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PubMed1994

Differentiation of selected members of the family Neisseriaceae (Alysiella, Eikenella, Kingella, Simonsiella and CDC groups EF-4 and M-5) by carbohydrate fingerprints and selected phenotypic features.

On the basis of nucleic acid relationships, the family Neisseriaceae consists of the genera Neisseria, Kingella, Simonsiella and of Alysiella filiformis, Eikenella corrodens, and the CDC groups EF-4 and M-5. Differentiation, especially of the new members of the family, by conventional phenotypic characteristics is difficult and in some cases leads to doubtful results. On the other hand, cellular components proved to be suitable for the characterization of bacterial taxa. We investigated the cellular carbohydrates derived from whole cell hydrolysates of the above mentioned taxa with the exception of Neisseria by gas chromatography/mass-spectrometry. The analysis revealed characteristic patterns for all taxa considered, although with some species of which only few strains were investigated so far only preliminary results could be established. With the method used, the carbohydrate analysis could be completed within six hours starting from a pure culture. All strains investigated exhibited a common pattern with ribose, arabinose, glucose, and galactose. Qualitative and quantitative differences in contents of fucose, sorbose, rhamnose, threose, heptose, galactosamine and an amino sugar similar to glucosamine discriminated members of the taxa investigated. To achieve a taxonomically precise differentiation of the species investigated by conventional phenotypic features as available in commercial rapid test kits, these tests should be completed by the carbohydrate analysis technique presented.

Carbohydrates↗
PubMed1995

Selection for high eight-week body weight in normal and dwarf chickens under high-protein and high-energy diets.

A study was conducted to examine genetic parameters associated with selection for high 8-wk BW in normal and dwarf Athens Canadian randombreds under two nutritional environments (high protein and high energy) and to determine the influence of the selection diet on subsequent water to feed ratios and abdominal fat levels. High-protein lines (HP) received a diet containing 25% CP and 3,100 kcal ME/kg, whereas high-energy lines (HE) received a 22% CP diet containing 3,200 kcal ME/kg. Body weights, selection gains, selection differentials, and heritabilities were similar for normal HP and HE lines. There was evidence, however, that the realized heritability was greater in the HE than the HP dwarf line. Similar selection differentials indicate that this difference was apparently due to the expression of greater genetic variation in the HE dwarf line. Water intake, feed intake, water to feed ratio, and abdominal fat data for the four lines under "selection" and "reciprocal" environments differed between experiments. In Experiment 1, when fed the HE diet, normal birds selected under the HP environment had higher (P < .05) water to feed ratios and lower abdominal fat levels than contemporaries selected under the HE diet. However, these differences were not significant in Experiment 2.

Adipose Tissue↗
PubMed2005

Dexamethasone selectively inhibits differentiation of cord blood stem cell derived-dendritic cell (DC) precursors into immature DCs.

Perinatal dexamethasone (Dx) alters the immune system leading to increased infections and developmental abnormalities. Dendritic cells (DCs) derived from cord-blood monocytes are especially Dx sensitive and we sought to determine the effects of Dx on cord-blood CD34+-DCs. Distinct stages of cord-blood CD34+-DC development were delineated: pre-DC, immature, and mature DCs. Dx added during development of pre-DCs did not suppress precursor number, or translocate the glucocorticoid receptor (GcR) from the cytoplasm to the nucleus. However, Dx added during pre-DCs differentiation into immature DCs, prompted GcR translocation to the nucleus, enhanced DC apoptosis, suppressed differentiation to CD1a+ cells, inhibited expression of CD86, reduced subsequent CD83 expression, maintained DC endocytic activity, suppressed IL-6 secretion, enhanced IL-10 secretion, and reduced DC-mediated T cell stimulation. Dx added during the maturation stage caused less dramatic effects. Thus, Dx stalled maturation, selectively induced apoptosis of developing DCs and the sensitivity peaked during pre-DCs differentiation into immature DCs.

Antigens, CD34↗
PubMedDate not supplied

Neuronal differentiation of mouse embryonic stem cells: lineage selection and forced differentiation paradigms.

