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C J Collins

Publications and source records attributed to C J Collins.

At least 19 recordsLinked to original sources
PubMed2001

Aminoborohydrides. 12. Novel tandem S(N)Ar amination-reduction reactions of 2-halobenzonitriles with lithium N,N-dialkylaminoborohydrides.

A novel tandem amination-reduction reaction has been developed in which 2-(N,N-dialkylamino)benzylamines are generated from 2-halobenzonitriles and lithium N,N-dialkylaminoborohydride (LAB) reagents. These reactions are believed to occur through a tandem S(N)Ar amination-reduction mechanism wherein the LAB reagent promotes halide displacement by the N,N-dialkylamino group, and the nitrile is subsequently reduced. This one-pot procedure is complimentary to existing synthetic methods and is an attractive synthetic tool for the nucleophilic aromatic substitution of halobenzenes with less nucleophilic amines. The (N,N-dialkylamino)benzylamine products of this reaction are easily isolated after a simple aqueous workup procedure in very good to excellent yields.

Journal Article↗
PubMed2001

Prolonged ex vivo culture of cord blood CD34(+) cells facilitates myeloid and megakaryocytic engraftment in the non-obese diabetic severe combined immunodeficient mouse model.

A clinical goal for ex vivo expansion of cord blood (CB) CD34(+) cells is to shorten the period of neutropenia and thrombocytopenia following myeloablative therapy and transplantation. Prolongation of cytokine expansion leads to the production of greater numbers of cells, and should have an impact on neutrophil and platelet recovery. Furthermore, expansion of CD34(+) cells should support the continued production of neutrophils and platelets in the 6-week period following transplantation. We tested these hypotheses by characterization of the kinetics (human CD45(+) cells in the blood) and phenotype (CD45, CD34, CD61, CD33, CD19 and CD3) of human engraftment in the non-obese diabetic severe combined immunodeficient mouse (NOD-SCID) following 7 or 14 d of ex vivo expansion of CB CD34(+) cells. Mice transplanted with 14 d cells showed greater percentages of human CD45(+) cells in the blood, bone marrow and spleen than mice transplanted with unexpanded cells or 7 d cells. Prolonging cytokine exposure of CD34(+) cells and transplantation with increasing numbers of input cells facilitated the production of absolute numbers of CD34(+), CD33(+), CD61(+) and CD19(+) cells in vivo. Furthermore, analysis of SCID engrafting potential showed that prolongation of culture duration facilitates in vivo expansion of CD45(+), CD34(+) and CD19(+) cells after transplantation. It is anticipated that prolonged (2 weeks) ex vivo culture of CB will have a beneficial clinical effect.

Analysis of Variance↗
PubMed2001

Prior cryopreservation of ex vivo-expanded cord blood cells is not detrimental to engraftment as measured in the NOD-SCID mouse model.

Cytokine-mediated expansion has been proposed and successfully used to facilitate engraftment post transplantation. This study examined whether cryopreservation following expansion has a detrimental effect on the ability of cells to engraft, using the NOD-SCID mouse model. Cord blood (CB) CD34(+) cells were incubated for 7 days with stem cell factor (SCF), flt-3 ligand (FL), and megakaryocyte growth and development factor (MGDF). Expanded CD34(+) cells were transplanted into NOD-SCID mice either fresh or following cryopreservation and thawing. After thawing, recovery of nucleated cells was 94%, of CD34 cells was 63%, and of day-14 progenitors was 17%. The loss of day-14 progenitor cells among the thawed expanded cells did not influence the kinetics of human engraftment in the mouse. Bone marrow (BM) of mice transplanted with thawed expanded CD34(+) cells (14 +/- 3.9%) showed significantly higher levels of human engraftment than mice transplanted with fresh expanded CD34(+) cells (1.5 +/- 0.5%, p = 0.0064). Thawed expanded CD34(+) cells had significantly higher SCID Engrafting Potential (SEP) than freshly expanded CD34(+) cells (p < 0.001). Results suggest that prior cryopreservation does not prevent expanded cells engrafting in NOD-SCID mice.

Animals↗
PubMed2000

Conditions that enable human hematopoietic stem cell engraftment in all NOD-SCID mice.

