Behavior of silica particles introduced into an isolated rat heart as potential drug carriers.

Silica powders consisting of small spherical particles (50-200 nm) have been obtained by the sol-gel method. A suspension of such particles in the Krebs-Hanseleit solution has been introduced into the coronary circulation of a beating perfused rat heart. The influence of the suspension on the heart muscle and the coronary vessels in the rat body has been histopathologically examined. The particles have not left the lumen of the vessels and have not caused any side effects. These observations suggest the possibility of using such silica particles as a carrier for selected drugs.

Exercise-induced apoptosis in rat kidney is mediated by both angiotensin II AT1 and AT2 receptors.

Excessive physical exercise may lead to disturbance of the entire homeostasis in the body, including damage not only in skeletal muscles but also in many distant organs. The mechanisms responsible for the exercise-induced changes could include oxidative stress or angiotensin II. We previously showed that acute exercise led to apoptosis in kidney but not as a result of oxidative stress. In this study, we examined the role of angiotensin II and its AT1 and AT2 receptors in mediation of exercise-induced apoptosis in kidney. We clearly demonstrated that acute physical exercise induced apoptosis in renal cells of distal convoluted tubuli and cortical and medullary collecting ducts. Moreover, the cells displayed an increased expression of both AT1 and AT2 angiotensin II receptors and of p53 protein. The results suggest that angiotensin II could upregulate p53 expression in renal distal convoluted tubular cells and in the cells collecting ducts via both AT1 and AT2 receptors, which might be the crucial apoptosis-mediating mechanism in kidneys after excessive exercise.

Antitumor activity of bacteriophages in murine experimental cancer models caused possibly by inhibition of beta3 integrin signaling pathway.

Bacteriophages (phages) as bacterial viruses are generally believed to have no intrinsic tropism for mammalian cells. In this study the interactions between phages and various eukaryotic cells were investigated. Binding of phages to the membranes of cancer and normal blood cells was observed. Moreover, it was shown that the wild-type phage T4 (wtT4) and its substrain HAP1 with enhanced affinity for melanoma cells inhibit markedly and significantly experimental lung metastasis of murine B16 melanoma cells by 47% and 80%, respectively. A possible molecular mechanism of these effects, namely a specific interaction between the Lys-Gly-Asp motif of the phage protein 24 and beta3-integrin receptors on target cells is proposed. It was also shown that anti-beta3 antibodies and synthetic peptides mimicking natural beta3 ligands inhibit the phage binding to cancer cells. This is in line with the well-described beta3 integrin-dependent mechanism of tumor metastasis. It is concluded that the blocking of beta3 integrins by phage preparations results in a significant decrease in tumor invasiveness.

The role of AT1 and AT2 angiotensin receptors in the mechanism of apoptosis in renal tubular cells after physical exercise.

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.

Histological and electron microscopic investigation of Reinke's edema.

Reinke's edema is a benign lesion of vocal fold affecting subepithelial space. This paper describes the histological features of Reinke's edema on the basis of an extensive number of cases (203 women and 58 men). In 10 cases the electron microscopic examination was performed. Edema of subepithelial tissue was present in 138(62%) cases. This phenomenon was observed more frequently in women than in men (p=0.01). In the subepithelial tissue there were a numerous wide vessels with oedematous endothelium. Leukoplakia and dysplasia of epithelium were present in 21(8%) and 16(6%) specimens, respectively. Leukoplakia was detected more often in men than in women. This relation was close to statistical significance (p=0.055). The presence of dysplastic lesions of the epithelium was correlated with the age of the patients and smoking habit (p=0.0042, p=0.0021). Electron microscopic investigations revealed loosened intercellular junctions and widening of intercellular spaces, especially in basal and spinous layers.

Sialosyl Le(a)-carrying gangliosides present on the surface of colon carcinoma cells are not directly involved in adhesion to E-selectin.

