The influence of strain, sex and age on selected biochemical parameters in blood serum of Buffalo and Wistar rats.

The objective of the study was to determine interval values of biochemical parameters in the most commonly applied experimental model among different species, i.e. rats. Blood analysis of experimental animals is done in different research fields. They are important especially in experiments in pharmacology, pathophysiology, experimental surgery, toxicology and for monitoring experimental disorders in laboratory animals. In this paper, basic biochemical markers in the blood serum of Buffalo and Wistar rats are also compared in relation to the animals' age and sex. The values were obtained using the latest available measurement methods and the above-listed checkpoints were considered. The biochemical markers show variability between the particular groups of animals related to their age, sex, and strain. The obtained data may be used to create a model of interval values of biochemical parameters for the Buffalo and Wistar rat strains. This study is necessary to enhance our understanding on basic parameters in these animals which are often used in different medical experiments.

Glycophorin A on normal and leukemia cells detected by monoclonal antibodies, including a new monoclonal antibody reactive with glycophorins A and B.

A new hemagglutinating monoclonal antibody, MoAb31, detected glycophorins A and B in Western blots. Results with enzyme-modified erythrocytes indicated the MoAb31 determinants were sialic acid dependent, and resided on glycophorin A on the trypsin-resistant, ficin-sensitive segment, and on glycophorin B on the ficin-sensitive segment. Another new monoclonal antibody, MoAb36, detected the Wrb antigen, located on the non-glycosylated segment of glycophorin A near its insertion into the lipid bilayer. Immunofluorescent staining of normal hematopoietic and leukemia cells with these and other monoclonal antibodies to glycophorin A demonstrated glycophorin A on erythroid cells only. Cytofluorograph analysis showed the majority of cells of the erythroleukemia cell lines K562 and HEL expressed glycophorin A, as indicated by reactivity with the monoclonal glycophorin A antibodies R10, R18, 6A7 and 10F7. However, reactivity with monoclonal antibodies to glycosylated determinants (MoAb31 and R1.3) and to the non-glycosylated segment near the membrane insertion (MoAb36, and R7.1) was reduced or absent. Expression of "missing" glycophorin A antigens on K562 and HEL could not be induced using a variety of chemical and biologically active modifiers. We conclude that glycophorin A of erythroleukemia cell lines K562 and HEL differs from glycophorin A at the surface of normal, mature erythrocytes with respect to reactivity with monoclonal glycophorin A antibodies.

Development and disappearance of tolerance to induction of interferon and tumor necrosis factor response in athletes treated with natural immunostimulant.

The effect of nonspecific immunostimulation was examined in 15 basketball players subjected to extensive physical effort. The Tolpa* Torf Preparation (TTP*), a natural immunostimulating drug, was applied orally, one 5 mg tablet daily, in two 21-day cycles, separated by 2-week hiatus. Blood samples were collected 4 times, after each of two TTP* cycles and after the first and second hiatus. Whole blood assay was used to determine the spontaneous and induced production of interferon (IFN) and tumor necrosis factor (TNF). The levels of the cytokines were measured by microbioassays. TTP* stimulated synthesis of IFN and TNF in the whole blood cultures. However, after the oral administraton of TTP* for 3 weeks the leukocytes of the athletes developed hyporeactivity to IFN induction by TTP* and to a lesser extent to another "superinducer"--a mixture of phytohemagglutinin and bacterial lipopolysaccharide. The hyporeactivity state disappeared spontaneously within 2 weeks. In contrast, the tolerance to TNF induction did not develop during the TTP* administration. The increase of immunoglobulins, mainly of IgM and IgG classes and an acute phase protein--alpha1-antitrypsin, was observed at the late phase of the treatment. We suggest that the cytokine levels may be early markers for immunoprophylaxis. Furthermore, high production of IFN and TNF may be associated with extensive physical effort.

Anaemia is an independent predictor of poor outcome in patients with chronic heart failure.

