Melatonin prevents mitochondrial dysfunctions and death in differentiated skeletal muscle cells

Oxidative stress increase induces cellular damage and apoptosis activation, a mechanism believed to represent a final common pathway correlated to sarcopenia and many skeletal muscle disorders. The goal of this study is to evaluate if melatonin, a ROS scavenger molecule, is able to counteract or modulate myotube death. Here, differentiated C2C12 skeletal muscle cells have been treated with melatonin before chemicals known to induce apoptotic death and oxidative stress, and its effect has been investigated by means of morpho‐functional analyses. Ultrastructural observations show melatonin protection against triggers by the reducing of membrane blebbing, chromatin condensation, myonuclei loss and in situ DNA cleavage. Moreover, melatonin is able to prevent mitochondrial dysfunctions which occur in myotubes exposed to the trigger alone. These findings demonstrate melatonin ability in preventing apoptotic cell death in skeletal muscle fibers in vitro, suggesting for this molecule a potential therapeutic role in the treatment of various muscle disorders.     

High passage numbers induce resistance to apoptosis in C2C12 muscle cells

Cell lines with high passage numbers exhibit alterations in cell morphology and functions. In the present work, C2C12 skeletal muscle cells with either low (<20) or high (>60) passage numbers (identified as lC2C12 or h-C2C12, respectively) were used to investigate the apoptotic response to H2 O2 as a function of culture age h-C2C12. We found that older cultures (h-C2C12 group) were depleted of mitochondrial DNA (mtDNA). When we analyzed the behavior of Bad, Bax, caspase-3 and mitochondrial transmembrane potential, we observed that cells in the h-C2C12 group were resistant to H2 O2 induction of apoptosis. We propose serially cultured C2C12 cells as a refractory model to H2 O2 -induced apoptosis. In addition, the data obtained in this work suggest that mtDNA is required for apoptotic cell death in skeletal muscle C2C12 cells.     

Exogenous melatonin alleviated growth inhibition and oxidative stress induced by drought stress in apple rootstock

Drought stress is one of the major environmental obstacles that limit the production and development of apples (Malus domestica Borkh.). The role of melatonin is well known in the protection of plants under environmental stresses. In this study, we investigated the effect of melatonin on apple rootstock M. hupehensis Rehd under drought stress. The results showed that drought inhibited the growth of M. hupehensis and dramatically reduced root surface area, root volume, the number of tips and forks, and root diameter. Drought-induced growth inhibition was significantly decreased by adding melatonin. Net photosynthetic rate (Pn), intercellular CO₂ concentration (Ci), stomatal conductance (Gs), transpiration rate (Tr), were markedly reduced under drought stress. However, the application of melatonin could mitigate the damage to the photosynthetic apparatus and increase the quantum yield of PSII photochemistry. Additionally, generation of hydrogen peroxide (H₂O₂) and superoxide radicals (O₂^•–) sharply increased in apple leaves after 4 days under drought stress, and the accumulation of electrolyte leakage (EL) represented oxidative stress, while by applying melatonin under drought stress, the generation of O₂^•– and H₂O₂ were significantly reduced and protected the membrane from drought damages. These results suggest that the adverse effects of drought can be minimized by applying melatonin to apples.

     

An extract of <i>Hypericum perforatum</i> induces wound healing through inhibitions of Ca²⁺ mobilizations,mitochondrial oxidative stress and cell death in epithelial cells: Involvement of TRPM2 channels

The wound is induced by several mechanical and metabolic factors. In the etiology of the wound recovery, excessive oxidative stress, calcium ion (Ca²⁺) influx, and apoptosis have important roles. Ca²⁺-permeable TRPM2 channel is activated by oxidative stress. Protective roles of Hypericum perforatum extract (HP) on the mechanical nerve injury-induced apoptosis and oxidative toxicity through regulation of TRPM2 in the experimental animals were recently reported. The potential protective roles in HP treatment were evaluated on the TRPM2-mediated cellular oxidative toxicity in the renal epithelium (MPK) cells. The cells were divided into three groups as control, wound, and wound + HP treatment (75 µM for 72 h). Wound diameters were more importantly decreased in the wound+HP group than in the wound group. In addition, the results of laser confocal microscopy analyses indicated protective roles of HP and TRPM2 antagonists (N-(p-Amylcinnamoyl) anthranilic acid and 2-aminoethyl diphenylborinate) against the wound-induced increase of Ca²⁺ influx and mitochondrial ROS production. The wound-induced increase of early (annexin V-FITC) apoptosis and late (propidium iodide) apoptosis were also decreased in the cells by the HP treatment. In conclusion, HP treatment acted protective effects against wound-mediated oxidative cell toxicity and apoptosis through TRPM2 inhibition. These effects may be attributed to their potent antioxidant effect.     

