Stimulation of the acute-phase response in simian virus 40-hepatocyte cell lines.

Seven simian virus 40 (SV40)-hepatocyte cell lines were characterized with respect to the ability to express eight liver acute-phase genes. cDNA clones corresponding to albumin, serum amyloid A, alpha 1-acid glycoprotein, haptoglobin, alpha-, beta-, and gamma-fibrinogen, and alpha 1-major-acute-phase protein mRNAs were used in Northern (RNA) or slot blot analyses. In the noninduced state, six of the seven cell lines showed significant (i.e., liverlike) levels of constitutive expression of all genes examined except that expression of haptoglobin mRNA was considerable lower than in the normal liver. To examine whether these immortalized liver cells can respond appropriately to inflammatory mediators, cells were treated with conditioned medium from activated human monocytes or mixed lymphocyte cultures. Results showed that these SV40-hepatocyte cell lines responded to the conditioned media in culture by down-regulating albumin gene expression and up-regulating other acute-phase genes in a time- and dose-dependent manner. These results indicate that the SV40-hepatocytes retained not only the ability to express a number of acute-phase genes but also the ability to respond to external stimuli. The usefulness of these cell lines for analysis of the molecular mechanisms involved in the regulation of these acute-phase genes is discussed.     

Comparative phylogenetic histories of two louse genera found on Catharus thrushes and other birds

The louse genera Brueelia (Ischnocera) and Myrsidea (Amblycera) are broadly codistributed on songbirds (Passeriformes), but differ in a variety of life history characteristics. We used mitochondrial and nuclear DNA sequences to assess levels of genetic divergence and reconstruct phylogenies of these 2 genera, focusing especially on Catharus thrushes in North America. We then qualitatively compared the phylogenies and levels of divergence within these 2 genera of codistributed parasites. Neither Brueelia nor Myrsidea appears to cospeciate with Catharus thrushes or passerine birds in general. The Myrsidea phylogeny exhibits significant levels of biogeographic structure, whereas the Brueelia phylogeny does not. Myrsidea and Brueelia also differ in their levels of intra-generic genetic divergence, with Myrsidea showing higher levels of genetic divergence and host specificity than Brueelia. Our genetic data support traditional morphology-based taxonomy in several instances in which the same species of Brueelia has been reported on multiple host taxa, e.g., all migrant Catharus spp. carry B. antiqua, with little haplotype divergence. Myrsidea found on each Catharus sp. are in general genetically distinct, except for M. incerta, which parasitizes both Catharus ustulatus and Catharus minimus. The strong biogeographic signal in the Myrsidea phylogeny and higher relative levels of host specificity of Myrsidea spp. suggest that infrequent host-switching, followed by speciation, is shaping the evolutionary history of this group. In contrast, the relatively lower host specificity of Brueelia spp. suggests that host-switching, combined with more frequent ongoing dispersal, has been more important in the evolutionary history of Brueelia.     

Sleep-Wake Cycle Dysfunction in the TgCRND8 Mouse Model of Alzheimer’s Disease: From Early to Advanced Pathological Stages

In addition to cognitive decline, individuals affected by Alzheimer’s disease (AD) can experience important neuropsychiatric symptoms including sleep disturbances. We characterized the sleep-wake cycle in the TgCRND8 mouse model of AD, which overexpresses a mutant human form of amyloid precursor protein resulting in high levels of β-amyloid and plaque formation by 3 months of age. Polysomnographic recordings in freely-moving mice were conducted to study sleep-wake cycle architecture at 3, 7 and 11 months of age and corresponding levels of β-amyloid in brain regions regulating sleep-wake states were measured. At all ages, TgCRND8 mice showed increased wakefulness and reduced non-rapid eye movement (NREM) sleep during the resting and active phases. Increased wakefulness in TgCRND8 mice was accompanied by a shift in the waking power spectrum towards fast frequency oscillations in the beta (14-20 Hz) and low gamma range (20-50 Hz). Given the phenotype of hyperarousal observed in TgCRND8 mice, the role of noradrenergic transmission in the promotion of arousal, and previous work reporting an early disruption of the noradrenergic system in TgCRND8, we tested the effects of the alpha-1-adrenoreceptor antagonist, prazosin, on sleep-wake patterns in TgCRND8 and non-transgenic (NTg) mice. We found that a lower dose (2 mg/kg) of prazosin increased NREM sleep in NTg but not in TgCRND8 mice, whereas a higher dose (5 mg/kg) increased NREM sleep in both genotypes, suggesting altered sensitivity to noradrenergic blockade in TgCRND8 mice. Collectively our results demonstrate that amyloidosis in TgCRND8 mice is associated with sleep-wake cycle dysfunction, characterized by hyperarousal, validating this model as a tool towards understanding the relationship between β-amyloid overproduction and disrupted sleep-wake patterns in AD.     

