Effects of a single bout of strenuous exercise on platelet activation in female ApoE/LDLR-/- mice.

Strenuous physical exercise leads to platelet activation that is normally counterbalanced by the production of endothelium-derived anti-platelet mediators, including prostacyclin (PGI2) and nitric oxide (NO). However, in the case of endothelial dysfunction, e.g. in atherosclerosis, there exists an increased risk for intravascular thrombosis during exercise that might be due to an impairment in endothelial anti-platelet mechanisms. In the present work, we evaluated platelet activation at rest and following a single bout of strenuous treadmill exercise in female ApoE/LDLR-/- mice with early (3-month-old) and advanced (7-month-old) atherosclerosis compared to female age-matched WT mice. In sedentary and post-exercise groups of animals, we analyzed TXB2 generation and the expression of platelet activation markers in the whole blood ex vivo assay. We also measured pre- and post-exercise plasma concentration of 6-keto-PGF1α, nitrite/nitrate, lipid profile, and blood cell count. Sedentary 3- and 7-month-old ApoE/LDLR-/- mice displayed significantly higher activation of platelets compared to age-matched wild-type (WT) mice, as evidenced by increased TXB2 production, expression of P-selectin, and activation of GPIIb/IIIa receptors, as well as increased fibrinogen and von Willebrand factor (vWf) binding. Interestingly, in ApoE/LDLR-/- but not in WT mice, strenuous exercise partially inhibited TXB2 production, the expression of activated GPIIb/IIIa receptors, and fibrinogen binding, with no effect on the P-selectin expression and vWf binding. Post-exercise down-regulation of the activated GPIIb/IIIa receptor expression and fibrinogen binding was not significantly different between 3- and 7-month-old ApoE/LDLR-/- mice; however, only 7-month-old ApoE/LDLR-/- mice showed lower TXB2 production after exercise. In female 4-6-month-old ApoE/LDLR-/- but not in WT mice, an elevated pre- and post-exercise plasma concentration of 6-keto-PGF1α was observed. In turn, the pre- and post-exercise plasma concentrations of nitrite (NO2-) and nitrate (NO3-) were decreased in ApoE/LDLR-/- as compared to that in age-matched WT mice. In conclusion, we demonstrated overactivation of platelets in ApoE/LDLR-/- as compared to WT mice. However, platelet activation in ApoE/LDLR-/- mice was not further increased by strenuous exercise, but was instead attenuated, a phenomenon not observed in WT mice. This phenomenon could be linked to compensatory up-regulation of PGI2-dependent anti-platelet mechanisms in ApoE/LDLR-/- mice.

Role of xanthine oxidoreductase in the anti-thrombotic effects of nitrite in rats in vivo.

The mechanisms underlying nitrite-induced effects on thrombosis and hemostasis in vivo are not clear. The goal of the work described here was to investigate the role of xanthine oxidoreductase (XOR) in the anti-platelet and anti-thrombotic activities of nitrite in rats in vivo. Arterial thrombosis was induced electrically in rats with renovascular hypertension by partial ligation of the left renal artery. Sodium nitrite (NaNO2, 0.17 mmol/kg twice daily for 3 days, p.o) was administered with or without one of the XOR-inhibitors: allopurinol (ALLO) and febuxostat (FEB) (100 and 5 mg/kg, p.o., for 3 days). Nitrite treatment (0.17 mmol/kg), which was associated with a significant increase in NOHb, nitrite/nitrate plasma concentration, resulted in a substantial decrease in thrombus weight (TW) (0.48 ± 0.03 mg vs. vehicle [VEH] 0.88 ± 0.08 mg, p < 0.001) without a significant hypotensive effect. The anti-thrombotic effect of nitrite was partially reversed by FEB (TW = 0.63 ± 0.06 mg, p < 0.05 vs. nitrites), but not by ALLO (TW = 0.43 ± 0.02 mg). In turn, profound anti-platelet effect of nitrite measured ex vivo using collagen-induced whole-blood platelet aggregation (70.5 ± 7.1% vs. VEH 100 ± 4.5%, p < 0.05) and dynamic thromboxaneB2 generation was fully reversed by both XOR-inhibitors. In addition, nitrite decreased plasminogen activator inhibitor-1 concentration (0.47 ± 0.13 ng/ml vs. VEH 0.62 ± 0.04 ng/ml, p < 0.05) and FEB/ALLO reversed this effect. In vitro the anti-platelet effect of nitrite (1 mM) was reversed by FEB (0.1 mM) under hypoxia (0.5%O2) and normoxia (20%O2). Nitrite treatment had no effect on coagulation parameters. In conclusion, the nitrite-induced anti-platelet effect in rats in vivo is mediated by XOR, but XOR does not fully account for the anti-thrombotic effects of nitrite.

