Prolactin and growth hormone response to intracerebroventricular administration of the food opioid peptide gluten exorphin B5 in rats

Although it has long been known that opioid peptides cause marked changes of pituitary hormone secretion in both animals and humans, little is known about the possible effect(s) of food-derived opioids (exorphins) on pituitary function. In order to investigate the possible role of exorphins derived from wheat gluten on pituitary function, we gave the following treatments to four groups of male rats: intracerebroventricular (ICV) vehicle, Gluten Exorphin B5 (GE-B5) 200 microg ICV, naloxone intraperitoneally (IP) followed by vehicle ICV, naloxone IP followed by GE-B5 ICV. Blood samples for Prolactin (PRL) and Growth Hormone (GH) were taken at intervals for 90 minutes after vehicle or GE-B5 administration. GE-B5 strongly stimulated PRL secretion; its effect was completely abolished by naloxone administration. GH secretion was unaffected by GE-B5 under these experimental conditions. The present study shows for the first time that an opioid peptide derived from wheat gluten, GE-B5, has an effect on pituitary function when administered ICV; its mechanism of action appears to be mediated via classical opioid receptors.     

Colonization of Female Watermelon Blossoms by Acidovorax avenae ssp. citrulli and the Relationship between Blossom Inoculum Dosage and Seed Infestation

The aim of this work was to investigate the ability of Acidovorax avenae ssp. citrulli, the causal agent of bacterial fruit blotch of cucurbits (BFB), to colonize female watermelon blossoms, and to explore the relationship between blossom inoculum dosage and seed infestation. Under greenhouse conditions A. avenae ssp. citrulli colonized stigmas and styles of female watermelon blossoms reaching populations of ≈10⁷ to 10⁸ colony‐forming units (CFU) per blossom for 96 h after inoculation. Acidovorax avenae ssp. citrulli growth on stigmas was slower than that of Pseudomonas syringae Cit7, a non‐pathogenic, foliar epiphyte of tomato. While pollination reduced growth of A. avenae ssp. citrulli, but P. syringae Cit7 was unaffected. Both bacteria colonized style tissues but bacterial growth in the style was significantly less than the stigma. Blossom inoculation with ≈1 × 10³A. avenae ssp. citrulli CFU/blossom led to 36–55% infested seedlots within symptomless fruits. On average 14% of the seedlings produced from these seedlots displayed BFB symptoms. There was a strong positive correlation between A. avenae ssp. citrulli inoculum concentration applied to blossoms and the percentage of infested seedlots, as determined by the seedling grow‐out assay (R² = 0.94). However, this relationship was weaker when seedlot infestation was determined by a polymerase chain reaction‐based assay (R² = 0.34). There was also a strong positive linear relationship between A. avenae ssp. citrulli blossom inoculum dose and the mean percentage of BFB‐infected seedlings (R² = 0.99) produced in seedling grow‐out assays. These data support the hypothesis that blossom colonization might be involved in seed infestation under field conditions.     

Increased prevalence of vulnerable atherosclerotic plaques and low levels of natural IgM antibodies against phosphorylcholine in patients with systemic lupus erythematosus

Introduction  

The risk of cardiovascular disease (CVD) and atherosclerosis is reported to be increased in systemic lupus erythematosus (SLE). We recently reported a negative association between natural IgM-antibodies against phosphorylcholine (anti-PC) in the general population, high anti-PC levels leading to decreased atherosclerosis development and low levels to increased risk of CVD. Potential mechanisms include anti-inflammatory properties and inhibition of uptake of oxidized low density lipoprotein (LDL) in macrophages. The objective herein was to study atherosclerosis in SLE in detail and in relation to traditional and non-traditional risk factors.     

