Reconstitution of a light-stimulated adenylate cyclase from retina and Neurospora crassa preparations. Characterization of the heterologous systems using normal and degenerative retinas

Adenylate cyclase catalytic subunits from Neurospora crassa membranes may interact with regulatory factors from membranes of bovine retinal rod outer segments (pretreated with N-ethylmaleimide), reconstituting a heterologous system which, in the presence of light, is catalytically active in assay mixtures containing MgATP. Maximal activation was observed at 550 nm. Transducin-depleted retinal membranes were not capable of reconstituting the heterologous light-stimulated adenylate cyclase system. Addition of a transducin preparation to depleted membranes restored the reconstitution capacity of these membranes. A similar heterologous adenylate cyclase system was reconstituted with Neurospora and mouse retinal whole membranes (pretreated with N-ethylmaleimide). Membranes from mice suffering photoreceptor degeneration (rd homozygotes) did not reconstitute an heterologous adenylate cyclase system.     

Observations on movements of yellow eels, Anguilla anguilla L., after displacement from coastal waters to sea

Four groups of yellow eels, three from three Po Delta brackish bays (Pila, Scardovari and Goro) and one from the French Atlantic coast, were marked by freeze-branding and released at an open-sea point 5 miles offshore. Of the 2137 eels released, 453 (21.2%) were recaptured over a 1-month period. The percentage recaptured is not independent of either fish size (the larger eels being recaptured in greater quantity than the smaller ones which are supposed to be less catchable and more easily overlooked by fishermen) or fish origin (the recapture rate of the French stock was both the lowest, at 9.8%, and the most dispersed in time and space). The highest multiple catches were for eels from Pila and Scardovari and were made in the first days of the experiment. Of the recaptured eels, 75% were caught in the inland water closest (9 km) to the release point; only 8.6% were recaptured at the mouth of the two branches of the River Po. There is no evidence of a real homing tendency for yellow eels previously adapted to living in brackish water: the massive return to the nearest coast is probably only an avoidance reaction to the unfavourable open-sea environment.     

Nuclear damage of colon epithelial cells by the food carcinogen 2-amino-3-methylimidazo[4,5-f]quinoline (IQ) is modulated by dietary lipids

Intestinal damage in C57BL/6J female mice was quantified by measuring the frequency of nuclear aberrations in colonic crypts. The animals were maintained on the following diets: standard (5% lipids, 5% cellulose); low- and high-cellulose (0-20% cellulose); high lipids (20% maize oil or 20% olive oil). All groups of animals were treated by gavage either with saline or 250 mg/kg of the dietary carcinogen 2-amino-3-methylimidazo[4,5-f]quinoline (IQ). After 24 h their colons were removed and stained and the nuclear aberrations scored under the microscope. The administration of IQ markedly increased the number of colon aberrations in all of the treated animals. Variations in dietary fiber did not modify the colon-damaging activity of this compound. Maize oil slightly increased the colon-damaging activity, whereas significant protection was observed in the animals on a high-lipid olive-oil diet. These results show that composition of the diet may vary the genotoxic effect of this dietary carcinogen.     

Novel variant Ph translocation t(9;22;11)(q34;q11.2;p15)inv(9)(p13q34) in chronic myeloid leukemia involving a one-step mechanism

Chronic myeloid leukemia (CML) is a clonal malignant disorder of a pluripotent hematopoietic stem cell characterized by the presence of a Philadelphia (Ph) chromosome. Less than 10% of patients present variant Ph chromosomes involving 1 or more additional chromosomes, other than chromosomes 9 and 22, with uncertain prognosis. There are mainly 1- or 2-step mechanisms proposed to explain the genesis of variant Ph chromosomes depending on whether the involved chromosomes are simultaneously broken and rejoined or if a standard t(9;22) occurs first. By combined standard cytogenetic and FISH analysis we detected a novel variant Ph translocation among chromosomes 9, 11 and 22 in a patient with CML without progression to an accelerated phase of the disease after 7 years, with the derivative chromosome 9 also having an acquired pericentric inversion. This novel case illustrates the use of FISH in metaphase to confirm a new rearrangement not previously described in variant Ph formation and that the present karyotype could have originated by a 1-step mechanism with 4 simultaneous breakages without deletion of ABL1.     