Cancer cell necrosis by autoschizis: Synergism of antitumor activity of vitamin C: Vitamin K3 on human bladder carcinoma T24 cells

Scanning and transmission electron microscopy and fluorescence light microscopy were employed to characterize the cytotoxic effects of vitamin C (VitC), vitamin K₃ (VitK₃) or a VitC:VK₃ combination on a human bladder carcinoma cell line (T24) following 1-h and 2-h vitamin treatment. T24 cells exposed to VitC alone exhibited membranous damage (blebs and endoplasmic extrusions, elongated microvilli). VitK₃-treated cells displayed greater membrane damage and enucleation than those treated with VitC as well as cytoplasmic defects characteristic of cytoskeletal damage. VitC:VitK₃-treated cells showed exaggerated membrane damage and an enucleation process in which the perikarya separate from the main cytoplasmic cell body by self-excision. Self-excisions continued for perikarya which contained an intact nucleus surrounded by damaged organelles. After further excisions of cytoplasm, the nuclei exhibited nucleolar segregation and chromatin decondensation followed by nuclear karryorhexis and karyolysis. This process of cell death induced by oxidative stress was named autoschizis because it showed both apoptotic and necrotic morphologic characteristics.     

Morphologic and temporal analysis of vascular smooth muscle cell apoptosis induced by c-Myc and E1A

Apoptosis is a physiologic form of cell death present in many disease conditions. When the balance of mitosis versus apoptosis is altered, tumor-like growth or degeneration of tissues may ensue. This appears to occur in several diseases, including those of the cardiovascular system, where apoptosis plays a key role in atherosclerosis and restenosis following angioplasty. Since c-myc is upregulated in the pathogenesis of these diseases, we chose to study the sequential morphologic features of programmed cell death in vascular smooth muscle cells induced by c-myc and by the adenovirus early gene E1A. Morphology and timed events in apoptotic cell cultures were analyzed by scanning electron microscopy, transmission electron microscopy, and time-lapse videomicroscopy. We observed that both c-myc-and E1A-induced apoptosis (in serum-free medium) resulted in numerous, tightly packed clusters of apoptotic blebs, as well as in one or two asymmetrically larger blebs. Transmission electron microscopy analysis revealed the larger blebs contained mostly nuclear chromatin, whereas the many smaller fragments often had little or no chromatin. Time-lapse studies showed that apoptosis was induced at a slower rate in cells stably transfected with c-myc versus those stably transfected with E1A. The early changes of apoptosis, including cell shrinkage and intense blebbing, occurred in under 5 min in both cells. Slight alterations such as cell size and further rounding occurred up to 8 h following the initial changes of apoptosis. Rather than being a part of the apoptotic response, release from the culture floor almost entirely resulted from movement of the culture flask. These studies provide a framework of timed morphologic events for future mechanistic investigation into the key aspects of myc-and E1A-induced apoptosis in vascular smooth muscle.     