Primitive embryonic stem cells are an ideal starting cell population for studies of gene expression and lineage segregation during development. Despite their potential, it has been difficult to determine culture conditions that cause single-lineage differentiation of these pluripotent cells. Both genetic and epigenetic approaches have been taken to promote neuronal differentiation of embryonic stem cells, including aggregation, exposure to the nonspecific teratogen/morphogen retinoic acid, low-density culture, exposure to growth/differentiation factors, and forced differentiation following expression of lineage-restricted "developmental control" genes. In the current investigation, a hybrid approach involving genetic techniques of "lineage selection" or "forced differentiation" has been employed to develop primitive neural progenitor cell lines. These lines form an important starting point to examine the cascades of gene expression (and inhibition) during neuronal and glial lineage segregation, to study growth factor effects on neural differentiation, and ultimately to provide a source of cells for transplantation to a damaged nervous system.

Animals↗
PubMed1993

Alternative methods of selection for litter size in mice: III. Response to 21 generations of selection.

Alternative methods of selection to increase litter size in mice have been practiced for 21 generations followed by six generations of relaxed selection. Three replicates were used with four selection criteria: index of components (IX:I = 1.21 x total ovulation rate + 9.05 x ova success), uterine capacity (UT), litter size (LS), and an unselected control (LC). In IX, ovulation rate and ova success were measured by number of corpora lutea and number of pups born/number of corpora lutea, respectively. In UT, uterine capacity was measured and defined as number of pups born to unilaterally ovariectomized (right ovary excised) females. Selection in LS was based on number born to unaltered dams. In all cases, number born was fully formed, live or dead pups. Pups from 16 randomly chosen LC dams and from the top 16 dams in IX, UT, and LS were selected to produce the next generation in each criterion-replicate line. Response in number born, selected criteria deviated from control, was regressed on generation number over the 21 generations of selection. Responses for the IX and LS criteria were quite similar (.14 +/- .01 and .16 +/- .01 pups per generation, respectively), whereas response in UT, with only one functional horn, was slightly lower (.09 +/- .01). The average cumulative selection differentials for IX, LS, and UT at Generation 21 were 32.78 index units, 36.38 pups, and 28.53 pups, respectively. The LC criterion had an unintentional cumulative selection differential of 3.3 pups.(ABSTRACT TRUNCATED AT 250 WORDS)

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↗
PubMed1992

Selective and differential binding of interleukin (IL)-1 alpha, IL-1 beta, IL-2 and IL-6 to glycosaminoglycans.

The binding of interleukin (IL)-1 alpha, IL-1 beta, IL-2 and IL-6 to acidic polysaccharides was investigated by affinity chromatography of the recombinant, radioiodinated interleukins on columns of immobilized polysaccharide. Each interleukin showed selective binding retention. Overall heparin bound all four interleukins significantly, whereas chondroitin sulfate provided little retention. IL-1 alpha and IL-1 beta showed differential binding, with only the latter binding to hyaluronic acid. IL-2 was virtually completely retained on fucoidan. Noniodinated recombinant IL-2 bound similarly to fucoidan, and fucoidan was found to sequester IL-2 activity in a bioassay employing IL-2-dependent CTLL cells. In all other cases tested, interleukin retention was partial, implying that interleukin binding sites are sparsely distributed along the polysaccharide chains. These findings suggest that during the immune response, interleukins will tend to be retained at sites of secretion by interaction with glycosaminoglycans in the extracellular matrix and on cell surfaces.

Glycosaminoglycans↗
PubMed2003

Osteogenic differentiation is selectively promoted by morphogenetic signals from chondrocytes and synergized by a nutrient rich growth environment.

