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Biomedical subjects

E A Reid

Publications and source records attributed to E A Reid.

12 recordsLinked to original sources
PubMed2001

The quantitative distribution of a putative PKC epsilon mRNA in Limulus central nervous system by modified competitive RT-PCR.

Recently, a full length cDNA for the epsilon (epsilon) isoform of protein kinase C (PKC) was cloned and sequenced from a cDNA library for the horseshoe crab, Limulus polyphemus. This multifunctional enzyme has been implicated in the modulation of the choline cotransporter in Limulus and the epsilon isoform has been identified in homogenates from its central nervous system (CNS). RT-PCR has proven to be a very useful method for quantifying even a few molecules of mRNA in tissue samples. A modified competitive RT-PCR was used here to quantify a putative PKC epsilon mRNA in Limulus CNS preparations. First, we replaced normally used oligo dT and random primers generated from mRNA with a PKC epsilon specific (3' end) primer P4. Then we used modified nucleotides to extend sample life in storage and finally, we used only annealing and denaturing temperatures during PCR to reduce background. The modified method was used for the first time to quantify PKC epsilon mRNA from three distinct areas of the CNS in Limulus. Results revealed high levels of PKC epsilon mRNA in the corpora pedunculata, in the abdominal ganglia and in the brain ring. These results indicate that PKC epsilon mRNA is broadly distributed throughout the Limulus CNS. Importantly, this modified competitive RT-PCR technique was successfully applied to the quantitation of specific mRNA from Limulus nervous tissue for which no internal standard is available commercially.

Animals↗
PubMed2001

Novel approaches to the analysis of the Maillard reaction of proteins.

The Maillard reaction comprises a complex network of reactions which has proven to be of great importance in both food science and medicine. The majority of methods developed for studying the Maillard reaction in food have focused on model systems containing amino acids and monosaccharides. In this study, a number of electrophoretic techniques, including two-dimensional gel electrophoresis and capillary electrophoresis, are presented. These have been developed specifically for the analysis of the Maillard reaction of food proteins, and are giving important insights into this complex process.

Electrophoresis, Gel, Two-Dimensional↗
PubMed2000

The use of competitive PCR mimic to evaluate a Limulus lambda phage genomic DNA library.

1. A lambda phage genomic DNA library for Limulus (L.) polyphemus brain was constructed using the AGEM-12 vector and the host strain KW251. 2. The primary library contained approximately 1.275 x 10(6) independent clones, increasing upon amplfication to 6.66 x 10(9) pfu/ml in a total volume of 58 ml. 3. A total of 28 clones was randomly chosen for a determination of the average size of inserts in the library. All clones contained inserts and the average size was 14.9 kb, ranging from 11.7 to 28.0 kb. The library provides a 10-fold equivalent of the L. polyphemus genome. 4. A new approach for evaluating a genomic DNA library was developed, in which competitive PCR MIMIC was employed to determine the target gene copy number in both constructed library and brain genomic DNA. The putative protein kinase C epsilon (PKCepsilon) was selected as the target gene because its partial sequence of cDNA was recently cloned from L. polyphemus brain in our laboratory (Cao et al., 1998). A 419-bp fragment of nonhomologous sequence derived from putative PKCepsilon and a 306-bp fragment from plasmid pUC 18 were generated for use as target and competitor in PCR MIMIC, respectively. 5. Within the genomic library DNA, a 0.8 value was obtained for the copy number of the putative PKCepsilon gene that was detected in 0.1 amol of one equivalent L. polyphemus genome in terms of the average recombinant molecular weight. In the genomic DNA, a single copy of putative PKCepsilon was found in 0.1 amol of one coverage for the L. polyphemus genome. Thus, it was implied that nearly 80% genetic resource was incorporated into the library. This percentage was termed the incorporation rate. 6. Based on these findings, we suggest that the incorporation rate is an essential factor for evaluating genomic libraries, particularly, when using partial digestion with restriction enzymes for library construction.

Animals↗
PubMed1996

Helen Bramson: treatment after sexual abuse by a mental health practitioner.

Patients who have had sexual contact or sexualized relationships with previous therapists present a unique set of subsequent treatment challenges. From the patient's perspective, the breach of trust experienced in the previous therapy has likely made the formation of a new therapeutic alliance both threatening and compelling. It is also likely that whatever caused the patient to enter therapy in the first place has never been adequately treated.

Adult↗
PubMed1977

Consultation in the college setting.

The author discusses the relationship between patient care and consulting and the rapport that contact between college health service psychiatrists and other college personnel can engender. People in the college community learn how psychiatrists approach problems and what they can or cannot do. Psychiatrists learn how people in a college setting function and are thus better able to understand and help both patients and consultees.

Confidentiality↗