Lipoyl synthase requires two equivalents of S-adenosyl-L-methionine to synthesize one equivalent of lipoic acid

Lipoyl synthase (LipA) catalyzes the formation of the lipoyl cofactor, which is employed by several multienzyme complexes for the oxidative decarboxylation of various alpha-keto acids, as well as the cleavage of glycine into CO(2) and NH(3), with concomitant transfer of its alpha-carbon to tetrahydrofolate, generating N(5),N(10)-methylenetetrahydrofolate. In each case, the lipoyl cofactor is tethered covalently in an amide linkage to a conserved lysine residue located on a designated lipoyl-bearing subunit of the complex. Genetic and biochemical studies suggest that lipoyl synthase is a member of a newly established class of metalloenzymes that use S-adenosyl-l-methionine (AdoMet) as a source of a 5'-deoxyadenosyl radical (5'-dA(*)), which is an obligate intermediate in each reaction. These enzymes contain iron-sulfur clusters, which provide an electron during the cleavage of AdoMet, forming l-methionine in addition to the primary radical. Recently, one substrate for lipoyl synthase has been shown to be the octanoylated derivative of the lipoyl-bearing subunit (E(2)) of the pyruvate dehydrogenase complex [Zhao, S., Miller, J. R., Jian, Y., Marletta, M. A., and Cronan, J. E., Jr. (2003) Chem. Biol. 10, 1293-1302]. Herein, we show that the octanoylated derivative of the lipoyl-bearing subunit of the glycine cleavage system (H-protein) is also a substrate for LipA, providing further evidence that the cofactor is synthesized on its target protein. Moreover, we show that the 5'-dA(*) acts directly on the octanoyl substrate, as evidenced by deuterium transfer from [octanoyl-d(15)]H-protein to 5'-deoxyadenosine. Last, our data indicate that 2 equiv of AdoMet are cleaved irreversibly in forming 1 equiv of [lipoyl]H-protein and are consistent with a model in which two LipA proteins are required to synthesize one lipoyl group.     

Prey choice and cannibalistic behaviour in the theropod Coelophysis

Direct evidence of prey choice in carnivorous dinosaurs is rare in the fossil record. The most celebrated example pertains to purported stomach contents in the carnivorous dinosaur Coelophysis bauri, which besides revealing prey choice, also points to cannibalistic behaviour as being commonplace (Colbert 1989, 1995). Here, we test this hypothesis by conducting the first comprehensive anatomical and histological examination of the famed Coelophysis 'cannibals'. The results unequivocally show that the gut contents derive from early crocodylomorphs rather than juveniles of Coelophysis. These findings suggest that this taxon is not cannibalistic and bring into question the commonality of this behaviour among non-avian dinosaurs.     

Measuring the maturity of the fast-spiking interneuron transcriptional program in autism, schizophrenia, and bipolar disorder

Background

Emerging evidence suggests that fast-spiking (FS) interneurons are disrupted in multiple neuropsychiatric disorders including autism, schizophrenia, and bipolar disorder. FS cells, which are the primary source of synaptic inhibition, are critical for temporally organizing brain activity, regulating brain maturation, and modulating critical developmental periods in multiple cortical systems. Reduced expression of parvalbumin, a marker of mature FS cells, has been reported in individuals with schizophrenia and bipolar disorder and in mouse models of schizophrenia and autism. Although these results suggest that FS cells may be immature in neuropsychiatric disease, this possibility had not previously been formally assessed.

Methods

This study used time-course global expression data from developing FS cells to create a maturation index that tracked with the developmental age of purified cortical FS cells. The FS cell maturation index was then applied to global gene expression data from human cortex to estimate the maturity of the FS cell developmental program in the context of various disease states. Specificity of the index for FS cells was supported by a highly significant correlation of maturation index measurements with parvalbumin expression levels that withstood correction for multiple covariates.

Conclusions

Results suggest the FS cell developmental gene expression program is immature in autism, schizophrenia, and bipolar disorder. More broadly, the current study indicates that cell-type specific maturation indices can be used to measure the maturity of developmental programs even in data from mixed cell types such as those found in brain homogenates.     

An Investigation on Platelet Transport during Thrombus Formation at Micro-Scale Stenosis

This paper reports on an investigation of mass transport of blood cells at micro-scale stenosis where local strain-rate micro-gradients trigger platelet aggregation. Using a microfluidic flow focusing platform we investigate the blood flow streams that principally contribute to platelet aggregation under shear micro-gradient conditions. We demonstrate that relatively thin surface streams located at the channel wall are the primary contributor of platelets to the developing aggregate under shear gradient conditions. Furthermore we delineate a role for red blood cell hydrodynamic lift forces in driving enhanced advection of platelets to the stenosis wall and surface of developing aggregates. We show that this novel microfluidic platform can be effectively used to study the role of mass transport phenomena driving platelet recruitment and aggregate formation and believe that this approach will lead to a greater understanding of the mechanisms underlying shear-gradient dependent discoid platelet aggregation in the context of cardiovascular diseases such as acute coronary syndromes and ischemic stroke.     

