A comparison of different pre-lysis methods and extraction kits for recovery of Stretococcus agalacticae (Lancefield group B Streptococcus) DNA from whole blood

Sub-optimal recovery of bacterial DNA from whole blood samples can limit the sensitivity of molecular assays to detect pathogenic bacteria. We compared 3 different pre-lysis protocols (none, mechanical pre-lysis and achromopeptidasepre-lysis) and 5 commercially available DNA extraction platforms for direct detection of Group B Streptococcus (GBS) in spiked whole blood samples, without enrichment culture. DNA was extracted using the QIAamp Blood Mini kit (Qiagen), UCP Pathogen Mini kit (Qiagen), QuickGene DNA Whole Blood kit S (Fuji), Speed Xtract Nucleic Acid Kit 200 (Qiagen) and MagNA Pure Compact Nucleic Acid Isolation Kit I (Roche Diagnostics Corp). Mechanical pre-lysis increased yields of bacterial genomic DNA by 51.3 fold (95% confidence interval; 31.6–85.1, p < 0.001) and pre-lysis with achromopeptidase by 6.1 fold (95% CI; 4.2–8.9, p < 0.001), compared with no pre-lysis. Differences in yield dueto pre-lysis were 2–3 fold larger than differences in yield between extraction methods. Including a pre-lysis step can improve the limits of detection of GBS using PCR or other molecular methods without need for culture.

Preservation in a growing city: a consideration of conservation districts for Washington, D.C. neighborhoods

Master final project submitted to the faculty of the Historic Preservation Program of the School of Architecture, Planning, and Preservation of the University of Maryland, College Park, in partial fulfillment of the requirements for the degree of Master of Historic Preservation, 2013.

Ancient Bruises: a Case of Skin Lesions due to Vitamin C Deficiency

Scurvy was a common 18th century disease caused by vitamin C deficiency. It presents with multiple non-specific symptoms and can lead to capillary fragility due to impaired collagen synthesis. We report the case of a 63-year-old woman who presented with fatigue, nausea and progressive skin lesions consisting of multiple ecchymoses on the legs as also described in the diary drawings of a navy doctor in the 19th century. The ascorbic acid level was undetectable low in the patient’s serum. However, treatment with 500 mg ascorbic acid daily dramatically improved the skin lesions within 5 days.

Expression of tumor-related Rac1b antagonizes B-Raf-induced senescence in colorectal cells

Mutations in the BRAF oncogene have been identified as a tumor-initiating genetic event in mainly melanoma, thyroid and colon cancer, resulting in an initial proliferative stimulus that is followed by a growth arrest period known as oncogene-induced senescence (OIS). It remains unknown what triggers subsequent escape from OIS to allow further tumor progression. A previous analysis revealed that overexpression of splice variant Rac1b occurs in around 80% of colorectal tumors carrying a mutation in BRAF. Using both BRaf-V600E-directed RNAi and overexpression we demonstrate that this mutation does not directly lead to Rac1b overexpression, indicating the latter as an independent event during tumor progression. Nonetheless, we observed that expression of oncogenic BRaf-V600E in non-transformed colonocytes (NCM460 cell line) increased both the transcript and protein levels of p14ARF, p15INK4b and p21CIP1 and led to increased expression of β-galactosidase, all indicators of OIS induction. Interestingly, whereas the protein levels of these markers were reduced upon Rac1b overexpression, the levels of their respective transcripts remained unchanged. Importantly, the co-expression of Rac1b with B-Raf-V600E reverted the OIS phenotype, reducing the expression levels of the cell-cycle inhibitors and β-galactosidase to those of control cells. These data identify increased Rac1b expression as one potential mechanism by which colorectal tumor cells can escape from B-Raf-induced OIS.

Profile of Washington & Fayette streets, Georgetown, D.C. /

Washington Street profile covers 30th Street between Q and R streets N.W. Fayette Street profile covers 35th Street between Volta Place and Wisconsin Avenue N.W.

[Map of the proposed enlargement and replanning of the Capitol grounds, Washington D.C.].

India ink on recto and orange wash on verso.

Guide to trees and shrubs in the grounds of the Executive Mansion : [Washington D.C.] /

Trees and shrubs numbered.