We have shown previously that human colon cancer CX-1 cells contain lipid- and protein-bound sialosyl Lewis(a) structures that support the adhesion of these cells to E-selectin. Treatment of cancer cells with O-sialoglycoprotease did not decrease either the binding of anti-sialosyl Le(a) antibodies or binding to E-selectin-expressing CHO cells. This suggested that cleavage of sialomucins uncovered cryptic sialosyl Le(a) gangliosides that support such interactions. In the present study, inhibitors of glycolipid and O-glycan biosynthesis, d,l-threo-PPPP and GalNAc-alpha-O-benzyl, respectively, were used to study whether the binding of anti-sialosyl Le(a) antibody and adhesion of CX-1 cells to E-selectin can be mediated by sialosyl Le(a) gangliosides. Treatment of cancer cells with each of the inhibitors decreased the expression of the respective glycoconjugates as shown by TLC-binding assay and immunoblotting with anti-sialosyl Le(a) antibody. However, only slight differences in binding of antisialosyl Le(a) antibody to the surfaces of control and inhibitor-treated CX-1 cells were found by flow cytometry, as well no differences were observed in binding of control and inhibitor-treated CX-1 cells to E-selectin-expressing CHO cells, supporting the earlier hypothesis on the involvement of gangliosides in binding of anti-sialosyl Lewis(a) in the partial absence of mucin O-glycans. This hypothesis was further proven by electron microscopy data. Both native CX-1 and d,l-threo-PPPP-treated cells were labelled with anti-sialosyl Lewis(a) antibody mostly at a distance 70-90 nm from cell surface, suggesting interaction with protein-bound carbohydrate structures only. In contrast, the cancer cells treated with GalNAc-alpha-O-benzyl showed most of the staining around 20 nm distance from the plasmalemma, implying that the antibody interacts with lipid-bound sialosyl Lewis(a) instead. The electron microscopy data in conjunction with other results described in this report strongly support the hypothesis that sialosyl Lea gangliosides are not involved in the adhesion of CX-1 cells to E-selectin when mucins are present on the cell surface, but they may be involved in binding to E-selectin in their absence.

[Secondary glaucoma in patients with lens subluxation or luxation]. / Jaskra wtórna u chorych z podwichnieciem lub zwichnieciem soczewki.

AIM: To estimate the frequency of secondary glaucoma appearance and the influence of surgical treatment on intraocular pressure (IOP) in patients with posterior lens dislocation. MATERIAL AND METHODS: The study concerned 152 patients (57 females and 95 males, age range: 19 to 91 years, mean--58 years). Secondary glaucoma was found in 62 patients (41%). Trauma was the cause of lens dislocation in 81% of these cases, while it was the cause in only 69% of cases in the whole material. Pars plana vitrectomy with limbal intracapsular lens extraction or lentectomy were performed in all cases. In 24 cases perfluorocarbon liquid was used. Scleral fixation PC IOLs were implanted in 85 eyes (group A), AC IOLs in 27 eyes (group B), and 40 eyes remained aphakic (group C). The follow-up time ranged from 6 months to 5 years, mean 22 months. RESULTS: Raised IOP was found preoperatively in 46 cases (30.3%). It was raised in 21 (24.7%) eyes in group A (24-60 mm Hg, mean 37.1 mm Hg), and it has normalized postoperatively in 14 eyes, while topical medication was necessary in 7 remaining cases. In group B raised IOP (26-60 mm Hg, mean 41.4 mm Hg) was found preoperatively in 8 (29.6%) eyes. It has normalized postoperatively in 5 eyes, while topical medication was necessary in 3 eyes. The respective values in group C were: 17 (42.5%), 24-80 mm Hg, mean 43.2 mm Hg. In 7 eyes IOP was normal after surgery, 10 patients required topical treatment, and in 3 of these cases trabeculectomy had to be performed. In 16 eyes with normal preoperative IOP it has raised postoperatively in different periods of time (from 1 week to 6 months). CONCLUSIONS: Secondary glaucoma is a frequent complication of the posterior lens dislocation, and it is more often related with traumatic cases. The dislocated lens removal with the use of vitrectomy causes IOP normalization in most of the cases. The periodical examination of IOP is necessary after the dislocated lens removal also in cases with normal preoperative IOP, because secondary glaucoma may appear in different time after surgery.

[Renovascular hypertension with high serum zinc in serum of a patient with ceroid-lipofuscinosis and genome 46,XY,9qh+]. / Nadcisnienie tetnicze naczyniowo-nerkowe z podwyzszonym stezeniem cynku w surowicy u chorego z ceroidolipofuscynoza i genomem 46,XY,9qh+.

A case of a 38-year-old patient with severe renovascular hypertension high serum zinc concentration and ceroidlipofuscinosis was presented. The diagnosis of ceroidlipofuscinosis was based on electron microscope picture of mucosus tissue of rectum where the secondary lisosoms, so characteristic of this disease, were found in cells of connective tissue.

Determination of blood serum oncoprotein NEU and antioncoprotein p-53--molecular biomarkers in various types of occupational exposure.

