Expression of Bcl-2 and Bax in Mouse Renal Tubules during Kidney Development

Bcl-2 and Bax play an important role in apoptosis regulation, as well as in cell adhesion and migration during kidney morphogenesis, which is structurally and functionally related to mitochondria. In order to elucidate the role of Bcl-2 and Bax during kidney development, it is essential to establish the exact location of their expression in the kidney. The present study localized their expression during kidney development. Kidneys from embryonic (E) 16-, 17-, 18-day-old mouse fetuses, and postnatal (P) 1-, 3-, 5-, 7-, 14-, 21-day-old pups were embedded in Epon. Semi-thin serial sections from two E17 kidneys underwent computer assisted 3D tubule tracing. The tracing was combined with a newly developed immunohistochemical technique, which enables immunohistochemistry on glutaraldehyde fixated plastic embedded sections. Thereby, the microstructure could be described in detail, and the immunochemistry can be performed using exactly the same sections. The study showed that Bcl-2 and Bax were strongly expressed in mature proximal convoluted tubules at all time points, less strongly expressed in proximal straight tubules, and only weakly in immature proximal tubules and distal tubules. No expression was detected in ureteric bud and other earlier developing structures, such as comma bodies, S shaped bodies, glomeruli, etc. Tubules expressing Bcl-2 only were occasionally observed. The present study showed that, during kidney development, Bcl-2 and Bax are expressed differently in the proximal and distal tubules, although these two tubule segments are almost equally equipped with mitochondria. The functional significance of the different expression of Bcl-2 and Bax in proximal and distal tubules is unknown. However, the findings of the present study suggest that the mitochondrial function differs between mature proximal tubules and in the rest of the tubules. The function of Bcl-2 and Bax during tubulogenesis still needs to be investigated

Comparing methods to estimate treatment effects on a continuous outcome in multicentre randomized controlled trials: A simulation study

<p>Abstract</p> <p>Background</p> <p>Multicentre randomized controlled trials (RCTs) routinely use randomization and analysis stratified by centre to control for differences between centres and to improve precision. No consensus has been reached on how to best analyze correlated continuous outcomes in such settings. Our objective was to investigate the properties of commonly used statistical models at various levels of clustering in the context of multicentre RCTs.</p> <p>Methods</p> <p>Assuming no treatment by centre interaction, we compared six methods (ignoring centre effects, including centres as fixed effects, including centres as random effects, generalized estimating equation (GEE), and fixed- and random-effects centre-level analysis) to analyze continuous outcomes in multicentre RCTs using simulations over a wide spectrum of intraclass correlation (ICC) values, and varying numbers of centres and centre size. The performance of models was evaluated in terms of bias, precision, mean squared error of the point estimator of treatment effect, empirical coverage of the 95% confidence interval, and statistical power of the procedure.</p> <p>Results</p> <p>While all methods yielded unbiased estimates of treatment effect, ignoring centres led to inflation of standard error and loss of statistical power when within centre correlation was present. Mixed-effects model was most efficient and attained nominal coverage of 95% and 90% power in almost all scenarios. Fixed-effects model was less precise when the number of centres was large and treatment allocation was subject to chance imbalance within centre. GEE approach underestimated standard error of the treatment effect when the number of centres was small. The two centre-level models led to more variable point estimates and relatively low interval coverage or statistical power depending on whether or not heterogeneity of treatment contrasts was considered in the analysis.</p> <p>Conclusions</p> <p>All six models produced unbiased estimates of treatment effect in the context of multicentre trials. Adjusting for centre as a random intercept led to the most efficient treatment effect estimation across all simulations under the normality assumption, when there was no treatment by centre interaction.</p

Large Scale Comparative Codon-Pair Context Analysis Unveils General Rules that Fine-Tune Evolution of mRNA Primary Structure

