Flow cytometry assessment of cytotoxicity and reactive oxygen species generation by single and binary mixtures of cadmium, zinc and copper on populations of the ciliated protozoan Tetrahymena thermophila

In the present study, we have assessed by flow cytometry, the cytotoxicity of the heavy metals Cd, Zn and Cu on populations of the ciliated protozoan Tetrahymena thermophila. The obtained LC(50) for these metals was estimated as 0.195mgCdl(-1), 3.58mgZnl(-1) and 0.47mgCul(-1), respectively. As a result, the toxicity rank for this eukaryotic microorganism is Cd>Cu>Zn. Using the same methodology and the Concentration Addition approach, the toxicity of binary mixtures of these metals was evaluated in order to detect the type of interaction between these metals. Results indicated that antagonism is the predominant interaction but it can change to additivity or even to synergism at high metal concentrations. Besides, the concentration ratio between metals plays also a crucial role in determining the type of metallic interaction, at least in Tetrahymena. Cytotoxicity data from single and bimetallic mixtures have been compared with those from selected microalgae, other species of ciliates, fish and mammalian cell lines and metazoan. By other hand, we have detected the mitochondrial generation of peroxides induced by both single and binary treatments with Cd, Zn and Cu on populations of Tetrahymena, using the specific fluorophore dyhidrorhodamine. The nature and concentrations of metal as well as the metallic ratio were important factors in reactive oxygen species production. All results found in T. thermophila are compared with previous reports in other organisms and, some explanations and hypothesis to support results are given, including the involvement of metallothioneins as antioxidants and their role in the binding of metal cations.     

Geographic and ontogenetic variation in the diet and daily ration of the bonnethead shark, <Emphasis Type="Italic">Sphyrna tiburo</Emphasis>, from the eastern Gulf of Mexico

To examine variation in diet and daily ration of the bonnethead shark, Sphyrna tiburo (Linnaeus 1758), animals were collected from three areas in the eastern Gulf of Mexico: northwest Florida (∼29°40′N, 85°13′W), Tampa Bay near Anclote Key (∼28°10′N, 82°42.5′W), and Florida Bay (∼24°50′N, 80°48′W) from March through September, 1998–2000. In each area, diet was assessed by life stage (young-of-the year, juveniles, and adults) and quantified using five indices: percent by number (%N), percent by weight (%W), frequency of occurrence (%O), index of relative importance expressed on a percent basis (%IRI), and %IRI based on diet category (%IRI_DC). Diet could not be assessed for young-of-the-year in Tampa Bay or Florida Bay owing to low sample size. Diet analysis showed an ontogenetic shift in northwest Florida. Young-of-the-year stomachs from northwest Florida (n = 68, 1 empty) contained a mix of seagrass and crustaceans while juvenile stomachs (n = 82, 0 empty) contained a mix of crabs and seagrass and adult stomachs (n = 39, 1 empty) contained almost exclusively crabs. Crabs made up the majority of both juvenile and adult diet in Tampa Bay (n = 79, 2 empty, and n = 88, 1 empty, respectively). Juvenile stomachs from Florida Bay (n = 72, 0 empty) contained seagrass and a mix of crustaceans while adult stomachs contained more shrimp and cephalopods (n = 82, 3 empty). Diets in northwest Florida and Tampa Bay were similar. The diet in Florida Bay was different from those in the other two areas, consisting of fewer crabs and more cephalopods and lobsters. Plant material was found in large quantities in all stomachs examined from all locations (>15 %IRI_DC in 6 of the 7 life stage-area combinations, >30 %IRI_DC in 4 of the 7 combinations, and 62 %IRI_DC in young-of-the-year diet in northwest Florida). Using species- and area-specific inputs, a bioenergetic model was constructed to estimate daily ration. Models were constructed under two scenarios: assuming plant material was and was not part of the diet. Overall, daily ration was significantly different by sex, life stage, and region. The bioenergetic model predicted increasing daily ration with decreasing latitude and decreasing daily ration with ontogeny regardless of the inclusion or exclusion of plant material. These results provide evidence that bonnetheads continuously exposed to warmer temperatures have elevated metabolism and require additional energy consumption to maintain growth and reproduction. Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Normal sonographic development of the central nervous system from the second trimester onwards using 2D, 3D and transvaginal sonography

The developmental changes of the fetal central nervous system (CNS) during the second and third trimesters, specifically the brain, relate mostly to changes in size. However, other changes do occur in the fetal brain during the second and third trimester such as: the union of the cerebellar hemispheres, development of the corpus callosum (CC), and increasing complexity of the cerebral cortex. These changes follow a well-defined developmental timeline recognizable by sonography. The fetal neuroscan can be divided into a ‘basic scan’ which is performed transabdominally and a ‘targeted Exam or neurosonogram’ which uses a multiplanar approach, which preferably should be performed transvaginally. During the ‘basic scan’, several brain structures are imaged in addition to obtaining important biometric measurements. The ‘neurosonogram’ is a more extensive or detailed fetal study during which the emphasis is on the addition of coronal and sagittal planes. The easiest way to obtain these planes, if the fetus is in a cephalic presentation, is the transvaginal route. Three-dimensional (3D) sonography should, if possible, be performed transvaginally using the multiplanar approach. An added benefit of 3D sonography is the ability to display and render the volume in a variety of ways which may enhance the detection of pathology. Copyright © 2008 John Wiley & Sons, Ltd.     

Rapid molecular prenatal diagnosis of ataxia-telangiectasia by direct mutational analysis

Mutations of the ataxia-telangiectasia-mutated (ATM) gene are responsible for the autosomal recessive disorder ataxia-telangiectasia (A-T). This study reports the first A-T prenatal diagnosis performed in Spain by direct molecular analysis. The pregnant woman had a previous child suffering from A-T due to a deletion in the ATM gene. The ATM coding region was sequenced in the A-T patient and her parents. Then, a specific polymerase chain reaction (PCR) to detect the deletion was performed for prenatal diagnosis. Additionally, polymorphic HLA loci were examined in order to exclude the possible contamination by maternal DNA. In this family of Gypsy origin, we carried out a rapid molecular diagnosis of A-T. Then, a prenatal diagnosis was carried out, identifying the deletion in the fetal DNA. Additionally, we performed a population study in unrelated Spanish Gypsies and in unrelated controls, showing that the deletion described could be a hotspot in the Spanish Gypsy population. The size of the coding region and the genomic structure, together with the absence of hotspots, make the mutation screening of the ATM gene difficult. The ability to identify ATM mutations provides a tool that can be applied in confirmatory diagnosis, genetic counselling, carrier prediction and prenatal diagnosis. Copyright © 2007 John Wiley & Sons, Ltd.     

Effect of combined nitrogen on dinitrogen fixation and productivity in Pisum sativum L. Inoculated with different strains of Rhizobium

This growth chamber study was designed to determine rates of nitrogen fixation, growth, productivity and concentrations of nitrogen and phosphorus during vegetative growth, flowering and the beginning of the fruiting and ripening period in Pisum sativum plants supplied with different concentrations of nitrate and nodulated by various strains of Rhizobium leguminosarum. Noninoculated control plants were grown under the same nutritional conditions. Nodulated and unnodulated plants responded differently to treatment. The effects of inoculation with Rhizobium and the nitrate treatments on the parameters of growth and nitrogen content varied with plant age. Although the presence of nitrate in the culture medium inhibited nodulation and acetylene reducing activity (ARA), the different Rhizobium strains used in the inoculations led to variations in tolerance to combined nitrogen.