Determination of DNA damage induced by oxidative stress in hyperlipidemic patients.

In the present paper, we report data on the genotoxic properties of hydrogen peroxide in polymorphonuclear neutrophils (PMNLs) separated from normolipidemic and type II/a hyperlipidemic patients. In all, 15 hyperlipidemic patients (11 female, 4 male, mean age 54.6+/-10.25 years) were involved in the study, and 7 normolipidemic patients (5 female, 2 male, mean age 53.4+/-8.07 years) served as controls. Using the comet assay, there was a significant difference in the degree of DNA damage between the two groups. The visual score characteristic of the degree of DNA damage was 350.97+/-31.31 in the hyperlipidemic group, while it was 289.5+/-29.49 in the control group (P<0.001). In the hyperlipidemic patients, a positive correlation was found between the degree of DNA damage and the basic oxidation of PMNLs (r=0.517), and the superoxide anion production of the cells stimulated with phorbolmiristate acetate (PMA) (r=0.326) and formyl-Met-Leu-Phe (FMLP) (r=0.525) as well. There was a negative correlation between DNA damage and HDL-associated antioxidant paraoxonase (PON) activity (r=-0.469), and the PON/HDL ratio (r=-0.631). No correlation was found between the degree of DNA damage and the plasma concentration of nitric oxide (NO) (r=0.098) and thiobarbituric acid-reactive substances (TBARS) (r=0.061) in hyperlipidemic patients. Our results show that in hyperlipidemic patients there is an increase in lymphocyte DNA damage caused by oxidative stress when compared to normolipidemic individuals as demonstrated by comet assay. Decreased antioxidant capacity in hyperlipidemic patients may play a significant role in this process.     

In field damage of high and low cyanogenic cassava due to a generalist insect herbivore Cyrtomenus bergi (Hemiptera: Cydnidae)

The hypothesis that cyanogenic potential in cassava roots deters polyphagous insects in the field is relevant to current efforts to reduce or eliminate the cyanogenic potential in cassava. To test this hypothesis, experiments were conducted in the field under natural selection pressure of the polyphagous root feeder Cyrtomenus bergi Froeschner (Hemiptera: Cydnidae). A number of cassava varieties (33) as well as 13 cassava siblings and their parental clone, each representing a determined level of cyanogenic potential (CNP), were scored for damage caused by C. bergi and related to CNP and nonglycosidic cyanogens, measured as hydrogen cyanide. Additionally, 161 low-CNP varieties (< 50 ppm hydrogen cyanide, fresh weight) from the cassava germplasm core collection at Centro Internacional de Agricultura Tropical (CIAT) were screened for resistance/tolerance to C. bergi. Low root damage scores were registered at all levels of CNP. Nevertheless, CNP and yield (or root size) partly explained the damage in cassava siblings (r2 = 0.82) and different cassava varieties (r2 = 0.42), but only when mean values of damage scores were used. This relation was only significant in one of two crop cycles. A logistic model describes the underlying negative relation between CNP and damage. An exponential model describes the underlying negative relation between root size and damage. Damage, caused by C. bergi feeding, released nonglycosidic cyanogens, and an exponential model fits the underlying positive relation. Fifteen low-CNP clones were selected for potential resistance/tolerance against C. bergi.     

Tolerance of several wild oat herbicides by a range of winter wheat varieties

Screening tests exposed varietal differences in the tolerance of winter wheat to high doses of the wild oat herbicides difenzoquat and diclofop-methyl. Subsequent yield trials at recommended and higher doses confirmed the sensitivity of Score and Sportsman to difenzoquat and showed Atou, Bouquet and Hobbit to be less tolerant than some other varieties. In most varieties, earlier spraying proved safer than treatment at or beyond the beginning of jointing. Varietal differences with diclofop-methyl were smaller and inconsistent. Crops damage from these two herbicides was more severe in another experiment the following season, in which isoproturon at double the recommended dose also caused severe damage but flamprop-methyl was fairly well tolerated by all varieties.     

