Telomere shortening exposes functions for the mouse Werner and Bloom syndrome genes

The Werner and Bloom syndromes are caused by loss-of-function mutations in WRN and BLM, respectively, which encode the RecQ family DNA helicases WRN and BLM, respectively. Persons with Werner syndrome displays premature aging of the skin, vasculature, reproductive system, and bone, and those with Bloom syndrome display more limited features of aging, including premature menopause; both syndromes involve genome instability and increased cancer. The proteins participate in recombinational repair of stalled replication forks or DNA breaks, but the precise functions of the proteins that prevent rapid aging are unknown. Accumulating evidence points to telomeres as targets of WRN and BLM, but the importance in vivo of the proteins in telomere biology has not been tested. We show that Wrn and Blm mutations each accentuate pathology in later-generation mice lacking the telomerase RNA template Terc, including acceleration of phenotypes characteristic of latest-generation Terc mutants. Furthermore, pathology not observed in Terc mutants but similar to that observed in Werner syndrome and Bloom syndrome, such as bone loss, was observed. The pathology was accompanied by enhanced telomere dysfunction, including end-to-end chromosome fusions and greater loss of telomere repeat DNA compared with Terc mutants. These findings indicate that telomere dysfunction may contribute to the pathogenesis of Werner syndrome and Bloom syndrome.     

Genetic structure of the LXS panel of recombinant inbred mouse strains: a powerful resource for complex trait analysis

The set of LXS recombinant inbred (RI) strains is a new and exceptionally large mapping panel that is suitable for the analysis of complex traits with comparatively high power. This panel consists of 77 strains—more than twice the size of other RI sets— and will typically provide sufficient statistical power (=0.8) to map quantitative trait loci (QTLs) that account for 25% of genetic variance with a genomewide p < 0.05. To characterize the genetic architecture of this new set of RI strains, we genotyped 330 MIT microsatellite markers distributed on all autosomes and the X Chromosome and assembled error-checked meiotic recombination maps that have an average F₂-adjusted marker spacing of 4 cM. The LXS panel has a genetic structure consistent with random segregation and subsequent fixation of alleles, the expected 3–4 × map expansion, a low level of nonsyntenic association among loci, and complete independence among all 77 strains. Although the parental inbred strains—Inbred Long-Sleep (ILS) and Inbred Short-Sleep (ISS)—were derived originally by selection from an 8-way heterogeneous stock selected for differential sensitivity to sedative effects of ethanol, the LXS panel is also segregating for many other traits. Thus, the LXS panel provides a powerful new resource for mapping complex traits across many systems and disciplines and should prove to be of great utility in modeling the genetics of complex diseases in human populations.(Robert W. Williams and Beth Bennett)These authors contributed equally to this work.     

The investigation of cytosolic phospholipase A2 using ELISA

Two studies of the behaviour of cytosolic phospholipase A(2) (cPLA(2)) in the red blood cell (RBC), as measured by ELISA, are described. In the first study we show a significant increase in cPLA(2) in patients with schizophrenia compared to controls and suggest that this measure, if corroborated, could be used as a diagnostic marker. In a second study we found that washing the RBC introduced an unknown confounding variable which led us to reject this study. A subsequent investigation of washing red cells showed that the washing effect may be due to a plasma factor likely to be more than 5kDa MW which can be removed from red cells by washing with buffers. When the cells are washed, the concentration of cPLA(2) in the red cell, as measured by ELISA, significantly increases. We advise against washing the red cell in any study that involves measuring cPLA(2) by ELISA.     

Utility of different gene enrichment approaches toward identifying and sequencing the maize gene space

Maize (Zea mays) possesses a large, highly repetitive genome, and subsequently a number of reduced-representation sequencing approaches have been used to try and enrich for gene space while eluding difficulties associated with repetitive DNA. This article documents the ability of publicly available maize expressed sequence tag and Genome Survey Sequences (GSSs; many of which were isolated through the use of reduced representation techniques) to recognize and provide coverage of 78 maize full-length cDNAs (FLCs). All 78 FLCs in the dataset were identified by at least three GSSs, indicating that the majority of maize genes have been identified by at least one currently available GSS. Both methyl-filtration and high-Cot enrichment methods provided a 7- to 8-fold increase in gene discovery rates as compared to random sequencing. The available maize GSSs aligned to 75% of the FLC nucleotides used to perform searches, while the expressed sequence tag sequences aligned to 73% of the nucleotides. Our data suggest that at least approximately 95% of maize genes have been tagged by at least one GSS. While the GSSs are very effective for gene identification, relatively few (18%) of the FLCs are completely represented by GSSs. Analysis of the overlap of coverage and bias due to position within a gene suggest that RescueMu, methyl-filtration, and high-Cot methods are at least partially nonredundant.     

