Protective effects of anise in prevention of cerebral ischemia reperfusion injury in rats.

The aim of this study is to determine the protective efficacy of anise in cerebral ischemia and reperfusion injury in rats. In this study, 28 Wistar Albino rats, weighing 250-300 grams (g), were used. Four groups were formed with 7 rats in each group. Group 1 (n=7): Control group, Group 2 (n=7): Anise group, 5 mL/kg/day of anise aqueous extract prepared according to Gamberini's protocol was given orally by gavage for 30 days. Group 3 (n=7): Cerebral ischemia reperfusion (CIR) group, at the beginning of the experiment, 30 minutes of cerebral ischemia and 1 hour of reperfusion were induced and the animals were sacrificed by exanguination. Group 4 (n=7): Anise+ CIR group, After administering 30 days of anise's aqueous extract, CIR was induced and the study was terminated. TOS values of the Anise+ CIR group was significantly lower than that of the CIR group (p<0.05). Il-6 and TNF-α values of the CIR group were significantly higher than the Anise+ CIR group (p<0,05). Our study revealed that anise ameliorates oxidative damage and inflammation due to cerebral ischemia/reperfusion, by reducing the levels of inflammatory cytokines (TNF-α, Il-6).

Enhancing Vicia faba's immunity against Rhizoctonia solani root rot diseases by arbuscular mycorrhizal fungi and nano chitosan.

BACKGROUND: The spreading of root rot disease of faba bean plant (Vichia faba L, VF) in Egypt is still of great challenge faced researchers since VF is an important legume in Egypt, because their seeds are used for human feeding. Fungicides are used for treatment of either seeds or soil; unfortunately they cause environmental pollution. Therefore, there is a need to continue research to find out safe natural solutions. In this regard, Arbuscular mycorrhizal fungi (AMF) and chitosan (micro or nanoform) were used as an inhibitory product against Rhizoctonia solani OM918223 (R.solani) either singly or in combinations. RESULTS: The results employed herein have exhibited that R.solani caused root rot disease of VF plants in more than 80% of the plants under investigation. Chitosan nanoparticles (Chitosan NPs) were prepared by ionic gelatin method and characterized by using dynamic light scattering (DLS), transmission electron microscopy (TEM) imaging and Fourier transform infra-red (FTIR). Chitosan NPs are spherical with a diameter of 78.5 nm and exhibited the presence of different functional groups. The inhibitory natural products against R.solani were arranged according to their ability to inhibit the pathogen used in the following descending manner; combination of AMF with Chitosan NPs, AMF with micro chitosan and single AMF, respectively. Where, Chitosan NPs showed a potent influence on R.solani pathogen and reduced the pre-and post-emergence of R. solani. In addition, Chitosan NPs reduced Disease Incidence (DI %) and Disease Severity (DS %) of root rot disease and are widely functional through mixing with AMF by about 88% and 89%. Further, Chitosan NPs and micro chitosan were proved to increase the growth parameters of VF plants such as nutritional status (mineral, soluble sugar, and pigment content), and defense mechanisms including total phenol, peroxidase, and polyphenol oxidase in mycorrhizal plants more than non-mycorrhizal one either in infected or healthy plants. Moreover, activity of AMF as an inhibitory against R.solani and improvement natural agent for VF growth parameters was enhanced through its fusing with Chitosan NPs. CONCLUSIONS: The use of AMF and Chitosan NPs increased faba bean plant resistance against the infection of root rot R. solani, with both prevention and cure together. Therefore, this research opens the door to choose natural and environmental friendly treatments with different mechanisms of plant resistance to disease.

Distribution of natural radionuclides in NORM samples from North Abu Rusheid area, Egypt.