BACKGROUND: Codon usage and codon-pair context are important gene primary structure features that influence mRNA decoding fidelity. In order to identify general rules that shape codon-pair context and minimize mRNA decoding error, we have carried out a large scale comparative codon-pair context analysis of 119 fully sequenced genomes. METHODOLOGIES/PRINCIPAL FINDINGS: We have developed mathematical and software tools for large scale comparative codon-pair context analysis. These methodologies unveiled general and species specific codon-pair context rules that govern evolution of mRNAs in the 3 domains of life. We show that evolution of bacterial and archeal mRNA primary structure is mainly dependent on constraints imposed by the translational machinery, while in eukaryotes DNA methylation and tri-nucleotide repeats impose strong biases on codon-pair context. CONCLUSIONS: The data highlight fundamental differences between prokaryotic and eukaryotic mRNA decoding rules, which are partially independent of codon usage

First beta-decay spectroscopy of In-135 and new beta-decay branches of In-134

The beta decay of the neutron-rich In-134 and In-135 was investigated experimentally in order to provide new insights into the nuclear structure of the tin isotopes with magic proton number Z = 50 above the N = 82 shell. The beta-delayed gamma-ray spectroscopy measurement was performed at the ISOLDE facility at CERN, where indium isotopes were selectively laser-ionized and on-line mass separated. Three beta-decay branches of In-134 were established, two of which were observed for the first time. Population of neutron-unbound states decaying via. rays was identified in the two daughter nuclei of In-134, Sn-134 and Sn-133, at excitation energies exceeding the neutron separation energy by 1 MeV. The beta-delayed one- and two-neutron emission branching ratios of In-134 were determined and compared with theoretical calculations. The beta-delayed one-neutron decay was observed to be dominant beta-decay branch of In-134 even though the Gamow-Teller resonance is located substantially above the two-neutron separation energy of Sn-134. Transitions following the beta decay of In-135 are reported for the first time, including. rays tentatively attributed to Sn-135. In total, six new levels were identified in Sn-134 on the basis of the beta.. coincidences observed in the In-134 and In-135 beta decays. A transition that might be a candidate for deexciting the missing neutron single-particle 13/2(+) state in Sn-133 was observed in both beta decays and its assignment is discussed. Experimental level schemes of Sn-134 and Sn-135 are compared with shell-model predictions. Using the fast timing technique, half-lives of the 2(+), 4(+), and 6(+) levels in Sn-134 were determined. From the lifetime of the 4(+) state measured for the first time, an unexpectedly large B(E2; 4(+)-> 2(+)) transition strength was deduced, which is not reproduced by the shell-model calculations.Peer reviewe

Proteins with Complex Architecture as Potential Targets for Drug Design: A Case Study of Mycobacterium tuberculosis

Lengthy co-evolution of Homo sapiens and Mycobacterium tuberculosis, the main causative agent of tuberculosis, resulted in a dramatically successful pathogen species that presents considerable challenge for modern medicine. The continuous and ever increasing appearance of multi-drug resistant mycobacteria necessitates the identification of novel drug targets and drugs with new mechanisms of action. However, further insights are needed to establish automated protocols for target selection based on the available complete genome sequences. In the present study, we perform complete proteome level comparisons between M. tuberculosis, mycobacteria, other prokaryotes and available eukaryotes based on protein domains, local sequence similarities and protein disorder. We show that the enrichment of certain domains in the genome can indicate an important function specific to M. tuberculosis. We identified two families, termed pkn and PE/PPE that stand out in this respect. The common property of these two protein families is a complex domain organization that combines species-specific regions, commonly occurring domains and disordered segments. Besides highlighting promising novel drug target candidates in M. tuberculosis, the presented analysis can also be viewed as a general protocol to identify proteins involved in species-specific functions in a given organism. We conclude that target selection protocols should be extended to include proteins with complex domain architectures instead of focusing on sequentially unique and essential proteins only

RNA Interference in Schistosoma mansoni Schistosomula: Selectivity, Sensitivity and Operation for Larger-Scale Screening