Three‐dimensional morphometric comparison of normal and apoptotic endothelial cells based on laser scanning confocal microscopy observation

Three‐dimensional (3D) morphometric analysis of cellular and subcellular structures provides an effective method for spatial cell biology. Here, 3D cellular and nuclear morphologies are reconstructed to quantify and compare morphometric differences between normal and apoptotic endothelial cells. Human umbilical vein endothelial cells (HUVECs) are treated with 60 μM H₂O₂ to get apoptotic cell model and then a series of sectional images are acquired from laser scanning confocal microscopy. The 3D cell model containing plasma membrane and cell nucleus is reconstructed and fused utilizing three sequential softwares or packages (Mimics, Geomagic, and VTK). The results reveal that H₂O₂ can induce apoptosis effectively by regulating the activity of apoptosis‐related biomolecules, including pro‐apoptotic factors p53 and Bax, and anti‐apoptotic factor Bcl‐2. Compared with the normal HUVECs, the apoptotic cells exhibit significant 3D morphometric parameters (height, volume and nucleus‐to‐cytoplasm ratio) variation. The present research provides a new perspective on comparative quantitative analysis associated with cell apoptosis and points to the value of LSCM as an objective tool for 3D cell reconstruction. Microsc. Res. Tech. 76:1154–1162, 2013. © 2013 Wiley Periodicals, Inc.     

Oxidative stress indicators in human and bottlenose dolphin leukocytes in response to a pro-inflammatory challenge

Marine mammals undergo cycles of tissue ischemia and reperfusion during the dive response. Reperfusion injury can result in oxidative tissue damage and the activation of a pro-inflammatory immune response. The risk of oxidative damage is reduced by antioxidants. Our hypothesis is that the reported higher antioxidant defenses within marine mammal tissues provide additional protection in situations that produce oxidative stress, like inflammation, in comparison to terrestrial mammal tissues. Leukocytes were isolated from the whole blood of Pacific bottlenose dolphins (Tursiops truncatus gilli) and humans (Homo sapiens) and were exposed to lipopolysaccharides (LPS, 10 µg/mL) in vitro to simulate a pro-inflammatory challenge. Oxidative stress indicators, including superoxide radical (O₂^•−) production, activities of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), glutathione reductase (GR), and glutathione S-transferase (GST), as well as oxidative protein damage, were quantified by spectrophotometry. Following 48 h under experimental conditions, bottlenose dolphin leukocytes produced 1.9 times more O₂^•− but displayed 2.0 times lower protein carbonyl concentrations compared to human leukocytes. Following 48 h under experimental conditions, bottlenose dolphin leukocytes displayed 7.9, 2.0, 11.1, and 3.3 times more activities of CAT, GPx, GR, and GST, respectively, compared to human leukocytes. These results suggest that, under cell culture conditions, the antioxidant defenses in bottlenose dolphin leukocytes provide additional protection against pro-inflammatory challenges in comparison to human leukocytes, likely as an adaptive advantage.     

Nitric oxide alleviates cadmium-impeded growth by limiting ROS accumulation in pea seedlings

Cadmium (Cd) causes oxidative stress, which leads to the oxidation of various biomolecules by the production of reactive oxygen species (ROS) to facilitate programmed cell death (PCD). The antioxidant defense system fails to detoxify ROS when it is produced in excess. Nitric oxide (NO), a gaseous free radical and a phytohormone, regulates various physiological processes of plants. Therefore, this work was undertaken to study the effects of the application of exogenous sodium nitroprusside (SNP, a NO donor) on growth parameters, oxidative stress, accumulation of secondary metabolites, and activities of antioxidant enzymes under Cd stress. Mild (50 µM) and severe (200 µM) Cd stress were applied to hydroponically grown pea (Pisum sativum L.) plants with or without 50 µM SNP. Severe Cd stress had a substantial impact on the plants. The effectiveness of NO in reducing Cd-induced negative effects on plant height, fresh weight, dry weight, protein content, nitrite content, nitrate reductase (NR) activity, catalase activity, and peroxidase activity were investigated. Seedling development, protein content, nitrite content, nitrate reductase (NR) activity, antioxidant defense systems disruption, overproduction of reactive oxygen species, and oxidative damage were observed. The antioxidant defense system (catalase and peroxidase activities) was activated by NO, which resulted in lower lipid peroxidation and lower hydrogen peroxide (H₂O₂) levels in Cd-exposed plants. SNP treatment boosted endogenous NO levels and NR activity in Cd-stressed plants while also enhanced proline levels to preserve osmotic equilibrium. The presence of total phenols and flavonoids increased after SNP treatment, indicating that SNP enhanced stress recovery and boosted plant development in Cd-stressed plants.