The North Abu Rusheid area in Egypt is a well-known high background natural radiation area (HBNRA) due to the existence of naturally occurring radioactive materials (NORMs) in mylonitic rocks. In this study, 27 rock samples were selected for dose estimation studies. 238U and 232Th were measured using inductively coupled plasma mass spectrometry (ICP-MS) and 40K was measured using sodium iodide (thallium) gamma-ray spectroscopy. The ranges of activity concentrations (Bq/kg) of 238U, 232Th and 40K in the samples varied from 270 ± 2 to 2120 ± 29, 350 ± 2 to 1840 ± 27 and 20 ± 2 to 1390 ± 35 with mean values of 980 ± 349, 770 ± 351, and 640 ± 402 Bq/kg, respectively. The radiological hazard parameters were estimated from activity concentrations of 238U, 232Th and 40K and compared to United Nations Scientific Committee on the Effects of Atomic Radiation (UNSCEAR) values. The present study revealed that the hazard parameters were several times higher than the worldwide averages. The U/Th concentration ratio ranged from 0.7 to 3 and could be attributed to the presence of kasolite, uranothorite, zircon, and columbite in mylonitic rocks. From the radiological protection viewpoint, it is necessary to monitor natural radionuclides in these rocks prior to their use in residential and commercial construction materials.

Enhancing the Potentiality of Trichoderma harzianum against Pythium Pathogen of Beans Using Chamomile (Matricaria chamomilla, L.) Flower Extract.

Our present study was designed to investigate the role of both Trichoderma harzianum and chamomile (Matricaria chamomilla L.) flower extract in mutual reaction against growth of Pythium ultimum. In vitro, the activity of chamomile extract was found to reduce the radial growth of Pythium ultimum up to 30% compared to the control. Whereas, the radial growth reduction effect of T. harzianum against P. ultimum reached 81.6% after 120 h. Data also showed the productivity of total phenolics and total flavonoids by T. harzianum, was 12.18 and 6.33 mg QE/100 mL culture filtrate, respectively. However, these compounds were determined in chamomile flower extract at concentrations of 75.33 and 24.29 mg QE/100 mL, respectively. The fractionation of aqueous extract of chamomile flower using HPLC provided several polyphenolic compounds such as pyrogallol, myricetin, rosemarinic acid, catechol, p-coumaric acid, benzoic acid, chlorogenic acid and other minor compounds. In vivo, the potentiality of T. harzianum with chamomile flower extract against Pythium pathogen of bean was investigated. Data obtained showed a reduction in the percentage of rotted seed and infected seedling up to 28 and 8%, respectively. Whereas, the survival increased up to 64% compared to other ones. There was also a significant promotion in growth features, total chlorophyll, carotenoids, total polyphenols and flavonoids, polyphenol-oxidase and peroxidase enzymes compared to other ones. To the best of our knowledge, there are no reported studies that included the mutual association of fungus, T. harzianum with the extract taken from the chamomile flower against P. ultimum, either in vitro or in vivo. In conclusion, the application of both T. harzianum and/or M. chamomilla extracts in the control of bean Pythium pathogen showed significant results.

Acute effect of cadmium treatment on the kidney of rats: biochemical and ultrastructural studies.

The present study was designed to explore the nephrotoxic effect of intraperitoneal acute administration of CdCl2 (2.5 and 5 mg kg(-1) b.w.) in rats. A number of toxicological parameters in kidney were examined including malondialdehyde (MDA) and endogenous antioxidants, e.g., catalase (CAT), superoxide dismutase (SOD) and Glutathione Peroxidase (GPx). The parameters that indicate tissue damage such as serum urea and creatinine were also determined, along with the ultrastructural changes of kidneys. A correlation was found between the dose and the intensity of changes. The results demonstrated that cadmium administration increased renal MDA but decreased CAT, SOD and GPx activities. In parallel, serum creatinine and urea elevated. The glomerular ultrastructural changes observed in cadmium-treated rats included narrowing of the capillary lumen and swelling of the capillary endothelium with occasional loss of fenestrae. The mesangium was wide with increased mesangial matrix. Loss of homogenous appearance of basement membrane displaying ondulation and thickening in many areas and deterioration of the slit membrane structures formed by the podocytes were also noted. The effects of cadmium on proximal cell ultrastructure were focal loss of brush border, nuclear membrane damage, chromatin condensation, swelling of the mitochondria with regression of mitochondrial cristae, degranulation and disintegration of protein-synthesizing structures such as rough endoplasmic reticulum, increased number of lysosomes and ultimately cell death.

Altered brain glucose metabolism in transgenic-PFKL mice with elevated L-phosphofructokinase: in vivo NMR studies.

