Search

Protein glycosylation is arguably the paramount post-translational modification on recombinant glycoproteins, and highly cited in the literature for affecting the physiochemical properties and the efficacy of recombinant glycoprotein therapeutics. Glycosylation of human immunoglobulins follows a reasonably well-understood metabolic pathway, which gives rise to a diverse range of asparagine-linked (N-linked), or serine/threonine-linked (O-linked) glycans. In N-linked glycans, fucose levels have been shown to have an inverse relationship with the degree of antibody-dependent cell-mediated cytotoxicity, and high mannose levels have been implicated in potentially increasing immunogenicity and contributing to less favorable pharmacokinetic profiles. Here, we demonstrate a novel approach to potentially reduce the presence of high-mannose species in recombinant human immunoglobulin preparations, as well as facilitate an approximate 100% replacement of fucosylation with arabinosylation in Chinese hamster ovary cell culture through media supplementation with D-arabinose, an uncommonly used mammalian cell culture sugar substrate. The replacement of fucose with arabinose was very effective and practical to implement, since no cell line engineering or cellular adaptation strategies were required. Arabinosylated recombinant IgGs and the accompanying reduction in high mannose glycans, facilitated a reduction in dendritic cell uptake, increased FcγRIIIa signaling, and significantly increased the levels of ADCC. These aforementioned effects were without any adverse changes to various structural or functional attributes of multiple recombinant human antibodies and a bispecific DVD-Ig. Protein arabinosylation represents an expansion of the N-glycan code in mammalian expressed glycoproteins.

Exudative age-related macular degeneration (AMD) is the most common cause of moderate and severe vision loss in developed countries. Intraocular injections of vascular endothelial growth factor (VEGF or VEGF-A)-neutralizing proteins provide substantial benefit, but frequent, long-term injections are needed. In addition, many patients experience initial visual gains that are ultimately lost due to subretinal fibrosis. Preclinical studies and early phase clinical trials suggest that combined suppression of VEGF and platelet-derived growth factor-BB (PDGF-BB) provides better outcomes than suppression of VEGF alone, due to more frequent regression of neovascularization (NV) and suppression of subretinal fibrosis. We generated a dual variable domain immunoglobulin molecule, ABBV642 that specifically and potently binds and neutralizes VEGF and PDGF-BB. ABBV642 has been optimized for treatment of exudative AMD based on the following design characteristics: 1) high affinity binding to all VEGF-A isoforms and both soluble and extracellular matrix (ECM)-associated PDGF-BB; 2) potential for extended residence time in the vitreous cavity to decrease the frequency of intraocular injections; 3) rapid clearance from systemic circulation compared with molecules with wild type Fc region for normal FcRn binding, which may reduce the risk of systemic complications; and 4) low risk of potential effector function. The bispecificity of ABBV642 allows for a single injection of a single therapeutic agent, and thus a more streamlined development and regulatory path compared with combination products. In a mouse model of exudative AMD, ABBV642 was observed to be more effective than aflibercept. ABBV642 has potential to improve efficacy with reduced injection frequency in patients with exudative AMD, thereby reducing the enormous disease burden for patients and society.

In the Canadian boreal forest, jack pine stands generally have a thin forest floor and occupy sites with coarsetextured soils and good drainage. Black spruce occurs more often on poorly drained sites and develops a thick mossdominated forest floor, but the common attribution of this development to poor quality of black spruce foliar litter has not been tested. We determined needle, twig, cone, and bark litter inputs during 10 y for black spruce and jack pine along the Boreal Forest Transect Case Study in northern Saskatchewan and Manitoba. Analysis of C, N, total phenolics, condensed tannins, and solid-state 13C NMR spectra from years 1–3 showed only small differences between species, notably higher tannins in black spruce cones. There was similarly little difference between area-based inputs, including classes of C structures determined by NMR. Condensed tannin input for black spruce was approximately twice that for jack pine, but both were in the very low range of reported values. Similar anal...

In Philosophy as a Way of Life, Pierre Hadot discusses the understanding of philosophy held by the Greco-Roman ancients. Philosophy was not understood only as an exegetical or analytical exercise, ...

