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R5 clade C SHIV strains with tier 1 or 2 neutralization sensitivity: tools to dissect env evolution and to develop AIDS vaccines in primate models.

BACKGROUND: HIV-1 clade C (HIV-C) predominates worldwide, and anti-HIV-C vaccines are urgently needed. Neutralizing antibody (nAb) responses are considered important but have proved difficult to elicit. Although some current immunogens elicit antibodies that neutralize highly neutralization-sensitive (tier 1) HIV strains, most circulating HIVs exhibiting a less sensitive (tier 2) phenotype are not neutralized. Thus, both tier 1 and 2 viruses are needed for vaccine discovery in nonhuman primate models. METHODOLOGY/PRINCIPAL FINDINGS: We constructed a tier 1 simian-human immunodeficiency virus, SHIV-1157ipEL, by inserting an "early," recently transmitted HIV-C env into the SHIV-1157ipd3N4 backbone [1] encoding a "late" form of the same env, which had evolved in a SHIV-infected rhesus monkey (RM) with AIDS. SHIV-1157ipEL was rapidly passaged to yield SHIV-1157ipEL-p, which remained exclusively R5-tropic and had a tier 1 phenotype, in contrast to "late" SHIV-1157ipd3N4 (tier 2). After 5 weekly low-dose intrarectal exposures, SHIV-1157ipEL-p systemically infected 16 out of 17 RM with high peak viral RNA loads and depleted gut CD4+ T cells. SHIV-1157ipEL-p and SHIV-1157ipd3N4 env genes diverge mostly in V1/V2. Molecular modeling revealed a possible mechanism for the increased neutralization resistance of SHIV-1157ipd3N4 Env: V2 loops hindering access to the CD4 binding site, shown experimentally with nAb b12. Similar mutations have been linked to decreased neutralization sensitivity in HIV-C strains isolated from humans over time, indicating parallel HIV-C Env evolution in humans and RM. CONCLUSIONS/SIGNIFICANCE: SHIV-1157ipEL-p, the first tier 1 R5 clade C SHIV, and SHIV-1157ipd3N4, its tier 2 counterpart, represent biologically relevant tools for anti-HIV-C vaccine development in primates.

EPA-R5.

Description based on: 72-001.

4-Carbamoylpiperidinium phenylacetate hemihydrate

In the structure of the title compound, C6H13N2O+ C8H7O2- . 0.5H2O, the asymmetric unit comprises two isonipecotamide cations, two phenylacetate anions and a water molecule of solvation. The hydrogen-bonding environments for both sets of ion pairs are essentially identical with the piperidinium and amide 'ends' of each cation involved in lateral heteromolecular hydrogen-bonded cyclic N---H...O associations [graph set R2/2(11)] which incorporate a single carboxyl O-atom acceptor. These cyclic motifs enclose larger R5/5(21) cyclic systems forming sheet substructures which lie parallel to (101) and are linked across b by the single water molecule via water O---H...O(carboxyl) associations to give a two-dimensional duplex-sheet structure

Default judgment set aside without costs

A recent District Court case is believed to be the first in Queensland in which UCPR r 5 has been used to support the setting aside of a regularly entered default judgment without a costs order.

Plain facts about pleading : how to work with r 166

The decision of Durward SC DCJ in OSM Group Pty Ltd v Holden [2013] QDC 151 involves a useful consideration of the requirements relating to the pleading of denials and non-admissions under the Uniform Civil Procedure Rules 1999 (Qld) (UCPR). In particular, the decision examines the extent of the obligations when pleading in response to allegations of law, or of mixed fact and law.

Looming aircraft threats : shape-based passive ranging of aircraft from monocular vision

This paper proposes new techniques for aircraft shape estimation, passive ranging, and shape-adaptive hidden Markov model filtering which are suitable for a monocular vision-based non-cooperative collision avoidance system. Vision-based passive ranging is an important missing technology that could play a significant role in resolving the sense-and-avoid problem in un-manned aerial vehicles (UAVs); a barrier hindering the wider adoption of UAVs for civilian applications. The feasibility of the pro- posed shape estimation, passive ranging and shape-adaptive filtering techniques is evaluated on flight test data.

