Both SEPT2 and MLL are down-regulated in MLL-SEPT2 therapy-related myeloid neoplasia

<p>Abstract</p> <p>Background</p> <p>A relevant role of septins in leukemogenesis has been uncovered by their involvement as fusion partners in <it>MLL</it>-related leukemia. Recently, we have established the <it>MLL-SEPT2 </it>gene fusion as the molecular abnormality subjacent to the translocation t(2;11)(q37;q23) in therapy-related acute myeloid leukemia. In this work we quantified <it>MLL </it>and <it>SEPT2 </it>gene expression in 58 acute myeloid leukemia patients selected to represent the major AML genetic subgroups, as well as in all three cases of <it>MLL-SEPT2</it>-associated myeloid neoplasms so far described in the literature.</p> <p>Methods</p> <p>Cytogenetics, fluorescence in situ hybridization (FISH) and molecular studies (RT-PCR, qRT-PCR and qMSP) were used to characterize 58 acute myeloid leukemia patients (AML) at diagnosis selected to represent the major AML genetic subgroups: <it>CBFB-MYH11 </it>(n = 13), <it>PML-RARA </it>(n = 12); <it>RUNX1-RUNX1T1 </it>(n = 12), normal karyotype (n = 11), and <it>MLL </it>gene fusions other than <it>MLL-SEPT2 </it>(n = 10). We also studied all three <it>MLL-SEPT2 </it>myeloid neoplasia cases reported in the literature, namely two AML patients and a t-MDS patient.</p> <p>Results</p> <p>When compared with normal controls, we found a 12.8-fold reduction of wild-type <it>SEPT2 </it>and <it>MLL-SEPT2 </it>combined expression in cases with the <it>MLL-SEPT2 </it>gene fusion (p = 0.007), which is accompanied by a 12.4-fold down-regulation of wild-type <it>MLL </it>and <it>MLL-SEPT2 </it>combined expression (p = 0.028). The down-regulation of <it>SEPT2 </it>in <it>MLL-SEPT2 </it>myeloid neoplasias was statistically significant when compared with all other leukemia genetic subgroups (including those with other <it>MLL </it>gene fusions). In addition, <it>MLL </it>expression was also down-regulated in the group of <it>MLL </it>fusions other than <it>MLL-SEPT2</it>, when compared with the normal control group (p = 0.023)</p> <p>Conclusion</p> <p>We found a significant down-regulation of both <it>SEPT2 </it>and <it>MLL </it>in <it>MLL-SEPT2 </it>myeloid neoplasias. In addition, we also found that <it>MLL </it>is under-expressed in AML patients with <it>MLL </it>fusions other than <it>MLL-SEPT2</it>.</p

Toxocariasis in children attending a Public Health Service Pneumology Unit in Paraná State, Brazil

The enzyme-linked immunosorbent assay (ELISA) is the most widely used tool to detect anti-Toxocara IgG antibodies for both serodiagnostic and seroepidemiological surveys on human toxocariasis. In the last eight years a high prevalence of toxocariasis (32.2-56.0%) has been reported in children attending public health units from municipalities in the state of Paraná, Brazil. Therefore, the aim of this work was to compare the frequency found among the general child population with that of children attending a public pneumology service in Maringá, Paraná, Brazil and describe the laboratorial, clinical and epidemiological findings. The research was conducted at the Consórcio Público Intermunicipal de Saúde do Setentrião Paranaense (CISAMUSEP) from July 2009 to July 2010 among children aged between one and 15 years. From a total of 167 children studied, only 4.2% (7/167) tested positive for anti-Toxocara spp. IgG antibodies and presented mild eosinophilia (2/7), increased serum IgE levels (6/7) and a positive allergy test for mites (5/7). The presence of pets (dogs or cats) at home did not correlate with the seroprevalence. In conclusion, cases of toxocariasis involving the respiratory tract are rare in children attending a public health pneumology unit in the northwestern region of Paraná State, despite the high prevalence of this type of toxocariasis among the infantile population attending Basic Health Units in the same geographical area

Physiology of Escherichia coli at high osmolarity and its use in industrial ethanol production

