Author Correction: Micronuclei in germ cells of hybrid frogs from Pelophylax esculentus complex contain gradually eliminated chromosomes.

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Micronuclei in germ cells of hybrid frogs from Pelophylax esculentus complex contain gradually eliminated chromosomes.

In most organisms, cells typically maintain genome integrity, as radical genome reorganization leads to dramatic consequences. However, certain organisms, ranging from unicellular ciliates to vertebrates, are able to selectively eliminate specific parts of their genome during certain stages of development. Moreover, partial or complete elimination of one of the parental genomes occurs in interspecies hybrids reproducing asexually. Although several examples of this phenomenon are known, the molecular and cellular processes involved in selective elimination of genetic material remain largely undescribed for the majority of such organisms. Here, we elucidate the process of selective genome elimination in water frog hybrids from the Pelophylax esculentus complex reproducing through hybridogenesis. Specifically, in the gonads of diploid and triploid hybrids, but not those of the parental species, we revealed micronuclei in the cytoplasm of germ cells. In each micronucleus, only one centromere was detected with antibodies against kinetochore proteins, suggesting that each micronucleus comprises a single chromosome. Using 3D-FISH with species-specific centromeric probe, we determined the role of micronuclei in selective genome elimination. We found that in triploid LLR hybrids, micronuclei preferentially contain P. ridibundus chromosomes, while in diploid hybrids, micronuclei preferentially contain P. lessonae chromosomes. The number of centromere signals in the nuclei suggested that germ cells were aneuploid until they eliminate the whole chromosomal set of one of the parental species. Furthermore, in diploid hybrids, misaligned P. lessonae chromosomes were observed during the metaphase stage of germ cells division, suggesting their possible elimination due to the inability to attach to the spindle and segregate properly. Additionally, we described gonocytes with an increased number of P. ridibundus centromeres, indicating duplication of the genetic material. We conclude that selective genome elimination from germ cells of diploid and triploid hybrids occurs via the gradual elimination of individual chromosomes of one of the parental genomes, which are enclosed within micronuclei.

Myocardial perfusion SPECT with dipyridamole stress test in cardiac syndrome X.

UNLABELLED: Cardiac syndrome X defines patients with typical anginal chest pain, a positive exercise ECG stress test and angiographically normal coronary arteries. AIM of this study was to evaluate the role of myocardial perfusion SPECT with dipyridamole stress in the diagnosis of cardiac syndrome X. PATIENTS, METHODS: 68 patients with syndrome X aged 32 to 60 years were subjected to myocardial imaging using (99m)Tc-MIBI according to the two-days protocol: at rest and after dipyridamole infusion. Semiquantitative evaluation of the images was based on the assessment of (99m)Tc-MIBI uptake in 17 myocardial segments using a 5-points scale (0 point -- normal uptake, 4 points -- no uptake). Scores obtained in each segment were summed up, constituting the summed rest score (SRS) and summed stress score (SSS). RESULTS: Mean SRS was 7.9 +/- 4.8 and mean SSS was 7.2 +/- 4.4 (non-significant difference). Individual comparison of SRS and SSS values revealed three patterns of scintigraphic images: 1) in 25 patients (36.8%), a paradoxical improvement of perfusion at stress images was found, 2) in 23 patients (33.8%), the myocardial perfusion deteriorated after dipyridamole, 3) in 20 patients (29.4%), no significant change of the myocardial perfusion between rest and stress images occurred. CONCLUSIONS: In cardiac syndrome X, myocardial SPECT with dipyridamole stress shows different patterns of myocardial perfusion that reflects heterogeneity of this pathology.

Metabolic control quality and free radical activity in diabetic patients.

The role of oxidative/reductive balance derangement in the pathogenesis of diabetic microangiopathy has often been discussed in the last few years. Therefore, we decided to evaluate the influence of intensive insulin therapy on selected indicators of free radical production. The levels of plasma hydrogen peroxide (H2O2) and serum malonyldialdehyde (MDA) were estimated in 15 patients with Type 1 and 15 with Type 2 diabetes before and after 2 weeks of intensive treatment. The initial H2O2 and MDA levels in all cases were significantly higher than in controls. After 2 weeks of treatment, the values for both estimated parameters were significantly lower; however, they were still higher than in the control group. Our results seem to confirm the previous suggestions concerning the relation between metabolic disturbances and oxidative stress in diabetic patients.