Parotid gland tumors in children - pre- and postoperative diagnostic difficulties.

Major salivary gland tumors are very rare in the developmental period. Confirming tumor changes in the salivary gland requires precise diagnostic imaging involving an ultrasonography scan, computed tomography and magnetic resonance. Needle aspiration biopsy (NAB) of the tumor is of high importance. Excision is the basic treatment method in cases of parotid gland tumor. The statistical data concerning tumors favor less invasive methods, which seems logical in the population of children. The surgical methods used in tumor treatment feature extracapsular excision of tumor, partial parotidectomy and total parotidectomy, sometimes followed by lymphatic node surgery. The clinical cases presented in the paper show difficulties with pre- and postoperative histopathological diagnosis in major salivary gland tumors in children. A core biopsy of the tumor may improve the accuracy of preoperative diagnosis but it does not exclude the possibility of misdiagnosis.

Cardiovascular and Renal Outcomes of Renin-Angiotensin System Blockade in Renal Transplant Recipients.

BACKGROUND: There is considerable controversy over the benefits of renin-angiotensin system (RAS) blockade in renal transplant recipients (RTRs). The aim of the study was to research the effects of RAS blockade on allograft and patient outcome. METHODS: A retrospective analysis of the effects of RAS blockade on allograft and patient outcome in 53 pairs of RTRs receiving grafts from the same donor was performed. The 106 RTRs (53 pairs), transplanted from 2002 to 2012, were included in the study when 1 patient from the pair used an angiotensin converting enzyme inhibitor (ACEI) or angiotensin receptor blocker (ARB) for a minimum period of 36 months (RAS[+]) and the second one did not use it (RAS[-]). RESULTS: There were no differences between RAS(+) and RAS(-) subjects in terms of age, body mass index, reason of end-stage renal disease, mismatches number, total ischemic time, episodes of cytomegalovirus infections, acute rejections, and immunosuppressive treatment. The mean time of observations was 66.28 months ± 24.39 months. RAS inhibitors were given in a mean dose of 23.1% (ACEI) and 27.08% (ARB) of the maximum recommended. The main reasons for the therapy were as follows: hypertension (39.62%), nephroprotection/proteinuria (39.62%), and polyglobulia (28.3%). The composite cardiorenal endpoint was reached by 6 (11.32%) and 7 (13.21%) patients in RAS(+) and RAS(-) group, respectively. There were no differences in changes of creatinine, potassium serum level, or estimated glomerular filtration rate between RAS(+) and RAS(-) patients in the early period after RAS blockade commencement. CONCLUSION: Agents inhibiting the RAS system neither improved nor deteriorated patients and graft survival in RTRs.

Influence of Renin-Angiotensin System Blockers on Graft Function in Retrospective Analysis of Pairs of Renal Transplant Recipients From the Same Donor.