Application of various diagnostic tests for occupational cancer risk monitoring is associated with the fact that gene mutations and changes in gene expression correspond to the earliest stages of carcinogenesis, namely early stages of the promotion process. The changes in protooncogenes and suppressor genes can be detected either at the genome level, at the level of transferring the genetic information from DNA to protein, or at the level of protein synthesis controlled by genes (oncogenes or antioncogenes). In the latter instance, as the concentrations of these proteins are considerably increased, their quantities in blood serum can be determined by the immunochemical methods. In our work, blood serum p-53 and NEU proteins were determined in 32 workers exposed to asbestos and in 57 workers exposed to PAHs. The proteins were also determined in 99 patients with overt cancer and in 47 controls. The data obtained in this work show positive values of oncoprotein NEU or antioncoprotein p-53 in 17.3% to 31.8% of workers exposed to PAHs or asbestos. The percentage of positive values for the examined proteins in the patients with overt cancer ranged from 12.5% to 42.5%. It should be noted that positive values of the oncoproteins detected in the biomaterial of the persons exposed do not mean that people must necessarily develop cancer, nevertheless the elevated values should be regarded as a warning and an implication for undertaking suitable preventive steps.

[Urinary cotinine as a biomarker of exposure to tobacco smoke]. / Kotynina w moczu jako biomarker ekspozycji na dym tytoniowy.

Several markers are used to monitor active or passive exposure to tobacco smoke. They include measurements of carboxyhaemoglobin in the blood, carbon dioxide in the expired air, thiocyanates and nicotine in the saliva, plasma (serum) or urine. The determination of cotinine, the main nicotine metabolite, in biological fluids is a biomarker which finds still wider application. This metabolite can be determined in the urine and saliva and plasma. Cotinine, as a biomarker of exposure to tobacco smoke, is used in epidemiological studies aimed to find out to what extent the exposure to occupational harmful factors affects the workers' health. The application of this biomarker helped to classify workers more effectively into smokers and non-smokers, and to provide better conditions for finding out whether other non-occupational factors such as smoking do not confound the evaluation of health threats induced by work-related hazards.

[Usefulness of different silicone oils for intraocular tamponade]. / Ocena przydatnosci róznych rodzajów oleju sylikonowego do tamponady wewnatrzgalkowej.

PURPOSE: This paper presents the influence of different silicone oils on surgical results achieved after pars plana vitrectomy and silicone oil tamponade. MATERIAL AND METHODS: Pars plana vitrectomy and silicone oil tamponade were used for the treatment of 141 eyes with proliferative Vitreoretinopathy, diabetic retinopathy and posttraumatic retinal detachment. In group I (23 eyes), group II (26 eyes), group III (32 eyes) and group IV (60 eyes) following silicone oils were used, respectively: highly purified silicone oil with viscosity 5000 mPa(x)s, 1000 mPa(x)s, cosmetically purified silicone oil with viscosity 3690 mPa(x)s and 1000 mPa(x)s. RESULTS: The achieved results did not show any statistically significant differences according to anatomical and functional results and early and late postoperative complications between the analysed groups. Compared to other groups, highly purified silicone oil with viscosity 5000 mPa(x)s did not show any emulsification. CONCLUSIONS: The achieved results show that different silicone oils can be successfully used for intraocular tamponade. Highly purified silicone oil with viscosity 5000 mPa(x)s is most biocompatible.

[Eosinophil cationic protein in persons with contact allergy to disinfectants]. / Kationowe bialko eozynofilów u osób z uczuleniem kontaktowym na srodki odkazajace.

The concentration of eosinophil cationic protein (ECP) in blood serum was determined in 53 health service workers (women) with contact allergy to disinfectants and in 16 healthy women using a radioimmunological method. The mean ECP concentration in the study group accounted for 7.518 micrograms/l, and in the control group 4.893 micrograms/l. The concentration of 8.403 micrograms/l (the mean concentration in the control group + 2 SD) was taken as a cut off concentration value. In 17 (32.1%) persons with contact dermatitis, pathological ECP values were found. An increased ECP was observed in persons with positive results of prick test, especially in the case of Dermatophagoides, pollens and latex as well as in those with increased total IgE concentration. Our results indicate the involvement of immediate allergy in the incidence of contact allergy to disinfectants.

[Biomarkers in diagnosis of kidney diseases caused by chemical toxins]. / Biomarkery w diagnostyce schorzen nerek powodowanych przez toksyny chemiczne.