1. This study used 4 wheat cultivars (Brigadier, Chaucer, Consort, Reaper) from three locations (Crossnacreevy, C; Downpatrick, D; Limavady, L), which had given rise to differences in wheat specific weight (SW), to examine the relationships between apparent metabolisable energy (AME) concentration, broiler performance and wheat SW. 2. The diets contained (g/kg): wheat 744, casein 142, blended vegetable fat 50, dicalcium phosphate 22, potassium bicarbonate 10.8, sodium bicarbonate 7.5, arginine 5, methionine 2, binder 8, trace minerals/vitamins 7.2, titanium dioxide 1.5. The diets were heat-treated (80 degrees C for 2 min) prior to pelleting (3 mm die). 3. SW ranged from 63 to 77 kg/hectolitre (hl), averaging 66, 69 and 76 kg/hl at D, C and L, respectively. In vitro viscosity of the wheat samples ranged from 5.2 to 17.5 cps and thousand grain weight (TGW) from 33.4 to 47.3 g. Mean TGW was similar at C and D (38.7, 37.0 g) but higher at L (43.1 g). In vitro viscosity was similar at C and L (11.2, 10.2 cps) but somewhat higher at D (14.4 cps). Crude protein (6.25 N) ranged from 116 to 147 g/kg and tended to be higher at D. Starch, which ranged from 612 to 656 g/kg, was least at D (617 g/kg) and greatest at L (641 g/kg). 4. Crude protein, crude fibre and total non-starch polysaccharide (NSP) were negatively correlated with SW, the R2 being respectively 0.38 (P

1. A 3x3x2 factorial experiment studied the interactions of fat source (tallow, soya, tallow:soya [2:1] blend), wheat level (700, 350, 0 g/kg) and enzyme inclusion (Avizyme 1300, absent, present) in diets for broilers fed ad libitum in individual cages from 7 to 35d. Bird performance, fat digestibility, viscosity of ileal contents and diet metabolisability (AME) were measured. 2. There were no significant effects of fat source on bird performance. However, there was a significant effect on fat digestibility, which was highest for soya and lowest for tallow. Diet AME content was also significantly affected by fat source and reflected differences in fat digestibility. 3. Dry matter (DM) intake, liveweight gain (LWG) and gain:food were all reduced at 700 g wheat/kg. Viscosity of ileal contents increased with increasing wheat inclusion. 4. There were no significant effects of enzyme on DM intake or LWG but gain:food was improved by 2%. Diet AME content was increased with enzyme addition, the effect being greatest (9%) with tallow at 700 g wheat/kg. 5. Viscosity of ileal contents was reduced and fat digestibility increased with enzyme addition and there were significant wheat enzyme interactions attributable to no differences with zero wheat but marked responses to enzyme at 700 g wheat/kg. 6. The results confirm important interactions between wheat content and fat composition in relation to fat digestibility, AME content and food efficiency.

Misuse Made Plain: Evaluating Concerns About Neuroscience in National Security Kelly Lowenberg a , Brenda M. Simon a , Amy Burns b , Libby Greismann c , Jennifer M. Halbleib b , Govind Persad b , David L.M. Preston c , Harker Rhodes b & Emily R. Murphy d a Stanford Law School Center for Law and the Biosciences , b Stanford Law School , c Stanford University , d Stanford Law School and Director of SIGNAL , Published online: 16 Apr 2010.

1. The effect of diet form (mash, cold-pelleted, steam-conditioned/pelleted, wet mash, whole wheat with balancer pellet, restricted pellet) and enzyme inclusion (Avizyme 1300, absent, present) was studied in 2 trials using individually caged, male broilers from 14 to 42 d. Bird performance, viscosity of ileal contents and diet metabolisability (AME) were measured. 2. The performance of mash-fed birds was significantly poorer than for the other treatments in relation to dry matter intake, liveweight gain and gain:food. This was not due to reduced diet AME content. 3. There was no significant effect of heat treatment on any of the variables measured, although viscosity of ileal contents was increased by 30% as compared to the cold-pelleted diet. 4. Gain:food was improved with wet-mash feeding in comparison to the dry mash treatment but it was concluded that this was not due to any intrinsic improvement in diet quality, but rather to voluntary food restriction on introduction of the wet food. 5. Whole wheat feeding improved gain:food and diet AME content by 3% as compared to the complete diets and caused approximately a 50% increase in gizzard weight as compared with the pelleted diets. 6. Food enzyme inclusion did not improve performance although a significant improvement in diet AME content was observed with enzyme inclusion in trial 1.

