Archive for March, 2010

Key role of the 3′ untranslated region in the cell cycle regulated expression of the Leishmania infantum histone H2A genes: minor synergistic effect of the 5′ untranslated region.

Tuesday, March 9th, 2010

Background:
Histone synthesis in Leishmania is tightly coupled to DNA replication by a post-transcriptional mechanism operating at the level of translation.
Results:
In this work we have analyzed the implication of the 5′ and 3′ untranslated regions (UTR) in the cell cycle regulated expression of the histone H2A in Leishmania infantum. For that purpose, L. infantum promastigotes were stably transfected with different plasmid constructs in which the CAT coding region used as a reporter was flanked by the 5′ and 3′ UTR regions of the different H2A genes. We report that in spite of their sequence differences, histone H2A 5′ and 3′ UTRs conferred a cell cycle dependent pattern of expression on the CAT reporter since de novo synthesis of CAT increased when parasites enter the S phase. Using one established L. infantum cell line we showed that CAT expression is controlled by the same regulatory events that control the endogenous histone gene expression. Thus, although we did not detect changes in the level of CAT mRNAs during cell cycle progression, a drastic change in the polysome profiles of CAT mRNAs was observed during the progression from G1 to S phase. In the S phase CAT mRNAs were on polyribosomal fractions, but in the G1 phase the association of CAT transcripts with ribosomes was impaired. Furthermore, it was determined that the addition of just the H2A 3′ UTR to the CAT reporter gene is sufficient to achieve a similar pattern of post-transcriptional regulation indicating that this region contains the major regulatory sequences involved in the cell cycle dependent expression of the H2A genes. On the other hand, although CAT transcripts bearing the H2A 5′ alone were translated both in the G1 and S phase, higher percentages of transcripts were detected on polyribosomes in the S phase correlating with an increase in the de novo synthesis of CAT. Thus, it can be concluded that this region also contributes, although to a minor extent than the 3′ UTR, in the enhancement of translation in the S phase relative to the G1 phase.
Conclusions:
Our findings indicate that both, the 5′ and the 3′ UTRs contain sequence elements that contribute to the cell cycle expression of L. infantum H2A. The 3′ UTR region is essential for cell cycle dependent translation of the L. infantum H2A transcripts whereas the 5′ UTR has a minor contribution in their S phase dependent translation.

Geographic distributions of Idh-1 alleles in a cricket are linked to differential enzyme kinetic performance across thermal environments

Tuesday, March 9th, 2010

Background:
Geographic clines within species are often interpreted as evidence of adaptation to varying environmental conditions. However, clines can also result from genetic drift, and these competing hypotheses must therefore be tested empirically. The striped ground cricket, Allonemobius socius, is widely-distributed in the eastern United States, and clines have been documented in both life-history traits and genetic alleles. One clinally-distributed locus, isocitrate dehydrogenase (Idh-1), has been shown previously to exhibit significant correlations between allele frequencies and environmental conditions (temperature and rainfall). Further, an empirical study revealed a significant genotype-by-environmental interaction (GxE) between Idh-1 genotype and temperature which affected fitness. Here, we use enzyme kinetics to further explore GxE between Idh-1 genotype and temperature, and test the predictions of kinetic activity expected under drift or selection.
Results:
We found significant GxE between temperature and three enzyme kinetic parameters, providing further evidence that the natural distributions of Idh-1 allele frequencies in A. socius are maintained by natural selection. Differences in enzyme kinetic activity across temperatures also mirror many of the geographic patterns observed in allele frequencies.
Conclusions:
This study further supports the hypothesis that the natural distribution of Idh-1 alleles in A. socius is driven by natural selection on differential enzymatic performance. This example is one of several which clearly document a functional basis for both the maintenance of common alleles and observed clines in allele frequencies, and provides further evidence for the non-neutrality of some allozyme alleles.