Ischemia stimulates the release of atrial natriuretic peptide from rat cardiac ventricular myocardium in vitro.

The effect of ischemia on atrial natriuretic peptide (ANP) release from heart ventricles was studied by exposing the perfused hearts of Wistar-Kyoto (WKY) and spontaneously hypertensive (SHR) rats to global ischemia after excision of the atria. Ischemia for 2, 5 and 20 min caused an increase of 0.3 +/- 1.1, 12.4 +/- 5.5 and 11.4 +/- 4.2 ng/g dry weight in ANP release of the WKY ventricles, respectively. ANP release increased 3.4 +/- 2.8 ng/g dry weight after 5 minutes' ischemia from the SHR ventricles. The increase was not caused by cell damage, as only processed form of the peptide was detected in the perfusates. The increase in ANP release in the WKY ventricles correlated positively with the tissue lactate/pyruvate ratio (r = 0.85) and adenosine (r = 0.99), and negatively with the phosphorylation potential (r = -0.70). The results indicate that ventricular ischemia increases ANP release, probably due to changes in myocardial energy metabolism.     

Recovery from terminal and fruit damage by dry season cotton crops in tropical Australia

We conducted a 2-yr field study on growth and yield compensation in cotton (Gossypium hirsutum L.) after artificial pest damage in Kununurra in NW Australia. The aim was to assess the responses of cotton grown in tropical Australia to simulated Helicoverpa damage early in the season (tip damage) and during fruiting (square removal). In this region, cotton is grown in an inverse temperature regimen to cotton grown in temperate regions, and it is possible that tropical cotton crops exhibit different responses to pest damage (Yeates 2001). To examine this, we imposed manual damage treatments by excising plant tips before squaring (flower bud production) and removing all large squares at 800 degree-days DD, (at base 12 degrees C; early square loss) or 1,200 DD (mid-square loss). We found high levels of tolerance in tropical cotton to simulated pest damage (particularly early square loss) equaling or possibly surpassing the compensatory response of cotton grown in temperate Australia. While yield did not differ between damage and control treatments, the trend suggests an increase in yield resulting from tip damage and early fruit loss. Compared with temperate crops, the delay in maturity caused by damage was minimal, owing to high rates of boll opening (up to 11% d(-1)). Early season tip damage resulted in a significant increase in lateral branch growth that later contributed to a higher fruiting potential. The altered canopy of tip-damaged plants also increased light interception, particularly in the upper canopy. Both of the above factors could confer a greater tolerance to subsequent fruit damage in tipped plants.     

Frequency and severity of western bean cutworm (Lepidoptera: Noctuidae) ear damage in transgenic corn hybrids expressing different Bacillus thuringiensis cry toxins

The frequency and severity of corn ear damage caused by western bean cutworm, Striacosta albicosta (Smith), were measured on transgenic corn, Zea mays L., hybrids expressing two different insecticidal Bacillus thuringiensis (Bt) (Berliner) Cry toxins (Bt) selected to protect against damage caused by larval European corn borer, Ostrinia nubilalis (Hübner). A field cage experiment deliberately infested with western bean cutworm egg masses resulted in less damage in the hybrid expressing the Cry1F protein and supported fewer western bean cutworm larvae than its non-Bt isoline. Corn hybrids expressing Cry1F, grown in small plot field experiments at three locations over two separate years and exposed to natural western bean cutworm infestations suffered less damage than non-Bt or Bt-hybrids expressing a Cry1Ab protein. Later maturing hybrids suffered more damage than shorter-season hybrids. Finally, corn ears observed in strip trials for several years in diverse agronomic conditions in farmer-cooperator fields corroborated the in-plant protection conferred by corn hybrids expressing the Cry1F protein in small plot field trials.     

