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PubMed1997

Divergent selection for heat loss in mice: I. Selection applied and direct response through fifteen generations.

Divergent selection for heat production/loss (kcal.kg-.75.d-1), measured in 9- to 11-wk-old male mice, was conducted for 15 generations. Heat loss was measured for 15 h on individual animals placed overnight in direct, gradient-layer calorimeters. Selection for high (MH) and low (ML) heat loss and unselected control (MC) occurred in each of three replicates for a total of nine unique lines. Repeatability of the heat loss measurement was .45 and the CV was 10.5%. Cumulative realized selection differentials, averaged for the three replicates, were 145.1 and -105.0 (kcal.kg-.75.d-1) and ranged from 136.9 to 149.2 and -17.1 to -101.3 for MH and ML selection, respectively. Cumulative standardized realized selection differentials, averaged for the three replicates, were 10.06 and -9.51 for MH and ML selection, respectively. Direct responses (kcal.kg-.75.d-1) in heat loss after 15 generations were 44.2 for MH and -27.4 for ML as deviations from MC. Asymmetry of response was evident (P = .03) by Generation 10. Realized heritability was .28 +/- .01 based on divergence of MH and ML selection. For selection for higher and lower heat loss, realized heritabilities were .31 +/- .01 and .26 +/- .01, respectively.

Animals↗
PubMed1985

Selection for increased weaning or yearling weight in Hereford cattle. I. Measurement of selection applied.

Selection was applied from 1964 to 1978 for increased weaning weight (WWL) or yearling weight (YWL) in two Hereford lines with an Angus line maintained as an unselected control line (CL). Each line was maintained with 50 cows and four sires (two sires selected each year and each used for 2 yr). Traits analyzed were birth weight (BW), preweaning daily gain (WDG), weaning weight (WW), weaning conformation grade (WG), weaning condition score (WC), weaning to yearling daily gain (YDG), yearling weight (YW), yearling conformation grade (YG) and yearling condition score (YC). After 15 yr of selection, a total of 3.22 generations of selection had occurred in both WWL and YWL. Average selection differentials in standard measure per generation for WWL, YWL and CL, respectively, were: BW, .44, .51, .0; WDG, .95, .81, .09; WW, .97, .85, .09; WG, .66, .57, .09; WC, .60, .38, -.02; YDG, .30, .79, .38; YW, .80, 1.05, .25; YG, .63, .62, .34 and YC, .45, .64, .24. The proportionate contribution of sire selection (delta S) to the average midparent selection differential per generation (delta M) was 70% in WWL and 76% in YWL. Selection indexes in retrospect were also calculated.

Animals↗
PubMed1988

Divergent selection for growth in Japanese quail under split and complete nutritional environments. 1. Genetic and correlated responses to selection.

Japanese quail were divergently selected for 4-wk body weight under two different nutritional environments. In one environment quail were divergently selected for high and low body weight (H and L) under a split diet (SD) with quail having the opportunity to self-select feed from high protein and high energy diets. In the other environment, quail were similarly selected (H and L) under a normal (28% protein) complete diet (CD). After 11 generations of selection, H-SD and H-CD lines were 48.9 and 49.7% higher in 4-wk body weight than controls. Quail from L-SD and L-CD lines were 46.5 and 45.4% lower in 4-wk body weight than controls. Realized heritabilities for 4-wk body weight were .36 +/- .03, .30 +/- .03, .52 +/- .02, and .47 +/- .03 for H-SD, L-SD, H-CD, and L-CD lines, respectively. Selection differentials were significantly higher in the SD than in the CD environment. Changes in 2-wk, adult body, and egg weight across generations paralleled that of the trait under selection (4-wk body weight). Selection also resulted in decreased hatchability and egg production with increased age to sexual maturity in both environments. The SD environment was determined to be a stress environment because of birds' inferior growth there compared with growth in the CD environment. Rate of growth, however, to reach adult body size was superior for L-SD quail once the stress environment was removed. There was also evidence that eggs were larger for quail in the SD than the CD environment.

Animals↗
PubMed1994

Selection for eight-week body weight in two randombred chicken populations under altered water to feed ratios. 1. Selection responses.