     

Inhibition of H₂O₂-induced apoptosis of GC2-spg cells by functionalized selenium nanoparticles with lentinan through ROS-mediated ERK/p53 signaling pathways

A H₂O₂-induced oxidative stress injury cell model was established to investigate the antioxidant effect of nano-selenium on mouse spermatocyte lines and the regulation mechanism of the expression level and activity of selenium-containing antioxidant enzymes induced by oxidative stress. A safe and effective nano-drug system of functionalized selenium-containing nanoparticles (SeNPs) was developed with lentinan (LNT) (SeNPs@LNT). Mice spermatocyte line GC2-spg cells were treated with SeNPs@LNT (1, 2, 4, 8, 16, 32 μM) for 24–72 h to evaluate the cytotoxicity of selenium. GC2-spg cells were randomly divided into the following groups: control, hydrogen peroxide (H₂O₂), SeNPs@LNT, and H₂O₂+SeNPs@LNT groups. H₂O₂+SeNPs@LNT group was pretreated with SeNPs@LNT 4 μM for 12 h, followed by H₂O₂ 600 μM for 8 h. The cell viability decreased in the H₂O₂ group and increased significantly in the SeNPs@LNT group. Compared with the H₂O₂ group, the SeNPs@LNT+H₂O₂ group exhibited obvious red fluorescence, indicating a higher level of mitochondrial membrane potential. The content of intracellular reactive oxygen species (ROS) in the SeNPs@LNT group reduced significantly, and the intensity of green fluorescence in the SeNPs@LNT+H₂O₂ group decreased significantly compared with the H₂O₂ group, indicating the inhibitory effect of SeNPs@LNT on the generation of ROS-induced oxidative stress. The activity of GPx and SOD increased significantly in the SeNPs@LNT group. The expression of p53 decreased significantly under the intervention of nano-selenium, and GPx1 expression increased. In the oxidative stress group, the expressions of DNA damage-related proteins and apoptosis-related proteins were higher than those in other groups. Thus, SeNPs@LNT can promote GC2-spg cell proliferation, improve GPx and SOD activities, remove intracellular ROS, and reduce mitochondrial damage and functional abnormalities caused by oxidative stress by regulating the ERK and p53 protein levels. SeNPs@LNT has good biological activity and antioxidant effect, which can be used to protect the male reproductive system from oxidative stress.     

Agmatine pretreatment protects retinal ganglion cells (RGC-5 cell line) from oxidative stress <i>in vitro</i>

Agmatine, 2-(4-aminobutyl)guanidine, has been reported to have neuroprotective effects against various neuronal damages. In this study it was investigated whether agmatine pretreatment rescues the retinal ganglion cells from oxidative injury in vitro. After differentiation of transformed rat retinal ganglion cells (RGC-5 cell line) with staurosporine, agmatine (0.0 to 100.0 μM) pretreatment was performed for 2 hours. Subsequently, they were exposed to hydrogen peroxide (0.0 to 2.5 mM) as an oxidative stress. Cell viability was monitored for up to 48 hours with the lactate dehydrogenase (LDH) assay and apoptosis was examined by the terminal deoxynucleotide transferase-mediated terminal uridine deoxynucleotidyl transferase nick end-labeling (TUNEL) method. As a result, differentiated RGC-5 cells were found to have decreased viability after addition of hydrogen peroxide in a dose-dependent manner. This hydrogen peroxide induced cytotoxicity caused apoptosis characterized by DNA fragmentation. Agmatine pretreatment not only increased cell viability but also attenuated DNA fragmentation. In conclusion, agmatine pretreatment demonstrated neuroprotective effects against oxidative stress induced by hydrogen peroxide in differentiated RGC-5 cells in vitro. This suggests a novel therapeutic strategy rescuing retinal ganglion cells from death caused by oxidative injury     