Testing associational resistance against pine weevils mediated by Lasius ants attending conifer seedlings

Natural enemies attracted to plants may provide those plants with protection against herbivores but may also protect neighbouring plants, that is through associational resistance. Ant attendance may be an important mechanism for the occurrence of such effects because ants can reduce the damage caused by a wide variety of herbivorous insects. Ants have been shown, in a previous field experiment, to decrease the damage caused by the pine weevil, Hylobius abietis (L.) (Coleoptera: Curculionidae), a pest species that causes high seedling mortality in forest regeneration areas. In this study, we specifically tested whether seedlings planted close to ant‐attended seedlings experience associational resistance. We did this under laboratory conditions using the ant species Lasius niger (L.) (Hymenoptera: Formicidae). The feeding damage by pine weevils was significantly reduced on seedlings attended by ants. The neighbouring seedlings, however, did not experience associational resistance. Nevertheless, some associational effects were observed as the number of weevils recorded on both ant‐attended and neighbouring seedlings was significantly lower compared with ant‐excluded seedlings.     

Comparison of DNA damage detected by plant comet assay in roadside and non-roadside environments

Although mixed air pollutants generated from traffic are suspected as one of the causes of DNA damage in living species, effects of the combination of these pollutants and other micro-environmental factors on urban biota have not been clarified yet. Thus, this study mainly aimed to detect the genetic damages in selected plant biomonitors, which were ginkgo (Ginkgo biloba), pohtos (Epipremnum aureum), and periwinkle (Vinca rosea), setting in roadside and non-roadside environments. Two monitoring positions were selected in the Hongo campus of the University of Tokyo, Japan. This area was categorized as the urban residential zone. Both roadside and non-roadside samples were analyzed by using comet assay protocol. Different distribution changes in DNA migration ratios of all species could be observed and further interpreted as percentages of DNA damage. For all test species, in the final stage of experiment, roadside samples showed significantly higher degrees of DNA damage than non-roadside one. Time-dependent response pattern of each species to the overall environmental stresses was performed. Increase in the percentages of DNA damage could be expressed by regression equations. In addition, ratio of percentage of DNA damage between roadside and non-roadside species (R/N ratio) was introduced in order to clarify the additional genetic effect caused by roadside air pollutants. Series of response phase of plant under stresses, including cell destabilization, damage intensification, and re-stabilization, were explained. This study might be applied as a preliminary method in urban air quality assessment for detecting the existing effects of air pollutants and micro-environmental stress in an urban ecosystem.     

Dds20 operates in cds1-independent mechanism of tolerance to UV-induced DNA damage in Schizosaccharomyces pombe cells

Repair of DNA double-stranded breaks caused by ionizing radiation or cellular metabolization, homologous recombination, is an evolutionary conserved process controlled by RAD52 group genes. Genes of recombinational repair also play a leading role in the response to DNA damage caused by UV light. Cells with deletion in gene dds20 of recombinational repair were shown to manifest hypersensitivity to the action of UV light at lowered incubation temperature. Epistatic analysis revealed that dds20+ is not a member of the NER and UVER gene groups responsible for the repair of DNA damage induced by UV light. The Dds protein has functions in the Cds1-independent mechanism of UV damage tolerance of DNA.     

Maternal dominance, maternal condition, and offspring sex ratio in ungulate mammals

Trivers and Willard's suggestion that natural selection favors maternal control of offspring sex ratio in relation to maternal condition has been much debated. The theoretical plausibility of the idea, under some conditions, is firmly established, and there is strong empirical support for conditional sex allocation in some taxa. However, the extent to which this hypothesis can be applied to mammals, particularly ungulates, has been more controversial. We used meta-analysis to review published studies of the Trivers-Willard hypothesis within ungulates and to assess the overall level of empirical support for the hypothesis. Overall, data from 37 studies of 18 species suggested a weak but significant positive correlation between maternal condition and sex ratio (r=+0.09). However, average effect size differed markedly between different categories of studies. Studies using measures of maternal condition that were taken preconception and on the basis of behavioral dominance provided strong evidence for a relationship between maternal condition and the sex ratio (r=+0.17-0.25). In contrast, studies that used morphological or physiological measures of condition that were measured postconception provided little or no evidence for a relationship between maternal condition and sex ratio (r=+0.05-0.06). There are several reasons to suggest that data collected postconception and relying on morphological measures of condition are less likely to capture variables that cause selection for biased sex allocation. In addition, we found that the relationship between sex ratio and maternal condition depended on life-history characteristics; relationships were stronger when sexual size dimorphism was more male biased and when gestation periods were longer. Overall, our analyses suggest that data from ungulates are consistent with the Trivers-Willard hypothesis but only when appropriate measures are used.     