A study was conducted to investigate the response to selection for 8-wk BW under selection environments that utilized different dietary salt levels to alter water to feed ratios. The Athens-Canadian (AC) and Athens Randombreds (ARB) served as base populations. A broiler diet consisting of 23% CP and 3,120 kcal of ME/kg was modified to contain .2, .4, and 1.6% dietary salt. Chicks from the AC and ARB populations were divided within full-sib families into three groups, with one group assigned to each of the three dietary salt selection environments. Individual phenotypic selection was conducted within the six lines for high 8-wk BW for six generations with unselected randombred controls included in each generation to measure environmental variation. No differences in 8-wk BW were observed among the three salt selection environments. Lines derived from the ARB population were initially approximately 75 g larger than those derived from the AC population. However, when BW were expressed as percentage deviation from the control in the sixth generation, selection response patterns were similar (AC lines, 28 to 32%; ARB lines, 32 to 36%). These data indicated that selection for 8-wk BW was as effective under high- and low-salt diets as under a normal-salt diet. Realized heritabilities, following correction for environmental variation, ranged from .38 to .46. Failure to correct for environmental variation resulted in large fluctuations in heritabilities across generations. These data provide strong support for inclusion of controls to accurately measure the heritability of BW in selection studies.

Age Factors↗
PubMed1995

Two-way selection for threshold body weight at first egg in broiler strain females. 5. Replication of results in a two-generation selection experiment.

Selection for high and low threshold weight for onset of lay (ThrWtLay), carried out in a broiler line over six generations, was previously found to increase reproductive performance in the low line (LL). Because of the potential importance of this effect, a two-generation replication of the original experiment was carried out using a modern broiler line. Selection was on the basis of ThrWtLay, measured as body weight at first egg, following gradual release from feed restriction. The total response to selection over two generations was 382 g, giving a realized heritability of .42. The difference in 6-wk body weight of the two lines was +19 g in favor (P > .05) of the high line (HL). Reproductive performance was measured in the S2 generation and in the first (R1) generation of relaxed selection. On the average, age at first egg of LL was 17.6 d earlier, and prepeak, postpeak, and total egg production of LL to a fixed age was 9.9, 1.1, and 11.1 eggs greater, respectively, than that of HL. The differences in age at first egg, prepeak, and total egg production were significant; those in postpeak production were not. There was a difference of 1.1 g in favor (P > .05) of HL for egg weight. Thus, the results of this short-term selection experiment are broadly consistent with those of the original, long-term selection. Namely, ThrWtLay responded strongly to selection, with a powerful accompanied response in age at onset of lay and in prepeak egg production, and only a small effect on 6-wk body weight. However, in contrast to the results obtained previously, LL in the present experiment showed only a small increase (P > .05) in postpeak production relative to HL. Nevertheless, although an effect on postpeak production was not obtained in the present experiment, the results do provide general support for the previously expressed view that selection for early onset of lay, on release from feed restriction, may provide a means of increasing photoperiodic drive and consequent reproductive performance of broiler breeders.

Animals↗
PubMed1981

[Selection of laboratory rats for fertility components in synchronized estrus using Suisychron-Praemix. 1. Effects of selection and synchronization].

The following results were obtained from experimental studies into effects of selection for fertility and into synchronised oestrus though eight generations of laboratory rats: -- The result of selection recordable from the population with selection only was better than that recorded from the population with selection and synchronised oestrus. This has been attributed to higher correlation between phenotype and genotype in the selection variant. -- Direct negative impact of synchronised oestrus on selection index was recorded from a population with random mating. -- The effects of synchronised oestrus regarding selection success were offset but not entirely ruled out by selection. -- Through the generations of both populations with synchronised oestrus the effects of synchronisation were lowered due to both physiological adaptation to Suisynchron application and contraselection with regard to suitability for synchronisation.

Animals↗
PubMed2001

Comparison of methods for erythroblast selection: application to selecting fetal erythroblasts from maternal blood.