BACKGROUND: Renin-angiotensin system (RAS) blocking agents efficiently control hypertension in renal transplant recipients (RTRs), and reduce proteinuria and post-transplant erythrocytosis. A beneficial effect on the retardation of the long-term decline in renal function has not yet been demonstrated. The aim of the study was to evaluate the effects of RAS blockade on allograft function. METHODS: In order to minimize donor variability and bias, 33 pairs of RTRs receiving grafts from the same donor were included into the retrospective analysis. A total of 66 RTRs were enrolled in which 1 patient from the pair used an angiotensin-converting enzyme inhibitor or angiotensin receptor blocker for a minimum period of 60 months (RAS[+]) and the second one did not use it at all (RAS[-]). RESULTS: There were no differences between RAS(+) and RAS(-) subjects in terms of age, body mass index, mismatches number, duration of total ischemia, episodes of cytomegalovirus infections, acute rejections, or immunosuppressive treatment. Significantly, more RAS(+) patients presented with diabetes and cardiovascular complications. Among RAS(+) patients, angiotensin-converting enzyme inhibitors and angiotensin receptor blockers were used in 28 (84.84%) and 5 (15.15%) patients in a mean dose of 23.03 ± 16.83% and 30 ± 11.18% of their maximum doses, respectively. There were no significant differences in estimated glomerular filtration rate changes (-0.37 ± 12.68 vs 2.54 ± 20.76 mL/min) and serum creatinine changes (0.05 ± 0.39 vs 0.14 ± 0.79 mg/dL) between RAS(+) and RAS(-) patients during the 60 months follow-up. CONCLUSION: Agents inhibiting the RAS did not significantly affect graft function in RTRs during 60 months of observation.

Differential effects of nitric oxide deficiency on primary tumour growth, pulmonary metastasis and prostacyclin/thromboxane A2 balance in orthotopic and intravenous murine models of 4T1 breast cancer.

The role of nitric oxide (NO) in tumour progression and metastasis is not clear, therefore the present work aimed to better characterise the effects of nitric oxide synthase (NOS) inhibition by L-Nω-nitroarginine methyl ester (L-NAME) on primary tumour growth, pulmonary metastasis, inflammatory state and prostacyclin (PGI2)/thromboxane A2 (TXA2) balance in a 4T1 murine model of breast cancer. To distinguish effects of NO deficiency on disease development, 4T1 cancer cells were administered orthotopically or intravenously to Balb/c mice. The systemic NO bioavailability, pulmonary inflammation and plasma levels of thromboxane B2 (TXB2) and 6-keto-prostaglandin F1α (6-keto-PGF1α) were assessed. The study shows that, in the orthotopic model of 4T1 breast cancer, L-NAME hampered primary tumour growth, reduced pulmonary metastases, delayed inflammatory response but did not alter biosynthesis of TXB2 and 6-keto-PGF1α as well as PGI2/TXA2 ratio in cancer-bearing mice. Interestingly, in the intravenous model of 4T1 breast cancer, NOS inhibition did not influence metastasis nor inflammation, but it increased both TXB2 and 6-keto-PGF1α biosynthesis without affecting PGI2/TXA2 ratio. In conclusion, in a 4T1 murine model of metastatic breast cancer, NO plays a major role in primary tumour development, while NO is not the key mediator of cancer cell extravasation to the lungs. Furthermore, NO-deficiency activates a PGI2-dependent compensatory mechanism only in the intravenous model of 4T1 breast cancer.

Chronic stress changes prepulse inhibition after amphetamine challenge: the role of the dopaminergic system.

The goal of this research was to examine the influence of chronic mild stress (CMS) on prepulse inhibition (PPI). We used an amphetamine challenge to study the role of the dopaminergic system in limbic structures. Chronic stress caused a reduction in both sucrose preference and body weight. It was found that the initially strong response to amphetamine in the control rats was weakened after stress on both the behavioural and biochemical levels: improved PPI, decreased dopamine D2 receptor expression in the central nucleus of amygdala (CeA) and nucleus accumbens (NAC), and decreased dopamine and 3-MT (3-methoxytyramine) levels in NAC. We observed that the stress-evoked attenuation of amphetamine-induced stimulation was also paralleled by changes in corticosterone level. These effects were accompanied by a decrease in both glutamate and the glutamate/gamma-aminobutric acid (GABA) ratio in the NAC. The interpretation of these results is that prolonged stress induces compensatory mechanisms in the mesolimbic system which are responsible for psychostimulant (amphetamine) effects.