Despite the growing interest in backyard composting as a means to reduce residential refuse at source, there has been little study of its chemical nature and state of maturity. We obtained samples of mature compost from backyard sources in Newfoundland, Ontario and British Columbia, Canada, as well as raw input and immature compost from one BC municipal source. Chemical analysis indicated that composting was effective, proceeding with loss of C, decrease in C:N ratio, increase in pH to near neutrality, and high available N and P. Contents of heavy metals were low. Solution 31P nuclear magnetic resonance (NMR) spectroscopy of extracts showed a high proportion of orthophosphate P, indicating vigorous microbial activity and high P availability. Solid state 13C NMR with cross-polarization and magic angle spinning (CPMAS NMR) of whole composts showed that decomposition proceeds to some extent with increasing ratio of alkyl to O-alkyl C, as observed in many studies of organic soils, forest floor and composts of...

Optical emission spectroscopy provides a rapid and precise method for determining 15N/14N ratios of 15N‐enriched plant and/or soil samples. The objective of this study was to determine the effect of an indicator added in the distillation solution on the success rate of tube lighting in the optical emission spectrometer over a large range of N concentration in 15N enriched plant samples. One‐hundred‐eighty plant samples with large ranges of N concentration (4 to 30 g.kg‐1 dry weight) and 15N atom enrichment (0.368 to 1.635%) were analyzed. Our data suggest that there was no difference in the success rate of tube lighting in emission spectroscopy and in 15N/14N ratio measured between samples prepared with and without addition of indicator in the distillation solution.

This paper considers the reasons for, and the repercussions of, the bearing and rearing of children outside marriage in certain sections of the black (African) community in South Africa. Family and domestic life based on what is loosely referred to as ‘non-marriage’ is occurring elsewhere in the world and it is noted that in some countries both welfare and legislative changes are underway to accommodate and destigmatize this phenomenon. The situation in South Africa has yet to recieve recognition in either the legal and welfare fields. Two points are made: first, that in a context where a high proportion of the population are born out of wedlock, the use of the prejudicial term ‘illegitimacy’ is inappropriate;second, that in contrast to the situation in much of the Western world where women (and for that matter men) are choosing to rear children outside marriage, many black South Africans have no choice in the matter. The strategies adopted by the women discussed in the paper are forced on them b...

Common parasite infections cause energy depletion and growth stunting in children in endemic areas, and there is a suggestion that mental processes and school achievement are affected. The efficacy of control and treatment programmes, as with other development programmes, will depend to a great extent on the acceptance and support of people in affected areas. This study focused on the views of women who have young children in their care with the object of understanding their perceptions, and the implications these would have for intervention programmes.

Research on the impact of television has shifted from an emphasis in the 1960s on the passive viewer to current approaches that investigate active processing of the television medium. Although evidence is accumulating about the cognitive processes used to mediate television, the nature of the individual's concept of television has not been studied. The present investigation employs a computerized version of Kelly's Repertory Grid Test with 40 male and 40 female university students. Using Bannisters measure of intensity to index integration, the results provide supporting evidence for Kelly's individuality corollary in terms of substantial individual differences. In addition, intensity tvas related to genre selection. Persons with low-intensity scores, reflecting less integration, tended to prefer television genres offering social interaction or action. High-intensity individuals, with more integrated constructions of the television medium, showed a greater preference for informational genres