BACKGROUND: Many methods have been employed to obtain fetal cells from maternal blood for prenatal diagnostics, but there has been little work done that compares the efficacy of different methods. This study presents a comparison of two commonly used methods for selecting erythroblasts with selection directly from whole blood. METHODS: Erythroblasts were isolated from maternal blood by either differential lysis or density separation, followed by selection with an antibody to the transferrin receptor. These methods were compared with antibody selection directly from whole blood. The total yield of erythroblasts was determined for each method. RESULTS: Red cell lysis is not recommended because the lysis step cannot be well controlled. Density separation followed by antibody selection works well. However, a faster and simpler method, antibody selection directly from whole blood using Immunicon Ferrofluid and magnetic separators, works as well and has the potential to yield even more cells. CONCLUSIONS: Considering the need for a simple and quick method for selecting fetal cells from maternal blood, we suggest selection directly from whole blood.

Acetazolamide↗
PubMed1993

CD8 is needed for positive selection but differentially required for negative selection of T cells during thymic ontogeny.

During thymic development, immature thymocytes expressing major histocompatibility complex (MHC) class I-restricted T cell receptors (TcR) differentiate into CD8+ T cells with cytolytic functions. To evaluate the role of CD8 in positive and negative selection during thymic ontogeny, mice rendered CD8-null by gene targeting were bred with three lines of transgenic mice expressing unique MHC class I-restricted TcR. In all three instances CD8 was required for positive selection of MHC class I-restricted transgenic T cells. The efficiency of positive selection decreased in accordance with a reduced level of CD8 expression on thymocytes. Surprisingly, there was a differential requirement for CD8 expression in negative selection of MHC class I-restricted thymocytes, depending on the antigen specificity of TcR. These observations show that CD8 is essential for positive selection but is differentially required for negative selection of MHC class I-restricted T cells. Thus thymic selection, at least for negative selection, can occur in the absence of the CD8 accessory molecule.

Animals↗
PubMed1998

The mode of movement selection. Movement-related cortical potentials prior to freely selected and repetitive movements.

In two previous studies, the readiness potential (RP) has been reported to be influenced by the mode of movement selection. Freely selected movements were found to have a higher RP amplitude than fixed repetitive movements. This was attributed to the higher demands on planning for the performance of freely selected movements. However, movements in the free mode are distinct from movements in the fixed mode in more than one respect. For example, they are also associated with a higher degree of alteration of the side and/or the finger of movement execution and hence serial "novelty" across blocks of trials. The aim of our study was to establish whether the greater novelty of movements in the free mode could also contribute to the enhanced RP amplitude of movements in the free mode of movement selection by comparing free versus fixed movements performed in long and short sequences that differ in terms of serial novelty. The RP was recorded in 31 healthy young subjects with electrodes placed over Fz, C3, Cz, C4 and Pz. Two types of movement were studied: randomly chosen button presses with right or left index or middle finger (free mode), and repetitive pressing of a predetermined button (fixed mode). We found that: (1) in confirmation of previous studies, the amplitude of the RP was higher for freely selected than free movements; (2) the effect of the mode of movement selection was present over central electrodes but was most pronounced for parietal electrode Pz, with movements in the free mode showing the earliest and greatest increase in negativity at this site; (3) this parietally enhanced negativity in free compared with the fixed mode was absent after the subjects had performed a block of long movement sequences, suggesting that serial novelty of movements also contributed to the effect of mode on the RP amplitude; (4) both the latency and the magnitude of the lateralized readiness potential (LRP) were altered by the mode of movement selection. Movements in the free mode showed an earlier onset of the LRP, which had a higher peak than the LRP prior to movements in the fixed mode. This effect was mainly due to an increased amplitude of the RP over the electrode contralateral to the side of movement prior to freely selected movements. These findings are discussed in relation to previous RP and positron emission tomography studies.

Adult↗
PubMed2005

Effective selection of transgenic papaya plants with the PMI/Man selection system.

The selectable marker gene phospho-mannose isomerase (pmi), which encodes the enzyme phospho-mannose isomerase (PMI) to enable selection of transformed cell lines on media containing mannose (Man), was evaluated for genetic transformation of papaya (Carica papaya L.). We found that papaya embryogenic calli have little or no PMI activity and cannot utilize Man as a carbon source; however, when calli were transformed with a pmi gene, the PMI activity was greatly increased and they could utilize Man as efficiently as sucrose. Plants regenerated from selected callus lines also exhibited PMI activity but at a lower specific activity level. Our transformation efficiency with Man selection was higher than that reported using antibiotic selection or with a visual marker. For papaya, the PMI/Man selection system for producing transgenic plants is a highly efficient addition to previously published methods for selection and may facilitate the stacking of multiple transgenes of interest. Additionally, since the PMI/Man selection system does not involve antibiotic or herbicide resistance genes, its use might reduce environmental concerns about the potential flow of those genes into related plant populations.