Monoclonal antibody screening of a phage-displayed random peptide library reveals mimotopes of chemokine receptor CCR5: Implications for the tertiary structure of the receptor and for an n-terminal binding site for HIV-1 gp120

The chemokine receptor CCR5 contains seven transmembrane-spanning domains. It binds chemokines and acts as co-receptor for macrophage (m)-tropic (or R5) strains of HIV-1. Monoclonal antibodies (mAb) to CCR5, 3A9 and 5C7, were used for biopanning a nonapeptide cysteine (C)-constrained phage-displayed random peptide library to ascertain contact residues and define tertiary structures of possible epitopes on CCR5. Reactivity of antibodies with phagotopes was established by enzyme-linked immunosorbent assay (ELISA). mAb 3A9 identified a phagotope C-HASIYDFGS-C (3A9/1), and 5C7 most frequently identified C-PHWLRDLRV-C (5C7/1). Corresponding peptides were synthesized. Phagotopes and synthetic peptides reacted in ELISA with corresponding antibodies and synthetic peptides inhibited antibody binding to the phagotopes. Reactivity by immunofluorescence of 3A9 with CCR5 was strongly inhibited by the corresponding peptide. Both mAb 3A9 and 5C7 reacted similarly with phagotopes and the corresponding peptide selected by the alternative mAb. The sequences of peptide inserts of phagotopes could be aligned as mimotopes of the sequence of CCR5. For phage 3A9/1, the motif SIYD aligned to residues at the N terminus and FG to residues on the first extracellular loop; for 5C7/1, residues at the N terminus, first extracellular loop, and possibly the third extracellular loop could be aligned and so would contribute to the mimotope. The synthetic peptides corresponding to the isolated phagotopes showed a CD4-dependent reactivity with gp120 of a primary, m-tropic HIV-1 isolate. Thus reactivity of antibodies raised to CCR5 against phage-displayed peptides defined mimotopes that reflect binding sites for these antibodies and reveal a part of the gp120 binding sites on CCR5.

The role of basic nutritional research in pediatric liver disease : an historical perspective

The advent of liver transplantation for end-stage liver disease (ESLD) in children has necessitated a major rethink in the preoperative preparation and management from simple palliative care to active directed intervention. This is particularly evident in the approach to the nutritional care of these patients with the historical understanding of the nutritional pertubations in ESLD being described from a single pediatric liver transplant center. ESLD in children is a hypermetabolic process adversely affecting nutritional status, metabolic, and non-metabolic body compartments. There is a complex dynamic process affecting metabolic activity within the metabolically active body cell mass, as well as lipid oxidation during fasting and at rest, with other factors operating in conjunction with daily activities. We have proposed that immediately ingested nutrients are a more important source of energy in patients with ESLD than in healthy children, among whom energy may be stored in various body compartments.

Mobile assessment of on-road air pollution and its sources along the East-West Highway in Bhutan

Human exposures in transportation microenvironments are poorly represented by ambient stationary monitoring. A number of on-road studies using vehicle-based mobile monitoring have been conducted to address this. Most previous studies were conducted on urban roads in developed countries where the primary emission source was vehicles. Few studies have examined on-road pollution in developing countries in urban settings. Currently, no study has been conducted for roadways in rural environments where a substantial proportion of the population live. This study aimed to characterize on-road air quality on the East-West Highway (EWH) in Bhutan and identify its principal sources. We conducted six mobile measurements of PM10, particle number (PN) count and CO along the entire 570 km length of the EWH. We divided the EWH into five segments, R1-R5, taking the road length between two district towns as a single road segment. The pollutant concentrations varied widely along the different road segments, with the highest concentrations for R5 compared with other road segments (PM10 = 149 µg/m3, PN = 5.74 × 104 particles/cm-3, CO = 0.19 ppm), which is the final segment of the road to the capital. Apart from vehicle emissions, the dominant sources were road works, unpaved roads and roadside combustion activities. Overall, the highest contributions above the background levels were made by unpaved roads for PM10 (6 times background), and vehicle emissions for PN and CO (5 and 15 times background, respectively). Notwithstanding the differences in instrumentation used and particle size range measured, the current study showed lower PN concentrations compared with similar on-road studies. However, concentrations were still high enough that commuters, road maintenance workers and residents living along the EWH, were potentially exposed to elevated pollutant concentrations from combustion and non-combustion sources. Future studies should focus on assessing the dispersion patterns of roadway pollutants and defining the short- and long-term health impacts of exposure in Bhutan, as well as in other developing countries with similar characteristics.