BACKGROUND: Transplantation of human hematopoietic stem cells is the only true test of their long-term repopulation potential. Models are readily available to evaluate murine hematopoietic stem cells, but few exist that allow reliable quantification of human stem cells. The non-obese diabetic-severe combined immunodeficient (NOD-SCID) mouse model enables quantification of human hematopoietic stem cells, but the conditions that permit human engraftment in all animals have yet to be defined. The aims of the project were, therefore, to describe the variables that allow human engraftment in the NOD-SCID mouse model and the techniques that accurately quantify this engraftment. METHODS: NOD-SCID mice that had or had not received 250, 325, or 400 cGy irradiation received cord blood (CB) mononuclear or CD34+ cells i.v. or i.p. Mice were killed 6 weeks after transplantation, and the bone marrow, spleen, and thymus were harvested. Four-color flow cytometric analysis, semi-quantitative PCR, myeloid and erythroid progenitor, and stem cell assays were used to monitor human engraftment. RESULTS: A 250 or 325 cGy and i.v. injection of CB mononuclear or CD34+ cells is required to detect multilineage human engraftment in the bone marrow, spleen, or thymus of NOD-SCID mice. Four-color flow cytometric analysis and semi-quantitative PCR enable accurate detection of 0.1% human cells. Progenitor and stem cell assays provide functional information about the engrafted cells. CONCLUSIONS: Successful development of the NOD-SCID mouse model and techniques to assess human engraftment now allow it to be used reliably to analyze the effects of short-term cytokine exposure on the long-term repopulating capacity of CB stem cells.

Animals↗
PubMed1999

Aminoborohydrides. 11. Facile reduction of N-alkyl lactams to the corresponding amines using lithium aminoborohydrides.

[reaction: see text] Various five- and six-membered N-alkyl lactams were reduced to the corresponding cyclic amines using lithium N,N-dialkylaminoborohydrides (LAB). Most of the reductions were essentially complete after refluxing in THF for 2 h. The cyclic amine products were easily isolated after an aqueous workup in very good to excellent yields. It is possible to selectively reduce most functional groups, such as esters, in the presence of a lactam using LAB reagents.

Amines↗
PubMed1999

Independent control of head and gaze movements during head-free pursuit in humans.

1. Head and gaze movements are usually highly co-ordinated. Here we demonstrate that under certain circumstances they can be controlled independently and we investigate the role of anticipatory activity in this process. 2. In experiment 1, subjects tracked, with head and eyes, a sinusoidally moving target. Overall, head and gaze trajectories were tightly coupled. From moment to moment, however, the trajectories could be very different and head movements were significantly more variable than gaze movements. 3. Predictive head and gaze responses can be elicited by repeated presentation of an intermittently illuminated, constant velocity target. In experiment 2 this protocol elicited a build-up of anticipatory head and gaze velocity, in opposing directions, when subjects made head movements in the opposite direction to target movement whilst maintaining gaze on target. 4. In experiment 3, head and gaze movements were completely uncoupled. Subjects followed, with head and gaze, respectively, two targets moving at different, harmonically unrelated frequencies. This was possible when both targets were visual, and also when gaze followed a visual target at one frequency whilst the head was oscillated in time with an auditory tone modulated at the second frequency. 5. We conclude that these results provide evidence of a visuomotor predictive mechanism that continuously samples visual feedback information and stores it such that it can be accessed by either the eye or the head to generate anticipatory movements. This overcomes time delays in visuomotor processing and facilitates time-sharing of motor activities, making possible the performance of two tasks simultaneously.

Acoustic Stimulation↗
PubMed1999

Progressive bradykinesia and hypokinesia of ocular pursuit in Parkinson's disease.