Agriculture↗
PubMed1990

Selective production of hybridoma cells: antigenic-based pre-selection of B lymphocytes for electrofusion with myeloma cells.

Methods for pre-selecting B lymphocytes were studied and investigated. First, biotinylated antigen was used for selecting B lymphocytes. These pre-selected B lymphocytes were then combined with biotinylated myeloma cells by adding streptavidin. The final formula of the selected B cell-myeloma cell was as follows: B cell-(antigen-biotin-strept-avidin-biotin)-myeloma cell. Then, this B cell-myeloma cell conjugate was fused by the pulsed electric field (PEF) method, which fused only those conjugated cells. The fusion efficiency obtained by this method was 3-15-times higher than that obtained by the non-specific poly(ethylene glycol) (PEG) fusion method. Second, avidin-antigen conjugate was used to select B lymphocytes. For this purpose, bifunctional cross-linkers such as N-succinimidyl 3-(2-pyridyldithio) propionate (SPDP) and m-maleimidobenzoyl N-hydroxysuccinimide (MBS) were chosen. Each reagent contains two heterofunctional groups which can make covalent bond with both Lys and Cys residues. Typical avidin-antigen conjugate is expressed as avidin-SPDP (or MBS)-antigen. Thus, final B cell-myeloma cell conjugate was B cell-antigen-SPDP (or MBS)-avidin-biotin-myeloma cell. The yield of this procedure was of the order of 10(-2). Here, we suggest that the pre-selection of B lymphocytes by biotinylated antigen or avidin-antigen conjugate is a new method of obtaining selected hybridoma cells which produce specific monoclonal antibodies against the antigen used for selecting B lymphocytes.

Animals↗
PubMed2004

Limited lentiviral transgene expression with increasing copy number in an MGMT selection model: lack of copy number selection by drug treatment.

Retroviral vector integration into the human genome carries increased risk of oncogenesis with increasing integrations. To boost transgene expression for gene therapy, multiple integrations are often sought. We studied the relationship between the number of vector integrations and transgene expression and the effect that drug selection in an MGMT-selection model would have on vector copy number. K562 cells were transduced using a lentiviral vector and a library of clones was generated. Median proviral copy number was 4 and a positive correlation with transgene expression was observed. Transgene expression increased at a linear rate between 1 and 4 vector copies/cell, but was unpredictable at >4 integrations/cell. When lentivirus MGMT(P140K)-transduced K562 cells were treated with O(6)-benzylguanine (BG)/BCNU, there was no selection for increased median copy number in colony-forming units, despite strong selection pressure and an increase in transgene expression and activity. These data show a direct and linear correlation between MGMT(P140K) transgene expression and vector copy number. Strong BG/BCNU selective pressure does not result in preferential survival of high-copy-number clones but does select for strong transgene expression. Thus drug selection would not be expected to increase the risk of oncogenesis due to exaggerated selection in favor of high-copy-number vector integration.

Alkyl and Aryl Transferases↗
PubMed2000

Cationic or anionic sites? Selectivity optimization of ion-selective electrodes based on charged ionophores