[Fibers glass induced cytotoxicity and genotoxicity]. / Citotossicità e genotossicità indotte da fibre di vetro.

Man-made vitrous fibers, have been widely used as a substitute for asbestos, as an insulation material. However the fibrous morphology of MMVFs raises concern about potential health hazard. The aim of our study was to assess cytotoxic and genotoxic effects induced on a human alveolar cell line A549 by exposure to glass wool fibers (GW). Cells were exposed for 72 h to 5, 50, 100 microg/ml of glass wool, after incubation the cell viability was determined by a MTT reduction assay. The genotoxic effect was studies by Comet test. An undamaged cell appeared as a nucleoid and a cell with damaged DNA as a comet. Measurement of Comet parameters: % DNA in the tail, tail length and tail momente (the product of relative tail intensity and lenght, that provides a parameter of DNA damage) were obtained from the analysis. A MTT assay indicated that glass wool caused a decrease in cell viability and this decrease was concentration-dependent. The results of the Comet test for DNA damage detection indicated in cell exposed to glass wool fibers a significant increase of mean TM value. All these results provide that the glass wool fibers can induce cytotoxicity and genotoxicity

[Evaluation of occupational exposure to biological agents, endotoxins and PNOC in a sewage treatment plant]. / Valutazione dell'esposizione professionale ad agenti biologici, endotossine e PNOC in un impianto di depurazione.

The aim of this work has been to determine the occupational exposure to the biological agents and airborne dust in a sewage treatment plant in south Italy. The air samplings were performed in a sewage treatment plant in Calabria, in two different seasons (spring and summer) at 5 sites associated with various phases of sewage treatment process. In addition we have estimated the concentration of airborne endotoxins and PNOC (Particles Not Otherwise Classified) by using personal samplers. The results showed a significant variation in exposure to bioaerosols, endotoxins and PNOC depending on the sampling season: the PNOC concentration increase as much as the endotoxins concentration in spring and decrease in summer

Effects of chronic nitric oxide synthase inhibition on V'O2max and exercise capacity in mice.

Acute inhibition of NOS by L-NAME (Nω-nitro-L-arginine methyl ester) is known to decrease maximal oxygen consumption (V'O2max) and impair maximal exercise capacity, whereas the effects of chronic L-NAME treatment on V'O2max and exercise performance have not been studied so far. In this study, we analysed the effect of L-NAME treatment, (LN2 and LN12, respectively) on V'O2max and exercise capacity (in maximal incremental running and prolonged sub-maximal incremental running tests), systemic NO bioavailability (plasma nitrite (NO2-) and nitrate (NO3-)) and prostacyclin (PGI2) production in C57BL6/J mice. Mice treated with L-NAME for 2 weeks (LN2) displayed higher V'O2max and better running capacity than age-matched control mice. In LN2 mice, NO bioavailability was preserved, as evidenced by maintained NO2- plasma concentration. PGI2 production was activated (increased 6-keto-PGF1α plasma concentration) and the number of circulating erythrocytes (RBC) and haemoglobin concentration were increased. In mice treated with L-NAME for 12 weeks (LN12), NO bioavailability was decreased (lower NO2- plasma concentration), and 6-keto-PGF1α plasma concentration and RBC number were not elevated compared to age-matched control mice. However, LN12 mice still performed better during the maximal incremental running test despite having lower V'O2max. Interestingly, the LN12 mice showed poorer running capacity during the prolonged sub-maximal incremental running test. To conclude, short-term (2 weeks) but not long-term (12 weeks) treatment with L-NAME activated robust compensatory mechanisms involving preservation of NO2- plasma concentration, overproduction of PGI2 and increased number of RBCs, which might explain the fully preserved exercise capacity despite the inhibition of NOS.