Adrenoceptor regulation of canine skeletal muscle blood flow during carbon monoxide hypoxia.

We questioned whether carbon monoxide hypoxia (COH) would affect peripheral blood flow by neural activation of adrenoceptors to the extent we had found in other forms of hypoxia. We studied this problem in hindlimb muscles of four groups of anesthetized dogs (untreated, alpha 1-blocked, alpha 1 + alpha 2-blocked, and beta 2-blocked). Cardiac output increased, but hindlimb blood flow (QL) and resistance (RL) remained at prehypoxic levels during COH (O2 content reduced 50%) in untreated animals. When activity in the sciatic nerve was reversibly cold blocked, QL doubled and RL decreased 50%. These changes with nerve block were the same during COH, suggesting that neural activity to hindlimb vasculature was not increased by COH. In animals treated with phenoxybenzamine (primarily alpha 1-blocked), RL dropped (approximately 50%) during COH, an indication that catecholamines played a significant role in maintaining tone to skeletal muscle. Animals with both alpha 1 + alpha 2-adrenergic blockade (phenoxybenzamine and yohimbine added) did not survive COH. RL was higher in beta 2-block than in the untreated group during COH, but nerve cooling indicated that beta 2-adrenoceptor vasodilation was accomplished primarily by humoral means. The above findings demonstrated that adrenergic receptors were important in the regulation of QL and RL during COH, but they were not activated by sympathetic nerve stimulation to the limb muscles.     

Female sex pheromone components of the cotton bollworm, Heliothis armigera

Detailed examination of abdominal tip extracts from adult female Heliothis armigera revealed the presence of two components which elicit electroantennographic responses from the male moth. These olfactory stimulants have been fully identified as (Z)-11-hexadecenal (I) and (Z)-11-hexadecen-1-ol(II), and detected in airborne volatiles from a ‘calling’ female moth. A third olfactory stimulant was detected only in female tip extracts from some moths of Malawi origin, and this was tentatively identified as (Z)-9-hexadecenal (III). No other olfactory stimulants could be found, although hexadecenal (IV) and 1-hexadecan-1-ol (V) were detected by gas chromatography. In field tests in Malawi, (Z)-11-hexadecenal (I) attracted a few male H. armigera moths to traps but was very much less attractive than the virgin female moth. The attractiveness of (I) was not consistently affected by addition of alcohol (II), aldehydes (III) and (IV), or (E)-11-hexadecenal. Significant numbers of male Earias biplaga moths were found to be attracted to (Z)-11-hexadecenal (I).     

Phosphorus nutrition of rice in relation to flooding and temporary loss of soil-water saturation in two lowland soils of Cambodia

In the rainfed lowlands, temporary loss of soil-water saturation during crop growth is a common factor limiting rice (Oryza sativa L.) yield but its effects on phosphorus (P) availability are poorly understood. Rice plants were transplanted into pots containing soils that were either continuously flooded, maintained at field capacity or flooded and then dried to field capacity for 3 weeks during the vegetative stage. A black clay soil (Kandic Plinthaquult) and a sandy soil (Plinthustalf) from south-east Cambodia were compared with or without amendments by rice straw and P fertilizer. Under continuously flooded conditions, the growth of rice was vigorous without straw addition and there was a strong response of rice growth to the addition of P fertilizer. The soil underwent reduction, which increased pH from 4.2 to 5.5 or 6.0, in the black clay or sandy soil, respectively. By contrast, a loss of soil-water saturation 3 weeks before panicle initiation (PI) markedly impaired the growth of rice. This was not through any effect of water stress, and the growth reductions were not as strong as with continued loss of soil-water saturation from transplanting to PI. Fluctuations in soil pH and Eh corresponded closely to changes in soil-water regimes. Growth reductions were attributed to reduced shoot P levels resulting from the decline in P availability during the loss of soil-water saturation. The addition of rice straw stimulated soil reduction and lessened changes in soil pH and Eh during the loss of soil-water saturation in both soils. Straw addition enhanced P uptake by the rice plants during loss of soil-water saturation, but its beneficial effects could not be attributed to the direct addition of P, N or K to the soils. Thus the application of rice straw may be effective in lessening the effects of temporary loss of soil-water saturation on rice growth in lowland rice soils by minimising the decline in P availability. This revised version was published online in August 2006 with corrections to the Cover Date.