The influence of ionic sites on the selectivities of ionophore-based ion-selective electrodes (ISEs) is described on the basis of a phase boundary potential model. The discussion presented here is significantly more general than previous ones. It is formulated for primary and interfering ions of any charges and it is valid for ISEs based on electrically charged or neutral ionophores. Furthermore, it also applies to membranes that contain more than one type of complex of the primary or interfering ion. It has been believed thus far that only ionic sites of the same charge sign as the primary ion improve the selectivities of ISEs based on charged ionophores. However, it is shown here that the charge sign of the ionic sites that give the highest potentiometric selectivities depends on the charge number of the primary and interfering ions and on the stoichiometry of their complexes with the ionophore. The validity of our model was confirmed experimentally with three ISEs based on different charged ionophores. ISEs based on lasalocid or 1-(N,N-dicyclohexylcarbamoyl)-2- (N,N-dioctadecylcarbamoyl)ethylphosphonic acid monomethyl ester (ETH 5639) as the ionophore responded selectively to Sr2+ or Mg2+, respectively, and discriminated well against other alkaline earth cations when their membranes contained anionic sites. These two electrodes are the first examples of ISEs based on charged ionophores for which maximum selectivities are obtained with membranes containing ionic sites with a charge sign opposite to that of the primary ion. On the other hand, the experimental F- selectivities of membranes based on oxo(5,10,15,20-tetraphenylporphyrinato)molybdenum-(V) improved gradually when the concentration of anionic sites was increased from 0 to 75 mol%. The selectivity-modifying influence of ionic sites for these three types of ISEs can be explained by considering the different stabilities of the 1:2 ion-ionophore complexes of the primary and of the interfering ions.

Journal Article↗
PubMed2004

Combinatorial selection and edited combinatorial selection of phosphorothioate aptamers targeting human nuclear factor-kappaB RelA/p50 and RelA/RelA.

Nuclear factor-kappaB (NF-kappaB) transcription factors are important in regulating the immune response and play critical roles in the pathogenesis of chronic inflammatory diseases and a variety of human cancers. Agents that target specific NF-kappaB dimers may serve as therapeutic agents for the prevention of pathogenic immune responses. We have selected monothiophosphate-modified aptamers, or "thioaptamers", to the NF-kappaB p50/RelA heterodimer using combinatorial selection techniques. We also utilized a "double sieve" or editing approach for the generation of thioaptamers with enhanced selectivity to the RelA/RelA homodimer. The thioaptamers from these selections and our previous selections on the p50/p50 and RelA/RelA homodimers all had unique sequences and bound tightly to the recombinant NF-kappaB dimers against which they were selected. The selected thioaptamers also appear to maintain their selectivity and specificity among other cellular proteins, because they have the ability to bind NF-kappaB proteins within nuclear extracts from lipopolysaccharide (LPS)-induced macrophages and B cells.

B-Lymphocytes↗
PubMed2003

Computer modeling of selective regions in the active site of nitric oxide synthases: implication for the design of isoform-selective inhibitors.

Selective inhibition of nitric oxide synthase (NOS) isoforms has great therapeutic potential in the treatment of certain disease states arising from the pathological overproduction of nitric oxide. In this study three structures of each NOS isoform were employed to examine selective regions in the active site using the GRID/CPCA approach. In the GRID calculations, 10 probes covering hydrophobic, steric, and hydrogen-bond-acceptor and -donor interactions were used to calculate the molecular interaction fields (MIFs) in the active site. The side chain flexibility of the residues and the grid spacings were considered at the same time. Consensus principal component analysis (CPCA) was applied to analyze the MIFs differences in the active site between the NOS isoforms. By combining the cutout tool with GRID/CPCA pseudofield differential plots, several selective regions in the active site were identified. The selectivity analysis showed that the most important determinants for NOS inhibitor selectivity are hydrophobic and charge-charge interactions. Twenty-five inhibitors of NOS were then docked into the active site using the program AutoDock3.0. The regions identified as being important for selectivity by this method are in excellent agreement with inhibitor structure-activity relationships. A rational usage of the selective region described in this work should make it possible to develop NOS isoform-selective inhibitors.

Binding Sites↗
PubMed1998

The xylose isomerase gene from Thermoanaerobacterium thermosulfurogenes allows effective selection of transgenic plant cells using D-xylose as the selection agent.

The xylose isomerase gene (xylA) from Thermoanaerobacterium thermosulfurogenes (formerly Clostridium thermosulfurogenes) has been expressed in three plant species (potato, tobacco, and tomato) and transgenic plants have been selected on xylose-containing medium. The xylose isomerase gene was transferred to the target plant by Agrobacterium-mediated transformation. The xylose isomerase gene was expressed using the enhanced cauliflower mosaic virus (CaMV) 35S promoter and the omega' translation enhancer sequence from tobacco mosaic virus. Unoptimized selection studies showed that, in potato and tomato, the xylose isomerase selection was more efficient than the established kanamycin resistance selection, whereas in tobacco the opposite was observed. Efficiency may be increased by optimization. The xylose isomerase system enables the transgenic cells to utilize xylose as a carbohydrate source. It is an example of a positive selection system because transgenic cells proliferate while non-transgenic cells are starved but still survive. This contrasts to antibiotic or herbicide resistance where transgenic cells survive on a selective medium but non-transgenic cells are killed. The results give access to a new selection method which is devoid of the disadvantages of antibiotic or herbicide selection.