Finite element modelling of skeletal muscles coupled with fatigue

In this study, Hill's muscle theory coupled with fatigue was proposed to describe mechanical behaviours of skeletal muscles. The force developed by a fatigued muscle was described by a muscle fatigue formula which was a time function of the activation α_a and the stretch λ. The modified Hill's muscle theory was hence incorporated into a three-dimensional (3D) finite element model using the PAK finite element code. In this paper, the theoretical derivation of the 3D muscle model was firstly described. After presenting the method of establishing the finite element programme, a case example of studying the mechanical response of a frog gastrocnemius muscle was used to illustrate the applicability of the proposed methodology. The effects of the muscle fatigue on the deformation as well as the stress and strain distribution of the frog muscle subject to a cyclic activation function have been determined. An experiment capturing the real-time shape change of a frog muscle was also conducted to assess the applicability of the proposed method. A comparison between the deformed shapes of the predictive model and the frog muscles was also made. It was shown that the method is capable of providing a reasonable model for describing the mechanical behaviour of skeletal muscles.     

Insight into 5-aminolevulinic acid-induced modulation of cellular antioxidant metabolism to confer salinity and drought tolerance in maize

The current study investigated the comparative oxidative damage in two maize seedlings induced by saline, drought, and combined stress and the ameliorative role of two different doses (20 and 80 µM) of 5-aminolevulinic acid (ALA) against the above-mentioned stresses. Hydroponically grown 10-day-old maize (Zea mays, var. BARI Hybrid Maize-7 (BHM-7) and BARI Hybrid Maize-9 (BHM-9)) seedlings were exposed to 12 dS/m of saline solution, 200 mM mannitol-induced drought stress alone and their combined stress for 7 days. Result revealed that individual stresses retard the plant growth to some degrees; however, their combined stress has more detrimental effects, which might be correlated with lipid peroxidation (MDA)-induced oxidative stress in seedlings, enhanced Na⁺ /K⁺ ratio, and augmented generation of superoxide (O₂^•− ) and hydrogen peroxide (H₂O₂). In contrast, exogenous ALA supplementation at 20 µM concentration markedly recovered from chlorosis and growth inhibition, substantially scavenged reactive oxygen species (ROS) and MDA by preserving ionhomeostasis and relaxing oxidative stress; also, by boosting catalase (CAT) and glutathione S-transferase (GST), and exclusively via depressing the activity of lipoxygenase (LOX) antioxidant enzyme. On the contrary, 80 µM ALA made things worse; nevertheless, higher activities shown by other antioxidant enzymes; like, superoxide dismutase (SOD), ascorbate peroxidase (APX), peroxidase (POD), and glutathione peroxidase (GPX), which were related to lessen the oxidative damage by highly produced O₂^•− and H₂O₂ under combined stress. Non-denaturing gel electrophoresis was done for further confirmation. However, ALA importantly increased the photosynthetic pigment contents in both genotypes irrespective of doses. Nevertheless, GST might have assisted the plants to escape from the herbicidal effect by detoxification. However, in the combined stress condition, high ALA concentration may have some positive role to play. Our findings also showed that BHM-9 performed better than BHM-7. Therefore, ALA at lower concentration was effective for single stress of saline and drought, while higher concentration can improve plant survival under combined stress.     

Imaging of reactive oxygen species burst from mitochondria using laser scanning confocal microscopy

Objective: Although several methods have been used to detect the intracellular reactive oxygen species (ROS) generation, it is still difficult to determine where ROS generate from. This study aimed to demonstrate whether ROS generate from mitochondria during oxidative stress induced mitochondria damage in cardiac H9c2 cells by laser scanning confocal microscopy (LSCM). Methods: Cardiac H9c2 cells were exposed to H₂O₂ (1200μM) to induce mitochondrial oxidant damage. Mitochondrial membrane potential (ΔΨm) was measured by staining cells with tetramethylrhodamine ethyl ester (TMRE); ROS generation was measured by staining cells with dichlorodihydrofluorescein diacetate (H₂DCFDA). Results: A rapid/transient ROS burst from mitochondria was induced in cardiac cells treated with H₂O₂ compared with the control group, suggesting that mitochondria are the main source of ROS induced by oxidative stress in H9c2 cells. Meanwhile, the TMRE fluorescence intensity of mitochondria which had produced a great deal of ROS decreased significantly, indicating that the burst of ROS induces the loss of ΔΨm. In addition, the structure of mitochondria was damaged seriously after ROS burst. However, we also demonstrated that the TMRE fluorescence intensity might be affected by H₂DCFDA. Conclusions: Mitochondria are the main source of ROS induced by oxidative stress in H9c2 cells and these findings provide a new method to observe whether ROS generate from mitochondria by LSCM. However, these observations also suggested that it is inaccurate to test the fluorescence intensities of cells stained with two or more different fluorescent dyes which should be paid more attention to. Microsc. Res. Tech. 76:612–617, 2013. © 2013 Wiley Periodicals, Inc.     

