Pure primary osteosarcoma of the breast: a case report.

INTRODUCTION: Mammary sarcomas are relatively uncommon and they represent less than one percent of all primary breast malignancies. Osteosarcoma of the breast, unassociated with other tumors, is distinctly rare, with published references generally limited to case reports and occasional cases in several series encompassing a heterogeneous group of mammary sarcomas and extraosseous osteosarcomas at various sites. The authors present a patient with pure osteosarcoma of the breast, osteoblastic type, with biologically aggressive pattern. CASE REPORT: A 79-year-old lady became aware of a rapidly enlarging lump in the lateral part of the right breast. Clinical examination revealed a firm to hard, mobile, irregular, and painful breast lump measuring about six by four cm. On examination there was no axillary or supraclavicular lymphadenopathy. After initial diagnosis, excisional biopsy without dissection of the axillary lymph nodes was performed. Therefore, the histological and immunohistochemical findings established the diagnosis of pure primary osteosarcoma of the breast. CONCLUSION: Pure osteosarcoma of the breast is extremely rare and needs to be distinguished from a variety of benign and malignant breast lesions producing metaplastic bone. Less than a hundred cases of pure osteosarcoma of the breast were reported, but diagnostic confirmation with immunohistochemistry has been performed in relatively few of these cases.

Tianeptine's effects on spontaneous and Ca2+-induced uterine smooth muscle contraction.

Tianeptine is a novel anti-depressant with an efficacy equivalent to that of classical anti-depressants. Additional beneficial effects include neuroprotection, anti-stress and anti-ulcer properties whose molecular mechanisms are still not completely understood but may involve changes in the anti-oxidant defence system. Herein, we have studied the effects of tianeptine on both contractile activity of isolated rat uteri and components of the endogenous anti-oxidative defence system. Tianeptine-induced dose-dependent inhibition of both spontaneous and Ca2+-induced contraction of uterine smooth muscle. The effect was more pronounced in the latter. Tianeptine treatment increased glutathione peroxidase (GSH-Px) and catalase (CAT) activities in spontaneous and Ca2+-stimulated uteri. A significant decrease in glutathione-reductase (GR) activity in both spontaneous and Ca2+-induced uterine contractions after tianeptine treatment indicated a reduction in reduced glutathione and consequently a shift toward a more oxidised state in the treated uteri. In spontaneously contracting uteri, tianeptine caused a decrease in copper-zinc SOD (CuZnSOD) activity. Tianeptine's anti-depressant effects may be accomplished by triggering a cascade of cellular adaptations including inhibition of smooth muscle contractility and an adequate anti-oxidative protection response.

Energy production and redox status of rat red blood cells after reticulocytosis induced by various treatments.

Stimulated erythropoiesis and reticulocytosis can be induced by daily bleeding, or by phenylhydrazine (PHZ) treatment. We compared the in vivo effects of PHZ and bleeding treatment on haematological, energy and redox status parameters in red blood cells (RBC) of rats. The results showed that all followed haematological parameters were significantly lower in bleeding, compared to PHZ-treated rats. PHZ induced even 2.58-fold higher reticulocytosis as compared to bleeding treatment. Although PHZ induced higher reticulocytosis, respiration intensity and energy production was lower than in bleeding-induced reticulocytes. These alterations were the consequence of increased superoxide anion and peroxynitrite concentrations in PHZ-treated rats. Bleeding treatment resulted in increased activity of an antioxidative enzyme, superoxide dismutase. In conclusion, differences in these two experimental models for reticulocytosis may be used as tools for appropriate pharmacological testing of redox-active substances considering energy and redox processes, as well as apoptosis pathways.

Hydrogen peroxide affects contractile activity and anti-oxidant enzymes in rat uterus.