Treatment of hairy cell leukemia patients with 2-chloro-2'-deoxyadenosine (2-CdA).

A preliminary study of six hairy cell leukemia patients treated with one course of 2-chloro-2'-deoxyadenosine (2-CdA) is presented. 2-CdA was administered 0.1 mg/kg/daily by intravenous infusion over 7 days. Two patients achieved CR and four PR.

Peroxide plasma level in patients with coronary heart disease as a possible indicator of ischemia during exercise test.

BACKGROUND: This study evaluated the possible changes in production of free oxygen species during myocardial ischemia in the course of exercise testing of ischemic coronary disease. METHODS: Exercise testing was performed on 58 patients with suspected ischemic coronary disease. Plasma hydrogen peroxide (H2O2) level was assayed according to Frew et al. (Anal Chim Acta 1983, 155:139-150) in venous blood samples obtained before the test, at submaximal and maximal work loads, and after 30 minutes of rest. RESULTS: Hydrogen peroxide plasma level significantly increased (P < 0.001) at maximal work load in patients with positive exercise test results. Patients with negative exercise test results initially manifested a higher H2O2 level that dropped during maximal work load. CONCLUSIONS: The rise in plasma H2O2 level during myocardial ischemia indicates that intensification of oxygen free radical production occurs. This may be the result of hypoxanthine conversion by xanthine oxidase, catecholamine auto-oxidation, polymorphonuclear neutrophil activation, and/or derangement within mitochondrial electron transfer. The noticed difference between patients with a positive exercise test and those diagnosed as without ischemia may be of clinical relevance.

Estimation of hydrogen peroxide plasma levels in patients evaluated for coronary heart disease using dipyridamole infusion followed by SPECT.

BACKGROUND: Prolonged ischemia leads to myocardial infarction and increased formation of toxic oxygen radicals. These substances exert deleterious effects on myocardial cells, contributing to reperfusion injury and generation of arrhythmia. Little information is available, however, concerning the toxic oxygen species generated during transient ischemia. The purpose of our study was to estimate hydrogen peroxide plasma levels in patients subjected to short-lasting ischemia induced by a dipyridamole stress test used for the diagnosis of coronary artery disease. Evaluation of the performed test was carried out with 99mTc-SestaMIBI followed by single photon emission computed tomography (SPECT). METHODS: Blood was obtained from peripheral veins of 53 patients (37 men and 16 women, mean age 49 +/- 11 years). Plasma hydrogen peroxide levels were estimated by spectrophotometric methods: immediately before a dipyridamole challenge and after drug infusion. RESULTS: Hydrogen peroxide plasma levels in patients with a negative SPECT test were 23.5 +/- 3.0 mumol/l (mean +/- SEM) and 21.0 +/- 2.9 after the dipyridamole infusion (P = 0.474). Plasma concentrations of hydrogen peroxide in patients with a positive SPECT test were 30.5 +/- 4.6 and increased after dipyridamole challenge to 50.3 +/- 5.4 (P = 0.004). Further analysis revealed that the observed difference cannot be attributed to previous history of myocardial infarction. CONCLUSIONS: Even transient myocardial ischemia can generate toxic oxygen derivatives. Evaluation of plasma levels of hydrogen peroxide may be of clinical relevance in patients with suspected coronary artery disease.

The effects of dipyridamole stress test on plasma levels of soluble adhesion molecules intracellular adhesion molecule-1, vascular cell adhesion molecule-1, E-selectin and L-selectin in patients with ischemic heart disease and patients with syndrome X.

