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The choice of methods for immunoglobulin IgG purification: yield and purity of antibody activity.

Six methods for the purification of immunoglobulin G (IgG) from serum were compared, using rabbit antiserum to Bacillus anthracis spores as a model. Antibody activity was monitored by a solid-phase immunoradiometric assay (IRMA). Salt precipitation/ion exchange chromatography and ethanol precipitation both resulted in IgG of high purity but there was considerable inactivation of antibody. Salt precipitation/affinity chromatography gave poor yields of antibody. PEG precipitation and gel filtration of Sephacryl S-300 gave moderate yields and purity of IgG, with little evidence of antibody inactivation. Salt precipitation was marginally more destructive than the last 2 methods, but is recommended for routine use on grounds of its simplicity. Should IgG prepared by salt precipitation prove inadequate for particular applications, gel filtration is recommended since it allows the balance of yield and purity to be altered at will.

Animals↗

Rapid method for the isolation of neutrophils in high yield without the use of dextran or density gradient polymers.

A simple, rapid and economical method for the isolation of polymorphonuclear leucocytes (PMNs) from whole blood is compared with dextran and dextran/Lymphoprep gradient techniques. The method eliminates the use of dextran and density gradient polymers such as Ficoll which have been shown to affect PMNs adversely. The technique is based on the lysis of red cells with isotonic ammonium chloride solution followed by differential centrifugation to separate the PMNs. This method gave a PMN yield of 73% (SD +/- 3.5) and a purity of 78% (SD +/- 2.5). Both morphology and functional activity were preserved, as assessed by bacterial phagocytosis and killing, chemotaxis, polarising response, superoxide production and adherence. In contrast, the dextran and dextran/Lymphoprep techniques gave yields of 50% and 15% with purities of 78% and 91% respectively. In a series of 14 PMN isolations, the differential centrifugation method gave an average yield of 63% with an average purity of 83%.

Ammonium Chloride↗

The chemical synthesis of ecdysone 22-long-chain fatty acyl esters in high yield.

Eight ecdysone 22-long-chain fatty-acyl esters (laurate, myristate, palmitate, stearate, oleate, linoleate, linolenate and arachidate) have been chemically synthesised in high yield. Ecdysone was first converted to the 2,3-acetonide derivative and then acylated selectively at C-22 with the appropriate acyl anhydride. The protecting acetonide group is then removed by mild acid treatment to yield the ecdysone 22-acyl ester. Reaction conditions have been optimised to maximise the overall yield (ca. 70%). The ecdysone acyl esters and their 2,3-acetonide derivatives have been characterised by 1H- and 13C-NMR and high-resolution FAB-mass-spectrometry.

Acylation↗

Structural identifiability of the yield coefficients in bioprocess models when the reaction rates are unknown.

The identification problem of bioprocesses in a stirred tank reactor essentially concerns the determination of the yield coefficients and the reaction rates involved in a general state-space model. This paper deals with the analysis of structural identifiability of the yield coefficients issuing from a two-step identification procedure. In fact, the identification of the yield coefficients can be completely decoupled from that of the reaction rates by means of an appropriate transformation of the dynamical model. It is shown that the identifiability properties of these coefficients can be drawn from the structure of the underlying reaction network. Necessary and sufficient identifiability conditions are given in the form of some simple algebraic tests.

Animals↗

The influence of growth medium on the yield of X-ray-induced chromatid exchanges in the presence and absence of aphidicolin.

The frequency of exchanges in JU56 cells irradiated in the G2 phase in the presence and absence of the polymerase inhibitor aphidicolin (APC), and in the presence of a range of concentrations of cysteine was measured. It was found that in the absence of cysteine, incubation for 2 h with APC had no effect on the yield. Addition of cysteine at concentrations of 50-250 mg/l reduced the frequency of exchanges, and at these concentrations the frequency was increased by incubation with APC. At higher concentrations, the yield was reduced and incubation with APC did not elevate it. In following experiments, it was found that incubation with cysteine for a period of longer than 10 minutes was necessary before APC affected the yield of exchanges.

Animals↗

The relationship between length of incubation, bacterial growth and tuberculin yield of a strain of Mycobacterium tuberculosis.