BACKGROUND AND PURPOSE: The effects of hydrogen peroxide (H(2)O(2)) on uterine smooth muscle are not well studied. We have investigated the effect and the mechanism of action of exogenous hydrogen peroxide on rat uteri contractile activity [spontaneous and calcium ion (Ca(2+))-induced] and the effect of such treatment on anti-oxidative enzyme activities. EXPERIMENTAL APPROACH: Uteri were isolated from virgin Wistar rats and suspended in an organ bath. Uteri were allowed to contract spontaneously or in the presence of Ca(2+) (6 mM) and treated with H(2)O(2) (2 microM-3 mM) over 2 h. Anti-oxidative enzyme activities (manganese superoxide dismutase-MnSOD, copper-zinc superoxide dismutase-CuZnSOD, catalase-CAT, glutathione peroxidase-GSHPx and glutathione reductase-GR) in H(2)O(2)-treated uteri were compared with those in uteri immediately frozen after isolation or undergoing spontaneous or Ca(2+)-induced contractions, without treatment with H(2)O(2). The effect of inhibitors (propranolol, methylene blue, L-NAME, tetraethylamonium, glibenclamide and 4-aminopyridine) on H(2)O(2)-mediated relaxation was explored. KEY RESULTS: H(2)O(2) caused concentration-dependent relaxation of both spontaneous and Ca(2+)-induced uterine contractions. After H(2)O(2) treatment, GSHPx and MnSOD activities were increased, while CuZnSOD and GR (In Ca(2+)-induced rat uteri) were decreased. N(omega)-nitro-L-arginine methyl ester antagonized the effect of H(2)O(2) on Ca(2+)-induced contractions. H(2)O(2)-induced relaxation was not affected by propranolol, potentiated by methylene blue and antagonized by tetraethylamonium, 4-aminopyridine and glibenclamide, with the last compound being the least effective. CONCLUSIONS AND IMPLICATIONS: H(2)O(2) induced dose-dependent relaxation of isolated rat uteri mainly via changes in voltage-dependent potassium channels. Decreasing generation of reactive oxygen species by stimulation of anti-oxidative pathways may lead to new approaches to the management of dysfunctional uteri.

Antioxidant defense in mitochondria during diapause and postdiapause development of European corn borer (Ostrinia nubilalis, Hubn.).

Antioxidant enzymes (CAT, catalase; GPx, selenium nondependent glutathione peroxidase; GST, glutathione-S-transferase; GR, glutathione reductase; DHAR, dehydroascorbate reductase) were determined in the mitochondria of diapausing and non-diapausing larvae and pupae of both diapausing and non-diapausing larvae of the European corn borer (Ostrinia nubilalis, Hubn., Lepidoptera: Pyralidae). CAT, GST, and DHAR activity in mitochondria of diapausing larvae were reduced compared to non-diapausing larvae. Pupae of diapaused-larvae possessed lower GST, but higher DHAR activities compared to pupae of non-diapaused individuals. Comparison between larvae and pupae revealed lower GPx activity in the mitochondria of pupae. CAT activity in the mitochondria of pupae was higher compared to diapausing larvae, but lower than in non-diapausing ones. Correlation and canonical discriminant analyses revealed different antioxidant enzyme compositions for a particular stage and developmental pattern. Our results show that antioxidant enzymes have a similar role in the regulation of energetics in mitochondria as that in diapause and metamorphosis.

The anti-oxidative defence system in the isolated rat uterus during spontaneous rhythmic activity.

Possible interactions between nitric oxide donors, reactive oxygen species and anti-oxidative defence enzymes led us to determine the activities of anti-oxidative defence enzymes in isolated uterine smooth muscle before and after spontaneous rhythmic activity ex vivo. For our experiments we used isolated uteri from female Wistar rats. Our results showed an increase in total superoxide dismutase (SOD) and Mn SOD activities in uterine smooth muscle after spontaneous contractions when compared with nonexercised uterine smooth muscle. The activity of catalase (CAT) and glutathione preoxidase (GSH-Px) were also increased. No statistically significant changes in the activities of glutathione reductase (GR) and CuZn SOD were found. It is known that an organism's anti-oxidative defence system (guarding against excessive reactive oxygen species generation) requires balanced increments in its individual anti-oxidative enzyme activities rather than increases in the activity of only some enzymes without increases in others. Thus, we may conclude that some adaptive responses are found in exercised uterine smooth muscle but are not complete. Therefore, our results indicate that changes in anti-oxidative enzyme activities may influence the results of the examination of substances ex vivo.

Effect of thyroxine on antioxidant defense system in the liver of different aged rats.