BACKGROUND: Adhesion of activated leukocytes to the endothelium as a result of myocardial ischemia/reperfusion has been shown to be involved in the development of tissue injury. Leukocyte adhesion to the endothelium occurs via adhesion molecules expressed on the surface of both cell types. Upon cell activation these proteins may be released into the circulation and measured in a soluble form. AIM: To verify whether the dipyridamole stress test, performed in patients with ischemic heart disease (IHD) and in patients with syndrome X, modifies plasma levels of the soluble adhesion molecules vascular cell adhesion molecule-1 (VCAM-1), intracellular adhesion molecule-1 (ICAM-1), E-selectin and L-selectin. METHODS: Plasma levels of the soluble endothelial adhesion molecules ICAM-1, VCAM-1 and E-selectin, as well as of the soluble leukocyte adhesion molecule L-selectin, were measured in venous blood samples taken before and 7 min after administration of dipyridamole in patients with IHD, patients with syndrome X and healthy individuals. Myocardial perfusion was evaluated using single photon emission tomography. The plasma levels of soluble VCAM-1, ICAM-1, E-selectin and L-selectin were all measured using enzyme-linked immunosorbent assays. RESULTS: After infusion of dipyridamole, plasma levels of ICAM-1 increased significantly in patients with IHD, whereas they remained unchanged in patients with syndrome X and in the control group. In patients with IHD, the initial plasma levels of VCAM-1, E-selectin and L-selectin, before administration of dipyridamole, were higher than those observed in patients with syndrome X and than those in the control group. Plasma levels of soluble VCAM-1, E-selectin and L-selectin decreased significantly in patients with IHD following the dipyridamole stress test, whereas they remained unchanged in patients with syndrome X, and in the control group. CONCLUSION: In patients with IHD, administration of dipyridamole induces myocardial ischemia resulting in modification of plasma levels of the soluble adhesion molecules.

Changes of acute phase proteins glycosylation profile as a possible prognostic marker in myocardial infarction.

In 24 consecutive patients with myocardial infarction, the concentrations of C-reactive protein, alpha 1-acid glycoprotein and alpha 1-antichymotrypsin, as well as acid alpha 1-glycoprotein and alpha 1-antichymotrypsin glycosylation profiles were estimated. Blood samples were taken at admission, after 4, 8, 12 and 24 h, on 2, 3, 6, 9, and 12 days of hospitalization. All studied patients were divided into 2 groups: 12 patients without clinical or radiological symptoms of acute heart failure and 12 patients with acute heart failure. The results of all investigations were tested statistically to appraise significance of differences between the two investigated groups. At admission, as well as after 6 and 12 h, C-reactive protein concentration was significantly higher in patients who developed heart failure. Similarly, the glycosylation profile of alpha 1-antichymotrypsin, reported as reactivity coefficient, was of good prognostic value from the first time-point on. Development of acute cardiac failure seemed to correlate more with the magnitude of inflammatory reaction (measured by changes in acute phase proteins) than with enzymatically estimated infarct size.

The influence of electrical cardioversion on superoxide anions (O2-) production by polymorphonuclear neutrophils, hydrogen peroxide (H2O2) plasma level and malondialdehyde serum concentration.

We studied the influence of electrical cardioversion on unstimulated and stimulated superoxide anion production by polymorphonuclear neutrophils in 22 patients with atrial flutter or atrial fibrillation. We also estimated hydrogen peroxide plasma level, as well as malondialdehyde serum concentration, in these subjects. We noted an increase in spontaneous production of superoxide anions from 14.9 +/- 1.8 nmol/10(6) neutrophils per 20 min to 21.37 +/- 2.7 nmol/10(6) neutrophils per 20 min (P = 0.002) in neutrophils obtained after electrical cardioversion. Similarly, stimulated production of O2- also increased after electrical cardioversion (41.8 +/ 3.4 nmol/10(6) neutrophils per 20 min vs. 59.0 +/- 5.9 nmol/10(6) neutrophils per 20 min, P = 0.0027). Moreover, hydrogen peroxide plasma level increased significantly after electrical cardioversion (39.9 +/- 6.2 mumol/l vs. 53.4 +/- 7.6 mol/l, P = 0.003). Serum malondialdehyde concentration also increased after countershock (2.56 +/- 0.26 nmol/ml vs. 2.94 +/- 0.26 nmol/ml, P = 0.023). These results seem to indicate that electrical cardioversion may lead to polymorphonuclear neutrophils activation, increased H2O2 production and lipid peroxidation.

