A Novel Experimental Scheme to Fully Separate Permethylated Glycans and Allow Online RP-LC-MS with Simplified ESI-MS Analysis

Abstract available for download

Identification and Localization of Myxococcus xanthus Porins and Lipoproteins

Myxococcus xanthus DK1622 contains inner (IM) and outer membranes (OM) separated by a peptidoglycan layer. Integral membrane, β-barrel proteins are found exclusively in the OM where they form pores allowing the passage of nutrients, waste products and signals. One porin, Oar, is required for intercellular communication of the C-signal. An oar mutant produces CsgA but is unable to ripple or stimulate csgA mutants to develop suggesting that it is the channel for C-signaling. Six prediction programs were evaluated for their ability to identify β-barrel proteins. No program was reliable unless the predicted proteins were first parsed using Signal P, Lipo P and TMHMM, after which TMBETA-SVM and TMBETADISC-RBF identified β-barrel proteins most accurately. 228 β-barrel proteins were predicted from among 7331 protein coding regions, representing 3.1% of total genes. Sucrose density gradients were used to separate vegetative cell IM and OM fractions, and LC-MS/MS of OM proteins identified 54 β-barrel proteins. Another class of membrane proteins, the lipoproteins, are anchored in the membrane via a lipid moiety at the N-terminus. 44 OM proteins identified by LC-MS/MS were predicted lipoproteins. Lipoproteins are distributed between the IM, OM and ECM according to an N-terminal sorting sequence that varies among species. Sequence analysis revealed conservation of alanine at the +7 position of mature ECM lipoproteins, lysine at the +2 position of IM lipoproteins, and no noticable conservation within the OM lipoproteins. Site directed mutagenesis and immuno transmission electron microscopy showed that alanine at the +7 position is essential for sorting of the lipoprotein FibA into the ECM. FibA appears at normal levels in the ECM even when a +2 lysine is added to the signal sequence. These results suggest that ECM proteins have a unique method of secretion. It is now possible to target lipoproteins to specific IM, OM and ECM locations by manipulating the amino acid sequence near the +1 cysteine processing site

Proteomic Analysis of the Acidocalcisome, an Organelle Conserved from Bacteria to Human Cells

Acidocalcisomes are acidic organelles present in a diverse range of organisms from bacteria to human cells. In this study acidocalcisomes were purified from the model organism Trypanosoma brucei, and their protein composition was determined by mass spectrometry. The results, along with those that we previously reported, show that acidocalcisomes are rich in pumps and transporters, involved in phosphate and cation homeostasis, and calcium signaling. We validated the acidocalcisome localization of seven new, putative, acidocalcisome proteins (phosphate transporter, vacuolar H+-ATPase subunits a and d, vacuolar iron transporter, zinc transporter, polyamine transporter, and acid phosphatase), confirmed the presence of six previously characterized acidocalcisome proteins, and validated the localization of five novel proteins to different subcellular compartments by expressing them fused to epitope tags in their endogenous loci or by immunofluorescence microscopy with specific antibodies. Knockdown of several newly identified acidocalcisome proteins by RNA interference (RNAi) revealed that they are essential for the survival of the parasites. These results provide a comprehensive insight into the unique composition of acidocalcisomes of T. brucei, an important eukaryotic pathogen, and direct evidence that acidocalcisomes are especially adapted for the accumulation of polyphosphate

Malaria Clusters among Illegal Chinese Immigrants to Europe through Africa

Between November 2002 and March 2003, 17 cases of malaria (1 fatal) were observed in illegal Chinese immigrants who traveled to Italy through Africa. A further cluster of 12 was reported in August, 2002. Several immigrants traveled by air, making the risk of introducing sudden acute respiratory syndrome a possibility should such illegal immigrations continue

The Impact of Dietary Analyzed Calcium to Phosphorus Ratios and Standardized Total Tract Digestible Phosphorus to Net Energy Ratios on Growth Performance, Bone, and Carcass Characteristics of Pigs

A total of 2,184 pigs (PIC 337 × 1050; initially 27.3 lb) were used to evaluate the effects of feeding varying analyzed calcium to phosphorus (Ca:P) ratios at two levels of standardized total tract digestible (STTD) P:NE. Pens of pigs (26 pigs per pen) were assigned to 6 dietary treatments in a randomized complete block design with 14 pens per treatment. Diets consisted of two levels of STTD P:NE, including the PIC recommendation (1.8, 1.62, 1.43, 1.25, 1.10, and 0.99 g STTD P/Mcal NE from 25 to 50, 50 to 90, 90 to 130, 130 to 180, 180 to 230, and 230 to 280 lb, respectively); or 75% of the PIC recommendation, and 3 analyzed Ca:P ratios: 0.90:1, 1.30:1, and 1.75:1. Diets were corn-soybean meal-based and contained phytase (Quantum Blue G, AB Vista, Marlborough, Wiltshire, UK); 500 to 210 FTU/kg with release values from 0.13 to 0.07% STTD P. There was a Ca:P × STTD P:NE interaction (P \u3c 0.05) observed for average daily gain (ADG), feed efficiency (F/G), and final body weight (BW). For ADG and final BW, when feeding 75% of PIC STTD P recommendation, increasing the analyzed Ca:P ratio decreased ADG and final BW (linear, P \u3c 0.001). However, when feeding the PIC STTD P recommendation, increasing the analyzed Ca:P ratio tended to improve ADG and final BW (linear, P \u3c 0.10). For F/G, when feeding 75% of the PIC STTD P recommendation, increasing the analyzed Ca:P ratio tended to worsen F/G (linear, P \u3c 0.10), whereas in pigs fed diets that met PIC STTD P recommendations, increasing the analyzed Ca:P ratio tended to improve F/G in a quadratic (P \u3c 0.10) manner. Despite the interactions, pigs fed the PIC STTD P recommendations had increased ADG, final BW, and improved F/G compared to pigs fed 75% of PIC STTD P recommendations (P \u3c 0.001). In summary, pigs fed at PIC STTD P recommendations had improved overall ADG and F/G compared to pigs fed diets at 75% of PIC STTD P recommendations. Additionally, increasing the analyzed Ca:P ratio worsened ADG and F/G when STTD P was below PIC recommendations but had marginal impacts when adequate STTD P was fed