The effects of altered thyroid state on the antioxidant defense system in the liver of differently aged rats were examined. Male rats aged 15, 45 and 75 days were treated with L-thyroxine, T(4) (40 microg/100 g body mass, s.c., one dose per day) for 14 days (finally aged 30, 60 and 90 days, respectively). The following antioxidant defense enzymes were measured: superoxide dismutases (both copper zinc, CuZn-SOD and manganese containing, Mn-SOD), catalase (CAT), glutathione peroxidase (GSH-Px), glutathione-S-transferase (GST), glutathione reductase (GR), as well as the content of low molecular mass antioxidant glutathione (GSH). The effect of T(4) on antioxidant defense system in the liver differs with respect to age. T(4) treatment decreased CAT and GST activities, as well as the content of GSH in animals aged 60 and 90 days. The same treatment elevated GR activity in rats at 30 days of age, this phenomenon was not observed in older animals. The different response of immature rats to thyroxine compared to older animals could be attributed to the differences in thyroxine metabolism and the developmental pattern. Direct effect of T(4) on mature rats can be considered as a part of its overall catabolic action.

The effects of nitroglycerine on the redox status of rat erythrocytes and reticulocytes.

The effects of nitroglycerine (NTG) are mediated by liberated nitric oxide (NO) after NTG enzymatic bio-transformation in cells. The aim of this study was to evaluate some products of NTG bio-transformation and their consequences on the redox status of rat erythrocytes and reticulocytes, considering the absence and presence of functional mitochondria in these cells, respectively. Rat erythrocyte and reticulocyte-rich red blood cell (RBC) suspensions were aerobically incubated (2 h, 37 degrees C) without (control) or in the presence of different concentrations of NTG (0.1, 0.25, 0.5, 1.0 and 1.5 mM). In rat erythrocytes, NTG did not elevate the concentrations of any reactive nitrogen species (RNS). However, NTG robustly increased concentration of methemoglobin (MetHb), suggesting that NTG bio-transformation was primarily connected with hemoglobin (Hb). NTG-induced MetHb formation was followed by the induction of lipid peroxidation. In rat reticulocytes, NTG caused an increase in the levels of nitrite, peroxinitrite, hydrogen peroxide, MetHb and lipid peroxide levels, but it decreased the level of the superoxide anion radical. Millimolar concentrations of NTG caused oxidative damage of both erythrocytes and reticulocytes. These data indicate that two pathways of NTG bio-transformation exist in reticulocytes: one generating RNS and the other connected with Hb (as in erythrocytes). In conclusion, NTG bio-transformation is different in erythrocytes and reticulocytes due to the presence of mitochondria in the latter.

Effects of exogenous donor of nitric oxide-sodium nitroprusside on energy production of rat reticulocytes.

The effects of the sodium nitroprusside (SNP), a nitric oxide (NO) donor clinically used in the treatment of hypertensive emergencies on the energy production of rat reticulocytes were investigated. Rat reticulocyte-rich red blood cell suspensions were aerobically incubated without (control) or in the presence of different concentrations of SNP (0.1, 0.25, 0.5, 1.0 mM). SNP decreased total and coupled, but increased uncoupled oxygen consumption. This was accompanied by the stimulation of glycolysis, as measured by increased glucose consumption and lactate accumulation. Levels of all glycolytic intermediates indicate stimulation of hexokinase-phosphofructo kinase (HK-PFK), glyceraldehyde 3-phosphate dehydrogenase (GAPD) and pyruvate kinase (PK) activities in the presence of SNP. Due to the decrease of coupled oxygen consumption in the presence of SNP, ATP production via oxidative phosphorylation was significantly diminished. Simultaneous increase of glycolytic ATP production was not enough to provide constant ATP production. In addition, SNP significantly decreased ATP level, which was accompanied with increased ADP and AMP levels. However, the level of total adenine nucleotides was significantly lower, which was the consequence of increased catabolism of adenine nucleotides (increased hypoxanthine level). ATP/ADP ratio and adenylate energy charge level were significantly decreased. In conclusion, SNP induced inhibition of oxidative phosphorylation, stimulation of glycolysis, but depletion of total energy production in rat reticulocytes. These alterations were accompanied with instability of energy status.

Role of antioxidant defense during different stages of preadult life cycle in European corn borer (Ostrinia nubilalis, Hubn.): Diapause and metamorphosis.