The gene for the liver-type subunit of phosphofructokinase (PFKL) resides on chromosome 21 and is overexpressed in Down syndrome (DS) patients. Transgenic PFKL (Tg-PFKL) mice with elevated levels of PFKL were used to determine whether, as in DS, overexpression of PFKL was also associated with altered sugar metabolism. We found that Tg-PFKL mice had an abnormal glucose metabolism with reduced clearance rate from blood and enhanced metabolic rate in brain. Transgenic-PFKL mice exhibited elevated activity of phosphofructokinase in both blood and brain, as compared to control non-transgenic (ntg) mice. Following glucose infusion, the rate of glucose clearance from the blood of Tg-PFKL mice was significantly slower than that of control ntg mice, although the basal blood glucose levels were similar. However, unlike the slower rate of glucose metabolism in blood, the initial rate of glucose utilization in the brain of the transgenic mice, was 58% faster than in control ntg mice. This was determined by infusion of [1-13C]-glucose followed by in vivo nuclear magnetic resonance (NMR) measurements of brain glucose metabolism. The faster utilization of glucose in Tg-PFKL brain is similar to the increased rate of cerebral glucose metabolism found in the brain of young adult DS patients, which may play a role in the etiology of their cognitive disabilities.

Transgenic mice with elevated level of CuZnSOD are highly susceptible to malaria infection.

Copper/zinc superoxide dismutase (CuZnSOD) catalyses the conversion of O2.- into H2O2. Constitutive overexpression of CuZnSOD in cells and animals creates an indigenous oxidative stress that predisposes them to added insults. In this study, we used transgenic CuZnSOD (Tg-CuZnSOD) mice with elevated levels of CuZnSOD to determine whether overexpression of CuZnSOD affected the susceptibility of these mice to plasmodium infection. Acute malaria is associated with oxidative stress, mediated by redox-active iron released from the infected RBC. Two independently derived Tg-CuZnSOD lines showed higher sensitivity than control mice to infection by Plasmodium berghei (P. berghei), reflected by an earlier onset and increased rate of mortality. Nevertheless, while Tg-CuZnSOD mice were more vulnerable than control mice, the levels of parasitemia were comparable in both strains. Moreover, treatment of infected red blood cells (RBC) with oxidative stress inducers, such as ascorbate or paraquat, reduced the viability of parasites equally in both transgenic and control RBC. This further confirms that increased CuZnSOD does not support plasmodia development. The data are consistent with the possibility that the combination of increased redox-active iron and elevated H2O2 in the plasmodium-infected Tg-CuZnSOD mice, led to an enhanced Fenton's reaction-mediated HO. production, and the resulting oxidative injury renders the transgenic mice more vulnerable to parasite infection.

Oxidative stress mediates impairment of muscle function in transgenic mice with elevated level of wild-type Cu/Zn superoxide dismutase.

Cases of familial amyotrophic lateral sclerosis (fALS; a neurodegenerative disorder) have been reported in which the gene for Cu/Zn superoxide dismutase (CuZnSOD) was mutated. Several studies with the fALS mutant CuZnSOD in transgenic mice and cells showed that the fALS mutations act through an as yet undefined dominant gain-of-function mechanism. Wild-type CuZnSOD catalyzes the dismutation of superoxide (O(2)(-).) but also produces hydroxyl radicals (.OH) with H(2)O(2) as substrate. Two laboratories have recently demonstrated that the .OH production ability was preferentially enhanced by the fALS mutant CuZnSOD, suggesting that this might be the function gained in fALS. In this study, we used transgenic CuZnSOD (Tg-CuZnSOD) mice with elevated levels of CuZnSOD to determine whether overexpression of wild-type CuZnSOD was also associated with increased .OH production and impaired muscle function. Enhanced formation of .OH was detected, by spin trapping, in brain and muscle extracts of the Tg-CuZnSOD mice. Three independently derived Tg-CuZnSOD lines showed muscle abnormalities, reflected by altered electromyography (EMG) and diminished performance in the rope grip test. After treatment with paraquat (PQ), a widely used herbicide and O(2)(-).-generating compound, muscle disability significantly deteriorated in Tg-CuZnSOD mice but not in control mice. The results indicate that elevated levels of CuZnSOD cause indigenous long-term oxidative stress leading to impairment of muscle function. These findings may provide valuable clues about the concurred role of indigenous oxidative stress and exogenous agents in the etiology of sporadic ALS and several other neurodegenerative diseases in which a specific subset of neurons is affected.