The relationship was studied between length of incubation in a liquid synthetic medium, amount of bacterial growth, pH of the culture filtrate and tuberculin yield of a single strain, DT, of Mycobacterium tuberculosis. No sinple relationship was found between the total crop of bacteria and the tuberculoprotein content of the culture filtrate. On the other hand, protein release and the pH of the culture filtrate were closely related. In each of two trials the bacterial crop increased rapidly during the early period of incubation, although maximum growth was not reached until 9 and 13 weeks respectively. The pH of the culture filtrate reached its maximum at a value of more than 9-0 after 5 weeks incubation and then fell until the 10th week; a second rise occurred until the 13th week when the pH dropped again and levelled off slightly on the alkaline side of neutrality. The rate of increase in tuberculin yield was most rapid during the first 7 weeks of incubation. After this, the rate of increase slowed down but the yield was still rising after 18 weeks. It is concluded that for the large-scale production of mammalian tuberculin by the method used in these trials, it is profitable to incubate production cultures for 14 weeks or longer.

Alanine↗

Biosynthesis of reovirus-specified polypeptides. Multiplication rate but not yield of reovirus serotypes 1 and 3 correlates with the level of virus-mediated inhibition of cellular protein synthesis.

Kinetic analysis of mouse L fibroblast cells infected with reovirus revealed that serotypes 1 (Lang strain) and 3 (Dearing strain) differ significantly from each other in terms of their rates of multiplication and their effects on cellular protein synthesis. Serotype 1 did not significantly affect the synthesis of cellular polypeptides in monolayer cultures of L cells at late times after infection when virus-specific protein synthesis was at a maximum. By contrast, under identical culture conditions, serotype 3 essentially completely inhibited the synthesis of cellular polypeptides at late times when viral protein synthesis was at a maximum. The kinetics of virus-specific polypeptide synthesis and the production of infectious progeny were considerably slower for the serotype 1 Lang strain as compared to the serotype 3 Dearing strain, both at 30 and 37 degrees. However, the relative pattern of viral polypeptide synthesis and the final yield of infectious progeny did not differ significantly between serotypes 1 and 3. For both serotypes, the maximum yield of infectious progeny was obtained shortly after the maximum rate of viral polypeptide synthesis was reached. These results suggest that the rate of multiplication, but not the final yield, of reovirus serotypes 1 and 3 is related to the extent of virus-mediated inhibition of cellular protein synthesis.

Animals↗

Reproductive disorders in dairy cattle: I. Respective influence of herds, seasons, milk yield and parity.

This study was performed in two large dairy units (with 130 and 213 calving cows each) during one year. The objectives were to investigate 1) epidemiological patterns of main post-partum reproductive disorders (metritis, post-partum and post-service anestrus, repeat breeding and embryonic death) and 2) the impact of herd, calving season, milk yield and parity on these patterns. Approximately 20% of the cows in both herds were not affected by any of the disorders. Prevalence of metritis was high (32 to 44%) and appeared influenced by the herds' conditions interacting with calving months and milk yield effects. Cyclic post-partum anestrus incidence was also essentially affected by the herd effect with an added seasonal interaction. Other disorders in both herds were also primarily subjected to the seasonal effect. Individual milk yield and parity only marginally affected the epidemiological patterns. We concluded that even in similar environmental conditions, no general patterns of incidence of reproductive disorders can be drawn and that they are essentially dependent on the characteristics of each herd management.

Journal Article↗

Relationship of declines in grain consumption and milk yield to estrus in dairy cattle.

A retrospective study was done to determine if a reduction in daily grain consumption or decline in daily milk yield were accurate indicators of estrus in 25 Holstein cows. A computer automatically identified all cows that left more than 10% of their daily grain allocation in the feed stall or that gave less than 85% of their most current 7-d daily average milk yeild. By retrospectively analyzing breeding records, grain refusal and milk declines were identified as either occurring in conjunction with estrus or nonestrus. Declines in grain consumption and milk yield were frequently associated with estrus, but many cows in estrus did not exhibit such declines. While such declines hold promise as estrous detection aids, it was concluded that refusals of daily grain allocation or declines in daily milk yield were not sufficiently accurate to supplant other methods of estrous detection in dairy cattle.

Journal Article↗

Some determinants of quality and yield in the isolation of adult heart cells from rat.

We investigated the effect of changes in perfusate substrate and Ca content on the quality and yield of isolated adult rat heart cells. When 1 mM Ca was added to the recirculating perfusate 15 min after collagenase addition, the ATP level of cells in the heart 15 min later, and their morphology in histological section, was no different from when no Ca was added back. The cells subsequently isolated were of similar yield, but a greater percentage were rod-shaped, compared with cells isolated without Ca restoration to the perfusate. Increased yield could be obtained by including substrates in the perfusate in addition to glucose. Either fatty acids or amino acids were effective. We conclude that: (1) all cells in the heart are Ca tolerant at the end of enzyme perfusion; (2) the presence of substrates in addition to glucose can help cells survive the isolation process.