Endothelial glycocalyx integrity is preserved in young, healthy men during a single bout of strenuous physical exercise.

In the present study we aimed to evaluate whether oxidative stress and inflammation induced by strenuous exercise affect glycocalyx integrity and endothelial function. Twenty one young, untrained healthy men performed a maximal incremental cycling exercise - until exhaustion. Markers of glycocalyx shedding (syndecan-1, heparan sulfate and hyaluronic acid), endothelial status (nitric oxide and prostacyclin metabolites - nitrate, nitrite, 6-keto-prostaglandin F(1alpha)), oxidative stress (8-oxo-2'-deoxyguanosine) and antioxidant capacity (uric acid, non-enzymatic antioxidant capacity) as well as markers of inflammation (sVCAM-1 and sICAM-1) were analyzed in venous blood samples taken at rest and at the end of exercise. The applied strenuous exercise caused a 5-fold increase in plasma lactate and hypoxanthine concentrations (p<0.001), a fall in plasma uric acid concentration and non-enzymatic antioxidant capacity (p<10(-4)), accompanied by an increase (p=0.003) in sVCAM-1 concentration. Plasma 6-keto-prostaglandin F(1alpha) concentration increased (p=0.006) at exhaustion, while nitrate and nitrite concentrations were not affected. Surprisingly, no significant changes in serum syndecan-1 and heparan sulfate concentrations were observed. We have concluded, that a single bout of severe-intensity exercise is well accommodated by endothelium in young, healthy men as it neither results in evident glycocalyx disruption nor in the impairment of nitric oxide and prostacyclin production.

Changes in age specific immunity to diphtheria in Poland in the past 40 years.

In recent years people in Poland have acquired diphtheria from eastern neighbours. It was considered important therefore to study the age specific immunity against diphtheria and to compare it with the results of seroepidemiological studies conducted in t

The influence of tonsillectomy on obstructive sleep apnea children with malocclusion.

Adeno-tonsillar hypertrophy is the most common cause of obstructive sleep apnea syndrome (OSAS) in childhood and may also play a role in the development of craniofacial abnormalities. The mode of breathing and the morphology of the dental arch are very closely connected. Most of the children who came to the ENT clinic had malocclusions. The early performance of surgical treatment on hypertrophied tonsils and/or adenoids influenced greatly the state of health and morphology of the dental arch. In the process of diagnosis and treatment of children with OSAS it is necessary to have multidisciplinary cooperation, especially between the otolaryngologist and the orthodontist.

Relationship between passive smoking, recurrent respiratory tract infections and otitis media in children.

UNLABELLED: The cause of upper respiratory tract infections (URTI) are multifactorial (enlarged adenoid, environmental conditions, staying at the care centers, smoking parents, allergy). Directly, viral infection causes damage to the ciliary cells and mucociliary clearance in the nasopharynx and Eustachian tube, promotes tubal occlusion and provokes otitis media. Enlarged adenoids reduce ventilation to the nasopharynx, increase accumulation of the secretion and provide a good condition for bacteria. AIMS OF THE STUDY: Evaluation of the factors playing a role in recurrent URTI and otitis media in children. Clinical and histopathological examination of adenoid tissue of children who were passive smokers and children who were not exposed to cigarette smoke. Evaluation of the difference between ciliary-mucous transport among passive smokers and children not exposed to cigarette smoke. METHODS: The analysis of interview questionnaires in 1000 children aged 3-14 years. Histopathological examinations of adenoid tissue excised in the group of children of recurrent upper respiratory tract infections and serous otitis media exposed and not exposed to cigarette smoke. CONCLUSIONS: Among the risk factors for URTI, the most important are: (1) socio-economic conditions; (2) staying at day care centers; and (3) passive smoking. Allergy was confirmed in 35-38% of URTI children. Surgical treatment was undertaken in 11.4-32.5% of URTI children (tonsilloadenoidectomy). Histopathological and ultrastructural evaluation of adenoid tissue in passive smoking children indicates significant differences to children not exposed to cigarette smoke.