Involution dependent changes in distribution and localization of bax,survivin, caspase‐3, and calpain‐1 in the rat endometrium

The endometrial layer of the uterus is characterized by continuous cycle of cell growth and apoptosis in response to hormonal changes. Apoptosis is regulated by several apoptotic regulators, but their significance in involuting uterus has not been well understood. For that reason, aim of this study was to investigate possible role of apoptosis‐related proteins (bax and survivin) and enzymes (caspase‐3 and calpain‐1) in the involuting uterus of the rat, using immunohistochemistry. Our results indicated cytoplasmic and nuclear immunostaining for bax, caspase‐3, calpain‐1 and survivin proteins were found in the endometrial epithelium and stromal cells such as fibroblasts, mast cells and macrophages, and blood vessels; however, calpain‐1 immunoreactivity in the endometrial fibroblast was quite weak or absent. Supranuclear punctate bax immunolabelling was also observed in the endometrial fibroblasts and luminal and glandular epithelial cells from days 1st and 3rd following parturition, respectively. Although survivin was localized in the apical cytoplasm underneath the apical membrane of the luminal epithelium on the 1st and 3rd days, it was also localized in the apicolateral membrane and basal cytoplasm on the 10th and 15th days of involution. Immunostainigs demonstrated that expression patterns of all examined proteins varied with structural changes in the luminal epithelium, and number of immunopositive fibroblasts for bax, caspase‐3 and survivin increased with advance of postpartum days and reached a maximum on postpartum days 10 and 15. These results suggest that the process of postpartum involution of endometrium may be regulated by apoptotic and non‐apoptotic activity of bax, caspase‐3, calpain‐1, and survivin. Microsc. Res. Tech. 79:285–297, 2016. © 2016 Wiley Periodicals, Inc.     

Application of ferrous sulfate alleviates negative impact of cadmium in rice (<i>Oryza sativa</i> L.)

Soil contamination with toxic heavy metals [such as cadmium (Cd)] is becoming a serious global problem due to rapid development of social economy. Iron (Fe), being an important element, has been found effective in enhancing plant tolerance against biotic and abiotic stresses. The present study investigated the extent to which different levels of Ferrous sulphate (FeSO₄) modulated the Cd tolerance of rice (Oryza sativa L.), when maintained in artificially Cd spiked regimes. A pot experiment was conducted under controlled conditions for 146 days, by using natural soil, mixed with different levels of CdCl₂ [0 (no Cd), 0.5 and 1 mg/kg] together with the exogenous application of FeSO₄ at [0 (no Fe), 1.5 and 3 mg/kg] levels to monitor different growth, gaseous exchange characteristics, oxidative stress, antioxidative responses, minerals accumulation, organic acid exudation patterns of O. sativa. Our results depicted that addition of Cd to the soil significantly (P < 0.05) decreased plant growth and biomass, gaseous exchange parameters, mineral uptake by the plants, sugars (soluble, reducing, and non-reducing sugar) and altered the ultrastructure of chloroplasts, plastoglobuli, mitochondria, and many other cellular organelles in Cd-stressed O. sativa compared to those plants which were grown without the addition of Cd in the soil. However, Cd toxicity boosted the production of reactive oxygen species (ROS) by increasing the contents of malondialdehyde (MDA), which is the indication of oxidative stress in O. sativa and was also manifested by hydrogen peroxide (H₂O₂) contents and electrolyte leakage to the membrane bounded organelles. Although, activities of various antioxidative enzymes like superoxidase dismutase (SOD), peroxidase (POD), catalase (CAT) and ascorbate peroxidase (APX) and non-enzymatic antioxidants like phenolics, flavonoid, ascorbic acid, anthocyanin and proline contents increased up to a Cd level of 0.5 mg/kg in the soil but were significantly diminished at the highest Cd level of 1 mg/kg in the soil compared to those plants which were grown without the addition of Cd in the soil. The negative impacts of Cd injury were reduced by the application of FeSO₄ which increased plant growth and biomass, improved photosynthetic apparatus, antioxidant enzymes, minerals uptake together with diminished exudation of organic acids as well as oxidative stress indicators in roots and shoots of O. sativa by decreasing Cd retention in different plant parts. These results shed light on the effectiveness of FeSO₄ in improving the growth and upregulation of antioxidant enzyme activities of O. sativa in response to Cd stress. However, further studies at field levels are required to explore the mechanisms of FeSO₄-mediated reduction of the toxicity of not only Cd, but possibly also other heavy metals in plants.     