OBJECTIVES: Patients with Parkinson's disease characteristically have difficulty in sustaining repetitive motor actions. The purpose of this study was to establish if parkinsonian difficulty with sustaining repetitive limb movements also applies to smooth ocular pursuit and to identify any pursuit abnormalities characteristic of Parkinson's disease. METHODS: Ocular pursuit in seven patients with moderate to severe bradykinesia predominant Parkinson's disease was compared with seven age matched controls. Predictive and non-predictive pursuit of constant velocity target ramps were examined. Subjects pursued intermittently illuminated 40(0)/s ramps sweeping to the left or right with an exposure duration of 480 ms and average interval of 1.728 s between presentations. To examine for any temporal changes in peak eye velocity, eye displacement or anticipatory smooth pursuit the 124 s duration of each record was divided into four epochs (E1, E2, E3, E4), each lasting 31 s and containing 18 ramp stimuli. Three test conditions were examined in each subject: predictive (PRD1), non-predictive (NPD), and predictive (PRD2) in that order. RESULTS: Both patients and controls initiated appropriate anticipatory pursuit before target onset in the PRD1 and PRD2 conditions that enhanced the response compared with the NPD condition. The distinctive findings in patients with Parkinson's disease were a reduction in response magnitude compared with controls and a progressive decline of response with stimulus repetition. The deficits were explained on the basis of easy fatiguability in Parkinson's disease. CONCLUSIONS: Ocular pursuit shows distinct anticipatory movements in Parkinson's disease but peak velocity and displacement are reduced and progressively decline with repetition as found with limb movements.

Aged↗
PubMed1998

Timing variability of repetitive saccadic eye movements.

We assessed the suitability of using the Wing and Kristofferson model for timing repetitive motor responses to analyse timing variability during repetitive saccadic eye movements. The model decomposes total timing variability (TV) into a central timing component (CV) and a peripheral motor delay component (MV). Eight normal subjects made voluntary horizontal saccades, in darkness, in synchrony with a regular auditory metronome. After 20 saccades had been produced, the metronome was switched off and subjects continued responding at the same frequency until 31 further saccades had been made. Inter-saccade intervals (ISIs) from the unpaced phase were used to calculate TV, CV and MV. Three different target intervals, paced by auditory cues, were used - 496 ms, 752 ms and 1000 ms. In the paced phase, subjects' ISIs closely matched the auditory cue intervals. In the unpaced phase, subjects were clearly able to respond at three different frequencies. As predicted by the Wing and Kristofferson model, the durations of successive ISIs tended to be negatively correlated. As expected, TV and CV increased with increasing ISI. Contrary to the expectation of the model that MV would remain constant, we found that it increased with increasing interval. Our results do not conclusively demonstrate the validity of applying the Wing and Kristofferson model to the analysis of timing variability during repetitive saccadic eye movements. However, comparison with previous studies shows that, at least in normal subjects, it is equally valid to apply the model to the analysis of repetitive saccadic eye movements as it is to apply it to the analysis of data from other effectors.

Adult↗
PubMed1998

A clinical evaluation of posterior composite resin restorations: 8-year findings.

OBJECTIVES: From a continuing investigation of the clinical performance of three different types of composite resin, the findings of the clinical evaluation at 8 years are presented. METHODS: One operator placed 330 restorations in Class I and Class II preparations in the posterior teeth of 72 patients. Every patient received at least one restoration of each type of material: a microfilled composite, a small particle hybrid, a relatively coarse particle hybrid, and an amalgam control. Restorations were evaluated using clinical criteria. RESULTS: Forty six patients attended the 8-year recall. Twenty-five of the 213 restorations (13.7% of the composites and 5.8% of the amalgams) originally placed in these 46 patients had previously failed or been lost from the study or were assessed as requiring replacement at the 8-year recall. Bulk fracture and secondary caries at the margin were the most common forms of failure in the composite restorations. Other failures or losses were associated with a non-margin defect in the composite, caries not associated with the restoration, pulpal considerations, extraction for orthodontics and reasons unknown. One-hundred and ninety-three restorations (including five that required replacement) were available for clinical evaluation at 8 years and these included 17 Class II restorations. Significantly fewer restorations placed with the coarse particle hybrid exhibited evidence of marginal deterioration. CONCLUSION: At 8 years, composite restorations in posterior teeth had failed at a rate two to three times that of amalgam restorations. The most common types of failure were bulk fracture and secondary caries at the margin.

Adolescent↗
PubMed1992

Finishing of amalgam restorations: a three-year clinical study.