The effects of salinity on acute and chronic nickel toxicity and bioaccumulation in two euryhaline crustaceans: Litopenaeus vannamei and Excirolana armata

We investigated the influence of salinity (5 ppt versus 25 ppt) on acute (96-h LC₅₀) and chronic toxicity (15–30 day LC₅₀) of Ni in two euryhaline crustaceans, the shrimp (Litopenaeus vannamei) and the isopod (Excirolana armata). 96-h LC50 values were 41 μmol L⁻¹ and 362 μmol L⁻¹ for L. vannamei and 278 μmol L⁻¹ and > 1000 μmol L⁻¹ for E. armata at 5 ppt and 25 ppt, respectively. Speciation analysis demonstrated that complexation with anions such as SO₄²⁻, HCO₃⁻ and Cl⁻ at 25 ppt had a negligible effect on reducing the free Ni²⁺ ion component in comparison to 5 ppt. The salinity-dependent differences in acute Ni toxicity could not be explained by differences in Ni bioaccumulation. Therefore, differences in physiology of the organisms at the two salinities may be the most likely factor contributing to differences in acute Ni toxicity. Chronic LC₅₀ values (2.7–23.2 μmol L⁻¹) were similar in the two species, but salinity had no significant effect, indicating that water chemistry and osmoregulatory strategy do not influence chronic toxicity. However chronic (15-day) mortality in both species could be predicted by acute (96-h) Ni bioaccumulation patterns.     

Assessment of Trace Elements in Scalp Hair of a Young Urban Population in Brazil

The aim of this study was to establish background values for toxic and essential elements in hair, interelement correlations and the differences in levels between genders in a healthy young population from Southern Brazil. Hair samples (n = 167) were collected from healthy students aged 12–18 years. Trace element concentrations in hair were determined by inductively coupled plasma mass spectrometry. The study provided relatively low values for toxic elements and balanced concentrations for the essential elements in the adolescents' hair with reliable reference data. Interestingly, this study also demonstrated statistical correlations considered newfound between the elements in hair. Hair mercury levels were influenced by gender; with males presenting higher values. The overall findings of the present study, with respect to the estimated chemical elements, are of prime importance in the evaluation of reference values for determining environmental effects on children living in urban areas.     

Genetic diversity and flooding survival in Aegiphila sellowiana (Lamiaceae), a typical tree species from upland riparian forests

Saplings of Aegiphila sellowiana were submitted to flooding and analysis of genetic diversity in order to investigate flooding tolerance as well as its genetic determination. This response is important because it means that some lines could be planted in degraded riparian areas. Leaves were sampled from each plant, and they were submitted to different flooding periods. Mortality of saplings was 40, 80, 50, 53.3, 33.3, and 33.3% in flooding for 15, 18, 25, 50, 80 days, and flooding for 50 days followed by re-aeration for 30 days, respectively. From the total number of flooded plants, 46.7% died in the first seven days of treatment, while 53.3% survived the flooding. The percentage of polymorphic loci (P(p)), Nei's genetic diversity (H) and the Shannon index (I) were slightly higher for the group that survived the stress of flooding (surviving: P(p) (%) = 67.48, H = 0.184, I = 0.287; not surviving: P(p) (%) = 66.67, H = 0.165, I = 0.261). Analysis of molecular variance showed that 5.88% of the genetic variability was due to the differences between groups of plants surviving and not surviving flooding, while 94.12% was due to genetic differences between individuals within these groups. Similar results were obtained by principal coordinate analysis. Based on these results, we can assume the existence of environment-specific genotypes and the genetic determination of flooding tolerance in A. sellowiana. Thus, some lines of A. sellowiana could be used in the reforestation of riparian habitats, especially in uplands along riverbanks.     

hERG1 Channels and Glut-1 as Independent Prognostic Indicators of Worse Outcome in Stage I and II Colorectal Cancer: A Pilot Study

BACKGROUND: There is a need to identify new markers to assess recurrence risk in early-stage colorectal cancer (CRC) patients. We explored the prognostic impact of ether-a-gò-gò-related gene 1 channels and some hypoxia markers, in patients with nonmetastatic (stage I, II, and III) CRC. METHODS: The expression of hERG1, vascular endothelial growth factor A (VEGF-A), glucose transporter 1, carbonic anhydrase IX (CA-IX), epidermal growth factor receptor (EGF-R), and p53 was tested by immunohistochemistry in 135 patients. The median follow-up was 35 months. Clinicopathologic parameters and overall survival were evaluated. RESULTS: hERG1 displayed a statistically significant association with Glut-1, VEGF-A, CA-IX, and EGF-R; p53 with VEGF-A and CA-IX; Glut-1 with the age of the patients; and EGF-R with TNM and mucin content. TNM and CA-IX were prognostic factors at the univariate analysis; TNM, hERG1, and Glut-1, at the multivariate analysis. Risk scores calculated from the final multivariate model allowed to stratify patients into four different risk groups: A) stage I-II, Glut-1 positivity, any hERG1; B) stage I-II, Glut-1 and hERG1 negativity; C) stage I-II, Glut-1 negativity, hERG1 positivity; D) stage III, any Glut-1 and any hERG1. CONCLUSIONS: hERG1 positivity with Glut-1 negativity identifies a patient group with poor prognosis within stage I-II CRC. The possibility that these patients might benefit from adjuvant therapy, independently from the TNM stage, is discussed. IMPACT: More robust prognostic and predictive markers, supplementing standard clinical and pathologic staging, are needed for node-negative patients.