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P S Nelson

Publications and source records attributed to P S Nelson.

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Oligonucleotide labeling methods. 3. Direct labeling of oligonucleotides employing a novel, non-nucleosidic, 2-aminobutyl-1,3-propanediol backbone.

Novel CE-phosphoramidite (7a-e) and CPG (8a, c, d, e) reagents have been prepared from a unique 2-aminobutyl-1,3-propanediol backbone. The reagents have been used to directly label oligonucleotides with fluorescein, acridine, and biotin via automated DNA synthesis. The versatile 2-aminobutyl-1,3-propanediol backbone allows for labeling at any position (5', internal, and 3') during solid phase oligonucleotide synthesis. Multiple labels can be achieved by repetitive coupling cycles. Furthermore, the 3-carbon atom internucleotide phosphate distance is retained when inserted internally. Using this method, individual oligonucleotides possessing two and three different reporter molecules have been prepared.

Automation

A new and versatile reagent for incorporating multiple primary aliphatic amines into synthetic oligonucleotides.

A novel and versatile phosphoramidite, N-Fmoc-O1-DMT-O2-cyanoethoxydiisopropylamino-phosphinyl-3-am ino-1,2-propanediol (1, Fig. 1), has been synthesized and used to incorporate primary aliphatic amines into synthetic oligonucleotides. Its convenient preparation and use in solid phase oligonucleotide synthesis is described. Using phosphoramidite 1, an amino-modified oligonucleotide probe complementary to M13mp18 DNA was constructed with five primary amines attached to the 5'-terminus. The amino-modified oligonucleotide was subsequently labeled with biotin and employed in a dot-blot hybridization assay. As little as 0.5 ng of M13mp18 target DNA was colorimetrically detected.

Amines

Bifunctional oligonucleotide probes synthesized using a novel CPG support are able to detect single base pair mutations.

A novel multifunctional controlled pore glass, MF-CPG (Fig. 1), has been synthesized and used to incorporate 3' terminal primary aliphatic amines into synthetic oligonucleotides. MF-CPG consists of a unique succinic acid linking arm which possesses both a masked primary amine for label attachment and a dimethoxytrityl protected hydroxyl for nucleotide chain elongation. Using MF-CPG, we have devised a simple and convenient technique to attach non-radioactive labels to the 3' terminus of oligonucleotides. Bifunctional probes can then be constructed by 32P labeling the 5' terminus with T4 kinase and gamma 32P-ATP. Using such bifunctional oligonucleotide probes in conjunction with polymerase chain reaction (PCR) amplification, we were able to detect single base substitutions in a target segment of the human H-ras protooncogene employing either functionality. Our technique thus expands the potential applications for oligonucleotides as hybridization probes.

Amines

Comparison of oral transmucosal fentanyl citrate and an oral solution of meperidine, diazepam, and atropine for premedication in children.

The safety and efficacy of premedication with oral transmucosal fentanyl citrate (OTFC) was compared with that of an orally administered solution of meperidine, diazepam, and atropine and no premedication in 59 children about to undergo elective operations. The patients were randomly assigned to receive no premedication (n = 19); 0.25 ml/kg of the oral solution (containing meperidine, 1.5 mg/kg, diazepam, 0.2 mg/kg, and atropine, 0.02 mg/kg, n = 20); or OTFC (15-20 micrograms/kg, n = 20). Children had activity (sedation) and anxiety scores, vital signs (including systolic and diastolic arterial blood pressures and heart and respiratory rates) and pulse oximetry determined oxygen saturation measured before and at 10-min intervals after premedication until they were taken to the operating room. Quality of induction and recovery was evaluated using scoring schedules; recovery times were measured and side effects noted. OTFC was readily accepted and provided significant reductions in preoperative activity (sedation) and anxiety starting after 30 min. After OTFC, sedation and anxiolysis were significantly greater than in children having no premedication but similar to children having the oral solution for premedication. Vital signs and oxygen saturations remained unchanged preoperatively in all groups. Induction and recovery evaluations and recovery times were similar in the three groups, although children having OTFC had the lowest requirements for narcotics in the recovery room. OTFC caused an 80% incidence of mild preoperative facial pruritus and a higher overall incidence of postoperative vomiting (37%) than premedication with the oral solution (5%) or no premedication (18%).(ABSTRACT TRUNCATED AT 250 WORDS)

Administration, Oral

Optometry in Britain.

The author spent seven months in Britain on sabbatical leave from the Illinois College of Optometry. One purpose of the time away was to learn about the optometric profession and education in the United Kingdom. The result is a view of the profession based on visits to optometric institutions and private practices and discussion with medical and optometric colleagues.

Curriculum