SerpinA3N is a novel hypothalamic gene upregulated by a high-fat diet and leptin in mice.

BACKGROUND: Energy homeostasis is regulated by the hypothalamus but fails when animals are fed a high-fat diet (HFD), and leptin insensitivity and obesity develops. To elucidate the possible mechanisms underlying these effects, a microarray-based transcriptomics approach was used to identify novel genes regulated by HFD and leptin in the mouse hypothalamus. RESULTS: Mouse global array data identified serpinA3N as a novel gene highly upregulated by both a HFD and leptin challenge. In situ hybridisation showed serpinA3N expression upregulation by HFD and leptin in all major hypothalamic nuclei in agreement with transcriptomic gene expression data. Immunohistochemistry and studies in the hypothalamic clonal neuronal cell line, mHypoE-N42 (N42), confirmed that alpha 1-antichymotrypsin (α1AC), the protein encoded by serpinA3, is localised to neurons and revealed that it is secreted into the media. SerpinA3N expression in N42 neurons is upregulated by palmitic acid and by leptin, together with IL-6 and TNFα, and all three genes are downregulated by the anti-inflammatory monounsaturated fat, oleic acid. Additionally, palmitate upregulation of serpinA3 in N42 neurons is blocked by the NFκB inhibitor, BAY11, and the upregulation of serpinA3N expression in the hypothalamus by HFD is blunted in IL-1 receptor 1 knockout (IL-1R1 -/- ) mice. CONCLUSIONS: These data demonstrate that serpinA3 expression is implicated in nutritionally mediated hypothalamic inflammation.

Breads Fortified with Freeze-Dried Vegetables: Quality and Nutritional Attributes. Part 1: Breads Containing Oil as an Ingredient.

There is increasing emphasis on reformulating processed foods to make them healthier. This study for the first time comprehensively investigated the effects of fortifying bread (containing oil as an ingredient) with freeze-dried vegetables on its nutritional and physico-chemical attributes. Breads fortified with carrot, tomato, beetroot or broccoli were assessed for nutrition, antioxidant potential, storage life, shelf stability, textural changes and macronutrient oxidation. Furthermore, using an in vitro model the study for the first time examined the impact of vegetable addition on the oxidative stability of macronutrients during human gastro-intestinal digestion. As expected, adding vegetables improved the nutritional and antioxidant properties of bread. Beetroot and broccoli significantly improved bread storage life. None of the vegetables significantly affected bread textural changes during storage compared to the control. Lipid oxidation in fresh bread was significantly reduced by all four types of vegetables whilst protein oxidation was lowered by beetroot, carrot and broccoli. The vegetables demonstrated varying effects on macronutrient oxidation during gastro-intestinal digestion. Beetroot consistently showed positive effects suggesting its addition to bread could be particularly beneficial.

Breads Fortified with Freeze-Dried Vegetables: Quality and Nutritional Attributes. Part II: Breads Not Containing Oil as an Ingredient.

The present article describes the second part of a study investigating the effect of adding vegetables on the nutritional, physico-chemical, and oxidative properties of wheat bread, and specifically focuses on bread that does not contain oil as an added ingredient. Wheat flour breads fortified with freeze-dried carrot, tomato, beetroot or broccoli were developed and assessed for their nutritional composition, antioxidant potential, oxidative stability, and storage properties. Using a simulated in vitro model, the study also examined the impact of vegetable addition on the oxidative stability of macronutrients during gastro-intestinal digestion. Adding vegetables improved the nutritional and functional attributes of the oil-free breads. However, they demonstrated a lower antioxidant potential compared to their oil-containing counterparts. Similarly, the textural and storage properties of the oil-free vegetable breads were poorer compared to the oil-containing breads. As expected, in the absence of oil the oil-free breads were associated with lower lipid oxidation both in their fresh form and during gastro-intestinal digestion. Adding vegetables reduced protein oxidation in the fresh oil-free breads but had no effect during gastro-intestinal digestion. The impact of vegetables on macronutrient oxidation in the oil-free breads during digestion appears to be vegetable-specific with broccoli exacerbating it and the others having no effect. Of the evaluated vegetables, beetroot showed the most promising nutritional and physico-chemical benefits when incorporated into bread that does not contain added oil.

Identification, expression, and tissue distribution of the three rat lysyl hydroxylase isoforms.

Lysyl hydroxylases (LH) (procollagen-lysine 2-oxoglutarate 5-dioxygenase; PLOD) catalyse the hydroxylation of lysine residues during the post-translational modification of collagenous proteins. In this paper, we describe the first identification and cloning of LH isoforms 2 and 3 from the rat, including both LH2 splice variants (LH2a and LH2b). The rat LHs are expressed in almost all tissue and cell types examined, indicating a probable lack of tissue specificity for LH function. All LH isoforms were stably transfected into CHO-K1 cells and this represents the first example of recombinant LH production in a eukaryotic cell line. Expression and production of all LH isoforms led to an increase in total collagen synthesis. LH1 and LH2a expression and production led to an increase in total pyridinium cross-link production. Evidence that LH2a possesses telopeptide lysyl hydroxylase activity, previously thought to be a novel enzyme, is presented.