Helicobacter pylori Infection Induces Anemia,Depletes Serum Iron Storage,and Alters Local Iron-Related and Adult Brain Gene Expression in Male INS-GAS Mice

Iron deficiency anemia (IDA) affects > 500 million people worldwide, and is linked to impaired cognitive development and function in children. Helicobacter pylori, a class 1 carcinogen, infects about half of the world’s population, thus creating a high likelihood of overlapping risk. This study determined the effect of H. pylori infection on iron homeostasis in INS-GAS mice. Two replicates of INS-GAS/FVB male mice (n = 9-12/group) were dosed with H. pylori (Hp) strain SS1 or sham dosed at 6–9 weeks of age, and were necropsied at 27–29 weeks of age. Hematologic and serum iron parameters were evaluated, as was gene expression in gastric and brain tissues. Serum ferritin was lower in Hp SS1-infected mice than uninfected mice (p < 0.0001). Infected mice had a lower red blood cell count (p<0.0001), hematocrit (p < 0.001), and hemoglobin concentration (p <0.0001) than uninfected mice. Relative expression of gastric hepcidin antimicrobial peptide (Hamp) was downregulated in mice infected with Hp SS1 compared to sham-dosed controls (p<0.001). Expression of bone morphogenic protein 4 (Bmp4), a growth factor upstream of hepcidin, was downregulated in gastric tissue of Hp SS1-infected mice (p<0.001). Hp SS1-infected mice had downregulated brain expression of tyrosine hydroxylase (Th) (p = 0.02). Expression of iron-responsive genes involved in myelination (myelin basic protein (Mbp) and proteolipid protein 2 (Plp2)) was downregulated in infected mice (p = 0.001 and p = 0.02). Expression of synaptic plasticity markers (brain derived neurotrophic factor 3 (Bdnf3), Psd95 (a membrane associated guanylate kinase), and insulin-like growth factor 1 (Igf1)) was also downregulated in Hp SS1-infected mice (p = 0.09, p = 0.04, p = 0.02 respectively). Infection of male INS-GAS mice with Hp SS1, without concurrent dietary iron deficiency, depleted serum ferritin, deregulated gastric and hepatic expression of iron regulatory genes, and altered iron-dependent neural processes. The use of Hp SS1-infected INS-GAS mice will be an appropriate animal model for further study of the effects of concurrent H. pylori infection and anemia on iron homeostasis and adult iron-dependent brain gene expression.     

Verkieste Amöben in Steinkohlen aus dem Ruhrgebiet — erster Nachweis vonArcella Ehrenberg im Paläozoikum

Within bituminous coals of Westphalian B age pyritized amoebae of the order Testacea (thecamoebians) were detected. They may belong to the genusArcella. The appearance of this genus, known until now only since the Pleistocene, is extended far into the Palaeozoic by this observation.     

A Matter of Timing: Identifying Significant Multi-Dose Radiotherapy Improvements by Numerical Simulation and Genetic Algorithm Search

Multi-dose radiotherapy protocols (fraction dose and timing) currently used in the clinic are the product of human selection based on habit, received wisdom, physician experience and intra-day patient timetabling. However, due to combinatorial considerations, the potential treatment protocol space for a given total dose or treatment length is enormous, even for relatively coarse search; well beyond the capacity of traditional in-vitro methods. In constrast, high fidelity numerical simulation of tumor development is well suited to the challenge. Building on our previous single-dose numerical simulation model of EMT6/Ro spheroids, a multi-dose irradiation response module is added and calibrated to the effective dose arising from 18 independent multi-dose treatment programs available in the experimental literature. With the developed model a constrained, non-linear, search for better performing cadidate protocols is conducted within the vicinity of two benchmarks by genetic algorithm (GA) techniques. After evaluating less than 0.01% of the potential benchmark protocol space, candidate protocols were identified by the GA which conferred an average of 9.4% (max benefit 16.5%) and 7.1% (13.3%) improvement (reduction) on tumour cell count compared to the two benchmarks, respectively. Noticing that a convergent phenomenon of the top performing protocols was their temporal synchronicity, a further series of numerical experiments was conducted with periodic time-gap protocols (10 h to 23 h), leading to the discovery that the performance of the GA search candidates could be replicated by 17–18 h periodic candidates. Further dynamic irradiation-response cell-phase analysis revealed that such periodicity cohered with latent EMT6/Ro cell-phase temporal patterning. Taken together, this study provides powerful evidence towards the hypothesis that even simple inter-fraction timing variations for a given fractional dose program may present a facile, and highly cost-effecitive means of significantly improving clinical efficacy.     

