Neuropeptides are important messenger molecules that influence nearly all physiological processes. In insects, they can be released as neuromodulators within the central nervous system (CNS) or as neurohormones into the hemolymph. We analyzed the peptidome of neurohormonal release sites and associated secretory peptidergic neurons of adult Drosophila melanogaster. MALDI-TOF mass spectrometric analyzes were performed on single organs or cell cluster from individual flies. This first peptidomic characterization in adult fruit flies revealed 32 different neuropeptides. Peptides not directly predictable from previously cloned or annotated precursor genes were sequenced by tandem mass spectrometry. These peptides turned out to be either intermediate products of neuropeptide processing or shorter versions of known peptides. We found that the peptidome of the CNS-associated neurohemal organs is tagma-specific in Drosophila. Abdominal neurohemal organs and their supplying peptidergic neurons contain the capa gene products periviscerokinins and pyrokinin-1, thoracic neurohemal organs contain FMRFamides, and the neurohemal release sites of the brain contain pyrokinin-1(2-15), pyrokinin-2, corazonin, myosuppressin, and sNPF as their major putative release products. Our results show that peptidomic approaches are well suited to study differential neuropeptide expression or posttranslational modifications in morphologically defined parts of the nervous system and in a developmental and physiological context in animals as small as Drosophila melanogaster.

Despite the importance of branched chain lipid oxidation in detoxification, almost nothing is known regarding factors regulating peroxisomal uptake, targeting, and metabolism. One peroxisomal protein, sterol carrier protein-x (SCP-x), is thought to catalyze a key thiolytic step in branched chain lipid oxidation. When mice with substantially lower hepatic levels of SCP-x were tested for susceptibility to dietary stress with phytol (a phytanic acid precursor and peroxisome proliferator), livers of phytol-fed female but not male mice i ) accumulated phytol metabolites (phytanic acid, pristanic acid, and ⌬ -2,3-pristanic acid); ii ) exhibited decreased fat tissue mass and increased liver mass/ body mass; iii ) displayed signs of histopathological lesions in the liver; and iv ) demonstrated significant alterations in hepatic lipid distributions. Moreover, both male and female mice exhibited phytol-induced peroxisomal proliferation, as demonstrated by liver morphology and upregulation of the peroxisomal protein catalase. In addition, levels of liver fatty acid binding protein, along with SCP-2 and SCP-x, increased, suggesting upregulation mediated by phytanic acid, a known ligand agonist of the peroxisomal proliferator-activated receptor ␣ . In summary, the present work establishes a role for SCP-x in branched chain lipid catabolism and demonstrates a sexual dimorphic response to phytol, a precursor of phytanic acid, in lipid parameters and hepatotoxicity.

Potatoes and other foods that have a high content of the amino acid asparagine and a high accumulation of reducing sugars are subject to the formation of acrylamide upon frying. The objectives of this research were (1) to analyze the level of acrylamide formed during deep-fat frying of potato chips and (2) to evaluate means of reducing acrylamide in potato chips by using different potato cultivars and vacuum frying. Several potato cultivars were used in this research, including Innovator (I), NDTX 4930-5W (N), ATX 854 04-8W (ATw), Atlantic (A), Shepody (S), ATX847806-2Ru (ATr), and White-Rose (W ). An electric bench-top (atmospheric conditions)-type fryer was used to fry the potatoes. Three temperatures were used: 150°C, 165°C, and 180°C. The vacuum frying experiments were performed at 118°C, 125°C, and 140°C and a vacuum pressure of 10 Torr. The potatoes were sliced (1.5-mm thick) and fried for different lengths of times. For potatoes fried at 165°C (for 4 min) at atmospheric conditions, the acrylamide contents were 5021 Ϯ Ϯ Ϯ Ϯ Ϯ 55 ppb (W), 552 Ϯ Ϯ Ϯ Ϯ Ϯ 25 ppb (I), 358 Ϯ Ϯ Ϯ Ϯ Ϯ 50 ppb (N), 397 Ϯ Ϯ Ϯ Ϯ Ϯ 25 ppb (ATw), 646 Ϯ Ϯ Ϯ Ϯ Ϯ 55 ppb (A), 466 Ϯ Ϯ Ϯ Ϯ Ϯ 15 ppb(S), and 537 Ϯ Ϯ Ϯ Ϯ Ϯ 14 ppb (ATr). Vacuum frying reduced acrylamide formation by 94%. Results showed that both cultivar and modified frying systems can play an important role in reducing acrylamide formation in fried potatoes. As the frying temperature decreased from 180°C to 165°C, acrylamide content in potato chips reduced by 51% during traditional frying and by 63% as the temperature decreased from 140°C to 125°C in vacuum frying. Increased frying time increased acrylamide formation during traditional frying for all temperatures and frying methods analyzed. However, the effect on acrylamide concentration was greater for the traditional frying than the vacuum frying.

Bovine trypsin (EC 3.4.21.4) is an enzyme that is widely used for commercial purposes to digest or process other proteins, including some therapeutic proteins. The biopharmaceutical industry is trying to eliminate animal-derived proteins from manufacturing processes due to the possible contamination of these products by human pathogens. Recombinant trypsin has been produced in a number of systems, including cell culture, bacteria and yeast. To date, these expression systems have not produced trypsin on a scale sufficient to fulfill the need of biopharmaceutical manufacturers where kilogram quantities are often required. The present paper describes commercial-level production of trypsin in transgenic maize (Zea mays) and its physical and functional characterization. This protease, the first enzyme to be produced on a large-scale using transgenic plant technology, is functionally equivalent to native bovine pancreatic trypsin. The availability of this reagent should allow for the replacement of animal-derived trypsin in the processing of pharmaceutical proteins.

The high-temperature requirement A (HtrA) family of serine proteases has been shown to play an important role in the environmental and cellular stress damage control system in Escherichia coli. Mycobacterium tuberculosis ( Mtb) has three putative HtrA-like proteases, HtrA1, HtrA2, and HtrA3. The deletion of htrA2 gives attenuated virulence in a mouse model of TB. Biochemical analysis reveals that HtrA2 can function both as a protease and as a chaperone. The three-dimensional structure of HtrA2 determined at 2.0 A resolution shows that the protease domains form the central core of the trimer and the PDZ domains extend to the periphery. Unlike E. coli DegS and DegP, the protease is naturally active due to the formation of the serine protease-like catalytic triad and its uniquely designed oxyanion hole. Both protease and PDZ binding pockets of each HtrA2 molecule are occupied by autoproteolytic peptide products and reveal clues for a novel autoregulatory mechanism that might have significant importance in HtrA-associated virulence of Mtb.