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Biomedical subjects

H M Davies

Publications and source records attributed to H M Davies.

7 recordsLinked to original sources

Fatty acid biosynthesis redirected to medium chains in transgenic oilseed plants.

Medium-chain fatty acids (FAs), found in storage lipids of certain plants, are an important renewable resource. Seeds of undomesticated California bay accumulate laurate (12:0), and a 12:0-acyl-carrier protein thioesterase (BTE) has been purified from this tissue. Sequencing of BTE enabled the cloning of a complementary DNA coding for a plastid-targeted preprotein. Expression of the complementary DNA in the seeds of Arabidopsis thaliana resulted in BTE activity, and medium chains accumulated at the expense of long-chain (greater than or equal to 16) FAs. Laurate became the most abundant FA species and was deposited in the storage triacylglycerols. These results demonstrate a mechanism for medium-chain FA synthesis in plants.

Acetyltransferases

A specific acyl-ACP thioesterase implicated in medium-chain fatty acid production in immature cotyledons of Umbellularia californica.

Umbellularia californica (California Bay) seeds accumulate 10:0 and 12:0 as principal reserve fatty acyl groups. An in vitro fatty acid synthesis system from the developing cotyledons produces chiefly 10:0 and 12:0, in approximately the same proportions as the intact tissue. The kinetics of acyl thioester and free fatty acid formation in this system suggest that a medium-chain specific acyl-acyl-carrier protein (ACP) hydrolysis mechanism is responsible for the preponderance of medium-chain products. A crude extract of the developing cotyledons exhibits hydrolytic activity toward acyl-ACPs, with marked preference for 12:0-ACP and 18:1-ACP in the test series 6:0, 8:0, 10:0, 11:0, 12:0, 14:0, 16:0, and 18:1-ACPs. Partial purification of the 12:0-ACP hydrolytic activity has resulted in its separation from the 18:1-ACP hydrolase(s) and the 12:0-coenzyme A hydrolase(s) that are also present, thereby demonstrating its specificity for the 12-carbon acyl chain length and the ACP derivative. During cotyledon development, as the proportion of medium-chain to other fatty acyl groups increases, the extractable yield of this activity also increases substantially. Collectively these results suggest a role for this 12-ACP thioesterase in medium-chain production in vivo.

Coenzymes

Developmental induction, purification, and further characterization of 12:0-ACP thioesterase from immature cotyledons of Umbellularia californica.

The fatty acyl content of developing cotyledons of Umbellularia californica (California Bay) changes from a long-chain composition to a predominance of 10:0 and 12:0 in just 4-5 days at the beginning of an approximately 100-day period of medium-chain deposition. This striking change occurs at the earliest appearance of 12:0-acyl-carrier protein (ACP) thioesterase activity. The coincidence of these rapid events is consistent with the hypothesis that the enzyme plays a major role in medium-chain biosynthesis. The 12:0-ACP thioesterase has been substantially purified; enzyme activity consistently comigrates in chromatographic and electrophoretic systems with a protein or pair of proteins having an apparent molecular weight of approximately 34 kDa. A native molecular weight of approximately 42 kDa has been estimated by gel filtration chromatography, suggesting that the enzyme is a monomer. Affinity chromatography on immobilized ACP is a critical step in the purification procedure, and resolves the 12:0-ACP and 18:1-ACP thioesterases sufficiently to confirm that the medium-chain enzyme has negligible action on 18:1-ACP.

Acyl Carrier Protein

Imaging microbubbles and tissues using a linear focussed scanner operating at 20 MHz: possible implications for the detection of cavitation thresholds.

B-Scan images have been used as a means of detecting the onset of cavitation in tissues. New echoes appearing during simultaneous sonication with therapeutic ultrasound have often been attributed to bubble formation. In this study temperature rises up to 15 degrees C W-1 cm2 in guinea pig leg (post-mortem) were found, which would itself increase the rate of growth of gas bubbles. It is also hypothesized that a temperature dependent attenuation coefficient predicted by Bamber and Hill may, at least in part, account for new echoes appearing in tissues. Possible artefactual echoes in the B-scan images arising from acoustically induced gas bubbles in the coupling medium are also discussed.

Animals

Evaluation of sequential plasma and urinary tumor necrosis factor alpha levels in renal allograft recipients.

The macrophage cytokine tumor necrosis factor-alpha is released early in immune activation and may be detected in the peripheral circulation. This study has investigated the occurrence of plasma and urinary TNF in 30 renal allograft recipients. Although circulating TNF may be detected in 20% of pretransplant or normal control samples, levels were significantly elevated during 65% of allograft rejection episodes. Plasma TNF levels did not rise in graft failure due to acute tubular necrosis, but were always highly raised in systemic infection. In contrast, urinary TNF was only detected in association with acute rejection (49%) or tubular necrosis (14%), and no controls had detectable urinary TNF. These findings indicate that evaluation of circulating and excreted TNF may give further insight into the immunobiology of graft rejection.

Communicable Diseases