Near-saturated and complete genetic linkage map of black spruce (Picea mariana)

<p>Abstract</p> <p>Background</p> <p>Genetic maps provide an important genomic resource for understanding genome organization and evolution, comparative genomics, mapping genes and quantitative trait loci, and associating genomic segments with phenotypic traits. Spruce (<it>Picea</it>) genomics work is quite challenging, mainly because of extremely large size and highly repetitive nature of its genome, unsequenced and poorly understood genome, and the general lack of advanced-generation pedigrees. Our goal was to construct a high-density genetic linkage map of black spruce (<it>Picea mariana</it>, 2n = 24), which is a predominant, transcontinental species of the North American boreal and temperate forests, with high ecological and economic importance.</p> <p>Results</p> <p>We have developed a near-saturated and complete genetic linkage map of black spruce using a three-generation outbred pedigree and amplified fragment length polymorphism (AFLP), selectively amplified microsatellite polymorphic loci (SAMPL), expressed sequence tag polymorphism (ESTP), and microsatellite (mostly cDNA based) markers. Maternal, paternal, and consensus genetic linkage maps were constructed. The maternal, paternal, and consensus maps in our study consistently coalesced into 12 linkage groups, corresponding to the haploid chromosome number (1n = 1x = 12) of 12 in the genus <it>Picea</it>. The maternal map had 816 and the paternal map 743 markers distributed over 12 linkage groups each. The consensus map consisted of 1,111 markers distributed over 12 linkage groups, and covered almost the entire (> 97%) black spruce genome. The mapped markers included 809 AFLPs, 255 SAMPL, 42 microsatellites, and 5 ESTPs. Total estimated length of the genetic map was 1,770 cM, with an average of one marker every 1.6 cM. The maternal, paternal and consensus genetic maps aligned almost perfectly.</p> <p>Conclusion</p> <p>We have constructed the first high density to near-saturated genetic linkage map of black spruce, with greater than 97% genome coverage. Also, this is the first genetic map based on a three-generation outbred pedigree in the genus <it>Picea</it>. The genome length in <it>P. mariana </it>is likely to be about 1,800 cM. The genetic maps developed in our study can serve as a reference map for various genomics studies and applications in <it>Picea a</it>nd Pinaceae.</p

Characteristics of Mechanical Ventilation Employed in Intensive Care Units: A Multicenter Survey of Hospitals

A 1D point-prevalence study was performed to describe the characteristics of conventional mechanical ventilation in intensive care units (ICUs). In addition, a survey was conducted to determine the characteristics of ICUs. A prospective, multicenter study was performed in ICUs at 24 university hospitals. The study population consisted of 223 patients who were receiving mechanical ventilation or had been weaned off mechanical ventilation within the past 24 hr. Common indications for the initiation of mechanical ventilation included acute respiratory failure (66%), acute exacerbation of chronic respiratory failure (15%) (including tuberculosis-destroyed lung [5%]), coma (13%), and neuromuscular disorders (6%). Mechanical ventilation was delivered via an endotracheal tube in 68% of the patients, tracheostomy in 28% and facial mask with noninvasive ventilation (NIV) in 4%. NIV was used in 2 centers. In patients who had undergone tracheostomy, the procedure had been performed 16.9±8.1 days after intubation. Intensivists treated 29% of the patients. A need for additional educational programs regarding clinical practice in the ICU was expressed by 62% of the staff and 42% of the nurses. Tuberculosis-destroyed lung is a common indication for mechanical ventilation in acute exacerbation of chronic respiratory failure, and noninvasive ventilation was used in a limited number of ICUs

Dual AAV/IL-10 Plus STAT3 Anti-Inflammatory Gene Delivery Lowers Atherosclerosis in LDLR KO Mice, but without Increased Benefit

Both IL-10 and STAT3 are in the same signal transduction pathway, with IL-10-bound IL10 receptor (R) acting through STAT3 for anti-inflammatory effect. To investigate possible therapeutic synergism, we delivered both full-length wild-type human (h) STAT3 and hIL-10 genes by separate adenoassociated virus type 8 (AAV8) tail vein injection into LDLR KO on HCD. Compared to control Neo gene-treated animals, individual hSTAT3 and hIL-10 delivery resulted in significant reduction in atherogenesis, as determined by larger aortic lumen size, thinner aortic wall thickness, and lower blood velocity (all statistically significant). However, dual hSTAT3/hIL-10 delivery offered no improvement in therapeutic effect. Plasma cholesterol levels in dual hSTAT3/hIL-10-treated animals were statistically higher compared to hIL-10 alone. While no advantage was seen in this case, we consider that the dual gene approach has intrinsic merit, but properly chosen partnered genes must be used