Aldose-Ketose Isomerases↗
PubMed2000

The effect of selective and non-selective phosphodiesterase inhibitors on allergen- and leukotriene C(4)-induced contractions in passively sensitized human airways.

Non-selective inhibitors of cyclic nucleotide phosphodiesterase (PDE) block allergen-induced contraction of passively sensitized human airways in vitro by a dual mechanism involving a direct relaxant effect on smooth muscle and inhibition of histamine and cysteinyl leukotriene (LT) release from airways. We investigated the effects of non-selective PDE inhibitors and selective inhibitors of PDE3 and PDE4 in order to determine the involvement of PDE isoenzymes in the suppression of allergic bronchoconstriction. Macroscopically normal airways from 76 patients were sensitized with IgE-rich sera (>250 u ml(-1)) containing specific antibodies against allergen (Dermatophagoides farinae). Contractile responses of bronchial rings were assessed using standard organ bath techniques. Passive sensitization caused increased contractile responses to allergen, histamine and LTC(4). Non-selective PDE inhibitors (theophylline, 3-isobutyl-1-methylxanthine [IBMX]), a PDE3-selective inhibitor (motapizone), PDE4-selective inhibitors (RP73401, rolipram, AWD 12-281) and a mixed PDE3/4 inhibitor (zardaverine) all significantly relaxed inherent bronchial tone at resting tension and to a similar degree. Theophylline, IBMX, zardaverine and the combination of motapizone and RP73401 inhibited the contractile responses to allergen and LTC(4). Pre-treatment with motapizone, RP73401, rolipram or the methylxanthine adenosine receptor antagonist, 8-phenyltheophylline, did not significantly decrease responses to either allergen or LTC(4). We conclude that combined inhibition of PDE3 and PDE4, but not selective inhibition of either isoenzyme or antagonism of adenosine receptors, is effective in suppressing allergen-induced contractions of passively sensitized human airways. The relationship between allergen- and LTC(4)-induced responses suggests that PDE inhibitors with PDE3 and PDE4 selectivity are likely to act in part through inhibition of mediator release and not simply through direct relaxant actions on airway smooth muscle.

1-Methyl-3-isobutylxanthine↗
PubMed1998

Selective versus non-selective antiarrhythmic approach for prevention of atrial fibrillation after coronary surgery: is there a need for pre-operative risk stratification? A prospective placebo-controlled study using low-dose sotalol.

AIM: This study evaluated the advantages of 'selective' over 'non-selective' antiarrhythmic prevention of atrial fibrillation after coronary surgery based on a new risk prediction algorithm. METHODS AND RESULTS: In a retrospective analysis of a prospective randomized trial, a model for risk prediction was determined based on clinical data of the control group (A; n = 107) and tested in a test group (B; n = 107, treated with low dose sotalol). Using this algorithm, the effect of a 'selective' antiarrhythmic approach in high-risk patients was compared to a 'non-selective' approach, where all patients were treated. In total, 75 (35%) patients developed atrial fibrillation and 14 (7%) side-effects led to discontinuation of study medication. Based on the risk prediction algorithm, 36% of group A patients were classified as high-risk patients with an incidence of atrial fibrillation of 76% compared to 26% in low-risk patients (P < 0.0001). The selective approach, i.e. treatment of high-risk patients only reduced the incidence of atrial fibrillation from 76% to 50% (P = 0.0295) compared to a reduction from 44% to 26% (P = 0.0065) when all patients were treated. More importantly, with the non-selective approach 100% of patients were exposed to the possible side-effects of sotalol and costs compared to 24% only with the selective approach (P < 0.0001). CONCLUSIONS: Thus, a selective approach based on a clinical risk prediction algorithm should improve the cost-effectiveness and safety of low-dose sotalol in the prevention of atrial fibrillation after coronary bypass surgery.

Adult↗