Class I and Class II amalgam restorations were placed in the permanent teeth of 66 patients, each of whom received a minimum of three restorations. At least one of these restorations was carved-only, at least one was immediately finished (using a prophylaxis paste on rubber cups at low speed, 8 or 10 min after trituration), and at least one restoration was polished (using finishing burs and polishing points, more than 24 h after placement). The number of restorations available for assessment of marginal fracture using the 11-unit photographic scale developed by Mahler was 253 at baseline, 249 at 1-year and 2-year recalls and 228 at the 3-year recall. Although polished restorations exhibited greater initial (baseline) marginal fracture, at 1-year, 2-year and 3-year recalls the restorations were found to exhibit a similar amount of marginal fracture regardless of the finishing technique. The amount of perceived marginal fracture increased steadily after 1 year. The immediate finishing of amalgam restorations showed no long-term benefit over the other techniques and has little to commend it. The clinical finishing of amalgam restorations is discussed in the light of this and other research.

Adolescent↗
PubMed1992

Finishing techniques for amalgam restorations: clinical assessment at three years.

Use of the optimum finishing technique for an amalgam restoration may enhance the marginal integrity of the restoration and discourage its unnecessary early replacement. Two hundred and twenty-eight high copper amalgam restorations in 56 patients were evaluated, using clinical assessment criteria, up to three years after placement. Each patient had received at least one carved-only amalgam, at least one immediately finished restoration, and at least one amalgam that was polished at a subsequent appointment. Regardless of the finishing technique, the restorations exhibited similar marginal integrity up to three years after placement. Polished restorations were found to have substantially superior surface texture and less likelihood of surface discoloration. No evidence was found to support the use of immediate finishing techniques. The clinical significance of these findings, with respect to the need to polish amalgam restorations, is discussed.

Color↗
PubMed1989

The finishing and early marginal fracture of clinical amalgam restorations.

One operator placed 249 Tytin amalgam restorations in Class I and II preparations in the permanent teeth of 60 patients each of whom received a minimum of three restorations. At least one of these restorations was carved-only, at least one was immediately finished (at 8 or 10 min after trituration), and at least one restoration was polished (more than 24 h after placement). Restorations were principally assessed for marginal fracture at baseline and 1 year using the 11-unit scale for the evaluation of photographs developed by Mahler. Five factors were also assessed clinically at 1 year. Amalgam restorations, polished using finishing burs and rubber points at least 24 h after placement, exhibited more initial marginal fracture than carved-only or immediately finished restorations. At 1 year, the three techniques were comparable except for the superior surface texture of the polished restorations. The technique of immediate finishing of amalgam restorations has nothing to commend it in preference to existing techniques.

Chemical Phenomena↗
PubMed1988

Stringent regulation of stably integrated chloramphenicol acetyl transferase genes by E. coli lac repressor in monkey cells.

Monkey cell lines that constitutively synthesize 38.6 kd lac repressor protein and bear stably integrated chloramphenicol acetyl transferase (CAT) genes linked to a lac operator-containing SV40 early promoter-enhancer were generated. When grown in medium containing isopropyl beta-D-thiogalactoside (IPTG), these cells acquired a CAT+ phenotype. In contrast, when grown in parallel in medium lacking IPTG, the cells remained CAT-. Maximum induction of CAT activity occurred after 4 days of IPTG exposure. Three days after removal of IPTG, induced cells had reverted to CAT-. Specific CAT activity increased up to 60-fold after induction, while background activity in uninduced cells was similar to or only slightly above that of parental, CAT- cells. CAT activity increased stepwise over a wide range of IPTG concentrations. Thus lac repressor-operator complexes can form on primate chromosomes and stringently block transcription from an adjoining promoter.

Acetyltransferases↗
PubMed1987

Molecular cloning of the human gene for von Willebrand factor and identification of the transcription initiation site.

A series of overlapping cosmid genomic clones have been isolated that contain the entire coding unit of the human gene for von Willebrand factor (vWf), a major component of the hemostatic system. The cloned segments span approximately 175 kilobases of human DNA sequence, and hybridization analysis suggests that the vWf coding unit is approximately 150 kilobases in length. Within one of these clones, the vWf transcription initiation site has been mapped and a portion of the vWf promoter region has been sequenced, revealing a typical "TATA box," a downstream "CCAAT box," and a perfect downstream repeat of the 8 base pairs containing the transcription start site. Sequencing of a segment of another genomic clone has revealed the vWf translation termination codon. Where tested, comparative restriction analysis of cloned and chromosomal DNA segments strongly suggests that no major alterations occurred during cloning and that there is only one complete copy of the vWf gene in the human haploid genome. Similar analyses of DNA from vWf-producing endothelial cells and nonexpressing leukocytes suggest that vWf gene expression is not accompanied by gross genomic rearrangements. In addition, there is significant homology of C-terminal coding sequences among the vWf genes of several vertebrate species.