Cellular oxidative stress and the control of apoptosis by wild-type p53, cytotoxic compounds, and cytokines.

Apoptosis induced by wild-type p53 or cytotoxic compounds in myeloid leukemic cells can be inhibited by the cytokines interleukin 6, interleukin 3, granulocyte-macrophage colony-stimulating factor, and interferon gamma and by antioxidants. The antioxidants and cytokines showed a cooperative protective effect against induction of apoptosis. Cells with a higher intrinsic level of peroxide production showed a higher sensitivity to induction of apoptosis and required a higher cytokine concentration to inhibit apoptosis. Decreasing the intrinsic oxidative stress in cells by antioxidants thus inhibited apoptosis, whereas increasing this intrinsic stress by adding H2O2 enhanced apoptosis. Induction of apoptosis by wild-type p53 was not preceded by increased peroxide production or lipid peroxidation and the protective effect of cytokines was not associated with a decrease in these properties. The results indicate that the intrinsic degree of oxidative stress can regulate cell susceptibility to wild-type p53-dependent and p53-independent induction of apoptosis and the ability of cytokines to protect cells against apoptosis.

Thymic abnormalities and enhanced apoptosis of thymocytes and bone marrow cells in transgenic mice overexpressing Cu/Zn-superoxide dismutase: implications for Down syndrome.

The copper-zinc superoxide dismutase (CuZnSOD) gene resides on chromosome 21 and is overexpressed in Down syndrome (DS) patients. Transgenic CuZnSOD mice with elevated levels of CuZnSOD were used to determine whether, as in DS, overexpression of CuZnSOD was also associated with thymus and bone marrow abnormalities. Three independently derived transgenic CuZnSOD strains had abnormal thymi showing diminution of the cortex and loss of corticomedullary demarcation, resembling thymic defects in children with DS. Transgenic CuZnSOD mice were also more sensitive than control mice to in vivo injection of lipopolysaccharide (LPS), reflected by an earlier onset and enhanced apoptotic cell death in the thymus. This higher susceptibility to LPS-induced apoptosis was associated with an increased production of hydrogen peroxide and a higher degree of lipid peroxidation. When cultured under suboptimal concentrations of interleukin 3 or in the presence of tumour necrosis factor, bone marrow cells from transgenic CuZnSOD mice produced 2- to 3-fold less granulocyte and macrophage colonies than control. The results indicate that transgenic CuZnSOD mice have certain thymus and bone marrow abnormalities which are similar to those found in DS patients, and that the defects are presumably due to an increased oxidative damage resulting in enhanced cell death by apoptosis.

Prevention of diabetes-associated embryopathy by overexpression of the free radical scavenger copper zinc superoxide dismutase in transgenic mouse embryos.

OBJECTIVES: It has recently been suggested that oxygen free radicals are involved in the high incidence of fetal dysmorphogenesis that is associated with diabetic pregnancies. The purpose of the current investigation was to study the effect of copper zinc superoxide dismutase, a free radical scavenging enzyme, on the prevention of diabetes-associated embryopathy in mice. STUDY DESIGN: Mice used in this study were either transgenic, bearing the human copper zinc superoxide dismutase gene, or nontransgenic controls. Diabetes was generated by streptozotocin administration on days 6 and 7 of gestation. Hyperglycemia developed on day 8 and was maintained through day 10 (critical period of organogenesis). On day 10 fetuses were examined for external anomalies, and their crown-rump lengths and deoxyribonucleic acid content were determined. RESULTS: Induction of maternal diabetes produced a significant reduction in mean crown-rump length of control embryos (4.48 +/- 0.7 mm vs 3.65 +/- 0.6 mm, p = 0.0001), whereas transgenic embryos were not affected (4.72 +/- 0.6 mm vs 4.45 +/- 0.8 mm, p > 0.05). After induction of diabetes fetal loss and malformation rates were significantly higher in control embryos (6.0% vs 23.8% and 8.4% vs 16.5%, respectively). Transgenic embryos were practically unaffected by diabetes and showed fetal loss and malformation rates of 5.9% and 4.4%, respectively, after induction of diabetes. CONCLUSIONS: Elevated levels of copper zinc superoxide dismutase, a key enzyme in the metabolism of free oxygen radicals, elicit a protective effect against diabetes-associated embryopathy.