Spring 1959

Seed testing - A Service for You by Miss Jessie L. Anderson (page 1) Increased Interest in Two-Year Turf Course by Fred P. Jeffrey - Director of Stockbridge (4) From the Editor (4) Message From Winter School President of 1959 (5) Turf club News (6) Number One Graduate (8) Liquid Fertilization by A.B. Longo (9) Public School Grounds by James Woodhouse (12) Comments on the 1959 Winter School (14) Picture - Stockbridge Turf Majors (16) Picture - Honorary Members of Turf Management Club (17) Letter on Chemical Compatibility (18) The Most Outstanding Turf Senior for 1958 (19) What it Means to be a Turf Manager by R. Russell (20) 10 Steps to a Better Lawn by P. Pedrazzi (24) A Scene to Remember (25) I switched from Hots to Cools by J. Spodnik (26) Why Attend Turfgrass Conferences (27) Picture - Winter School for Turf Managers - 1959 (29) Picture - University of Masssachusetts Annual Turfgrass Conference (30) Organic Fertilizers by O.J. Noer (A-1) Inorganic Fertilizers by Charles Winchell (A-1) Urea Formaldehyde by G.F. Stewart (A-2) Phosphorus and Potash Fertilization by Raph Donaldson (A-3) Questions on Fertilization to the Panel (A-4) Cemetery Maintenance by S.E. Robbins (A-6) Lime by Anson Brewer (A-6) Limited Budgets by R.W. Sharkey (A-7) Fertilization of Park Turf by E.J. Pyle (A-7) Disease and Insect Control by Orlando Capizzi (A-8) Cost of Establishing Turf by Victor Taricano (A-9) Question and Answers (A-10) Control of Pests of Ornamentals and Turf Occuring on Golf Courses by John C. Schread (A-12) Behind the Scenes in Soil Testing and What it Means to You Bertram Gersten and Wm. G. Colby (A-19) Lessons Learned from the 1958 Season as Applied to Golf Course Maintenance by A.M. Radko (A-21) The Outlook in Chemical Weed Control on Fine Turf by John Gallagher (A-24) New Developments in Turfgrass Disease Diagnosis and Control by Frank Howard (A-26

An integrated transcriptomic and computational analysis for biomarker identification in gastric cancer

This report describes an integrated study on identification of potential markers for gastric cancer in patients’ cancer tissues and sera based on: (i) genome-scale transcriptomic analyses of 80 paired gastric cancer/reference tissues and (ii) computational prediction of blood-secretory proteins supported by experimental validation. Our findings show that: (i) 715 and 150 genes exhibit significantly differential expressions in all cancers and early-stage cancers versus reference tissues, respectively; and a substantial percentage of the alteration is found to be influenced by age and/or by gender; (ii) 21 co-expressed gene clusters have been identified, some of which are specific to certain subtypes or stages of the cancer; (iii) the top-ranked gene signatures give better than 94% classification accuracy between cancer and the reference tissues, some of which are gender-specific; and (iv) 136 of the differentially expressed genes were predicted to have their proteins secreted into blood, 81 of which were detected experimentally in the sera of 13 validation samples and 29 found to have differential abundances in the sera of cancer patients versus controls. Overall, the novel information obtained in this study has led to identification of promising diagnostic markers for gastric cancer and can benefit further analyses of the key (early) abnormalities during its development

Grey wolf genomic history reveals a dual ancestry of dogs

The grey wolf (Canis lupus) was the first species to give rise to a domestic population, and they remained widespread throughout the last Ice Age when many other large mammal species went extinct. Little is known, however, about the history and possible extinction of past wolf populations or when and where the wolf progenitors of the present-day dog lineage (Canisfamiliaris) lived(1-8). Here we analysed 72 ancient wolf genomes spanning the last 100,000 years from Europe, Siberia and North America. We found that wolf populations were highly connected throughout the Late Pleistocene, with levels of differentiation an order of magnitude lower than they are today. This population connectivity allowed us to detect natural selection across the time series, including rapid fixation of mutations in the gene IFT8840,000-30,000 years ago. We show that dogs are overall more closely related to ancient wolves from eastern Eurasia than to those from western Eurasia, suggesting a domestication process in the east. However, we also found that dogs in the Near East and Africa derive up to half of their ancestry from a distinct population related to modern southwest Eurasian wolves, reflecting either an independent domestication process or admixture from local wolves. None of the analysed ancient wolf genomes is a direct match for either of these dog ancestries, meaning that the exact progenitor populations remain to be located.Peer reviewe