An experiment on apoptosis induced by polyamine adducts produced in the presence of serum amine oxidase

A two-session experiment is proposed to train students with an easy, economic and educative procedure to detect the typical DNA ladder produced in many apoptotic events. The procedure is accurate enough to provide an easy way to compare degrees of damage in DNA caused by different treatments.     

Monitoring and modeling of field infestation and damage by the maize ear borer Mussidia nigrivenella (Lepidoptera: Pyralidae) in Benin, West Africa

In many countries in West Africa, the pyralid ear borer Mussidia nigrivenella Ragonot occasionally causes severe damage to pre- and postharvest maize. Between 1992 and 1995, the distribution of and damage caused by M. nigrivenella were studied in Benin using survey data and an on-station field experiment. The borer was distributed across the whole country, and at maturity an average 25% of the ears sampled in maize fields were infested. Damage levels varied with agro-ecological zones and were highest in the Guinea Savannas. However, borer-related yield losses were comparatively low. Three applications of cypermethrin over the growing season did not provide sufficient control in the on-station field experiment. A model was developed to estimate maize losses caused by M. nigrivenella, using the percentage of infested ears, which explained 93% of the variance. Extrapolation of field data indicated a 25% yield loss once a 100% infestation of maize ears was reached. For surveys in maize fields the model is a valid tool for a rapid assessment of crop losses caused by M. nigrivenella.     

Photoneutron-induced damage reduction for cardiac implantable electronic devices using neutron-shielding sheets in high-energy X-ray radiotherapy: A phantom study

PurposeTo evaluate damage reduction in cardiac implantable electronic devices (CIEDs) caused by photoneutrons in high-energy X-ray radiotherapy using a neutron-shielding sheet (NSS).MethodsThe NSS consists of a bolus with a thickness of 1 or 2 cm (Bls1 or Bls2) as a moderator and several absorbers (20%, 50%, or 80% B₄C silicone sheet [B₄C20, B₄C50, or B₄C80] or a 40% LiF silicone sheet [LiF40]). First, a linear accelerator (LINAC) with a water-equivalent phantom was modeled in the simulation and measured experimentally. Several NSSs were placed on the phantom, a Eu:LiCaAlF₆ scintillator was placed between the phantom and the NSS, and X-rays were irradiated. The relative counts (Cr = counts when placing the NSS or Bls2) were compared between the experiment and simulation. Second, CIED damage was evaluated in the simulation. The relative damage (Dr = damage when placing or not placing the NSS) was compared among all the NSSs. In addition, the γ-ray and leaking X-ray dose from B₄C was measured using a dosimetric film. After determining the optimal NSS combination, Dr value analysis was performed by changing the length of one side and the thickness.ResultsThe Cr values of the simulation and experiment agreed within a 30% percentage difference, except for Bare or LiF40-only. The Dr value was reduced by 43% when Bls2 + B₄C80 was applied. The photon dose was less than 5 cGy/1500 MU. The Dr values were smaller for the smaller lengths of one side of B₄C80 and decreased as the M-layer thickness increased.ConclusionsThe CIED damage induced by photoneutrons generated by a LINAC was effectively reduced by applying the optimal NSS.     

