Ecotoxicological studies with the freshwater rotifer Brachionus calyciflorus II. An assessment of the chronic toxicity of lindane and 3,4-dichloroaniline using life tables

The effects of chronic exposure of the freshwater rotifer Brachionus calyciflorus to the toxicants lindane and 3,4-dichloroaniline (DCA), were evaluated. The parameters used to determine the toxicity on these compounds were the age-specific and fertility, and the demographic parameters: intrinsic rate of natural increase (r), generation time (T), net reproductive rate (R _o), reproductive value (V _x) and life expectancy (e _o). All the demographic parameters studied decreased with increasing toxicant concentrations. The use of life tables techniques with B. calyciflorus as a test method for the determination of chronic toxicity of xenobiotics is discussed.     

Chronic toxicity of methylparathion to the rotifer Brachionus calyciflorus fed on Nannochloris oculata and Chlorella pyrenoidosa

The effect of sublethal levels of methylparathion (0, 1, 3, 5, 7 mg l^–1) on the freshwater rotifer, Brachionus calyciflorus, during their entire life cycle was studied. Rotifers were fed on two species of unicellular algae: Nannochloris oculata and Chlorella pyrenoidosa; both algal concentrations were 5 × 10⁵ cell ml^–1.The parameters used to determine the toxicity of this compound were survival, fecundity, net reproductive rate (R)_o, generation time (T), intrinsic rate of natural increase (r), reproductive value (V _x/V_o) and life expectancy at hatching (eo). All the demographic parameters studied were affected by methyl-parathion exposure on rotifers fed on both species of algae, but the toxic effect was larger when animals were fed on Chlorella pyrenoidosa; in this case, animals showed a decreased in fertility and also a delayed first reproduction. Sublethal methylparathion levels produced a reduction in most of the parameters selected, especially after exposure to 7 mg l^–1, where the animals died before reproducing.     

Demographic parameters of Brachionus calyciflorus Pallas (Rotifers) exposed to sublethal endosulfan concentrations

The effects of sublethal levels of endosulfan (0, 1, 1.5, 2.5 and 3.3 mg 1^–1) on the demography of the rotifer Brachionus calyciorus were studied. Life expectancy at birth (e _o), net reproductive rate (Ro), generation time (T) and intrinsic rate of natural increase (r) were significant differentes between blank controls and controls with acetone. The effective endosulfan concentration at which a given parameter value was reduced to 50% of the controls (EC50) was calculated for life expectancy.     

QSAR modeling and transmission electron microscopy stereology of altered mitochondrial ultrastructure of white blood cells in patients diagnosed as schizophrenic and treated with antipsychotic drugs.

Treatment of patients diagnosed as schizophrenic with antipsychotic drugs (neuroleptics) is known to cause occasional unexplained depletion of white blood cells, especially neutrophil granulocytes. It has been known for many years that neuroleptics can interfere with the mitochondrial respiratory chain in vitro. Because there has been a growing interest recently in mitochondrial targeting of drugs, and since a quantitative structure-activity relationship (QSAR) model that predicts mitochondrial accumulation of neuroleptics has been published, we investigated the effects of neuroleptics on white blood cell mitochondria. Venous blood samples were collected from both patients undergoing treatment with neuroleptics and healthy volunteers. The samples were processed for transmission electron microscopy. The resulting images of white blood cells were analyzed using stereology to compare quantitatively mitochondrial morphology in the patient and control groups. We found that in patients, but not in controls, there was swelling of mitochondria and fragmentation of the mitochondrial cristae. There also were fewer mitochondria in patients than in controls, although due to the swelling of the organelles, the volume density of mitochondria in the two groups was not significantly different. Such changes are typical of a toxic insult. Consequently, it seems plausible that, since schizophrenia is not a disease considered to affect white blood cells per se, these changes probably are due to the medication.     

Heme Oxygenase-1 (HO-1) Expression in Prostate Cancer Cells Modulates the Oxidative Response in Bone Cells

Prostate cancer (PCa) is a leading cause of death among males. It is currently estimated that inflammatory responses are linked to 15-20% of all deaths from cancer worldwide. PCa is dominated by complications arising from metastasis to the bone where the tumor cells interact with the bone microenvironment impairing the balance between bone formation and degradation. However, the molecular nature of this interaction is not completely understood. Heme oxygenase-1 (HO-1) counteracts oxidative damage and inflammation. Previous studies from our laboratory showed that HO-1 is implicated in PCa, demonstrating that endogenous HO-1 inhibits bone derived-prostate cancer cells proliferation, invasion and migration and decreases tumor growth and angiogenesis in vivo. The aim of this work was to analyze the impact of HO-1 modulated PCa cells on osteoblasts proliferation in vitro and on bone remodeling in vivo. Using a co-culture system of PC3 cells with primary mice osteoblasts (PMOs), we demonstrated that HO-1 pharmacological induction (hemin treatment) abrogated the diminution of PMOs proliferation induced by PCa cells and decreased the expression of osteoclast-modulating factors in osteoblasts. No changes were detected in the expression of genes involved in osteoblasts differentiation. However, co-culture of hemin pre-treated PC3 cells (PC3 Hem) with PMOs provoked an oxidative status and activated FoxO signaling in osteoblasts. The percentage of active osteoblasts positive for HO-1 increased in calvarias explants co-cultured with PC3 Hem cells. Nuclear HO-1 expression was detected in tumors generated by in vivo bone injection of HO-1 stable transfected PC3 (PC3HO-1) cells in the femur of SCID mice. These results suggest that HO-1 has the potential to modify the bone microenvironment impacting on PCa bone metastasis.     