Biofuels are becoming increasingly important in the light of climate change, increasing energy demands and higher fuel prices. Their production must be carefully balanced against the production of foods and use of fresh water, both of which are consumed by crop based biofuels such as corn ethanol. One proposed solution is to instead use waste materials such as plant matter including wood offcuts and plant trimmings. This waste can be turned into syngas (a mix of CO and H₂) and converted to ethanol using microorganisms. Production of ethanol using microorganisms however, is complicated as the ethanol produced by the cells becomes toxic at higher concentrations, inhibiting their growth and further production. The usual method of keeping the toxicity down to allow further production is to continuously distil ethanol off at low concentrations and consequently, a high cost. Since the mechanisms of ethanol damage to microbes are similar to those that occur during osmotic challenge: damage to the membrane, cytoplasmic dehydration, and protein unfolding, I hypothesized that we can use knowledge of osmoregulatory mechanisms to increase the resistance of cells to ethanol damage and decrease distillation costs. While working under this hypothesis I had to address some of the challenges one faces when understanding the physiology and growth of microbes, and for the purpose I have developed a number of useful techniques; a method for calibrating optical densities to cell number, a neural network for identifying cells and determining their concentrations via microscope imaging and a simple particle diffusion simulation for correcting errors due to confinement of particles within cells. In addition, I have produced a simplified model of industrial production to help evaluate economic impacts that changes to the growth of microbes and the plant process may have. To study any useful links between osmolarity and ethanol resistance, I chose to use Escherichia coli as the model organism due to the large amount of data available on its osmoregulatory mechanisms. It has been long known that when bacteria do grow at high but not lethal osmolarity, they grow at a reduced rate which, even if it increases the ethanol resistance, may have a detrimental effect on the desired production rates. So therefore, in addition to testing the ethanol tolerance of the bacteria under different osmotic conditions, and as a second focus of this project, I have tried to understand why the reduction in growth rates occurs, with the hope of mitigating this effect. This will offer a better understanding of osmotic growth and provide useful insights for industrial bio-production. To this end, I have tried to discern some of the possible reasons for this slower growth by measuring various cell physiological parameters such as batch-culture yield, cytoplasmic diffusion and proteome allocation using my newly developed techniques. I have found a reduction in the cell yield with increasing osmolarity of 50% with an increase of 1Osm of osmotic agent, a slight decrease in cytoplasmic diffusion and a slight decrease in RNA content at high osmolarity. I have also proposed a coarse-grained model of proteome partitioning to help integrate these results and explain growth at high osmolarity. It is still to be determined if, as a whole, the changes observed explain fully the reduction in growth. When it comes to ethanol resistance, and contrary to my hypothesis, I found that increasing the osmolarity of the medium with sucrose or NaCl reduced the ethanol resistance. However, I found that the proW gene provides significant ethanol resistance, indicating glycine betaine, or another substrate for this transporter, is highly useful as a protectant. And this transporter is a potential candidate for overexpression. A reduction in growth temperature also provides significant solvent tolerance at the expense of a reduction in growth rate and hence production.Restricted Acces

Effects of instrumentation, irrigation and dressing with calcium hydroxide on infection in pulpless teeth with periapical bone lesions

Aim The aim of this study was to evaluate the fate of microorganisms in root canals of teeth with infected pulps and periapical bone lesions with and without the use of calcium hydroxide medication. Methodology Endodontic samples were cultured and microorganisms were counted and identified in 4 3 teeth before (sample 1) and after (sample 2) treatment during the first visit and before (sample 3) and after (sample 4) treatment during the second visit. In the first visit teeth were instrumented and half of the teeth were filled with a thick slurry of calcium hydroxide in sterile saline, The other teeth were obturated with gutta-percha and AH-26 seater. After 4 weeks the teeth with calcium-hydroxide were accessed again and after microbiological sampling they were obturated with gutta-percha and AH-26 sealer. Results The mean total colony forming unit (CFU) counts of positive samples dropped significantly as a result or canal preparation during the first visit from 1.0 X 10(6) to 1.8 x 10(3) (between samples 1 and 2) but increased to 9.3 x 10(3) in the period between the two visits (sample 2 and 3). There was no difference in mean total CFU counts of positive samples between the end of the first (sample 21) and the end of the second visit (sample 4). The most frequently isolated species were Prevotella intermedia, Capnocytophaga spp.. Actinomyces odontolyticus, Propionibacterium acnes and Peptostreptococcus micros. Conclusions Although a calcium hydroxide paste was placed in the prepared canals. the number of positive canals had increased in the period between visits. However, the number of microorganisms had only increased to 0.93% of the original number of CFU (sample 1). It is concluded that a calcium hydroxide and sterile saline slurry limits but does not totally prevent regrowth of endodontic bacteria