Cartilage formation always precedes that of bone during endochondral skeletal development. To determine if chondrocytes provide inductive signals for osteogenesis, C3H10T(1/2) mesenchymal stem cells were co-cultured in membrane separated transwell culture chambers with chondrocytes, osteoblasts, or fibroblasts. Osteogenesis, as assessed by the expression of osteocalcin mRNAs, was strongly induced in the C3H10T(1/2) cells co-cultured with chondrocytes but not induced by co-culture with either osteoblasts or fibroblasts. Interestingly, while only osteogenic differentiation was observed in the C3H10T(1/2) cells co-cultured with chondrocytes, bone morphogenetic protein (BMP)-7 treatment induced an ordered endochondral progression of skeletal cell differentiation in which chondrogenic differentiation preceded osteogenesis by 2 to 4 days. A nutrient enriched growth environment enhanced osteogenic differentiation induced by either co-culture or BMP-7 treatment 2- to 5-fold. Nutrient enhanced osteogenic differentiation was associated with an activation of the retinoblastoma-mediated signal transduction pathways. In summary, these results show that osteogenesis is selectively induced by morphogenetic signals produced by chondrocytes and that a nutrient rich environment enhances both BMP-7- and co-culture-induced osteogenic differentiation.

Animals↗
PubMed1998

Comparison of selection by independent culling levels for below-average birth weight and high yearling weight with mass selection for high yearling weight in line 1 Hereford cattle.

Mass selection by independent culling levels (YB subline) for below-average birth weight (BWT) and high yearling weight (YWT) was compared with single-trait mass selection (YW subline) for high YWT in the inbred population of Line 1 Hereford cattle at Miles City, Montana. There were 4.2 generations of selection in YB and YW. Heritability estimates for the base population derived from multiple-trait REML were .28 and .31 for direct effects and .16 and .06 for maternal effects on BWT and YWT, respectively. Mid-parent cumulative selection differentials for BWT of YB and YW diverged (-2.9 vs 8.2 kg, respectively), as did the associated genetic trends for direct effects (-.014 kg/yr vs .105 kg/yr, respectively). Mid-parent cumulative selection differential for YWT of YB (102.1 kg) was 64% of that attained in YW (160.7 kg). Likewise, response in YWT of YB (.91 kg/yr) was 61% of response attained in YW (1.5 kg/yr). For BWT and YWT, maternal genetic trends were similar across selection lines. Assistance at parturition of first-parity 2-yr-old heifers was consistently less frequent in YB than in YW.

Aging↗
PubMed2002

Generation of amplifiable genome-specific oligonucleotide probes and libraries.

Here we describe a process for the generation of oligonucleotide libraries representative of a given nucleic acid. Starting from at random pool of DNA oligonucleotides, the technique selects only those that hybridize to the nucleic acid template. This selection yields a highly specific library that represents an oligonucleotide image of the chosen template. The novel quality of this approach is the generation of amplifiable oligonucleotide probes that are of unique length and are easily subjected to differential selection. Here we apply this technique to produce different genomic oligonucleotide libraries and show that these genomic oligonucleotide libraries do not cross-hybridize. Differential selection of these genomic oligonucleotide libraries produces oligonucleotides that can be used in the identification, characterzation, and isolation of nucleic acids.

Adenoviridae↗
PubMed1991

A new plate medium for rapid presumptive identification and differentiation of Enterobacteriaceae.

A new selective differential agar medium for rapid presumptive identification of Enterobacteriaceae from water and food samples is described (EMX ID agar). By a combination of fluorogenic and chromogenic substrates, the medium detects the presence of beta-D-glucuronidase, beta-D-galactosidase, beta-D-xylosidase, tryptophane deaminase and H2S; additionally, cytochrome-oxidase and indole production can be demonstrated. This medium provides an inexpensive means for simple and rapid presumptive identification of E. coli and coliforms and for the differentiation within the Klebsiella-Enterobacter and the Proteus-Providencia-Morganella group. Furthermore, it allows to distinguish between the H2S-positive Enterobacteriaceae Citrobacter freundii, Salmonella spp., S. arizonae, Edwardsiella, Proteus mirabilis, P. vulgaris and some oxidase-positive bacteria.

Agar↗
PubMed1999

A recombinase-based selection of differentially expressed bacterial genes.