As yet there has been no evidence produced that nephrotoxic chemicals damage nephron in sites, characteristic of a given toxin. A number of nephrotoxic effects result rather from changes induced by these substances mostly in the peripheral circulatory system than from a direct effect of chemical toxin on the kidneys. In many cases the morbid process in the kidneys does not emerge until a considerable number of nephrons is already damaged. Therefore, it is essential that doctors, attending patients exposed to nephrotoxins, have at their disposal, during periodical examinations, appropriate laboratory tests (biomarkers) able to detect subclinical forms of chemical damage of the kidneys. These biomarkers should also help to identify which of the functional parts of the kidneys have been damaged. It seems that determination of protein excreted with urine is one of the best biomarkers most frequently used to detect the dysfunction of renal glomerulus. It is recommended that the relationship between urine concentration of total protein and urine concentration of creatinine be determined. In the group of biomarkers of tubule dysfunction the measurement of substances usually reabsorbed from glomal filtrate (low-molecular protein) in proximal tubule or determination of enzymes' activity in urine (e.g. N-acetylo-glucosoaminidase) and those cellular components which are not excreted with normal urine are recommended. In the assessment of distal renal tubule dysfunction it is advised to examine urine osmolarity and/or determination of Thamm-Horsfall glycoprotein.

Determination of tissue polypeptide antigens (TPA) and carcinoembryonic antigen (CEA) in serum: its value in the preliminary cancer risk assessment in asbestos exposed workers.

Seeking the changes at the cellular level or at the level of cellular metabolism products, present in the biological fluids, in order to detect early stages of the carcinogenic process is an essential step in preventing cancer development among asbestos exposed workers. Carcinogenic biomarkers such as tissue polypeptide antigens (TPA) and carcinoembryonic antigen (CEA) were found very useful in this attempt. The objective of this work was to identify individuals at critical cancer risk in the population of workers exposed to asbestos and to evaluate the value of TPA and CEA determinations for this particular purpose. The study was carried out in the group of workers exposed to asbestos (n = 274). Age, exposure duration, smoking habits and the kind of job performed, were considered in the analysis of the results. To sum up, it should be concluded that in 22 persons exposed to asbestos TPA values exceeded the cut off concentrations, established on the basis of the studies performed in the control group, and CEA value accounted for 10 ng/lm. Statistically significant differences in the percentage of TPA increased values between two groups under study were indicated. Such a relationship did not apply to CEA. In the exposed group, an evident effect of the age and exposure duration on the number of persons with TPA concentrations above the cut off, was also revealed. These changes show a growing tendency and statistical significance for TPA only. Smoking had a great impact on the occurrence of TPA increased concentrations. Three kinds of jobs were considered: operation of the production line, white collar workers and miscellaneous'. The significant differences in TPA concentrations between the operators and miscellaneous, and between white collar workers and miscellaneous were found. Therefore, it may be concluded that a similar percentage of TPA increased values was observed in the group of operators and white collar workers. The study allowed to identify, among those exposed to asbestos, 22 persons who should be covered with target medical care. It also indicated that TPA determination was more useful than that of CEA in this kind of investigations.

Nasal response to allergen challenge in patients with immediate asthmatic reaction.

Ten patients with bronchial asthma and allergy to house-dust mite (HDM) and ten normal, nonatopic control subjects underwent a bronchial challenge with flour. Before and 24 hr after the allergen provocation with flour, the levels of eosinophil cationic protein (ECP) and myeloperoxidase (MPO) were determined in the serum and nasal lavage fluid. All allergics showed an isolated immediate asthmatic reaction (IAR). After the flour challenge only in asthmatic patients the increase was detected in the mean values of: 1) eosinophils (mean value before 16.7 x 103/mm3; mean after: 10 min 132.9 x 103/ml; 3 hr 183.6 x 103/mm3; 24 hr 110.6 x 103/mm3, p < 0.05), 2) basophils (mean before 1.2 x 103/mm3; mean after: 10 min 5.3 x 103/ml; 3 hr s 14.1 x 103/mm3 24 hr was 18.3 x 103/mm3, p < 0.05), 3) neutrophils (mean before 9.2 x 103/mm3; mean after 24 h 18.2 x 103/mm3, p < 0.05) in the nasal lavage fluid. In contrast to a group of normal subjects, asthmatics were found to have higher postchallenge levels of ECP and MPO in the nasal secretions as compared with the prechallenge levels (ECP-mean 3.85 ug/l compared with 32.17 ug/l, p < 0.05; MPO-mean 120-02 ug/l compared with 1313.2 ug/l, p < 0.05). The authors did not find any significant difference between pre- and postchallenge levels of ECP and MPO in the serum of asthmatics and controls. The higher levels of MPO as well as higher count of neutrophils observed in asthmatic patients 24 hr after allergen challenge support the neutrophil involvement in the allergic inflammation. Our results indicate that both neutrophils and eosinophils take part in allergic reaction in the mucosa.

