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Biomedical subjects

L Crocker

Publications and source records attributed to L Crocker.

12 recordsLinked to original sources

Giving up or finding a solution? The experience of attempted suicide in later life.

Older people constitute one of the highest risk groups for suicide. Existing research in this area has been largely dominated by a risk factor approach. This is of limited usefulness since only a minority of those at risk go on to make an attempt. Therefore, prediction, prevention and the management of risk remain challenging. The present study aimed to capture the subjective experience of older people who had recently made a suicide attempt through exploring their understanding of the pathway to and from this attempt, within the context of ageing. Fifteen participants were interviewed. Transcripts were analysed using Interpretative Phenomenological Analysis. Three broad themes emerged--Struggle (experiencing life as a struggle before and after the attempt, and in relation to growing older), Control (trying to maintain control over life before the attempt, and following it either failing or succeeding to regain control) and Visibility (feeling invisible or disconnected from others and trying to fight against this before the attempt and either becoming more or less connected afterwards). Risk factors identified in the literature were often absent or construed by participants as not relevant to their attempt. Individual accounts highlight the diversity and complexity of experience of older people who attempt suicide.

Aged↗

Development of a nanofiltration process to improve the stability of a novel anti-MRSA carbapenem drug candidate.

The benzenesulfonate salt of an anti-methicillin-resistant Staphylococcus aureus carbapenem antibiotic studied is a crystalline, nonhygroscopic powder which is stable at room temperature, making it an ideal compound for long-term storage. However, the limited aqueous solubility of this salt prohibits parenteral administration. Conversely, the chloride salt of this carbapenem demonstrates opposing characteristics; it is quantitatively soluble in water, however is amorphous and subject to significant hydrolytic degradation in the solid state. Given two such extreme alternatives for pharmaceutical salt selection, a common approach taken is to develop the bioavailable salt and devise manufacturing and storage conditions that minimize degradation. This report describes a different approach to this manufacturing dilemma via the application of a simple and efficient nanofiltration process to convert the benzenesulfonate salt (storage entity) to the chloride salt (formulated drug product). Such an approach combines the positive attributes of these two salt forms into a single scalable process that reduces processing cycle times via elimination of redundant unit operations, increases the flexibility in manufacturing schedule, and improves overall product quality.

Benzenesulfonates↗

In vitro development of porcine nuclear transfer embryos constructed using fetal fibroblasts.

The in vitro development of porcine nuclear transfer embryos constructed using primary cultures from day 25 fetal fibroblasts which were either rapidly dividing (cycling) or had their cell-cycle synchronized in G0/G1 using serum starvation (serum-starved) was examined. Oocyte-karyoplast complexes were fused and activated simultaneously and then cultured in vitro for seven days to assess development. Fusion rates were not different for either cell population. The proportion of reconstructed embryos that cleaved was higher in the cycling group compared to the serum-starved group (79 vs. 56% respectively; P < 0.05). Development to the 4-cell stage was not different using either population. Both treatments supported similar rates of development to the morula (1.5 vs. 7%, cycling vs. serum-starved) and blastocyst stage (1.5 vs. 3%, cycling vs. serum-starved). The blastocyst produced using cycling cells had a total cell number of 10. Total cell numbers for the three blastocysts produced serum-starved cells were 22, 24, and 33. These blastocysts had inner cell mass numbers of 0, 15, and 4, respectively. Six hundred and thirty-five nuclear transfer embryos reconstructed using serum-starved cells were transferred to 15 temporarily mated recipients for 3-4 days. Of these, 486 were recovered (77% recovery rate) of which 106 (22%) had developed to the 4-cell stage or later. These were transferred to a total of 15 recipients which were either unmated or mated. Seven recipients farrowed a total of 51 piglets. Microsatellite analysis revealed that none of these were derived from the nuclear transfer embryos transferred.

Animals↗

Secretion of eukaryotic growth hormones in Escherichia coli is influenced by the sequence of the mature proteins.