We're Gonna Crush It! Sediment Creation through Destruction

Why are there only crumbs left at the bottom of the cereal box? Many factors, such as package handling, have caused the cereal pieces to break down into crumbs. This explanation is also related to the process of creating sediment from rocks. Sediment is created by weathering over millions of years, and it is deposited all over the world by erosion. The We're Gonna Crush It! activity serves as a brief introduction to sediment composition. Students learn how sediment is created by demonstrating weathering at a much quicker pace than what occurs under natural conditions. They see firsthand how weathering affects rocks by mirroring the process using cereal to make “sediment.” The students learn how to calculate grain size percentages to determine the overall composition of the sediment while becoming more aware of the organisms and organic matter that are also present. In less than 60 min, students can experience a large-scale process that generally occurs over millions of years. Several Next Generation Science Standards, Common Core Math Standards, and Ocean Literacy Standards are addressed in this activity.     

PR3-ANCA: A Promising Biomarker in Primary Sclerosing Cholangitis (PSC)

Background and Aims

The only recognized biomarker for primary sclerosing cholangitis (PSC) is atypical anti-neutrophil cytoplasmic antibodies (aANCA), which, in addition to having low sensitivity and specificity, is an indirect immunofluorescence (IIF) test lacking the advantages of high throughput and objectivity. Recent reports have shown that antibodies to proteinase-3 (PR3-ANCA) might add diagnostic value in inflammatory bowel disease (IBD), specifically in ulcerative colitis (UC). As PSC is associated with IBD, the objective of this study was to evaluate the frequency and clinical significance of PR3-ANCA in a large cohort of patients.

Methods

A total of 244 PSC and 254 control [autoimmune hepatitis (AIH), primary biliary cirrhosis (PBC), hepatitis C viral infection (HCV), hepatitis B viral infection (HBV), and healthy controls] sera and their clinical correlations were retrospectively analyzed for PR3-ANCA determined by ELISA and a new chemiluminescence immunoassay (CIA). Testing was also performed for aANCA by IIF.

Results

When measured by CIA, PR3-ANCA was detected in 38.5% (94/244) of PSC patients compared to 10.6% (27/254) controls (p<0.0001). By ELISA, PR3-ANCA was detected in 23.4% (57/244) of PSC patients compared to 2.7% (6/254) controls (p<0.0001). PR3-ANCA in PSC patients was not associated with the presence or type of underlying IBD, and, in fact, it was more frequent in Crohn''s disease (CD) patients with PSC than previously reported in CD alone. PR3-ANCA in PSC measured by CIA correlated with higher liver enzymes.

Conclusion

PR3-ANCA is detected in a significant proportion of PSC patients compared to other liver diseases including PBC and AIH. PR3-ANCA is associated with higher liver enzyme levels in PSC, and is not solely related to underlying IBD.     

Phylogenetic signal in matK vs. trnK: a case study in early diverging eudicots (angiosperms)

The matK gene has been among the most useful loci for resolving plant phylogenetic relationships at different evolutionary time-scales, but much less is known about the phylogenetic utility of the flanking trnK intron, especially for deep level phylogenetics. We compared the relative performance of matK and trnK intron regions for resolving the relationships of the early diverging eudicots (angiosperms). The two regions display similar nucleotide compositions and distributions of rate variation among sites. The trnK intron sequences also provide similar levels of phylogenetic information per-site as matK. Combining the trnK intron sequences with matK increases overall bootstrap support for the early diverging eudicots compared to analyses of matK alone. MP, ML and Bayesian analyses provide strong support for eudicots, the sister group relationship of Ranunculales to remaining eudicots, and a Buxales+Trochodendraceae+core eudicots clade. matK and the trnK intron support conflicting positions for Buxales and Trochodendrales in relation to the core eudicots.