[Cadastral map of part of the West End, Washington D.C.].

Cadastral map showing unidentified tinted lots, lot numbers, and block numbers.

Automated morphometry provides accurate and reproducible virtual staging of liver fibrosis in chronic hepatitis C

International audience

Chemical composition of cultivated seaweed ulva clathrata (Roth) C. Agardh

Samples of cultivated Ulva clathrata were collected from a medium scale system (MSS, 1.5 1.5 m tank), or from a large scale system (LSS, 0.8 ha earthen pond). MSS samples were dried directly while the LSS sample was washed in freshwater and pressed before drying. Crude protein content ranged 20–26%, essential amino acids accounting for 32–36% of crude protein. The main analysed monosaccharides were rhamnose (36–40%), uronic acids (27–29%), xylose (10–13%) and glucose (10–16%). Some notable variations between MSS and LSS samples were observed for total dietary fibre (26% vs 41%), saturated fatty acids (31% vs 51%), PUFAS (33% vs 13%), carotenoids (358 vs 169 mg kg1 dw) and for Ca (9 vs 19 g kg1 ), Fe (0.6 vs 4.2 g kg1 ), Cu (44 vs 14 mg kg1 ), Zn (93 vs 17 mg kg1 ) and As (2 vs 9 mg kg1 ). The chemical composition of U. clathrata indicates that it has a good potential for its use in human and animal food.

Protective effect of (-)-epigallocatechin gallate on ultraviolet b-induced skin damage in hairless mice

Purpose: To investigate the protective effect of green tea (-)-epigallocatechin gallate (EGCg) on ultraviolet B (UV-B)-induced skin damages in hairless mice in order to develop a natural sunscreen compound for use in skin care products and cosmetics. Methods: EGCg was dissolved in acetone at concentrations of 1.0, 10.0 and 50.0 mg/mL, and topically applied to the skin of hairless mice at doses of 0.2 mL/cm2, with acetone as control. The mice were then irradiatd m2 UV-B for 30 min daily. EGCg treatment and UV-B irradiation were carried out daily for 28 consecutive days. The mice were then sacrificed and their dorsal skin examined by transmission electron microscopy (TEM) on the 28th day. Results: UV-B irradiation induced severe macroscopic skin damage including chapping, cracking and abnormal desquamation in the treated hairless mice. EGCg showed dose-dependent protective effects against UV-B induced damage on the skin. Treatments with 10.0 and 50.0 mg/mL EGCg alleviated UVB-induced skin damage by suppressing both keratinocyte apoptosis and mitochondrial dysfunction, along with inhibiting the production of melanin pigment. Conclusion: Topical application of green tea EGCg shows dose-dependent protective effect against UV-B-induced damage on hairless mouse skin. Thus, the plant compound can potentially be used as an alternative agent for photoprotection against UV-B exposure.

Regulation of electrical transmission by protein kinase C in Aplysia bag cell neurons

Electrical synapses are composed of gap junctions, made from paired hemi-channels that allow for the transfer of current from one neuron to another. Gap junctions mediate electrical transmission in neurons, where they synchronize spiking and promote rapid transmission, thereby influencing the coordination, pattern, and frequency of firing. In the marine snail, Aplysia calfornica, two clusters of neuroendocrine bag cell neurons use electrical synapses to synchronize a 30-min burst of action potentials, known as the afterdischarge, which releases egg-laying hormone and induces reproduction. In culture, paired bag cell neurons present a junctional conductance that is non-rectifying and largely voltage-independent. During the afterdischarge, PKC is activated, which is known to increase voltage-gated Ca2+ current; yet, little is understood as to how this pathway impacts electrical transmission. The transfer of presynaptic spike-like waveforms (generated in voltage-clamp) to the postsynaptic cell (measured in current-clamp) was monitored with or without PKC activation. It was found that pretreatment with the PKC activator, phorbol-12-myristate-13-acetate (PMA), enhanced junctional conductance between bag cell neurons. Furthermore, in control, presynaptic action potential waveforms mainly evoked postsynaptic electrotonic potentials at both -60 and -40 mV. However, with PKC activation the presynaptic stimulus consistently elicited postsynaptic action potentials from resting potentials of -40 mV, and would occasionally result in firing from repetitive input at -60 mV. Moreover, to assess whether this enhanced electrical transmission genuinely reflects a greater junctional conductance or a change in postsynaptic responsiveness, a fast-phase junctional-like current was applied to single bag cell neurons. Neurons in PMA always fired action potentials in response to current injection as opposed to control, which were less likely to spike. This outcome did not change when the junctional-like current was artificially enhanced in control conditions. Also, in response to fast- and slow-phase electrotonic potential (ETP) waveforms, Ca2+ current was markedly larger in single PMA-treated neurons. These findings suggest that PKC activation may contribute to afterdischarge fidelity by recruiting postsynaptic Ca2+ current to promote synchronous network firing. Finally, Aplysia gap junction genes (innexins) were transfected into mouse N2A cells and characterized. This revealed a biophysical and pharmacological profile similar to native gap junctions.