Cervical cancer isolate PT3, super-permissive for adeno-associated virus replication, over-expresses DNA polymerase δ, PCNA, RFC and RPA

<p>Abstract</p> <p>Background</p> <p>Adeno-associated virus (AAV) type 2 is an important virus due to its use as a safe and effective human gene therapy vector and its negative association with certain malignancies. AAV, a dependo-parvovirus, autonomously replicates in stratified squamous epithelium. Such tissue occurs in the nasopharynx and anogenitals, from which AAV has been clinically isolated. Related autonomous parvoviruses also demonstrate cell tropism and preferentially replicate in oncogenically transformed cells. Combining these two attributes of parvovirus tropism, squamous and malignant, we assayed if AAV might replicate in squamous cervical carcinoma cell isolates.</p> <p>Results</p> <p>Three primary isolates (PT1-3) and two established cervical cancer cell lines were compared to normal keratinocytes (NK) for their ability to replicate AAV. One isolate, PT3, allowed for high levels of AAV DNA replication and virion production compared to others. In research by others, four cellular components are known required for <it>in vitro </it>AAV DNA replication: replication protein A (RPA), replication factor C (RFC), proliferating cell nuclear antigen (PCNA), and DNA polymerase delta (POLD1). Thus, we examined PT3 cells for expression of these components by DNA microarray and real-time quantitative PCR. All four components were over-expressed in PT3 over two representative low-permissive cell isolates (NK and PT1). However, this super-permissiveness did not result in PT3 cell death by AAV infection.</p> <p>Conclusion</p> <p>These data, for the first time, provide evidence that these four cellular components are likely important for AAV <it>in vivo </it>DNA replication as well as <it>in vitro</it>. These data also suggest that PT3 will be a useful reagent for investigating the AAV-permissive transcriptome and AAV anti-cancer effect.</p

Effect of intradialytic change in blood pressure and ultrafiltration volume on the variation in access flow measured by ultrasound dilution

AbstractBackgroundProspective access flow measurement is the preferred method for vascular access surveillance in hemodialysis (HD) patients. We studied the effect of intradialytic change in blood pressure and ultrafiltration volume on the variation in access flow measured by ultrasound dilution.MethodsAccess flow was measured 30minutes, 120minutes, and 240minutes after the start of HD by ultrasound dilution in 30 patients during 89 HD sessions and evaluated for variation.ResultsThe mean age of the 30 patients was 62±11 years: 19 were male. The accesses comprised 16 fistulae and 14 grafts. The mean access flow over all sessions decreased by 6.1% over time (1265±568mL/min after 30minutes, 1260±599mL/min after 120minutes, and 1197±576mL/min after 240minutes, P<0.01 by repeated measures ANOVA). In addition, a≥5% decrease in mean arterial pressure during HD significantly reduced access flow (P=0.014). However, no other variable (ultrafiltration volume, sex, age, presence of diabetes, type or location of access, body surface area, hemoglobin, serum albumin level) interacted significantly with the effect of time on access flow. Furthermore, mean arterial pressure did not correlate with ultrafiltration volume.ConclusionWe conclude that the variation in access flow during HD is relatively small. Decreased blood pressure is a risk factor for variation in access flow measured by ultrasound dilution. In most patients whose blood pressures are stable during HD, the access flow can be measured at any time during the HD treatment

Microstructural evolution induced by micro-cracking during fast lithiation of single-crystalline silicon

h i g h l i g h t s Lithiation of Si results in various microstructures depending of crystal orientation. A complex vein-like microstructure of Li x Si was observed in {100} oriented Si. Micro-cracks provide a fast path for Li diffusion and cause a non-uniform lithiation. Crystalline Li x Si plays an important role in micro-crack generation. a r t i c l e i n f o t r a c t We report observations of microstructural changes in {100} and {110} oriented silicon wafers during initial lithiation under relatively high current densities. Evolution of the microstructure during lithiation was found to depend on the crystallographic orientation of the silicon wafers. In {110} silicon wafers, the phase boundary between silicon and Li x Si remained flat and parallel to the surface. In contrast, lithiation of the {100} oriented substrate resulted in a complex vein-like microstructure of Li x Si in a crystalline silicon matrix. A simple calculation demonstrates that the formation of such structures is energetically unfavorable in the absence of defects due to the large hydrostatic stresses that develop. However, TEM observations revealed micro-cracks in the {100} silicon wafer, which can create fast diffusion paths for lithium and contribute to the formation of a complex vein-like Li x Si network. This defect-induced microstructure can significantly affect the subsequent delithiation and following cycles, resulting in degradation of the electrode