Plasma chemotactic activity during dipyridamole induced myocardial ischemia.

BACKGROUND: We have previously reported that transient myocardial ischemia induced during exercise or dipyridamole challenge leads to the release of increased amounts of hydrogen peroxide into circulating blood. It would indicate that the temporary functional changes within myocardial cells may constitute there a sterile inflammatory area. Therefore we decided to evaluate the chemotactic properties of plasma in patients undergoing dipyridamole provocative test, as a sign of released inflammatory mediators. The ischemia occurrence was evaluated with 99mTc-SestaMIBI followed by single photon emission computed tomography (SPECT). METHODS: Blood samples were obtained from the peripheral vein of 42 patients (18 men and 24 women, mean age 61 years). Plasma chemotactic activity was determined by the use of the Boyden chamber method: immediately before dipyridamole challenge (time 0), 7, and 30 min after drug infusion. The migration of control polymorphonuclear neutrophils towards evaluated plasma samples was estimated. RESULTS: Chemotaxis of control PMNs towards plasma isolated from patients without signs of myocardial ischemia 7 min after dipyridamole administration was significantly diminished in comparison with baseline values (p=0.003). Plasma obtained 7 min after dipyridamole infusion from patients manifesting signs of myocardial ischemia by SPECT attracted control PMNs significantly more intensively in comparison to plasma isolated at time 0 (p=0.0005). CONCLUSIONS: The obtained results indicate that transient myocardial ischemia induced by dipyridamole challenge leads to generation of chemotactic factors detectable in peripheral blood plasma.

Intense acute phase response in ischemic patients.

The aim of this study was to estimate the qualitative and quantitative changes of acute phase proteins in patients suffering from coronary heart disease. The study was carried out on 74 patients and 12 healthy volunteers. The patients were divided into three groups as follows: patients with myocardial infarction (n=37), Group I--without heart failure, Group II--with heart failure (II-III NYHA), Group III--patients with unstable angina pectoris (n=35); controls-healthy volunteers (n=12). The immunological measurements were performed at the beginning of hospitalisation (point 0), after 4, 8, 12 and 72 h, and after 6, 9 and 12 days of hospitalisation. The concentrations of C-reactive protein (CRP), alpha1-acid glycoprotein (AGP) and alpha1-antichymotrypsin (ACT) were measured using rocket immunoelectrophoresis according to Laurell. Glycosylation profiles of AGP and ACT were determined using crossed affinity immunoelectrophoresis with Con A as ligand according to Bøg-Hansen. Between Groups I and II statistically significant differences were observed for all investigated parameters. Highest concentration values were observed for Groups II and III; for Group II they appeared earlier than for Group III. The maximal values for reactivity coefficients (AGP-RC and ACT-RC) were observed earlier than the respective maximal values of concentrations. Continuous activation occurring in unstable angina leads to a more rapid increase in the concentrations of acute phase proteins and more marked alterations in their glycosylation profiles. In a way these patients seem to be 'primed' with constant stimulation, so that they respond dramatically to the stimulus of ischemia.

An alternative to dental floss in a personal dental hygiene program.

The concept of gingival massage on maintaining the health of the gingiva may be of value as an adjunct in the periodontal maintenance phase of treatment. This study compared dental floss to a test product, a massage device, in a randomized population of 20 male and female subjects. Clinical measurements were taken at baseline, week 3, and week 6. Patients were instructed to brush twice daily using their normal hygiene methods and to use the assigned product once daily. Plaque index (PI), bleeding index (BI) and pocket probing depths did improve in both the control and the test groups, but the differences did not favor either group over the other. The gingival index (GI) did, however, significantly decrease in the massage group, suggesting that this product may not only be an alternative to dental floss, but may provide added benefits from gingival massaging.

Clinical evaluation of the effect of a hydrogen peroxide mouth rinse, sodium bicarbonate dentifrice, and mouth moisturizer on oral health.