通过第一性原理计算研究内地核的结构与成分

The determination of the composition and structure of the Earth’s inner core has long been the major subject in the study of the Earth’s deep interior. It’s widely believed that the Earth’s core is formed by iron with a fraction of nickel. However, light elements must exist in the inner core because the earth core is less dense than pure iron-nickel alloy (~2-3% in the solid inner core and ~6-7% in the liquid outer core). The questions are what and how much light element is there in the iron-nickel alloy. Besides the composition, the crystal structure of the iron with or without light element is also not well known. According to the seismological observations, the sound waves propagate 3-4% faster along the spin axis than in the equatorial plane. That means the inner core is anisotropic. The densest structure of iron-nickel alloy should be h.c.p structure under the very high pressures. However, the h,c,p structure does not propagate waves anisotropic ally. Then what is the structure of the iron-nickel alloy or the iron-nickle-light element alloy. In this study, we tried to predict the composition and the structure of the inner core through ab initio calculation of the Gibbs free energy, which is a function of internal energy, density and entropy. We conclude that the h.c.p structure is more stable than the b.c.c structure under high pressure and 0 K, but with the increase of temperature, the free energy of the b.c.c structure is decreasing much faster than the h.c.p structure caused by the vibration of the atomics, so the b.c.c structure is more stable at high temperatures. With the addition of light elements (S or Si or both), the free energy of b.c.c. decreases even faster, about 3at% of Si not only explains why the inner core is about 2-3 % lighter than the iron-nickle alloy, but also reasons why the inner core is anisotropic, since the b.c.c. structure becomes more stable than the h.c.p structure at 5500-6000K and b.c.c. is anisotropic in propagating seismic waves. Therefore, we infer that the inner core of the earth is formed by b.c.c iron and a fraction of nickel plus ~3at.% Si, with a temperature higher than 5500K, which is consistent with the studies from other approaches.

Finding ‘Good Friday’ An explorative study into the sedimentary structures left from the Good Friday tsunami

Senior thesis written for Oceanography 444

Nanosized iron based permeable reactive barriers for nitrate removal - Systematic review

It is unquestionable that an effective decision concerning the usage of a certain environmental clean-up technology should be conveniently supported. Significant amount of scientific work focussing on the reduction of nitrate concentration in drinking water by both metallic iron and nanomaterials and their usage in permeable reactive barriers has been worldwide published over the last two decades. This work aims to present in a systematic review of the most relevant research done on the removal of nitrate from groundwater using nanosized iron based permeable reactive barriers. The research was based on scientific papers published between 2004 and June 2014. It was performed using 16 combinations of keywords in 34 databases, according to PRISMA statement guidelines. Independent reviewers validated the selection criteria. From the 4161 records filtered, 45 met the selection criteria and were selected to be included in this review. This study's outcomes show that the permeable reactive barriers are, indeed, a suitable technology for denitrification and with good performance record but the long-term impact of the use of nanosized zero valent iron in this remediation process, in both on the environment and on the human health, is far to be conveniently known. As a consequence, further work is required on this matter, so that nanosized iron based permeable reactive barriers for the removal of nitrate from drinking water can be genuinely considered an eco-efficient technology.