BACKGROUND: Mild anaemia frequently occurs in patients with chronic heart failure (CHF), particularly in the advanced stages of the disease. The correction of anaemia with erythropoietin is a therapeutic possibility. The aim of this study was to assess prospectively the relationship between the prevalence of anaemia (haemoglobin level18 months in all survivors), and the end-point of the study was all-cause mortality. RESULTS: A total of 176 patients were enrolled (mean age: 63 years, New York Heart Association (NYHA) classification I/II/III/IV: 15/81/51/29; left ventricular ejection fraction (LVEF): 42%, ischaemic aetiology in 62%). In the whole population the mean haemoglobin level was 140+/-15 g/l. Anaemia was found in 18 (10%) patients, and was significantly more common in women than in men (18 vs. 7%, respectively, P=0.02) and in those with most severe CHF symptoms (frequency in NYHA I/II/III/IV: 0/9/10/21%, respectively; NYHA IV vs. I-III, P=0.03), but not related to the other clinical indices. Univariate analysis revealed NYHA class III-IV (hazard ratio 3.8, 95% CI: 1.6-8.9, P=0.003), low LVEF <35% (hazard ratio 2.3, 95% CI: 1.0-4.9, P=0.04) and anaemia (hazard ratio 2.9, 95% CI: 1.2-7.2, P=0.02) as predictors of 18-month mortality. In multivariate analysis, anaemia remained an independent predictor of death when adjusted for NYHA class and LVEF (hazard ratio: 2.6, 95% CI: 1.0-6.5, P=0.04). In anaemic patients, 18-month survival was 67% (95% CI: 45-89%) compared to 87% (81-92%) in patients with a normal haemoglobin level (P=0.016). CONCLUSIONS: Mild anaemia is a significant and independent predictor of poor outcome in unselected patients with CHF. Correction of low haemoglobin level may become an interesting therapeutic option for CHF patients.

Hormone plasma levels from pituitary-gonadal axis in performance athletes after the 400 m run.

BACKGROUND: The aim of this study was to compare the concentration changes in hormones from pituitary - gonadal axis, induced by the 400 m run in the well-trained athletes (vice-champions in the Hall and Summer Athletic World Championship in 1999) to the changes observed in the competitors with shorter training period and achieving worse final results. METHODS: This research was conducted on 6 males - members of the Polish Olympic Team, who won vice-championship in the Hall and Summer World Championships 1999 and 6 athletes trained in the academic sport clubs. In the recent investigation, the 400 m run was assumed to be a stimulating impulse for evoking hormonal changes. The blood samples were taken from the elbow vein before the run, immediately after the effort and after the 24-hour rest. In the serum, the luteinizing hormone (LH), follicle-stimulating hormone (FSH), total testosterone (TT), free testosterone (FT) as well as the sex hormones-binding globulin (SHBG) concentrations were determined. RESULTS: During our research, immediately after the 400 m run in group I - the top class sportsmen - the statistically significant increase in both gonadotrophins (LH, FSH) was determined as well as the decrease in the total and free testosterone. In the group II - the athletes with the lower training level - the increase in FSH and the total and free testosterone concentrations was noticed. There were no statistically significant differences in the SHBG concentration. After-effort increase in the lactic acid concentration was observed in both groups. In the master group I, the increase in lactic acid concentration was higher than in group II. In both groups after the 24-hour restitution, the examined parameters, except LH levels in the group I, showed the concentrations similar to those before the effort. Analysis of the time needed to cover the distance of the race showed that the athletes from group I covered the distance of 400 m in the shorter time. CONCLUSIONS: The group of master class athletes, whose average intensive training period was 8 years, had higher VO(2max) and higher after-effort increase in the lactic acid concentration than in the group of sportsmen with the shorter training period (4 years), who had lower VO(2max), worse sport results and lower after-effort increase in the lactic acid concentration, gave different hormonal response (particularly TT, FT concentration) for the same exercise impulse. The difference based on the fact, that after the run in group I the decrease in the total and free testosterone levels and in group II the increase in the same parameters were observed. The observed hormonal changes in the master class athletes induced by the years-long anaerobic training might provide evidence for the reduction of functional reserves in gonads when compared to the group of less trained sportsmen.

[Influence of materials with different hydrophobic properties on selected parameters of blood coagulation]. / Wplyw materialów o róznym stopniu zwilzalnosci na wybrane parametry ukladu krzepniecia.