The rat prolactin gene is expressed at a high basal level in the pituitary tumor GH3 cell line. Culturing GH3 cells in a low-Ca2+, serum-free medium (SFM) depresses prolactin mRNA levels, and subsequent addition of Ca2+ to the SFM results in a specific, gradual, and sustained increase in prolactin mRNA levels. We have now examined whether the observed increase in prolactin mRNA levels can be attributed solely to an increase in the transcriptional rate of the prolactin gene. Treatment of GH3 cells in SFM with 0.4 mM CaCl2 for 24 to 48 h increased cytoplasmic prolactin mRNA levels by 5- to 10-fold, whereas the transcriptional rate of the prolactin gene was increased by less than twofold over values for SFM controls. Prolactin mRNA levels increased progressively during the 24-h period after Ca2+ addition, whereas prolactin gene transcription never exceeded a twofold increase over values for SFM controls. The activities of nuclear extracts from control and Ca2(+)-induced cells were examined in an in vitro transcription assay. The two extracts directed transcription from the prolactin promoter and the adenovirus major late promoter equally well. Cycloheximide had no effect on the ability of Ca2+ to increase or maintain prolactin mRNA levels. In dactinomycin mRNA clearance experiments, prolactin mRNA was cleared at the same rate in the absence and presence of Ca2+. These results demonstrate that although Ca2+ has a small effect on the transcriptional rate of the prolactin gene, Ca2+ produces a significant increase in prolactin mRNA levels by acting at a posttranscriptional site(s). Furthermore, Ca(2+) appears to increase prolactin mRNA levels by posttranslational modification of a stable protein, probably at a nuclear site.

Oogenesis within the hologonic ovary of the trichuroid nematode, Trichuris muris, was observed by light and electron microscopy. Early germinal stages in the form of oogonia and young primary oocytes were characterised by a high nuclear-cytoplasmic ratio, numerous ribosomes and several mitochondrial clusters. Previtellogenic primary oocytes contained a prominent nucleus with a nuclear envelope punctuated by pores. They also contained increased amounts of granular endoplasmic reticulum (GER), often arranged as annulate lamellae, several Golgi complexes and limited amounts of lipid. The appearance of three types of cytoplasmic inclusion, in the form of lipid, dense yolk granules and reticulate granules, indicated the onset of vitellogenesis. At this stage of oogenesis, all three types were distributed throughout the ooplasm. The possible role of the granules is discussed. During passage along the oviduct the oocyte was coated by an additional unit membrane and associated fibrillar layer external to the oole...

(1989). The evacuation of schoolchildren from Newcastle upon Tyne, 1939‐1942: an assessment of the factors which influenced the nature of educational provision in Newcastle and its reception areas. History of Education: Vol. 18, No. 3, pp. 231-241.

Early during the course of Trypanosoma congolense infection in cattle decreases in PCV occurred and coincided with increases in both MCV and MCH. The indices reached highest levels between eight and 12 weeks post-infection. By week 20 of infection MCV and MCH had decreased to pre-infection levels even though a substantial anemia persisted. Serum iron levels were elevated at eight weeks postinfection. (Infected 271 mg dl-1 v. control 140 mg dl-1) but decreased to low levels in infected animals by Week 28 (63 mg dl-1). At eight weeks post-infection PITR was elevated in infected animals (infected 2.6 mg dl-1 day-1 v. control 0.82 mg dl-1 day-1). By Week 28, however, PITR had fallen in infected animals to 0.62 mg dl-1 day-1 indicating a severe dyshaemopoesis since PCV levels averaged only 22%. Cattle which were treated with Berenil or that self-cured the infection had normal iron parameters when tested 61 weeks post-infection. Cattle treated early during the course of infection showed a significantly greater PCV response that those treated later during infection.

(1976). Race attitudes and behaviour: the case of domestic employment in white South African homes. African Studies: Vol. 35, No. 2, pp. 71-90.

This paper reviews the history of land allocation, land use and also some of the major attempts made over the last century to introduce change and what is now popularly referred to as development at Indaleni, a black community which is predominantly Zulu, but where there has been a strong white mission presence for over a hundred years. It is argued that the missionaries and colonial administrators of the last century were the forerunners of many of the interest groups active in the development field at Indaleni and elsewhere in KwaZulu/Natal today. Although the aims of intervention have changed, the high‐handed methods and above all the certainty with which many outsiders approach ‘development’ has not. Chronicled also are white attitudes to granting freehold land tenure to blacks in Natal and consideration is given to the reasons why the first missionaries to work at Indaleni failed to achieve their aim of not only converting local people to Christianity, but of equipping them with skills to compete wit...