[Investigation of immunogenic properties of diphtheria antitoxin by intradermal active test of potency]. / Badanie wlasciwosci uodparniajacych anatoksyny bloniczej w sródskórnym tescie czynnym mocy.

Usefulness of four variants of intradermal test for routine testing of protective potency and diphtheria anatoxin was studied. Comparing with classical active test it was found that full intradermal test possesses data system and results comparable to the classic test and may replace it. This in effect results in four-fold reduction of guinea pig number necessary for performance of the investigation. As orientating test, estimate determining AnaDi potency a single dose performed on six guinea pigs was selected. In the intradermal test the time of investigation, as compared with the classic test, may be shortened by one week.

[Simplified active test of estimating tetanus toxoid]. / Skrócony test czynny do oceny mocy toksoidu tezcowego.

The main purpose of these investigations was the comparison of two potency tests of Tetanus Toxoid. It was found that two doses test with reduced number of animals can be used in assay of different vaccines containing Tetanus component. One dose test is agreeable with requirements of WHO. It is credible and not complicated method of estimation Diphtheria and Tetanus Toxoids. This test required minimal number of animals.

[Usefulness of the cytotoxic test for testing levels of diphtheria antibodies in sera of immunized animals]. / Przydatnosc testu cytotoksycznego do badania poziomu przeciwcial bloniczych w surowicach uodparnianych zwierzat.

Vero cells method was used for determination of antibodies level in humans vaccinated with Td vaccine and also in animals immunized with Di, DiTe, Td and DTP vaccines. We have found quite high levels of antibodies in both groups of animals. There was also a significant increase of antibody level in the human sera after Td booster dose in the CCV and HA tests, but the correlation with antitoxin titre was low. CCV test could be useful for determination of immunogenicity of diphtheria toxid.

[Diphtheria and tetanus immunity of blood donors]. / Ocena poziomu przeciwcial bloniczych i tezcowych u dawców krwi.

Immunity to diphtheria and tetanus was determined in serum samples from 108 blood donors. Antitoxin concentration was evaluated by ELISA assay. 65% of donors were protected to diptheria and 100% to tetanus. Significant trend of decreasing immunity was observed with increasing age.

[Evaluation of antitoxin and antibacterial activity of human preparations of normal immunoglobulin. I. Level of tetanus antitoxin in preparations of IMIG and IVIG]. / Ocena aktywnosci antytoksycznej i antybakteryjnej preparatów ludzkiej normalnej immunologlobuliny. I. Poziom antytoksyn tezcowych w preparatach IMIG i IVIG.

The level of anti-tetanus antibodies were measured in 791 lots of IMIG and 330 lots of IVIG of human normal gamma globulin. The antibody levels varied 15-80 IU/ml of IMIG lots and 5-20 IU/ml of IVIG lots. Every lot of human normal gamma globulin (IMIG or IVIG) having high anti-tetanus activity may be an acceptable alternative to specific tetanus immune globulin.

[Pilot study of the safety and immonogenicity of the diphtheria adsorbed toxoid "d"]. / Badanie pilotowe bezpieczenstwa i immunogennosci anatoksyny bloniczej "d" o zmniejszonej zawartosci antygenu.

The absorbed vaccine d recommended as the booster dose contained in 0.5 ml 2 Lf diphtheria toxoid and was safety and stable, the potency was 30 JO/ml. Eight adults volunteers 25-65 years received toxoid d. The tolerance and the serological response was very good.