Three correlations between extracellular calcium concentration and myocardial cell injury induced by drugs

Primary cultures of newborn rat myocardial cells were treated in various extracellular calcium concentrations (0, 1.35, 2.7, 4.05, and 5.4 mM) with three different drugs; namely, ouabain, sulmazole, and chlorpromazine. Lactate dehydrogenase (LDH) release was used as an indicator of damage. The results showed 10(-3) M ouabain caused apparent damage of the cells and the damage was increased by an increased extracellular calcium concentration. Sulmazole (10(-3)M) caused damage of the cells in the absence of calcium; but it did not cause damage of the cells in the presence of calcium; it protected the cells from damage caused by high calcium concentrations (4.05 and 5.4 mM). Chlorpromazine (1.6 X 10(-4)M) caused severe damage of the cells. The various calcium concentrations had no influence on the degree of the damage. Correlation coefficients showed that correlations between the calcium concentrations and the cell damage caused by ouabain, sulmazole and chlorpromazine were positive correlation, negative correlation, and no correlation, respectively. It is suggested that influx of extracellular calcium is not a final common pathway of drug-induced myocardial cell injury, although it plays an important role in cell injury.     

Mechanism of metal-mediated DNA damage and apoptosis induced by 6-hydroxydopamine in neuroblastoma SH-SY5Y cells

6-Hydroxydopamine (6-OHDA) is a neurotoxin to produce an animal model of Parkinson's disease. 6-OHDA increased the formation of 8-oxo-7, 8-dihydro-2'-deoxyguanosine (8-oxodG), a biomarker of oxidatively damaged DNA, and induced apoptosis in human neuroblastoma SH-SY5Y cells. Iron or copper chelators inhibited 6-OHDA-induced 8-oxodG formation and apoptosis. Thus, iron and copper are involved in the intracellular oxidatively generated damage to DNA, a stimulus for initiating apoptosis. This study examined DNA damage caused by 6-OHDA plus metal ions using (32)P-5'-end-labelled DNA fragments. 6-OHDA increased levels of oxidatively damaged DNA in the presence of Fe(III)EDTA or Cu(II). Cu(II)-mediated DNA damage was stronger than Fe(III)-mediated DNA damage. The spectrophotometric detection of p-quinone and the scopoletin method showed that Cu(II) more effectively accelerated the 6-OHDA auto-oxidation and H(2)O(2) generation than Fe(III)EDTA. This study suggests that copper, as well as iron, may play an important role in 6-OHDA-induced neuronal cell death.     

Population Growth and Damage Caused by Rhopalosiphum padi (L.) (Hemiptera, Aphididae) on Different Cultivars and Phenological Stages of Wheat

Among the aphids associated with wheat and other winter cereals, Rhopalosiphum padi (L.) is currently the predominant species in the wheat growing region of southern Brazil. The damage caused by this aphid occurs by direct feeding and/or by the transmission of pathogenic viruses, such as the Barley/Cereal yellow dwarf virus. In order to estimate the direct damage caused by R. padi on wheat, we evaluated the population growth of this aphid during the tillering and elongation stages and its effects on grain yield components. The experiment was conducted in a screenhouse with three wheat cultivars (BRS Guabiju, BRS Timbaúva, and Embrapa 16). The effect of a period of 16 days, starting from an infestation of 40 aviruliferous aphids/plant, was evaluated and compared to non-infested plants. In both stages, the population growth of R. padi was lower on the BRS Timbaúva. Although infestation caused a reduction in the grain yield of the three cultivars, this effect was lower for BRS Timbaúva. The cultivar Embrapa 16 supported higher infestations and was more tolerant to damage than the BRS Guabiju.     

Additive and non-additive effects of simulated leaf and inflorescence damage on survival, growth and reproduction of the perennial herb Arabidopsis lyrata

Herbivores may damage both leaves and reproductive structures, and although such combined damage may affect plant fitness non-additively, this has received little attention. We conducted a 2-year field experiment with a factorial design to examine the effects of simulated leaf (0, 12.5, 25, or 50% of leaf area removed) and inflorescence damage (0 vs. 50% of inflorescences removed) on survival, growth and reproduction in the perennial herb Arabidopsis lyrata. Leaf and inflorescence damage negatively and independently reduced flower, fruit and seed production in the year of damage; leaf damage also reduced rosette size by the end of the first season and flower production in the second year. Leaf damage alone reduced the proportion of flowers forming a fruit and fruit production per plant the second year, but when combined with inflorescence damage no such effect was observed (significant leaf × inflorescence damage interaction). Damage to leaves (sources) caused a greater reduction in future reproduction than did simultaneous damage to leaves and inflorescences (sinks). This demonstrates that a full understanding of the effects of herbivore damage on plant fitness requires that consequences of damage to vegetative and reproductive structures are evaluated over more than 1 year and that non-additive effects are considered.     