Animals↗

Decreased Coomassie brilliant blue colour yield with glycated proteins.

Collagen, myosin and albumin were incubated for 7 days at 20 degrees C with fructose, ribose or glyceraldehyde. For thus-formed glycated proteins, quantities were determined by the Conway microdiffusion technique and by the colorimetric method based on Coomassie brilliant blue G-250 colour yield. It was found that when albumin was glycated with increasing amounts of glyceraldehyde, the colour yield was decreased by 7-33%. In collagen, myosin and albumin incubated with 0.5 mol/l fructose, 0.5 mol/l ribose or 0.1 mol/l glyceraldehyde, protein concentration was not changed, as proved by the Conway microdiffusion technique; the Coomassie brilliant blue G-250 colour yield was up to 50% lower, depending on the protein used, and was decreased much less when proteins were incubated with less sugar.

Albumins↗

Age-related increase of porcine natural interferon alpha producing cell frequency and of interferon yield per cell.

Porcine blood mononuclear cells (PBMC) were shown to secrete interferon alpha (IFN-alpha) after induction by a coronavirus, the transmissible gastroenteritis virus (TGEV). IFN-alpha producing cells, referred to as natural interferon alpha producing (NIP) cells, were detected by an ELISPOT assay using anti-porcine IFN-alpha monoclonal antibodies. The frequency of NIP cells among blood cells is low, at most 40-110 per 10(5) PBMC and each NIP cell was found to produce several units of IFN. We have shown that NIP cell frequency and IFN yield per cell gradually increased with the age of the donor animals, from the neonatal period to the adult age, with a significant increase around puberty. Our present results also indicate that NIP cells may be influenced by physiological and genetic factors; thus (1) NIP cell frequency and IFN yield per cell were decreased during lactation; (2) Chinese (Meishan) pigs were found to have higher NIP cell frequency and IFN yield per cell than European (Large White) animals.

Aging↗

Leukocyte concentration and the yields of metaphase cells in cultured peripheral blood of leukemic patients with hyperleukocytosis.

The optimal concentration of leukocytes for maximizing the yields of metaphase cells was examined using PHA-stimulated and unstimulated peripheral blood cells of various leukemic patients with hyperleukocytosis, including 5 acute nonlymphocytic leukemias, 4 Ph-positive chronic myelocytic leukemias, one acute lymphoblastic leukemia, and one malignant lymphoma at a leukemic phase. The yields of metaphases reached maximum at leukocyte concentrations of either 1 X 10(6) or 2 X 10(6)/ml in both stimulated and unstimulated blood cultures of each patient, except for one patient with malignant lymphoma who exhibited no metaphases at any cell concentration level in the unstimulated culture. Metaphase yields in the stimulated and unstimulated cultures correlated with neither the percentages of blast cells and lymphocytes in the peripheral blood of the leukemia patients nor with the type of leukemia studied.

Adult↗

Enhanced yields of measles virus from cultured cells.

Conditions for increasing the yield of measles virus from cultured cells have been investigated. Important factors for high virus yields were found to be cell type, incubation at 33 degrees C, concentration of serum in the medium, use of virus as inoculum within five passages of plaque purification, and repeated harvesting of the virus-containing medium at 12 h intervals after extensive virus-induced cell fusion had occurred. Under these conditions, Vero cells infected with the Edmonston strain of measles virus yielded a total of 2.98 X 10(9) plaque-forming units (p.f.u.) of released virus, which is equivalent to 270 p.f.u./cell. A similar amount of cell-associated virus was produced.

Animals↗

Effect of SO2 on the reproduction of pea aphids, Acyrthosiphon pisum, and the impact of SO2 and aphids on the growth and yield of peas.

Pea aphids feeding from birth to maturity on pea plants (Pisum sativum) exposed to SO(2) concentrations of 50 nl litre(-1) or 80 nl litre(-1) showed a significant 19% increase in the rate of nymph production during the reproductive period, compared to control aphids feeding on plants in charcoal-filtered air. The higher nymph production resulted in a mean 4.6% increase in the intrinsic rate of population increase (rm). In longer term glasshouse fumigation experiments pea aphid populations were, on average, 1.8 times greater on pea plants in ambient air plus 45 nl litre(-1) SO(2) than in ambient air alone. Aphid infestation in ambient air caused a 42% reduction in pea yield and affected most plant parameters adversely. Ambient air plus SO(2) had no direct effect on yield, but, in combination with aphid infestation, a further 10% reduction in yield was recorded.