Influence of roots and climate on mineral and trace element storage and flux in tropical mangrove soils

The storage and flux of various mineral and trace elements in soils (0–30cm depth) were examined in relation to monsoonal rains and fine root biomass in four mangrove forests of different age and type in southern Thailand. The onset of the wet SW monsoon resulted in the percolation and dilution of porewater solutes by rainwater and by less saline tidal water, as indicated by shifts in Eh, pH and porewater SO₄/Cl ratios. This is contrary to temperate intertidal environments where seasonal patterns of porewater constituents, and biological and biogeochemical activities, are strongly cued to temperature. Fluxes across the soil–water interface were most often not statistically significant. Concentration of dissolved porewater metals were dominated by Fe, Mn, Al, Mo and Zn. The decreasing order of solid-phase element inventories in these soils, on average, was: Al, S, Fe, Na, Mg, K, Ca, N, P, Mn, V, Zn, Cr, Ni, As, Co, Cu, Pb, Mo, Cd and Hg. There were no gradients in concentrations of dissolved or solid-phase elements with increasing soil depth. This phenomenon was attributed to physical and biological processes, including the presence and activities of roots and tidal recharge of soil water. Fine dead roots were storage sites for most mineral and trace elements, as some elements in roots composed a significant fraction (5%) of the total soil pool. Analysis of S and Fe concentration differences between live and dead roots suggested extensive formation of pyrite associated with dead roots; correlation analysis suggested that trace metals coprecipitated with pyrite. An analysis of inventories and release/uptake rates indicate turnover of the N, P, Na and Ca soil pools equivalent to other tropical forests; turnover was slow (decades to centuries) for S, Fe, K and trace elements. Our results indicate that mineral and trace element cycling in these soils are characterized by net storage, with net accumulation of most elements much greater than uptake and release by tree roots.     

Pain and compassion in the neonatal unit -- a neonatologist's view

Research has shown that even extremely premature babies are sufficiently developed, anatomically and physiologically, to be capable of experiencing and responding to pain. All newborn infants and especially those who require intensive care in the first days of life are exposed to some painful procedures. Part of the neonatologist's role is the detection and management of pain in these infants. Difficult challenges come with this role. All medications carry known or potential adverse effects and limited research has been done in this vulnerable population. The benefits and risks of all available pain-relieving measures should be balanced when planning management. Compassion is no excuse for a high incidence of undesirable or dangerous side effects. We must proceed with great care.     

Regulation of protein kinases; controlling activity through activation segment conformation

There are currently at least forty-six unique protein kinase crystal structures, twenty-four of which are available in an active state. Here we examine these structures using a structural bioinformatics approach to understand how the conformation of the activation segment controls kinase activity.     

SeqExpress: desktop analysis and visualization tool for gene expression experiments

SUMMARY: SeqExpress is a stand-alone desktop application for the identification of relevant genes within collections of microarray or SAGE experiments. A number of analysis, filtering and visualization tools are provided to aid in the selection of groups of genes. If R is installed then the application can use this to provide further analysis. AVAILABILITY: SeqExpress is available at: http://www.seqexpress.com     

Predicting function from structure: 3D structure studies of the mammalian Golgi complex

3D electron tomography studies of the structure of the mammalian Golgi complex have led to four functional predictions (1). The sorting and exit site from the Golgi comprises two or three distinct trans-cisternae (2). The docking of vesicular-tubular clusters at the cis-face and the fragmentation of trans-cisternae are coordinated (3). The mechanisms of transport through, and exit from, the Golgi vary with physiological state, and in different cells and tissues (4). Specialized trans-ER functions in the delivery of ceramide to sphingomyelin synthase in the trans-Golgi membrane, for the regulated sorting via sphingolipid-cholesterol-rich domains. These structure-based predictions can now be tested using a variety of powerful cell and molecular tools.