Quantitative comparison of errors in 15N transverse relaxation rates measured using various CPMG phasing schemes

Nitrogen-15 Carr-Purcell-Meiboom-Gill (CPMG) transverse relaxation experiment are widely used to characterize protein backbone dynamics and chemical exchange parameters. Although an accurate value of the transverse relaxation rate, R(2), is needed for accurate characterization of dynamics, the uncertainty in the R(2) value depends on the experimental settings and the details of the data analysis itself. Here, we present an analysis of the impact of CPMG pulse phase alternation on the accuracy of the (15)N CPMG R(2). Our simulations show that R(2) can be obtained accurately for a relatively wide spectral width, either using the conventional phase cycle or using phase alternation when the r.f. pulse power is accurately calibrated. However, when the r.f. pulse is miscalibrated, the conventional CPMG experiment exhibits more significant uncertainties in R(2) caused by the off-resonance effect than does the phase alternation experiment. Our experiments show that this effect becomes manifest under the circumstance that the systematic error exceeds that arising from experimental noise. Furthermore, our results provide the means to estimate practical parameter settings that yield accurate values of (15)N transverse relaxation rates in the both CPMG experiments.     

Increased oxidative damage to all DNA bases in patients with type II diabetes mellitus

Gas chromatography-mass spectrometry was used to measure the oxidative DNA damage in diabetic subjects and controls. Levels of multiple DNA base oxidation products, but not DNA base de-amination or chlorination products, were found to be elevated in white blood cell DNA from patients with type II diabetes as compared with age-matched controls. The chemical pattern of base damage is characteristic of that caused by an attack on DNA by hydroxyl radical. An increased formation of the highly reactive hydroxyl radical could account for many of the reports of oxidative stress in diabetic subjects. There was no evidence of an increased DNA damage by reactive nitrogen or chlorine species.     

Organ-specific DNA damage of tris(2,3-dibromopropyl)-phosphate and its diester metabolite in the rat.

The organ specificity of tris(2,3-dibromopropyl)phosphate(Tris-BP)-induced DNA damage was investigated in the rat 2 h after a single i.p. injection of 350 mumol/kg. Extensive DNA damage, measured with the alkaline elution method, was found in the kidney, liver and small intestine. Less, but significant DNA damage was detected in the brain, lung, spleen, large intestine and testis. The role of different pathways in the activation of Tris-BP to DNA damaging products was studied in isolated liver and testicular cells. Concentrations as low as 2.5-5 microM Tris-BP caused DNA damage in the hepatocytes, whereas an approximately 10-fold higher concentration was needed in testicular cells to produce a similar amount of DNA damage. Depletion of GSH by diethyl maleate (DEM) did not affect the extent of DNA damage caused by Tris-BP in the liver cells, but blocked the genotoxic effect in testicular cells. Two specifically deuterated Tris-BP analogs, C3D2-Tris-BP and C2D1-Tris-BP, were significantly less potent in causing DNA damage than the protio compound in isolated liver cells and were somewhat less potent in testicular cells. The major urinary metabolite of Tris-BP, bis(2,3-dibromopropyl)phosphate (Bis-BP), was less potent than Tris-BP in causing kidney DNA damage after in vivo exposure. Furthermore, Bis-BP induced substantially less DNA damage in isolated liver and testicular cells. Similar to the effect of DEM on the DNA damage caused by Tris-BP, the DNA damage caused by Bis-BP could be decreased by DEM-pretreatment in testicular cells but not in liver cells. The present study shows that Tris-BP is a potent multiorgan genotoxic agent in vivo. The in vitro data indicate that P-450 mediated metabolism of Tris-BP is more important than activation by glutathione S-transferases of Tris-BP in liver cells, whereas the latter activation pathway seems to be most important in testicular cells.