Antioxidant enzymes, total glutathione (GSH), and ascorbic acid (ASA) were determined in whole body homogenates of nondiapausing larvae, diapausing larvae during the diapausing period (October, December, and February), and in pupae emerged from both diapausing and nondiapausing larvae of the European corn borer (Ostrinia nubilalis, Hubn., Lepidoptera: Pyralidae). The activities of catalase, selenium nondependent glutathione peroxidase (GPx), and glutathione-S-transferase (GST), as well as the content of GSH and ASA, were found to vary throughout the larval diapause. Compared to diapausing larvae, nondiapausing larvae were higher in levels of catalase, GPx, GST, and dehydroascorbate reductase (DHAR) activity. GSH content was also increased. However, nondiapausing larvae contained less ASA than diapausing ones. Pupae had higher GPx and GST activity and an increased ASA content compared to larvae. The pupae emerged from nondiapausing larvae had higher GST, glutathione reductase (GR), and DHAR activities, but lower GPx activity and ASA content than those emerged from diapausing larvae. Correlation analysis revealed differences in the way the antioxidant level is equilibrated for a particular stage and developmental pattern. The results suggest that cellular antioxidants are involved in both the protection of cells and the regulation of redox levels during the pre-adult stages of Ostrinia nubilalis. Arch. Insect Biochem. Physiol. 55:79-89, 2004.

The effect of ovarian hormones on antioxidant enzyme activities in the brain of male rats.

The brain is widely responsive to gonadal hormones. The functional significance of ovarian hormones in the brain is evident from biochemical studies indicating that estradiol or progesterone treatment of testectomized rats produces changes of antioxidant enzyme activities. The effect of estradiol benzoate (EB) and progesterone (P) in the control of antioxidant (AO) enzyme activities was studied in the brain of adult male Wistar rats. The activities of catalase (CAT), glutathione peroxidase (GSH-Px), glutathione-S-transferase (GST) and glutathione reductase (GR) were measured in appropriate subcellular fractions, prepared from brains of animals belonging to various experimental groups. These groups were designed with the intention to follow changes in enzyme activities 2 h or 24 h after systemic administration of 5 microg EB or 2 mg P to testectomized (TX) animals. The obtained results show that both EB and P increase CAT activity, whereas EB decreases GSH-Px, GST and GR activities. These findings clearly show the modulatory role of EB and P in the control of enzymes responsible for the protection of rat nerve cells against oxidative damage caused by free oxygen radicals.

Effects of progesterone and estradiol benzoate on glutathione dependent antioxidant enzyme activities in the brain of female rats.

The activities of glutathione dependent antioxidant enzymes were measured in subcellular fractions of whole brain homogenates prepared from ovariectomized (OVX) female rats, untreated or treated 2 h or 24 h prior to sacrifice with a single dose of 2 mg progesterone (P) or 5 micrograms estradiol benzoate (EB). Glutathione peroxidase (GSH-Px) activity was not changed following systemic administration of EB, but P increased GSH-Px in the brain of OVX rats 24 h after the treatment. The activity of glutathione reductase (GR) was suppressed by EB short time, only 2 h following treatment, whereas P increased the enzyme activity 24 h after treatment. On the other hand, the activities of catalase (CAT) and glutathione-S-transferase (GST) were not changed following systemic administration of EB or P. The present work was carried out to study the involvement of ovarian steroids, especially P, in the control of GSH-Px and GR activities, and our results suggest that oxidative stress in the brain of female rats may be modulated by the level of progesterone.

Seasonal changes in the antioxidative defense in ground squirrels (Citellus citellus): possible role of GSH-Px.

As seasonal hibernators, ground squirrels decrease their body temperature to 7 degrees C and hibernate during the winter. Maintenance at 30 degrees C prevents seasonal changes of body temperature and animals remain euthermic and active. We measured selenium (Se)-dependent glutathione peroxidase (GSH-Px), as well as the activity of other antioxidative components such as superoxide dismutase (SOD), catalase (CAT), glutathione reductase (GR), and glutathione-S-transferase (GST) and the amount of low-molecular-weight antioxidants glutathione (GSH), ascorbic acid (AsA), and vitamin E (vit E) in spring, summer, and winter in ground squirrels continuously kept at a temperature of 30 degrees C. We examined liver and interscapular brown adipose tissue (IBAT) as thermogenic tissues, as well as the brain and the kidneys. During the winter, we found a decrease in enzymatic activity and an increase in the level of low molecular antioxidants in all tissues. Correlation analysis revealed a similarity in the composition of antioxidative defense (AD) among the tissues examined. The results obtained clearly demonstrated numerous correlative expressions of antioxidative components in this experimental model, especially of GSH-Px, suggesting the complexity of the system responsible for the maintenance of physiological homeostasis.