The objective of this 60-day single-blind, parallel trial, using 150 subjects, was to evaluate the effect of a 20% sodium bicarbonate dentifrice, a 1.5% hydrogen peroxide solution and a mouth moisturizer on oral tissues and microflora. Subjects were randomly assigned to one of five groups. The treatments were: 1) Sage dentifrice (sodium bicarbonate). Toothette Plus containing baking soda saturated with the hydrogen peroxide solution and use of a mouth moisturizer, 2) Crest dentifrice, Toothette Plus containing baking soda saturated with the hydrogen peroxide solution and use of a mouth moisturizer, 3) Crest dentifrice, Toothette Plus containing baking soda with a control solution and no mouth moisturizer, 4) Crest dentifrice, Toothette (without baking soda), saturated with a control solution and no mouth moisturizer, and 5) Crest dentifrice, Toothette saturated with 1.5% flavored H2O2 and no mouth moisturizer. From a subgroup of 35 patients (seven from each group) buccal smears for exfoliative cytology were taken as were supragingival microbiological samples from the mesial aspect of first molars (pooled). Buccal smears were evaluated for signs of histopathological changes. Microbiological samples from supra- and subgingival plaque for P. gingivalis, P. intermedia, A. actinomycetemcomitans. A viscosus, F. nucleatum, F. sanguis and C. albicans were evaluated. Clinical parameters measured were a stain index (SI), the modified gingival index (MGI), and a plaque index (PI). There were no adverse changes in the oral microflora and no anaplastic or other pathological changes in any subjects. Clinical parameters showed a statistically significant reduction in the MGI ranging from 26.7-29.9% with no significant differences among the groups (p > 0.05). The PI showed small reductions in all groups except group 2, but the differences were not statistically significant from each other or baseline (p > 0.05). The SI revealed slight increases in all groups and no differences among the groups. It can be concluded that use of Sage dentifrice, Toothette Plus saturated with Perox-A-Mint and Sage Mouth Moisturizer are safe to oral tissues. Using these components did not result in clinically noticeable stain formation, promote plaque formation, or produce any significant adverse changes in the oral microflora.

[Platelets and the fibrinolytic system]. / Plytki a uklad fibrynolityczny.

The authors present interactions between fibrinolysis and platelets. Platelets may exert both stimulatory and inhibitory effect on fibrinolysis. Also the problem of fibrinolysis activation on platelets function is considered.

Standard heparin and low molecular weight heparin effect on spontaneous platelet aggregation measured in whole blood and in platelet-rich plasma.

The evaluation of both standard heparin (SH) and low molecular weight heparin (LMWH) (Fraxiparine) impact on spontaneous platelet aggregation assessed in the whole blood and in platelet-rich plasma (PRP). The results obtained reveal that SH enhance spontaneous aggregation in a similar degree both in the whole blood (34.5 +/- 3.0, control 21.3 +/- 4.1) and in PRP (33.5 +/- 2.7, control 13.8 +/- 3.2). LMWH significantly intensifies spontaneous aggregation in the whole blood (46.9 +/- 3.2, control 21.3 +/- 4.1) but not in PRP (19.4 +/- 4.3, control 13.8 +/- 3.2).

[Non-Hodgkin's lymphoma in a patient after many years of remission of Hodgkin's disease]. / Nieziarniczy chloniak zlosliwy u chorej z wieloletnia remisja choroby Hodgkina.

Authors present 38-years old woman with 11-years lasting complete remission of Hodgkin's disease who was recognized to have non-Hodgkin's lymphoma of the base of the tongue and also of stomach three months later. After polychemotherapy complete remission was achieved.

[Effect of thrombin stimulated blood platelets on plasma fibrinolytic activity]. / Wplyw krwinek plytkowych stymulowanych trombina na aktywnosc fibrynolityczna osocza.

The influence of thrombin stimulated-blood platelets on plasma fibrinolytic activity was evaluated. Thrombin-activated blood platelets have been shown to significantly inhibit plasma fibrinolytic activity before and after venous stasis. This was expressed by a reduction of the digestion area of fibrin dish from 10.3 +/- 3.3 cm2 to 3.7 +/- 1.5 cm2 and from 15.6 +/- 6.8 cm2 to 3.7 +/- 1.7 cm2, respectively.