Journal Article↗

Effects of air filtration at small SO2 and NO2 concentrations on the yield of barley.

Two cultivars of Igri and Gerbel winter barley Horteum vulgare L. were grown in open-top chambers in filtered and unfiltered air at a site with approximately 10 nl litre(-1) SO2 and 12 nl litre(-1) NO2 (seasonal mean). The experiment ran for three consecutive seasons 1982-1983, 1983-1984, 1984-1985, and significant effects of filtration were observed for each crop. In years 1982-1983 and 1984-1985, the crops in unfiltered air yielded larger grain dry matter, 9% in 1982-1983, and 8% in 1984-1985. For both crops, the differences were statistically significant at the 5% level. Differences were also observed for the remaining above-ground dry matter, and these were consistent in direction in each year but statistically significant only in 1984-1985. In both growing seasons (1982-1983 and 1984-1985), there were no major pest infestations and no long-term water stress or photochemical ozone episodes. In the remaining experiment (1983-1984) similar air concentrations of SO2 and NO2 produced effects of the opposite sign to those observed in 1982-1983 and 1984-1985. Significant reductions in grain yield (13%) were obtained in unfiltered air. The only major environmental difference for the 1983-1984 crop was a notable dry period in May and June 1984 with marked water stress in the crop, requiring irrigation. These results suggest that the relationship between yield and pollutant concentration may be confounded by additional stresses, many of which are a common component of the growing season for major crops.

Journal Article↗

Influence of ozone stress on growth processes, yields and grain quality characteristics among soybean cultivars.

Field studies were conducted at USDA Beltsville Agricultural Research Center, Beltsville, Maryland, in 1984 and 1985 using open-top chambers to acquire information on the responses of 12 soybean (Glycine max L. Merr.) cultivars to O3 stress and to examine the interactions between maturity groups and O3 stress. Cultivars representing Groups III, IV, and V were exposed for approximately 3 months to charcoal-filtered air (CF) and nonfiltered air plus 40 nl litre(-1) O3 (NF + O3). Ozone was added 6 h d(-1), 5 d week(-1) for 13 weeks. The CF effectively reduced the accumulative oxidant exposure (AOX) to less than 1.0 microl litre(-1) h and the NF + O3 treatment approximately doubled the ambient AOX (16.7 microl litre(-1) h) to about 30 microl litre(-1) h. The AOX estimates the total O3 exposure above 30 nl litre(-1) during an entire growing season. Plant growth rates and relative growth rates were reduced by 17.0 and 14.4%, respectively, when averaged over cultivars. Based on growth rates, the Group III cultivars were the most affected by O3 stress. Averaged over cultivars, leaf expansion rates, leaf conductance, and transpiration rates were lower in the NF + O3 treatment compared to the CF control; however, wide variation was found with the stomatal results from field observations. Combined over years and cultivars, grain yield was reduced by an average of 12.5% by O3 stress with 3 of 12 cultivars showing significant reductions. Grain protein content was increased by 0.7% by O3 stress, but cultivar differences were equal to the differences caused by the O3 treatments. Grain oil content was unchanged by the O3 treatments. Group IV cultivars showed the greatest decrease in grain yield due to O3 stress. Multiple regression analyses were calculated using the difference between the CF and NF + O3 treatment as a measure of O3 stress. Significant positive relationships were found among net assimilation rates, plant growth rates, relative growth rates, and leaf expansion rates, which suggest that growth analysis characteristics would be useful in addition to yield in air pollution tolerance improvement studies with soybeans.

Journal Article↗

The joint action of sulphur dioxide and hydrogen fluoride on the yield and quality of wheat and barley.

The effects of joint action of SO(2) and HF on the yield and quality of wheat and barley were studied by exposing them to combinations of <13,130 or 267 microg m(-3) SO(2) and 0.03 or 0.38 microg m(-3) HF in open top chambers for 90 days. At the concentrations used, SO(2) had greater effects than HF. All responses were marked by compensatory changes. The treatments had no effect on wheat yield, although SO(2) reduced shoot weight. SO(2) increased the growth and yield of barley, and HF or SO(2) increased the grain protein concentration of barley and wheat. The effects of mixtures of SO(2) and HF were complex, but often antagonistic, as the addition of HF counteracted the effect of SO(2) alone.

Journal Article↗