). There was also a significant positive correlation between surface rotation (+max) (°), General Stability Index (r = 0.3526, p = 0.0206), and Anteroposterior Stability Index (r = 0.3873, p = 0.0285). Metabolic age also presented a significant positive correlation between metabolic age and General Stability Index (r = 0.4057, p = 0.0212), as well as Anteroposterior Stability Index (r = 0.3507, p = 0.0490).

Development of pulmonary oxygen toxicity in rats after hyperoxic exposure

Abstract The aim of the study was to examine the effects of hyperbaric oxygen on lung aeration on an animal experimental model and compare the obtained results with the anticipated scope of damage to pulmonary parenchyma in humans under the same exposure conditions. The research was carried out on Black Hood rats that were kept in a hyperbaric chamber designed for animals in an atmosphere of pure oxygen and at overpressures of 0.15, 0.2, 0.3, 0.4, and 0.5 MPa for 1, 2 or 4 h. After sacrificing the animals, histopathological specimens were obtained encompassing cross-sections of entire lungs, which were subjected to qualitative and quantitative examination with the use of the 121-point Haug grid. A statistically significant decrease in pulmonary parenchyma was observed as a result of an increasing oxygen partial pressure as well as with prolonged exposure time. The intensification of changes observed was much higher than expected on the basis of calculations performed with the use of tables.

Ultrastructure of Renal Tubular Epithelial Cells Of Rat’s Kidneys after Administration of L-Arginine

Abstract Sixteen white Wistar female rats were divided into two equal groups. Experimental group received per os 40 mg/kg b.w. of L-arginine, every other day for 2 weeks and were decapitated after 3 weeks of the experiment. Control rats received in the same manner 2 ml of distilled water and were decapitated after 3 weeks of the experiment. The renal lesions observed under electron microscope were of focal character and concerned only the experimental group. The tubules with necrotic cells were observed among normal tubules or single normal epithelial cells of the tubular wall. The boundaries between epithelial cells of the tubule wall were blurred. The mitochondria indicated abnormal structure. Numerous lysosomes and peroxysomes with dark, homogenous content were observed. The rough endoplasmic reticulum had widened channels and was focally completely destroyed. The nucleus of damaged cells was most commonly located in one of the cell poles; its shape was changed and visibly smaller than the nuclei of normal cells. Condensation and peripherally located chromatin were noticed. The lesions observed were characteristic for apoptotic cells.

The Use of Saliva in Medical Diagnostics

For approximately 15 years saliva has been an important element of laboratory diagnostics. Scientists are still looking for modern ways to make use of it in medical diagnostics and the monitoring of therapeutic effects. Advantages associated with the use of saliva in medical diagnostics are multiple. Using saliva as a diagnostic material is non-invasive and painless, and convenient for professionally active adults, elderly patients and young children alike. Saliva may be the perfect diagnostic material and an alternative to routinely used serum, plasma and urine. It is one of the regularly produced biological secretions, containing numerous substances similarly to other fluids, e.g. blood and urine. It seems, however, that its practical use remains underestimated. The availability of saliva as a biological material, and its non-invasive collection method, makes assays of various salivary components increasingly important. ABSTRACT

Assessment of L-Arginine as an Effect of Exogenous Nitric Oxide (No) on Expression of Markers of Cellular Stress in Rats’ Hepatocytes

Abstract The aim of this study was immunohistochemical evaluation of heat shock protein (Hsp70) and p-53 proteins in the Larginine- induced cellular stress in hepatocytes of rats. Sixteen white Wistar female rats were divided into two equal groups. The rats from the experimental group received per os 40 mg/kg b.w. of L-arginine every day for 2 weeks and were decapitated after 3 weeks of the experiment. The rats from the control group received in the same manner 2 ml of distilled water and were decapitated after 3 weeks of the experiment. After decapitation specimens from the liver were collected, fixed in 10% formalin, and then embedded in paraffin blocks. Proteins Hsp70 and p-53 were detected on slides using the standard three step immunohistochemical method. The quantitative evaluation of Hsp70 and p-53 expression showed that the area of positive staining in the liver of the experimental rats (Hsp70 305,763.00 μm² +/-58,289.66, p-53 9,551.42 μm² +/-1,078.86) was comparable to that in the control groups (Hsp70 291,636.80 μm² +/-34,492.31, p-53 14,104.67 μm² +/-3,571.35). Our experiment showed, that L-arginine as a precursor of exogenous nitric oxide given to rats in dose similar to that used in pregnant women treated for hypertension did not exhibit an influence on hepatocytes.