Bacterial genes are often differentially expressed in response to specific environmental conditions. We have devised a method to identify regulated bacterial promoters, such that transient promoter expression leads to a permanent and selectable change in bacterial phenotype. This system consists of a promoterless derivative of cre, the phage P1 recombinase, carried on a plasmid, and two chromosomal loxP sites, the targets of the Cre recombinase. The loxP sites flank npt, conferring kanamycin resistance, and sacB, which confers sensitivity to sucrose, allowing positive selection for both the presence and absence of this chromosomal cassette. Fusion of active promoters to cre induces recombination of the loxP sites and deletion of intervening DNA, allowing selection on media containing sucrose, while inactive promoters fail to induce recombination and so remain resistant to kanamycin. We tested the system in Salmonella typhimurium using a known regulated promoter, that from the araBAD operon, and found it to be a sensitive indicator of gene expression over a wide range of promoter induction. We then used this system to identify S. typhimurium genes that are specifically expressed when bacteria interact with cultured epithelial cells and identified a novel DNA fragment, not found in E. coli, which might represent part of a new pathogenicity island.

DNA, Bacterial↗
PubMed1995

Modeling positive and negative selection and differentiation processes in the thymus.

T cells begin their development as precursor cells in the bone marrow. These cells migrate to the thymus, where they further divide, differentiate, and mature into functional T cells. Most thymocytes (95-99%) die in the course of this process, and only relatively few exit the thymus as mature cells. Here we develop a differential equation model of cell proliferation, differentiation and death in the thymus that can account for both the total number of thymus cells and the fractions of various types of immature and mature thymocytes. Our model suggests that positive and negative selection may have more complex effects than simply deleting some cells and allowing others to survive.

Animals↗
PubMed1971

Modified selective and differential isolation medium for Vibrio parahaemolyticus.

The semiselective salt-starch-agar formulation of Baross and Liston was modified as the result of a systematic study of the effect of each constituent on the growth of Vibrio parahaemolyticus and competitive species characteristic of the marine environment. The selection of nutrient constituents depended on an analysis of their effect on generation time. The addition of inhibitors depended on an analysis of minimal inhibitory concentrations. The modified formulation included: peptone, 2.0%; yeast extract, 0.2%; corn starch, 0.5%; NaCl, 3.0%; agar, 1.5% (pH 8.0). Penicillin at 2 to 5 units/ml increased selectivity without significantly inhibiting Vibrio in pure suspensions. Over 62% of the most sensitive strain (YM-K33) was recovered at a concentration of 5 units of penicillin per ml. The per cent recovery of V. parahaemolyticus from fish homogenate compared favorably with other selective formulations. At an initial concentration of 10(5) cells/ml, recovery varied with the strain used from 60 to 119%, whereas at 10(2) cells/ml the range was 36 to 94%. Applications of the medium for Vibrio quantification are discussed.

Agar↗
PubMed1993

V-region directed selection in differentiating B lymphocytes.

We here analyse the repertoire of VH7183 rearrangements isolated from different stages of B cell differentiation in adult mice. The nucleotide sequence analyses of VH7183-D-JH rearrangements derived from large pre-B cells (B220+, mu-), small pre-B cells (B220+, mu-) and mature B cells (B220+, mu+) isolated from adult bone marrow revealed a sequential accumulation, among functional rearrangements, of D segments of the FL16 family and a depletion of D segments using the second and the third reading frame (RF). One member (VH7183.1) of the VH7183 gene family was utilized in 60-80% of the rearrangements of all populations analysed. In neonates the majority of the rearrangements utilizing this gene was found to be functional. In contrast, > 96% of the VH7183 rearrangements isolated from adult spleen were non-functional. These data provide evidence for cellular selection of VH regions acting at different points of the B cell differentiation pathway and at the transition of B cells from the bone marrow to the periphery.

Animals↗
PubMed1986

Determination of NADPH-specific dihydropteridine reductase in extract from human, monkey, and bovine livers by single radial immunodiffusion: selective assay differentiating NADPH- and NADH-specific enzymes.