We employed a protein gel blotting procedure to search for nuclear proteins from rat pituitary cells that bind preferentially to the 5'-flanking region of the rat prolactin gene. By gel blots of chromatin proteins from GH3 rat pituitary tumor cells with a 32P-labeled prolactin genomic clone, we detected two major binding proteins with molecular weights of approximately 44,000 and 48,000, designated NP44 and NP48, respectively. Both NP44 and NP48 are minor chromatin proteins which are extracted at low salt concentrations (0.4 M NaCl) and exhibit a range of slightly acidic isoelectric variants. NP44 and NP48 were detected at similar levels in chromatin extracts of GH3 cells, the prolactin-negative GC cell variant of the GH3 cells, and normal rat pituitary tissue. Considerably lower levels of these two proteins were found in chromatin extracts from rat liver and rat C6 glial cells. NP44 and NP48 exhibit DNA sequence specificity, as evidenced by their strong binding to the upstream flanking region of the prolactin gene, but only very weak binding to plasmid DNA, rat prolactin or growth hormone cDNAs, or upstream flanking regions of two other rat genes. By analyzing subclones of a rat prolactin genomic clone, we established that NP44 and NP48 bind to at least two sites, which are located between 0.4 and 2.0 kilobases (region I) and between 2.0 and 4.8 kilobases (region II) upstream of the transcription initiation site. These findings are discussed in the context of a possible functional association between the strong binding of NP44 and NP48 to the prolactin 5'-flanking region and pituitary-specific expression of the prolactin gene.

Oats (Avena sativa L. cv. 3C‐W) were grown on a 2‐ha field of an organic soil, the 0–20 cm layer of which varied in Cu content from 91 to 248 ppm. Analyses of samples of the soil and the mature crop revealed that soil Cu content did not correlate with Cu concentrations in the grains or straw. The EDTA‐soluble or ‘available’ Cu content of the soil was ‘abundant’ all over the field, ranging from 200 to 1100 times the total uptake of Cu by the crop. In spite of this, soil concentration of Cu was positively correlated with grain yield, and consequently with grain uptakes of Cu, Fe and P. The results suggest that (a) large proportions of native and residual fertilizer Cu in organic soils are not available to plants, and (b) that there is little luxury consumption of Cu by oats growing on an organic soil even when sufficient soil Cu is available and its concentration correlates with grain yield.

Examination of the moisture in fibres by the rate-of-cooling method shows evidence of freezing only when the moisture is above a limiting value with any fibres. The limiting value has been correlated (i) with the chemical composition of the fibres, e.g., the degree of substitution of cellulose acetate, (ii) with the physical characteristics, e.g., with the crystallinity and (iii) with other properties, e.g., absorption of water vapour.

Some effects of urea solutions on cellulose fibres have been studied. Swelling of native and regenerated cellulose fibres, loss of strength of the latter and the ease of stripping acid dyes all increase with the concentration of the urea solutions.

(1949). MEASURING THE SWELLING OF FIBRES IN WATER. Journal of the Textile Institute Proceedings: Vol. 40, No. 7, pp. P674-P688.

(1952). 31—CAPILLARY PHENOMENA IN ASSEMBLIES OF FIBRES. Journal of the Textile Institute Transactions: Vol. 43, No. 8, pp. T402-T422.

(1972). Studies on leishmaniasis in Ethiopia. Annals of Tropical Medicine & Parasitology: Vol. 66, No. 3, pp. 313-328.

(1951). 4—Capillary and Imbibed Water in Assemblies of Moist Fibres. Journal of the Textile Institute Transactions: Vol. 42, No. 2, pp. T79-T90.

The centrifuge method has been used to measure the effect of temperature on the swelling of viscose rayon and cotton. It is shown that a minimum occurs in the temperature-swelling curve for both fibres in the region of 50–60°C. Results are also given for the irreversible change in the water-imbibition of viscose rayon which occurs when these fibres are heated in water for short periods.