RNA interference (RNAi) is a technique to selectively suppress mRNA of individual genes and, consequently, their cognate proteins. RNAi using double-stranded (ds) RNA has been used to interrogate the function of mainly single genes in the flatworm, Schistosoma mansoni, one of a number of schistosome species causing schistosomiasis. In consideration of large-scale screens to identify candidate drug targets, we examined the selectivity and sensitivity (the degree of suppression) of RNAi for 11 genes produced in different tissues of the parasite: the gut, tegument (surface) and otherwise. We used the schistosomulum stage prepared from infective cercariae larvae which are accessible in large numbers and adaptable to automated screening platforms. We found that RNAi suppresses transcripts selectively, however, the sensitivity of suppression varies (40%–>75%). No obvious changes in the parasite occurred post-RNAi, including after targeting the mRNA of genes that had been computationally predicted to be essential for survival. Additionally, we defined operational parameters to facilitate large-scale RNAi, including choice of culture medium, transfection strategy to deliver dsRNA, dose- and time-dependency, and dosing limits. Finally, using fluorescent probes, we show that the developing gut allows rapid entrance of dsRNA into the parasite to initiate RNAi

New ß-decaying state in 214Bi

A new β-decaying state in 214Bi has been identified at the ISOLDE Decay Station at the CERN-ISOLDE facility. A preferred Iπ = (8−) assignment was suggested for this state based on the β-decay feeding pattern to levels in 214Po and shell-model calculations. The half-life of the Iπ = (8−) state was deduced to be T1/2 = 9.39(10) min. The deexcitation of the levels populated in 214Po by the β decay of this state was investigated via γ -γ coincidences and a number of new levels and transitions was identified. Shell-model calculations for excited states in 214Bi and 214Po were performed using two different effective interactions: the H208 and the modified Kuo-Herling particle interaction. Both calculations agree on the interpretation of the new β-decaying state as an Iπ = 8− isomer and allow for tentative assignment of shell-model states to several high-spin states in 214Po.peerReviewe

New ß-decaying state in 214Bi

A new β-decaying state in 214Bi has been identified at the ISOLDE Decay Station at the CERN-ISOLDE facility. A preferred Iπ = (8−) assignment was suggested for this state based on the β-decay feeding pattern to levels in 214Po and shell-model calculations. The half-life of the Iπ = (8−) state was deduced to be T1/2 = 9.39(10) min. The deexcitation of the levels populated in 214Po by the β decay of this state was investigated via γ -γ coincidences and a number of new levels and transitions was identified. Shell-model calculations for excited states in 214Bi and 214Po were performed using two different effective interactions: the H208 and the modified Kuo-Herling particle interaction. Both calculations agree on the interpretation of the new β-decaying state as an Iπ = 8− isomer and allow for tentative assignment of shell-model states to several high-spin states in 214Po.peerReviewe

Change in structure between the I = 1/2 states in 181Tl and 177,179Au

The first accurate measurements of the α-decay branching ratio and half-life of the Iπ=1/2+ ground state in 181Tl have been made, along with the first determination of the magnetic moments and I=1/2 spin assignments of the ground states in 177,179Au. The results are discussed within the complementary systematics of the reduced α-decay widths and nuclear g factors of low-lying, Iπ=1/2+ states in the neutron-deficient lead region. The findings shed light on the unexpected hindrance of the 1/2+→1/2+, 181Tl→g177Aug α decay, which is explained by a mixing of π3s1/2 and π2d3/2 configurations in 177Aug, whilst 181Tlg remains a near-pure π3s1/2. This conclusion is inferred from the g factor of 177Aug which has an intermediate value between those of π3s1/2 and π2d3/2 states. A similar mixed configuration is proposed for the Iπ=1/2+ ground state of 179Au. This mixing may provide evidence for triaxial shapes in the ground states in these nuclei