We report the construction of secretion plasmids expressing the fusion proteins, OmpA::pGH (pSpGH.01) and OmpA::hGH (phGH.01), and compare the secretion of mature porcine growth hormone (pGH) and human growth hormone (hGH) employing Escherichia coli. E. coli [phGH.01] secreted 10-15 micrograms hGH/ml/A600 cells into the periplasmic space, representing 30% of total periplasmic proteins. E. coli [pSpGH.01], however, secreted 30-fold less mature pGH. On the basis that both pSpGH.01 and phGH.01 are stably maintained in E. coli and in vitro transcription/translation data showed equivalent expression of OmpA::pGH and OmpA::hGH precursors, we attribute the higher secretion of hGH to the translocation-competent OmpA::hGH protein configuration. Two OmpA::GHF (growth hormone fusion) precursors, OmpA::GHF.02 and OmpA::GHF.03, both with hGH helix 3/helix 4 together instead of the pGH equivalent, secreted mature proteins as efficiently as OmpA::hGH. We propose that hGH helices 3 and 4 in these OmpA::GHF precursors play a major role in the folding of the precursor to a translocation-competent state, mimicking the translocation-competent nature of the OmpA::hGH precursor.

Animals↗

Site-specific transgene insertion: an approach.

Methods to improve the production of transgenic animals are being developed. Conventional transgenesis, involving microinjection of DNA into fertilized eggs, has a number of limitations. These result from the inability to control both the site of transgene insertion and the number of gene copies inserted. The approach described seeks to overcome these problems and to allow single copy insertion of transgenes into a defined site in animal genomes. The method involves the use of embryonic stem cells, gene targeting and the FLP recombinase system.

Animals↗

Hair follicle differentiation: expression, structure and evolutionary conservation of the hair type II keratin intermediate filament gene family.

During hair follicle development several cell streams are programmed to differentiate from the cell population of the follicle bulb. In the hair cells, a number of keratin gene families are transcriptionally activated. We describe the characterization of the type II keratin intermediate filament (IF) gene family which is expressed early in follicle differentiation. In sheep wool, four type II IF proteins are expressed. One gene has been completely sequenced and the expression of three of the genes examined in detail. The sequenced gene encodes a 55 x 10(3) Mr protein of the type II keratin IF protein family, designated KII-9 in the new nomenclature we have adopted and described in the Introduction. The gene has a similar exon/intron structure to the epidermal type II keratin IF genes. In situ hybridization experiments show that the genes are expressed in the hair cortical cells but not in the cells of the outer root sheath, inner root sheath or medulla. During hair keratinocyte differentiation the type II IF genes are sequentially activated and coexpressed in the same cells. Expression is first detected in cells in the middle of the follicle bulb located near the dermal papilla and, subsequently, two of the genes are transcriptionally activated in the differentiating keratinocytes as they migrate upwards, in the upper part of the bulb. A fourth type II IF gene is activated later. The genes with the same expression pattern are also closely related in sequence and a number of conserved elements are present in the promoters of those genes, including a novel element which is also found in the promoter of a coexpressed type I IF gene and three other hair keratin genes.

Amino Acid Sequence↗

Molecular cloning and sequencing of Australian black bream Acanthopagrus butcheri and barramundi Lates calcarifer fish growth hormone cDNA using polymerase chain reaction.

The complementary DNA (cDNA) encoding the preprotein growth hormone (pre-GH) from two Australian marine fish species, namely Acanthopagrus butcheri and Lates calcarifer, have been isolated, cloned and sequenced. The sequences were amplified from reverse transcribed total RNA of whole brains using Polymerase Chain Reaction (PCR) and oligonucleotide primers corresponding to the 5' and 3' regions of Pagrus major. Use of PCR offers a rapid method of isolating fish GH cDNA sequences for commercial and taxonomic applications. Sequence comparison indicates a high degree of conservation for GH cDNAs within the family Sparidae.

Amino Acid Sequence↗

The role of the interview in student selection.

Four types of data are commonly considered in selecting applicants for academic programs: 1) test scores; 2) grade-point averages; 3) personal data; and 4) interview results. Limitations and advantages of using interview data in the selection decision are discussed. Fifteen suggestions are offered for improving the validity and reliability of interview data. Strategies for data collection and analysis are suggested for validation studies of selection interviews.

Education, Medical↗