It has been difficult to determine exactly NADPH-specific dihydropteridine reductase [EC 1.6.99.10] in samples which also contain NADH-specific dihydropteridine reductase [EC 1.6.99.7], because the latter enzyme interferes with the activity measurement of the former. We have devised a method to measure selectively the NADPH-specific reductase in crude extracts of bovine, human and monkey livers by the single radial immunodiffusion method using specific antiserum against the enzyme. This method makes it possible to determine the enzyme amount in 5 microliters of the 3-volume extracts of the livers. The amounts of NADPH-specific dihydropteridine reductase were calculated to be 0.252, 0.296, and 0.583 munits/5 microliter of the extracts of bovine, human, and monkey livers, respectively.

Animals↗
PubMed1982

Abnormal differentiation of selected nuclear centers in the brain of a duck embryo associated with partial duplication of the primitive streak.

In a control set of duck embryos, an abnormal duck embryo of 16 days incubation was found which had two beaks as the only outward sign of duplication. The beaks were of equal size, each with upper and lower bills. Bill-clapping movements were absent. The embryo had two normal eyes placed one on either side of the head, and the rest of the body was normal in external appearance. Sections through the head revealed further duplication of the fore-, mid-, and hindbrain divisions. The medial half of each of the embryonic brain divisions, however, was greatly reduced. Two additional optic primordia were also noticed in sections, each of which was reduced to a mass of tissue representing a lens and a much-folded pigment epithelium. The orbital tissues associated with the rudimentary eyes were greatly disorganized. Abnormal differentiation associated with duplication of the brain divisions was determined by cell counts in selected nuclear centers. Cell numbers in each case appeared to be remarkably proportional to the size of the innervation field. Our data, based on cell counts in the nuclear centers chosen for this study in the abnormal embryo and normal control embryos of the same age, are consistent with the hypothesis that cell survival in related parts of the nervous system may be regulated by their peripheral field of innervation.

Animals↗
PubMed1996

Quantitative structure-activity relationships studies with micellar electrokinetic chromatography. Influence of surfactant type and mixed micelles on estimation of hydrophobicity and bioavailability.

Applications of micellar electrokinetic chromatography (MEKC) in quantitative structure-activity relationships (QSAR) were studied. First, quantitative structure-retention relationships (QSRR), which describe the correlation between logarithm of capacity factor (log k') in MEKC and logarithm of distribution coefficient between 1-octanol and water (log P(ow)), were investigated for 60 aromatic compounds and 9 corticosteroids using three different anionic surfactants [e.g., sodium dodecyl sulfate (SDS), sodium cholate (SC), and lithium perfluorooctane sulfonate (LiPFOS)], one cationic surfactant (C14TAB), and mixed anionic micellar systems. Linear solvation energy relationships (LSER) and solvatochromic parameters were used to shed light on the different log k' vs. log P(ow) relationships of the various surfactants. It was concluded that hydrogen bonding interactions have a great influence on retention behavior in MEKC and its relationships with hydrophobicity. Interestingly, bile salt surfactants (e.g., SC) and mixed bile salt micellar systems provide better correlations for log k' vs. log P(ow) than SDS and/or SDS with buffer additives (e.g., beta-cyclodextrin, urea, and acetonitrile). Using SC micelles, only one line was adequate to describe the relationship between retention in MEKC and hydrophobicity for a group of 60 aromatic compounds. The existence of higher correlation for the SC system was attributed to a similar hydrogen bonding pattern between SC micelles and 1-octanol. In the SDS system, however, three lines were recognized for the congeneric subgroups of compounds. This is due to the hydrogen bond door (HBD) characteristic of SDS micelles that selectively differentiate between the solutes with different hydrogen bond acceptor (HBA) strength, thus demonstrating that retention is not solely based on hydrophobicity. A similar result was observed for a C14TAB-MEKC system, however, the HBA characteristic of C14TAB selectively differentiates between the solutes with different HBD strength. In addition, quantitative retention-activity relationships in MEKC were also investigated for 9 corticosteroids. Two types of biological activities [small intestinal absorption in the rat (log A/NA) and protein binding to human serum albumin (log B/F) were examined in this work. High correlations were observed between bioactivity and log k' in MEKC using bile salt surfactants and mixed bile salt systems.

Adrenal Cortex Hormones↗