Cytotoxicity of half sandwich ruthenium(II) complexes with strong hydrogen bond acceptor ligands and their mechanism of action

Neutral and cationic organometallic ruthenium(II) piano stool complexes of the type [(eta(6)-cymene)R-uCl(X)(Y)] (complexes R1-R8) has been synthesized and characterized. In cationic complexes, X, Y is either a eta(2) phosphorus ligand such as 1,1-bis(diphenylphosphino)methane (DPPM) and 1,2-bis(diphenylphosphino)ethane (DPPE) or partially oxidized ligands such as 1,2-bis(diphenylphosphino)methane monooxide (DPPMO) and 1,2-bis(diphenylphosphino)ethane monooxide (DPPEO) which are strong hydrogen bond acceptors. In neutral complexes. X is chloride and Y is a monodentate phosphorous donor. Complexes with DPPM and DPPMO ligands ([(eta(6)-cymene)Ru(eta(2)-DPPM)Cl]PF6 (R2), [(eta(6)-cymene)Ru(eta(2)-DPPMO)Cl]PF6 (R3), [(eta(6)-cymene)Ru(eta(1)-DPPM)Cl-2] (R5) and [(eta(6)-cymene)Ru(eta(1)-DPPMO)Cl-2] (R6) show good cytotoxicity. Growth inhibition study of several human cancer cell lines by these complexes has been carried out. Mechanistic studies for R5 and R6 show that inhibition of cancer cell growth involves both cell cycle arrest and apoptosis induction. Using an apoptosis PCR array, we identified the sets of antiapoptotic genes that were down regulated and pro-apoptotic genes that were up regulated. These complexes were also found to be potent metastasis inhibitors as they prevented cell invasion through matrigel. The complexes were shown to bind DNA in a non intercalative fashion and cause unwinding of plasmid DNA in cell-free medium by competitive ethidium bromide binding, viscosity measurements, thermal denaturation and gel mobility shift assays.

Aspergillus niger -proliiniendopeptidaasi prolamiinien eliminoinnissa

Tutkielman kirjallisuusosassa perehdyttiin vehnän, rukiin ja ohran, eli Triticeaeprolamiinien erityisasemaan keliakianäkökulmasta tarkasteltuna ja prolamiinien hydrolyysiin proliinispesifeillä entsyymeillä. Lisäksi tarkasteltiin prolamiinien immunologisia määritysmenetelmiä. Keliakiassa haitalliset gluteenipeptidit sisältävät runsaasti proliinia ja ovat hankalia pilkkoa muilla kuin proliinispesifeillä peptidaaseilla. Suurin osa immunologisen reaktion aiheuttavista gluteenilähtöisistä peptideistä voidaan pilkkoa idätetyn viljan endogeenisilla entsyymeillä happamissa olosuhteissa, mutta jäljellejäävä prolamiinipitoisuus ylittää edelleen gluteenittomille tuotteille sallitun rajan. Kokeellisen työn tavoitteena oli eliminoida happamalla mallasinkubaatiolla valmistettujen vehnä-, ohra- ja ruismallasautolysaattien sisältämä jäännösprolamiini Aspergillus niger -homeen tuottamalla proliinispesifillä endopeptidaasilla (AN-PEP) siten, että hydrolysaattia voitaisiin käyttää gluteenittomissa leivontasovelluksissa. Proteiinien hydrolyysiä tarkkailtiin kokoekskluusiokromatografialla (SEC), vapaan aminotypen (FAN) muodostumisena ja SDS-PAGE-elektroforeesilla. Jäännösprolamiinien pilkkoutumista seurattiin immunologisella R5-ELISA-menetelmällä. AN-PEP-inkubaatiolla saatiin aikaan voimakasta prolamiinien pilkkoutumista; mallasautolysaattien jäännösprolamiinista pilkkoutui yli 96 %. SEC- ja FAN-analyysien perusteella inkubaatioaikaa kannatti jatkaa yli 4 h, jolloin polypeptidit pilkkoutuivat edelleen pienemmiksi hydrolyysituotteiksi. Vehnä- ja ruismallashydrolysaattien prolamiinipitoisuuden todettiin laskevan 22 h inkubaation aikana alle tason 100 mg/kg R5-ELISA-menetelmällä määritettynä. Matalimmat prolamiinipitoisuudet saavutettiin AN-PEP-pitoisuudella 35 ?l / g mallasautolysaattia. Codex Alimentarius -komission säädöksen mukaan keliakiaruokavalioon soveltuvat ns. erittäin vähägluteeniset tuotteet saavat sisältää gluteenia enintään 100 mg/kg. Erityisesti AN-PEP-käsiteltyä ruismallasraaka-ainetta voitaisiin mahdollisesti käyttää tuomaan rukiista aromia gluteenittomiin leipiin. Ennen kuin mallashydrolysaatit ovat valmiita kaupallisiin sovelluksiin, on tarkasteltava niiden todellisia mahdollisuuksia parantaa elintarvikkeiden makua ja aromia sekä todettava uuden teknologian turvallisuus keliaakikoille.