In this paper we presented the evaluation and assessment of the influence of the knitted polyester materials with the different hydrophobic properties of theirs surfaces on the blood coagulation and fibrinolysis investigated on dynamic "in vitro" model. On the basis of the received results we can stated that the material with hydrophobic surface do not change the coagulation time in the internal-factors model nor in the external-factor model. It also has no influence for the concentration of fibrinogen, activity of the factors XII and VIII, antithrombin III, protein C nor for plasminogen. The knitted material with the hydrophilic surface increased the coagulation time while use in internal-factors model but do not change the coagulation time in the model with external-factors. It also has the influence in decreasing the activity of the factors XII and VIII. The activity of blood coagulations' inhibitors and plasminogen remained not changed.

[Changes in the level of selected laboratory parameters in blood after implantation of chitosan fibers]. / Zmiany w poziomie wybranych parametrów laboratoryjnych krwi po implantacji wlókien chitozanowych.

The tests were carried out on rats of the Wistar tribe to the peritoneal cavity of which chitosan fibres were implanted. The blood for the tests was taken in 3, 7, 14, 30 and 60 days after the implantation. In the blood hematological, biochemical and clotting system parameters were marked. Moreover they made pathomorphological tests of the tissues surrounding the implanted material and laboratory and biological tests of aqueous extracts from chitosan fibres. The shown quantitative changes in the level of the marked parameters of blood to the 14th day of observation are connected mainly with toxicity of chitosan fibres, whereas to the 30th and 60th day-with the process of biodegradation and resorption of collagenous fibres.

[Usefulness of selected parameters from the diagnostic laboratory for evaluating absorbed grafting materials]. / Przydatnosc wybranych parametrów z diagnostyki laboratoryjnej do oceny wchlanialnych materialów implantacyjnych.

The results of hematologic, biochemical research and research of clotting system after intraperitoneal implantation of absorbed synthetic threads Dexon S are presented in this work. The research was made on rats of Wistar type. Blood for the research was taken 3, 7, 14, 30 and 60 days after the implantation. Morphology (Ht, Hb, MCH, WBC, the number of platelets), activity of aspartic and alanine aminotransferase and antithrombin activity, concentration of comolete protein and its fraction and concentration of glucose, urea, creatinine and also concentration of ions Na+, K+, Mg2+, Ca2+ and fibrinogen, protein C3 and C4 of complement system as well as kaolin-kephalin time and prothrombin time of plasma were marked. On the basis of the obtained results of the research it was noticed that the used methods of research constitute supplement of biological estimation of absorbed grafting materials.

Monoclonal antibodies to murine leukemia virus gp 70 recognize the same T lymphoma cell surface molecules as anti-immunoglobulin.

Chicken antisera to mouse immunoglobulin (anti-Ig) detect molecules on T lymphocytes of mice consisting of "heavy chains" similar in size to IgM mu chains and "light chains" similar in size to lambda or kappa chains. Because these T cell-derived proteins react with anti-Ig and are composed of Ig-like heavy and light chains, these molecules have been viewed as candidates for "IgT," the putative T lymphocyte receptor for antigen. We have demonstrated, by immunoprecipitation and two-dimensional gel analysis, that the lymphoma surface molecule precipitated by chicken anti-Ig is identical to the viral 70,000 dalton glycoprotein (gp 70) expressed by that lymphoma and is unassociated with any "light chain" or equivalent. For each of the three lymphomas analyzed, the gp 70 two-dimensional gel pattern was individually distinctive, and, in each case, the molecules precipitated by anti-Ig exhibited the same pattern as the gp 70. The viral gp 70 does have cross-reacting determinants with mouse IgM as seen by the chicken antiserum. We have substantiated these results by demonstrating that monoclonal antibodies to viral gp 70 precipitate the same molecules as chicken anti-Ig. These findings demonstrate that these T lymphoma cell surface molecules do not represent the T cell immunoglobulin receptor for antigen unless gp 70 itself is an antigen receptor.

Heat shock-induced shedding of cell surface integrins in A549 human lung tumor cells in culture.