Antioxidant capacity of vitamin C in mouse liver and kidney tissues

In the present study, the antioxidant capacity of vitamin C was examined in the liver and the kidney tissues of mice with or without ciprofloxacin (CFX) treatment. The antioxidant capacity of the vitamin was evaluated in terms of lipid hydroperoxides (LOOH) and thiobarbituric acid reactive substances (TBARs). The experimental design was 15 days of water (control and CFX groups) or vitamin C (vitamin C and vitamin C plus CFX groups) in drinking water. One dose of CFX was injected, 15 minutes before sacrifice, in the corresponding mice. The initial nmol of lipid hydroperoxides/g of tissue were 137 +/- 11 in the kidney and 145 +/- 15 in the liver, and the nmol of TBARs were 13 +/- 0.7 and 12 +/- 0.6, respectively. Pre-treatment with vitamin C reduced the levels of LOOH in the liver to 45 +/- 11 (p < 0.01) and vitamin C with CFX injection to 54 +/- 9 (p < 0.01). Vitamin C treatment also reduced the LOOH levels in the kidney roughly duplicated by CFX. Through the TBARs method we have not observed these effects. Quantification of LOOH is more sensitive than that of TBARs for estimating lipid peroxidation. CFX is used especially for urinary infections and can produce oxidative stress in the kidney. Pre-treatment with vitamin C may ameliorate this stress and also may improve the oxidative balance in the liver.     

Oxides and apoptosis in inflammatory myopathies.

Reactive oxygen intermediates (ROI) and nitric oxide (NO(.)) are produced in abundance in the inflammatory muscle diseases of autoimmune origin polymyositis (PM), dermatomyositis (DM), and inclusion body myositis (IBM). However, their role in the pathogenesis of these diseases is so far not clear. In contrast to demyelinating neuropathies, there is no convincing evidence for oxide-induced apoptosis either in myocytes or in lymphocytes and phagocytes in inflammatory myopathies. On the contrary, NO(.) released at low concentrations at target sites may even have cell-protective effects. A major mechanism of protection from apoptosis in both myocytes and inflammatory cells seems to be the upregulation of anti-apoptotic proteins like Bcl-2. Caution is warranted to apply antioxidative and anti-apoptotic agents to patients with inflammatory myopathies as long as the pathogenic role of oxides and apoptosis in the individual case is not resolved.     

In vitro antitumor activity of broccolini seeds extracts

Broccolini (Brassica oleracea Italica × Alboglabra) is a hybrid of broccoli and kai‐lan, Chinese broccoli. To date, no report on antitumor activity of Broccolini (NOT Broccoli) is available. In this study, we evaluated the antiproliferative effects of broccolini seeds extract (BSE) on human lung and ovarian cancer cells. It was found that BSE induces A549 and OVCAR‐3 cells apoptosis in a dose‐dependent manner by using MTT assay. The IC₅₀ values of BSE in A549 and OVCAR‐3 cells were estimated to be 81.94 and 78.6 µg/ml, respectively. Furthermore, the phase contrast microscope showed that in high‐dose group (90～120 µg/ml), the morphology structure of OVCAR‐3 cells become irregular and exhibited characteristics of apoptosis such as cell membrane shrinkage, condensation and fragmentation of nuclear chromatin as well as formation of apoptotic bodies. SCANNING 33:402–404, 2011. © 2011 Wiley Periodicals, Inc.