Seasonal changes in the activity of antioxidative defense in the kidneys of the euthermic ground squirrel (Citellus citellus).

The aim of this work was to determine the activity of the antioxidant enzymes: superoxide dismutase (EC 1.15.1.1; SOD), catalase (EC 1.11.1.6; CAT), glutathione peroxidase (EC 1.11.1.9; GSH-Px), glutathione-S-transferase (EC 2.5.1.18; GST), glutathione reductase (EC 1.6.4.2; GR) and the low molecular mass antioxidants: ascorbic acid (ASA) and vitamin E (vit E) in the kidney of ground squirrels during circannual changes. Keeping the ground squirrel at the temperature of thermic neutrality (30 degrees C) provides a stable euthermic state during the whole year and thus any change is due to the circannual rhythm. The highest specific activity of all examined antioxidative defense enzymes in the kidney was found in the spring, when ground squirrels are seasonally the most active. In the summer, lower specific activity of GSH-Px as well as of SOD and CAT were noted and, when expressed per g wet mass, only a decrease in GSH-Px activity was recorded. In the kidney of ground squirrels kept at 30 degrees C, the lowest specific activity of all examined enzymes was found during the winter and, when expressed per g wet mass, only the SOD activity was lower than in the spring and summer. Higher amounts of vitamins C and E were found in the ground squirrel kidneys in the summer. The results obtained in this work demonstrate that circannual regulation of metabolic activity, which is inherent to seasonal hibernators, is also expressed at the level of antioxidative defense in the kidneys.

Antioxidative defense in human myocardial reperfusion injury.

The activity of glutathione peroxidase (GSH-Px) as well as the activities of other antioxidative enzymes such as CuZn superoxide dismutase (CuZn SOD), catalase (CAT), glutathione reductase (GR) in erythrocytes, the plasma activity of glutathione-S-transferase (GST), and the plasma levels of vitamin E and vitamin C were evaluated in nine patients with acute myocardial infarction (AMI). Blood samples were taken before and 1, 3, 6, and 24 hours after the institution of thrombolytic therapy. The results were compared with those in 30 healthy volunteers. A significant decrease in catalase (CAT) activity and vitamin E content in patients before and after thrombolytic therapy as compared with controls was recorded. Our results confirmed that a disturbed oxidative/antioxidative balance is present after AMI and after thrombolytic therapy.

Glutathione peroxidase in amyotrophic lateral sclerosis: the effects of selenium supplementation.

The activity of glutathione peroxidase (GSH-Px) as well as the activities of other antioxidative enzymes: CuZn superoxide dismutase (CuZn SOD), catalase (CAT), glutathione reductase (GR) in erythrocytes, as well as the activity of plasma glutathione transferase (GST), and the plasma content of vitamins E and C were evaluated in 35 sporadic amyotrophic lateral sclerosis (sALS) patients. The results revealed significantly decreased activity of both GSH-Px and CuZn SOD in sALS patients compared with the control. These data showed that a disturbed oxidative/antioxidative balance in sALS patients exists not only in motoneurons but also in the blood. The effect of exogenously administered selenium (Se), antioxidants, amino acids, a Ca2+ channel blocker such as nimodipine, and their combination in Alsamin was evaluated by screening parameter levels after 9 weeks of treatment. Only the use of all components together enhanced the activity of GSH-Px and the amount of vitamin E in sALS patients. Judging by the results of clinical trials, this treatment slowed the course of the disease.

Glutathione-dependent antioxidant enzyme activities and glutathione content in the rat brain at different stages of oestrous cycle.

Enzymatic activities of glutathione peroxidase, glutathione-S-transferase, glutathione reductase and catalase, as well as the glutathione content were measured in the brain tissue of regularly cycling rats at dioestrus, proestrus and estrus. The activity of glutathione peroxidase was found to be suppressed at proestrus, whereas that of catalase was increased at dioestrus. Glutathione transferase and glutathione reductase activities, as well as the glutathione content appeared to be stable during the oestrous cycle. These results suggest that, in the female rat, glutathione peroxidase and catalase activities in the brain tissue are influenced by the ovarian hormone status.