Impact of experimental diabetes and chronic hypoxia on rat fetal body weight

OBJECTIVES The aim of the study is to determine the impact of the experimental diabetes and the chronic hypoxia on pregnancy development and rat fetal body weight. MATERIAL AND METHODS The experiment was performed on female Wistar rats. Animals were divided into the experimen-tal groups. I - Controls, II - Untreated diabetes, III - Insulin-treated diabetes, IV - No diabetes with chronic hypoxia, V - Untreated diabetes and chronic hypoxia, VI - Insulin- treated diabetes and chronic hypoxia. Diabetes was induced in groups II, III, V and VI with intraperitoneal injection of streptozocin (STZ) at a dose of 40 mg/kg. Chronic hypoxia was induced by placing dams (groups IV, V and VI) in conditions of 10.5% oxygen and 89.5%. Insulin was administered subcutaneously at the dose of 9 IU/kg. Starting from the 6th day after STZ injection and chronic hypoxia conditions animals were caged together for 12 hours for 3 consecutive days to ensure fertilization. On day 21 of gestation the animals were decapitated, the fetuses were removed and weighted. RESULTS Mean fetal body weight in separate groups were: I - 5.38 g, II - 6.04g, III - 5.32g, IV- 5.56 g, V - 3.45 g, VI - 6.23 g. CONCLUSIONS Pre-existing type 1 diabetes does not affect fetal body weight compared to healthy newborn control rats. Pro-longed hypoxia does not impact on fetal body weight. Chronic hypoxia during pregnancy complicated with untreated type 1 diabetes mellitus leads to significant reduction of fetal body weight. Insulin treatment reversed the detrimental effect of chronic hypoxia on fetal development.

Follow-up monitoring of physical activity after rehabilitation by means of a mobile application: Effectiveness of measurements in different age groups

BACKGROUND Active monitoring of the level of daily physical activity seems to be a useful element for secondary prevention in public health. Low physical activity increases the incidence of cardiovascular diseases, obesity, diabetes, musculoskeletal diseases, and causes loss of the previously achieved effects of rehabilitation. OBJECTIVES The purpose of this study was to assess the level of physical activity in different age groups of adults with the use of the telemedical system based on a mobile application. MATERIAL AND METHODS The research covered data collected remotely from 927 individuals of both genders, aged 20-80 years (group I: 20-40 years, group II: 41-60 years, group III: 61-80 years). A monitoring system (Activity Measurement Tool) developed in the Department of General Rehabilitation at the Witold Chodźko Institute of Rural Health (Lublin, Poland) was used to measure home physical activity in the examined group. The system uses a dedicated mobile application, cellular data transmission and web data-showing software. Home physical activity was assessed using the International Physical Activity Questionnaire - Short Form (IPAQ-SF) and simultaneous processing of data from a smartphone accelerometer. RESULTS The mean level of physical activity in the group of active application users (≥2 days, n = 494), expressed as Metabolic Equivalent of Task (MET)-min/week (IPAQ-SF), was as follows: group I (female participants (F): 5,767.9, n = 73; male participants (M): 4,888.4 n = 251), group II (F: 3,468.7, n = 24; M: 4,053.5, n = 119) and in group III (M: 5,769.3, n = 27; no female participants were involved). In 72.3% of users, the registered physical activity was smaller in relation to IPAQ-assessed/7 days physical activity (sign test: n = 494; percentage of negative differences: 72.3%; Z = 9.9; p = 0.00). CONCLUSIONS The research findings indicate a high level of self-reported physical activity among the users in all age and gender groups, although it is not reflected in the level of registered activity. Although the level of daily physical activity was evaluated, it was mainly among young and middle-aged men who gladly and regularly made use of the measurement possibilities offered by mobile technology.