Male rats were subjected to 1 h testicular torsion of the spermatic cord or 1 h torsion followed by detorsion and recovery up to 4 weeks. The extent of tissue damage was evaluated by a testicular biopsy score count and mitochondrial function. Torsion for 1 h followed by detorsion induced significant morphological damage, which became more severe with longer periods of recovery. This morphological damage could not be correlated with mitochondrial damage as assessed by measuring the 4834 bp mitochondrial DNA 'common deletion' using a quantitative competitive polymerase chain reaction (PCR) assay. Mitochondrial respiratory chain activity, as measured by mitochondrial oxygen consumption using an oxygen electrode, did not vary between the treated animals and the controls. We conclude that the common mitochondrial DNA deletion and oxygen consumption are not good indicators of testicular damage induced by torsion.
A fluorescent-based quantitative PCR assay to measure the percentage of the common deletion in rat mitochondria is reported. The amount of the common deletion is measured relative to the amount of total mitochondrial (mt)DNA. The use of a heterologous competitor construct allows the reaction to be monitored to ensure that exponential accumulation of products occurs. The use of fluorescence-labeled primers provides a safe and sensitive means to quantify products without any adjustment for size. This assay will allow the quantitative determination of the common deletion with one dilution range.
The goal of this study is to assess the acute and sublethal potential hazards on tilapia fish (Oreochromis niloticus) exposed to two different commercial formulations of abamectin (ABM, 5%): emulsifiable concentration (EC) and microemulsions (ME). The median lethal concentration (LC50 -96 h) to each formulation was determined, as well as the adverse effects of sublethal concentrations (½ LC50) on physiological and oxidative stress biomarkers after 96hs of exposure. The LC50 (96 h) values for EC and ME were 10 and 16.6 µg/L, respectively. Furthermore, in both EC and ME, the findings of physiological and oxidative stress aspects revealed significant markedly increases in serum total protein, albumin, globulin, and urea, as well as a significant decline in aminotransferases activity (ALT and AST), and creatinine levels (Cre). Moreover, superoxide dismutase (SOD), malondialdehyde (MDA), catalase (CAT), and glutathione peroxidase (GPx) were greatly increased, whereas glutathione-S-transferase (GST) and glutathione (GSH) were significantly reduced in the liver and brain.
MATERIALS AND METHODS
Pesticide Used:Abamectin (ABM) as two formulations: 5% EC (Profery) and 5% ME (Spider gold) were supplied by the central agricultural pesticide laboratory (CAPL), Agricultural
The "common" 4977 bp deletion in mitochondrial DNA (Delta4977) is commonly used as an indicator of tissue deterioration in ageing and bioenergetic diseases. Deletion levels are normally measured by a serial dilution polymerase chain reaction (PCR) approach, where test reactions are compared with dilutions of control amplifications of DNA from a similar sized stable region of the mitochondrial genome. The end-point of this assay is the dilution that can just detect any PCR product; however, this is an inherently unstable measure. We constructed a chimaeric DNA construct that binds to both control and deletion primers with similar annealing properties. This was used in a competitive PCR assay to quantify Delta4977 in human testicular tissues that had been well-characterized using the serial dilution approach. We found the competitive assay to be highly replicable as it compares the PCR product of the construct with that of test DNA samples during the linear growth phase of the PCR reaction. Moreover, the serial dilution assay was shown to significantly overestimate the amounts of deleted mitochondrial DNA present. The assay promises to throw new light on the role of mitochondrial DNA deletions in tissue dysfunction and ageing, as such deletions can now be determined with high accuracy and repeatability and is much cheaper to apply than real-time fluorescent quantitative PCR.
In the present study, four botanical oils of Melissa officinalis, Borago officinalis, Laurus nobilis and Carapichea ipecacuanha were tested against newly emerged adult males and females of the red palm weevil, Rhynchophorus ferrugineus (Oliver) for investigating their insecticidal activity, their effects on some bilogical aspects and morphogenetic abnormalities in the all stages.The results revealed that the males of Rhynchophorus ferrugineus were more susceptible to the four botanical oils than the females, followed by the immature larvae. The larvae showed different degrees of susceptibility to the four botanical oils, they showed the highest susceptibility to C. ipecacuanha and the lowest to B. officinalis oil at LC 50 levels. Both adult males and females showed the highest susceptibility to C. ipecacuanha and the lowest to L. nobilis oil at LC 50 levels. Also, the obtained results showed that the four tested oils disrupted some biological aspects of the tested insects. These oils markedly reduced male and female longevity, female fecundity and hatchability of the laid eggs. Morphogenetic abnormalities such as darkness of pupae, incomplete adult eclusion , dwarfism and crumpled wings and legs of adults appeared in the different stages of the insect as a result of treatment with the four tested oils. Group Tested plants Family name Used parts Common name Scientific name Group (A) Fixed oils Lemon balm اﻟﺘﺮﻧﺠﺎن Melissa officinalis Lamiaceae Leaf Borage or Star flower اﻟﺤﻤﺤﻢ Borago officinalis Boraginaceae Leaf Group (B)Volatile oils Bay Laurel or Bay leaf اﻟﻐﺎر ﺣﺐ Laurus nobilis Lauraceae Leaf & fruit Rio ipecac اﻟﺬھﺐ ﻋﺮق Carapichea ipecacuanha Rubiaceae Root&leaf
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