Electromyographic analysis of selected shoulder muscles during a rugby football tackle

Tuesday, March 9th, 2010

Background:
Epidemiological studies have shown that the incidence of shoulder injuries is increasing in rugby and the majority are related to the contact/tackle phase of play. However, no data currently exists that describes preparatory muscle activity during tackle. This information could aid in guiding training and rehabilitation, if available. The purpose of the study was to assess the sequence of onset of EMG activity of selected scapulohumeral muscles during rugby tackle. 15 healthy professional rugby players participated in the study. Surface EMG activity was assessed for timing of onset relative to time of impact during a modified tackle activity in pectorialis major, biceps brachii, latissimus dorsi, serratus anterior and infraspinatus muscles.
Results:
Onset of activity occurred in all muscles prior to impact. Factorial ANOVA showed significant differences between muscles in activation timing (p=0.0001), paired t-tests revealed that serratus anterior was activated prior to all other muscles tested (p0.05), except pectorialis major on all comparisons showed significantly later activation timing than all other muscles (p<0.001).
Conclusion:
Muscle activation timing may if not properly balanced around the shoulder girdle expose the glenohumeral joint to excessive load and stress. This paper demonstrates a simple method which sets out some preliminary normative data in healthy players. Further studies relating these data to injured players are required.

Alterations in transcriptional responses associated with vascular aging

Tuesday, March 9th, 2010

Vascular aging is an independent risk factor for cardiovascular disease that can occur in the absence of other traditional risk factors. A hallmark of vascular aging is inflammation that ultimately leads to structural changes in the vessel wall including an increase in medial thickness and perivascular fibrosis. Several classes of transcription factors have been identified that participate in the regulation of cellular responses associated with vascular aging. Nuclear factor (NF)-kappaB is the prototypic example of a transcriptional activator in the setting of inflammation, being activated in response to multiple inflammatory mediators including pro-inflammatory cytokines and bacterial endotoxin. In contrast, the activation of the nuclear hormone receptor and transcription factor peroxisome proliferator-activated receptor-alpha (PPAR-alpha) results in its translocation from the cell surface to the nucleus where it exerts anti-inflammatory effects. Vascular aging is also associated with endothelial dysfunction. One important repair mechanism for improving endothelial function is the recruitment of endothelial progenitor cells (EPCs). In the setting of aging the number of EPCs diminishes which has been linked to a decrease in the activity and/or expression of the transcription factor hypoxia inducible factor (HIF)-1 alpha. A change in the balance of the activity of pro-inflammatory transcription factors versus those that inhibit inflammation likely contributes to the process of vascular aging. The purpose of this review is to summarize our current knowledge of these age-related changes in transcriptional responses, and to discuss the therapeutic potential of targeting some of these factors.

Laboratory-Based Evaluation of Legionellosis Epidemiology in Ontario, Canada, 1978 to 2006

Tuesday, March 9th, 2010

Background:
Legionellosis is a common cause of severe community acquired pneumonia and respiratory disease outbreaks. The Ontario Public Health Laboratory (OPHL) has conducted most testing for Legionella species in the Canadian province of Ontario since 1978, and represents a multi-decade repository of population-based data on legionellosis epidemiology. We sought to provide a laboratory-based review of the epidemiology of legionellosis in Ontario over the past 3 decades, with a focus on changing rates of disease and species associated with legionellosis during that time period.
Methods:
We analyzed cases that were submitted and tested positive for legionellosis from 1978 to 2006 using Poisson regression models incorporating temporal, spatial, and demographic covariates. Predictors of infection with culture-confirmed L. pneumophila serogroup 1 (LP1) were evaluated with logistic regression models.
Results:
1,401 cases of legionellosis tested positive from 1978 to 2006. As in other studies, we found a late summer to early autumn seasonality in disease occurrence with disease risk increasing with age and in males. In contrast to other studies, we found a decreasing trend in cases in the recent decade (IRR 0.93, 95% CI 0.91 to 0.95, P-value = 0.001); only 66% of culture-confirmed isolates were found to be LP1.
Conclusions:
Despite similarities with disease epidemiology in other regions, legionellosis appears to have declined in the past decade in Ontario, in contrast to trends observed in the United States and parts of Europe. Furthermore, a different range of Legionella species is responsible for illness, suggesting a distinctive legionellosis epidemiology in this North American region.