Performance of the eclipse monitor unit objective tool utilizing volumetric modulated arc therapy for rectal cancer

AimTo assess the performance of the monitor unit (MU) Objective tool in Eclipse treatment planning system (TPS) utilizing volumetric modulated arc therapy (VMAT) for rectal cancer.BackgroundEclipse VMAT planning module includes a tool to control the number of MUs delivered: the MU Objective tool. This tool could be utilized to reduce the total number of MUs in rectal cancer treatments.Materials and methods20 rectal cancer patients were retrospectively studied using VMAT and the MU Objective tool. The baseline plan for each patient was selected as the one with no usage of the MU Objective tool. The number of MUs of this plan was set to be the reference number of MUs (MU_ref). Five plans were re-optimized for each patient only varying the Max MU parameter. The selected values were 30%, 60%, 90%, 120% and 150% of MU_ref for each patient. Differences with respect to the baseline plan were evaluated regarding MU number and parameters for PTVs coverage evaluation, PTVs homogeneity and OARs doses assessment. A two-tailed, paired-samples t-test was used to quantify these differences.ResultsAverage relative differences in MU number obtained was 10% for Max MU values of 30% and 60% of MU_ref, respectively (p < 0.03). PTVs coverage and homogeneity were not compromised and discrepancies obtained with respect to baseline plans were not significant. Furthermore, maximum OARs doses deviations were also not significant.ConclusionsA 10% reduction in the MU number could be obtained without an alteration of PTV coverage and OARs doses for rectal cancer.     

1064 nm Nd:YAG laser light affects transmembrane mitochondria respiratory chain complexes

Photobiomodulation (PBM) is a non‐plant‐cell manipulation through a transfer of energy by means of light sources at the non‐ablative or thermal intensity. Authors showed that cytochrome‐c‐oxidase (complex IV) is the specific chromophore's target of PBM at the red (600‐700 nm) and NIR (760‐900 nm) wavelength regions. Recently, it was suggested that the infrared region of the spectrum could influence other chromospheres, despite the interaction by wavelengths higher than 900 nm with mitochondrial chromophores was not clearly demonstrated. We characterized the interaction between mitochondria respiratory chain, malate dehydrogenase, a key enzyme of Krebs cycle, and 3‐hydroxyacyl‐CoA dehydrogenase, an enzyme involved in the β‐oxidation (two mitochondrial matrix enzymes) with the 1064 nm Nd:YAG (100mps and 10 Hz frequency mode) irradiated at the average power density of 0.50, 0.75, 1.00, 1.25 and 1.50 W/cm² to generate the respective fluences of 30, 45, 60, 75 and 90 J/cm². Our results show the effect of laser light on the transmembrane mitochondrial complexes I, III, IV and V (adenosine triphosphate synthase) (window effects), but not on the extrinsic mitochondrial membrane complex II and mitochondria matrix enzymes. The effect is not due to macroscopical thermal change. An interaction of this wavelength with the Fe‐S proteins and Cu‐centers of respiratory complexes and with the water molecules could be supposed.      

Molecular cloning and developmental expression of Tlx (Hox11) genes in zebrafish (Danio rerio)

Tlx (Hox11) genes are orphan homeobox genes that play critical roles in the regulation of early developmental processes in vertebrates. Here, we report the identification and expression patterns of three members of the zebrafish Tlx family. These genes share similar, but not identical, expression patterns with other vertebrate Tlx-1 and Tlx-3 genes. Tlx-1 is expressed early in the developing hindbrain and pharyngeal arches, and later in the putative splenic primordium. However, unlike its orthologues, zebrafish Tlx-1 is not expressed in the cranial sensory ganglia or spinal cord. Two homologues of Tlx-3 were identified: Tlx-3a and Tlx-3b, which are both expressed in discrete regions of the developing nervous system, including the cranial sensory ganglia and Rohon-Beard neurons. However, only Tlx-3a is expressed in the statoacoustic cranial ganglia, enteric neurons and non-neural tissues such as the fin bud and pharyngeal arches and Tlx-3b is only expressed in the dorsal root ganglia.     

Rapid identification of Arabidopsis insertion mutants by non-radioactive detection of T-DNA tagged genes

To assist in the analysis of plant gene functions we have generated a new Arabidopsis insertion mutant collection of 90 000 lines that carry the T-DNA of Agrobacterium gene fusion vector pPCV6NFHyg. Segregation analysis indicates that the average frequency of insertion sites is 1.29 per line, predicting about 116 100 independent tagged loci in the collection. The average T-DNA copy number estimated by Southern DNA hybridization is 2.4, as over 50% of the insertion loci contain tandem T-DNA copies. The collection is pooled in two arrays providing 40 PCR templates, each containing DNA from either 4000 or 5000 individual plants. A rapid and sensitive PCR technique using high-quality template DNA accelerates the identification of T-DNA tagged genes without DNA hybridization. The PCR screening is performed by agarose gel electrophoresis followed by isolation and direct sequencing of DNA fragments of amplified T-DNA insert junctions. To estimate the mutation recovery rate, 39 700 lines have been screened for T-DNA tags in 154 genes yielding 87 confirmed mutations in 73 target genes. Screening the whole collection with both T-DNA border primers requires 170 PCR reactions that are expected to detect a mutation in a gene with at least twofold redundancy and an estimated probability of 77%. Using this technique, an M2 family segregating a characterized gene mutation can be identified within 4 weeks.