Amino Acid Sequence↗
PubMed1985

Molecular cloning of cDNA for human von Willebrand factor: authentication by a new method.

We have identified a 2.4 kb partial cDNA clone (pDL34) for human von Willebrand factor (vWf) mRNA. pDL34 was selected by screening an endothelial cell cDNA library with a radiolabeled reverse transcript of mRNA obtained by specific immunoisolation of vWf polysomes from endothelial cells. pDL34 selectively hybridized to an endothelial cell-associated 9.5 kb mRNA. To confirm its identity, SP6 RNA polymerase was used to generate in vitro transcripts of the cDNA. This synthetic RNA, truncated at its 5' end, directed the synthesis of several unique polypeptides in rabbit reticulocyte lysates. These polypeptides were immunoprecipitated by polyclonal and monoclonal anti-vWf antibodies. These results indicate that pDL34 contains an authentic partial copy of vWf mRNA. In vitro transcription of partial cDNA clones and translation of the resulting RNAs may be a useful general method of verifying the identity of various cDNA clones in circumstances where antibodies are available and protein sequence is not.

Antibodies, Monoclonal↗
PubMed1983

External quality assessment of assays for cortisol in plasma: use of target data obtained by gas chromatography/mass spectrometry.

We describe procedures for measuring cortisol in plasma and serum by isotope dilution and mass spectrometry. A method that incorporated solvent extraction, derivatization, and gas chromatography/high-resolution mass spectrometry provided data of good precision; interassay CVs were generally 3 to 4% for the concentration range of 100-650 nmol/L. Replacing solvent extraction with extraction on a column of Lipidex 1000 or extraction by immunoadsorption yielded data in excellent agreement with the first method. Plasma and serum pools were analyzed to provide target data for use in the U.K. National External Quality Assessment Scheme for cortisol assays. Routine laboratory assays, as judged by comparison with mass-spectrometric data, were generally positively biased except for analysis of a charcoal-stripped plasma supplemented with cortisol. The results emphasize the importance of using unadulterated plasma or serum pools in assessments of steroid assay procedures.

Charcoal↗
PubMed1980

Arrangement of integrated avian sarcoma virus DNA sequences within the cellular genomes of transformed and revertant mammalian cells.

We have examined the arrangement of integrated avian sarcoma virus (ASV) DNA sequences in several different avian sarcoma virus transformed mammalian cell lines, in independently isolated clones of avian sarcoma virus transformed rat liver cells, and in morphologically normal revertants of avian sarcoma virus transformed rat embryo cells. By using restriction endonuclease digestion, agarose gel electrophoresis, Southern blotting, and hybridization with labeled avian sarcoma virus complementary DNA probes, we have compared the restriction enzyme cleavage maps of integrated viral DNA and adjacent cellular DNA sequences in four different mouse and rat cell lines transformed with either Bratislava 77 or Schmidt-Ruppin strains of avian sarcoma virus. The results of these experiments indicated that the integrated viral DNA resided at a different site within the host cell genome in each transformed cell line. A similar analysis of several independently derived clones of Schmidt-Ruppin transformed rat liver cells also revealed that each clone contained a unique cellular site for the integration of proviral DNA. Examination of several morphologically normal revertants and spontaneous retransformants of Schmidt-Ruppin transformed rat embryo cells revealed that the internal arrangement and cellular integration site of viral DNA sequences was identical with that of the transformed parent cell line. The loss of the transformed phenotype in these revertant cell lines, therefore, does not appear to be the result of rearrangement or deletions either within the viral genome or in adjacent cellular DNA sequences. The data presented support a model for ASV proviral DNA integration in which recombination can occur at multiple sites within the mammalian cell genome. The integration and maintenance of at least one complete copy of the viral genome appear to be required for continuous expression of the transformed phenotype in mammalian cells.

Animals↗