The human lung carcinoma-derived cell line A549 attaches to plastic and vitronectin-coated substrates in a manner dependent upon the specific cell surface integrin alpha v beta 3. Exposure to hyperthermic temperatures (42-45 degrees C) causes these cells to detach from the substrates. In heat-shocked cultures, the alpha v, alpha 5, and beta 3 integrin subunits remain attached to the substrate. Analysis of individual cells by fluorescence-activated cell sorting shows that cell surface levels of alpha v beta 3 decrease by up to 10-fold in response to heat shock, while the abundance of another integrin found on the surface of A549 cells, alpha 3 beta 1, is only minimally affected by this stress. Heat shock-induced decreases in alpha v beta 3 also occur in cells growing in suspension cultures, showing that physical attachment onto an extracellular substrate is not required for the hyperthermia-induced loss of this integrin. The heat shock-induced detachment of the cells and the shedding of alpha v beta 3 from the cell surface can be inhibited by fetal bovine serum and alpha 2 macroglobulin. Reattachment of A549 cells to substrate is reduced by heat shock. These results demonstrate that heat shock can reduce the cell surface abundance of specific integrin subunits, some of which are involved at sites of cellular attachment to extracellular substrates. These findings may be relevant to the heterogeneous patterns of invasion and metastasis of human tumors following fevers or hyperthermia therapy.

Human T cell antigens defined by monoclonal antibodies: the 65,000-dalton antigen of T cells (T65) is also found on chronic lymphocytic leukemia cells bearing surface immunoglobulin.

Peripheral blood lymphocytes from 15 patients with chronic lymphocytic leukemia (CLL), four patients with lymphosarcoma cell leukemia (LCL), four patients with hairy cell leukemia, and three patients with a monoclonal gammopathy (two with IgM, one with IgA) were tested for reactivity with the anti-T cell monoclonal antibody, T101. By immunofluorescent staining, all of the patients had circulating monoclonal surface Ig+ (sIg+) lymphocytes except for three CLL patients whose leukemia cells were sIg-. The leukemia cells of all of the sIg+ CLL cases were reactive with T101 antibody by indirect immunofluorescence; however, the abnormal cells in all of the remaining cases were unreactive. Reactivity of sIg+ CLL cells with T101 was confirmed by a radioactive binding assay, absorption analysis, and complement-dependent cytotoxicity. Moreover, a 65,000-dalton protein (T65), similar to that found on T cells, was precipitated by T101 antibody from the surface of sIg+ CLL cells. The fluorescent staining of sIg+ CLL cells by T101 antibody was weak as was the staining of the sIg. This was in contrast to the LCL cells, which had intense staining sIg and absence of staining with T101 antibody. These data demonstrate the existence of two major subtypes of CLL that have phenotypes sIg+ and T101+ and sIg-T101-. The implication of the finding of dual T and B markers on the major type of CLL, but not other B cell malignancies is discussed.

Alteration of cellular adhesion by heat shock.

Chinese hamster ovary (CHO) cells were analyzed for their ability to reassemble microfilament bundles, to remain attached to a tissue culture surface, or to initiate and complete attachment onto a substrate after heat shock (45 degrees C/10 min). The cells remained attached to the tissue culture surface during and after the heat shock while the actin microfilament bundles were reversibly disrupted. Heat shock inhibited the ability of the cells to initiate and complete attachment onto a new tissue culture surface or onto a plastic surface coated with vitronectin. An inspection of the proteins present in substrate-attached material (SAM) revealed 11 major proteins containing glucosamine whose apparent Mr values were 250,000, 200,000, 150,000, 140,000, 90,000, 86,000, 82,000, 68,000, 54,000, 47,000, and 46,000. Three of the proteins (p200, p150, and p46) bound to wheat germ agglutinin while p150 and p140 bound to concanavalin A. The composition of the 11 proteins of the SAM fraction synthesized previous to the heat shock was not altered during heat shock. However, the appearance of the newly synthesized proteins in the SAM fraction was delayed by heat shock (0.5 h for p150 and 6 h for p82). The ability of heat-shocked cells to reattach onto a vitronectin-coated surface correlated with the appearance of newly synthesized p150 and p82 in the SAM fraction. Our results suggest that in addition to the microfilament bundles, heat shock may reversibly disrupt the cellular adhesion site. Further, p150 and p82, proteins whose appearance in the SAM fraction is delayed by heat shock, may be involved in the cellular attachment onto substrates, including vitronectin.