Applying PyRosetta molecular energies to separate properly oriented protein models from mirror models,obtained from contact maps

Reconstructing protein structure based on contact maps leads to two types of models: properly oriented models and mirror models. This is due to the fact that contact maps do not include information on protein chirality. Therefore, both types of model orientations share the same contact map and are geometrically allowed. In this work, we verified the hypothesis that some of the energy terms calculated by PyRosetta could be useful to distinguish between properly oriented and mirror models. We studied 440 models of all-alpha protein domains reconstructed manually from their contact maps, where 50 % of the models were properly oriented and 50 % had mirror orientation. We showed that dihedral angles and energy terms, based on the probability of specific geometrical arrangement of the residues, differed significantly for properly oriented and mirror models.     

Gene mapping of 28S rDNA sites in allotriploid Cobitis females (Pisces: Cobitidae) from a diploid-polyploid population

We performed cytogenetic analysis on triploid Cobitis hybrid specimens from a natural diploid-polyploid population in the Bug River (Poland). Chromosomes were examined by fluorescence in situ hybridization method (FISH) with 28S rDNA as a probe. The rDNA sites were commonly identified in the terminal position of p arms of two small submetacentric chromosomes, in the centromere area of two acrocentric chromosomes and in the terminal position of q arm of one acrocentric chromosome. Some of the observed signals were stronger than others. In few cases, one small acrocentric chromosome showing rDNA sites located terminally on both p and a q arm was observed. This study brings useful information about polymorphism of 28S rDNA sites regarding their number, location and size in triploid specimens. Moreover, it gives preliminary data for comparison with similar researches on the parental species of Cobitis hybrid females, viz. C. taenia, C. taurica and C. tanaitica, and other related Cobitis taxa of different ploidy level. The present study increases our knowledge about ribosomal genes inheritance between Cobitis species and their polyploid hybrids.     

Microarray-based genetic identification of beneficial organisms as a new tool for quality control of laboratory cultures

The use of beneficial organisms to help control pests and pathogens in field and greenhouse crops is constantly increasing. Insects and mites are commonly used as beneficial organisms and, nowadays, rearing companies have to produce them in large quantities. Because of the peculiarities of laboratory culture conditions, the quality of lab-reared organisms generally degrades over time. To maintain high fitness levels, cultures are refreshed with field specimens at regular intervals. However, this bears the risk of contaminating laboratory cultures with species or strains other than the intended natural enemy. To ensure that the correct species is produced and also to facilitate surveys after field release, we have developed a diagnostic microarray for identification of beneficial species. Probes have been designed from the different haplotypes of a fragment of the mitochondrial cytochrome oxidase I (COI) gene of each species. Hybridization of labeled PCR amplicons of COI on the microarray chip allows precise identification of 28 economically relevant arthropod species.     

Seasonal Changes of Body Iron Status Determine Cadmium Accumulation in the Wild Bank Voles

The objective of this study was to examine relations between body iron (Fe) status and cadmium (Cd) accumulation in a small rodent, the bank vole, caught from the wild population in late autumn (November) and early spring (March). The concentrations of Fe in the liver, kidneys, and duodenum in the bank voles from the spring were only 30%, 60%, and 70%, respectively, of those found in the animals from the autumn. An analysis of hematocrit and hemoglobin content of blood showed no significant effect of the season, suggesting that the animals from the spring were not anemic. The exposure to dietary Cd (10 μg/g) for 7 days resulted in 70% higher accumulation of Cd in the liver and kidneys of the spring than autumn bank voles, and the concentration of Cd in the duodenum was 3.5 times higher in the spring animals, despite the fact that relative Cd intake was significantly higher in the autumn bank voles. The data indicate that seasonal changes of body Fe status occurring in the wild bank voles may influence tissue accumulation of Cd.     

Molecular markers from a BAC contig spanning the Rdr1 locus: a tool for marker-assisted selection in roses

We constructed a BAC contig of about 300 kb spanning the Rdr1 locus for black spot resistance in Rosa multiflora hybrids, using a new BIBAC library from DNA of this species. From this contig, we developed broadly applicable simple sequence repeat (SSR) markers tightly linked to Rdr1, which are suitable for genetic analyses and marker-assisted selection in roses. As a source for the high molecular weight DNA, we chose the homozygous resistant R. multiflora hybrid 88/124-46. For the assembly of the BAC contig, we made use of molecular markers derived from a previously established R. rugosa contig. In order to increase the resolution for fine mapping, the size of the population was increased to 974 plants. The genomic region spanning Rdr1 is now genetically restricted to 0.2 cM, corresponding to a physical distance of about 300 kb. One single-stranded conformational polymorphism (SSCP) and one SSR marker cosegregate with the Rdr1-mediated black spot resistance, while one SSR and several cleaved amplified polymorphic sequence or SSCP markers are very tightly linked with one to three recombinants among the 974 plants. The benefits of the molecular markers developed from the R. multiflora contig for the genetic analysis of roses and the integration of rose genetic maps are discussed.