Seasonal variation in the antioxidant defense system of the brain of the ground squirrel (Citellus citellus) and response to low temperature compared with rat.

Seasonal variation in the activity of antioxidant enzymes (superoxide dismutase (EC 1.15.1.1.; SOD), catalase (EC 1.11.1.6; CAT), glutathione peroxidase (EC 1.11.1.9; GSH-Px), glutathione reductase (EC 1.6.4.2; GR), glutathione-S-transferase (EC 2.5.1.18; GST) and low-molecular-weight antioxidants: ascorbic acid (AsA), vitamin E (VIT E) and glutathione (CSH+GSSG) were examined in the brain of the ground squirrels (Citellus citellus) maintained at 30 degrees C during the whole year. The highest activity (per mg protein) of antioxidant defense (AD) enzymes was found in the spring and was much lower in the summer. A further decrease in activity of CAT, GSH-Px and GST was observed in the winter. The highest levels of AsA and glutathione were recorded in winter in comparison with spring and summer. AD system in the brain of the ground squirrel and rates (maintained at thermoneutrality) exposed to low temperature (4 degrees C) for 3, 6 or 24 hr during the summer was studied as well. Summer was chosen as a period of stable euthermia for ground squirrels and in thermoregulation similar to rats. Consumption of free fatty acid and glucose during the acute exposure to low temperature was found to be species specific. In the ground squirrel, an increase in the specific activities of SOD, after 3, 6 and 24 hr, CAT after 3 and 6 hr and GR after 6 hr of exposure to low temperature was detected. When activities were expressed in U/g wet mass, an increase of SOD after 3, 6 and 24 hr (P < 0.02, P < 0.02, P < 0.005) and CAT and GSH-Px 3 hr (P < 0.01) upon exposure to low temperature was observed. In the rats, no changes in the specific activities of these enzymes after exposure to low temperature were recorded and only an increase in GST activity (U/g wet mass) after 6 hr exposure was registered. Low-molecular-weight AD components in both animal species were unchanged upon short-term exposure to low temperature. The species-specific differences in brain AD between the rats and the ground squirrels after short exposure to low temperature may be ascribed to seasonal changes of the brain activity in the latter.

Activity of antioxidant defense enzymes and glutathione content in some tissues of the Belgrade (b/b) laboratory rat.

The activity of antioxidant defense (AD) enzymes--superoxide dismutase (SOD, EC 1.15.1.1.), catalase (CAT, EC 1.11.1.6.), glutathione peroxidase (GSH-Px, EC 1.11.1.9.), glutathione-S-transferase (GST, EC 2.5.1.18), glutathione reductase (GR, EC 1.6.4.2) and glutathione (GSH) content of the anemic Belgrade (b/b) laboratory rats--were measured and analyzed in liver, spleen, lung, heart, brain and testes in comparison with nonanemic controls. The activities of hepatic Mn SOD, CAT, GSH-Px and GST (P < 0.02, P < 0.01 and P < 0.005) were decreased in anemic, comparing with nonanemic animals, whereas the spleen CuZn SOD, Mn SOD, CAT and GSH-Px (P < 0.005, P < 0.02, P < 0.005 and P < 0.01) activities were increased. In the lung of anemic rats, Mn SOD, GSH-Px and GR (P < 0.005, P < 0.01, P < 0.05) activities were higher, whereas GST (P < 0.01) activity was lower in relation to nonanemic ones. In anemic rats, heart Mn SOD (P < 0.05) activity was increased, brain GSH-Px (P < 0.005) activity was lower, whereas GR (P < 0.02) activity was higher compared with nonanemic controls. CuZn SOD (P < 0.05) activity in the testes was elevated and GSH-Px (P < 0.05) reduced in anemic animals. GSH content was decreased in the liver (P < 0.01), lung and brain (P < 0.005) and increased in the spleen (P < 0.02) of anemic rats in relation to the controls. Our data suggest phenotype specific differences in the AD system of the Belgrade (b/b) rat tissues in comparison with nonanemic controls.