Nasal response to allergen challenge in patients with immediate asthmatic reaction.

Ten patients with bronchial asthma and allergy to house-dust mite (HDM) and ten normal, nonatopic control subjects underwent a bronchial challenge with flour. Before and 24 h after the allergen provocation with flour, the levels of eosinophil cationic protein (ECP) and myeloperoxidase (MPO) were determined in the serum and nasal lavage fluid. All allergics showed an isolated immediate asthmatic reaction (IAR). After the flour challenge only in asthmatic patients the increase was detected in the mean values of: 1/eosinophils (mean value before 16.7 x 10(3)/mm3; mean after; 10 min 132.9 x 10(3)/ml; 3 hr 183.6 x 10(3)/mm3; 24 h 110.6 x 103/mm3, p < 0.05), 2/basophils (mean before 1.2 x 10(3)/mm3; mean after: 10 min 5.3 x 10(3)/ml; 3 h s 14.1 x 10(3)/mm3 24 h was 18.3/mm3, p < 0.05), 3/neutrophils (mean before 9.2 x 10(3)/mm3; mean after 24 h 18.2 x 10(3)/mm3, p < 0.05) in the nasal lavage fluid. In contrast to a group of normal subjects, asthmatics were found to have higher postchallenge levels of ECP and MPO in the nasal secretions as compared with the prechallenge levels (ECP-mean 3.85 ug/l compared with 32.17 ug/l p < 0.05; MPO-mean 120.02 ug/l compared with 1313.2 ug/l, p < 0.05). The authors did not find any significant difference between pre- and postchallenge levels of ECP and MPO in the serum of asthmatics and controls. The higher levels of MPO as well as higher count of neutrophils observed in asthmatic patients 24 h after allergen challenge support the neutrophil involvement in the allergic inflammation. Our results indicate that both neutrophils and eosinophils take part in allergic reaction in the mucosa.

[Exposure to asbestos and levels of selected tumor biomarkers]. / Ekspozycja na azbest a stezenia wybranych biomarkerów nowotworowych.

Occupational exposure to asbestos, a recognised carcinogen, poses a risk for such diseases as asbestosis, lung cancer and mesothelioma. It is thought that asbestos fibres may damage microphages which undergo neoplastic transformation as well as fibroblast, while partial phagocytosis may generate free oxygenic radicals which induce cellular peroxidase and damage macromolecules. A search for cellular changes or changes in cellular metabolism products, present in biological fluids, in order to detect early stages of a neoplastic process is an important factor in the prophylaxis of workers exposed to asbestos. Neoplastic biomarkers such as tissue polypeptide antigen (TPA) or carcinoembryonic antigen (CEA) are now used for this purpose. The aim of the work was to identify workers exposed to asbestos in the population, especially high risk groups neoplastic diseases and to evaluate the usefulness of TPA and CEA determinations. The study covered a group of asbestos exposed workers (n = 4000 and the control group of workers (n = 135) nonexposed to any toxic factor at work. Age, exposure time, smoking habits and workpost characteristics were taken into consideration in the analysis of the results. It was revealed that in 38 persons exposed to asbestos, TPA values were above the concentration limit set on the basis of studies carried out in the control group, and elevated CEA values applied to 13 persons. Significant differences between groups under study were found in the proportion of pathological TPA values. Such a relationship was not observed in regard to CEA values. In the exposed group the results also indicated an evident effect of age and exposure time on the number of persons with TPA values above concentration limit. There is a growing tendency in those changes but only in regard to TPA values. The effect of smoking on the frequency of pathological TPA values was also clear-cut in workers exposed to asbestos. Taking into account three types of employment: blue collar workers, white collar workers and other personnel, the analysis indicated significant differences in TPA values between blue collar workers and other personnel; and between white collar workers and other personnel. This means a similar percentage of pathological TPA values in the group of blue collar and white collar workers. The study carried out allowed to identify persons exposed to asbestos who should be covered with targeted medical care. They also proved that TPA biomarker is better than CEA one for this kind of studies.