Gluten exorphin B5 stimulates prolactin secretion through opioid receptors located outside the blood-brain barrier

Gluten exorphin B5 (GE-B5) is a food-derived opioid peptide identified in digests of wheat gluten. We have recently shown that GE-B5 stimulates prolactin (PRL) secretion in rats; this effect is abolished by preadministration of the opioid receptor antagonist naloxone. However, since the structure of naloxone allows it to cross the blood-brain barrier (BBB) and antagonize opioid effects centrally as well as peripherally, it could not established, on the basis of those data, if GE-B5-induced PRL release is exerted through sites located inside or outside the BBB. In this study, we sought to determine the site of action of GE-B5 on PRL secretion, by pretreating male rats with naloxone methobromide (NMB), an opioid antagonist that does not cross the BBB. Four groups of rats were given the following treatments: 1) intravenous vehicle; 2) intravenous GE-B5 (3 mg kg(-1) body weight); 3) intraperitoneal NMB (5 mg kg(-1) body weight), followed by vehicle; 4) NMB, followed by GE-B5. Blood samples for PRL were taken at intervals for 40 minutes after vehicle or GE-B5 administration. GE-B5 stimulated PRL secretion; the effect was statistically significant at time 20. NMB preadministration completely abolished PRL response. Our experiment indicates that GE-B5 stimulates PRL secretion through opioid receptors located outside the BBB. Since opioid peptides do not exert their effect on PRL secretion directly, but via a reduced dopaminergic tone, our data suggest that GE-B5 can modify brain neurotransmitter release without crossing the BBB.     

Glp-1 analog, liraglutide, ameliorates hepatic steatosis and cardiac hypertrophy in C57BL/6J mice fed a Western diet

The aims of this study were designed to determine whether liraglutide, a long-acting glucagon-like peptide, could reverse the adverse effects of a diet high in fat that also contained trans-fat and high-fructose corn syrup (ALIOS diet). Specifically, we examined whether treatment with liraglutide could reduce hepatic insulin resistance and steatosis as well as improve cardiac function. Male C57BL/6J mice were pair fed or fed ad libitum either standard chow or the ALIOS diet. After 8 wk the mice were further subdivided and received daily injections of either liraglutide or saline for 4 wk. Hyperinsulinemic-euglycemic clamp studies were performed after 6 wk, revealing hepatic insulin resistance. Glucose tolerance and insulin resistance tests were performed at 8 and 12 wk prior to and following liraglutide treatment. Liver pathology, cardiac measurements, blood chemistry, and RNA and protein analyses were performed. Clamp studies revealed hepatic insulin resistance after 6 wk of ALIOS diet. Liraglutide reduced visceral adiposity and liver weight (P < 0.001). As expected, liraglutide improved glucose and insulin tolerance. Liraglutide improved hypertension (P < 0.05) and reduced cardiac hypertrophy. Surprisingly, liver from liraglutide-treated mice had significantly higher levels of fatty acid binding protein, acyl-CoA oxidase II, very long-chain acyl-CoA dehydrogenase, and microsomal triglyceride transfer protein. We conclude that liraglutide reduces the harmful effects of an ALIOS diet by improving insulin sensitivity and by reducing lipid accumulation in liver through multiple mechanisms including, transport, and increase β-oxidation.     

Seasonal changes in photosynthesis, antioxidant systems and ELIP expression in a thermonastic and non-thermonastic Rhododendron species: A comparison of photoprotective strategies in overwintering plants

Leaves of overwintering evergreen rhododendrons are typically exposed to freezing temperatures and high light during winters which can potentially result in photon flux exceeding that required for photochemistry. This excess energy, if not dissipated as heat or fluorescence, may cause photooxidative damage to PSII. The goal of this study is to compare the photoprotection strategies during seasonal cold acclimation (CA) in two Rhododendron species (R. catawbiense Michx. and R. ponticum L.) that are divergent in their leaf freezing tolerance and thermonastic behaviour (temperature-induced leaf movement). R. catawbiense exhibits thermonasty while R. ponticum does not. Differences in leaf freezing tolerance (LT₅₀), photosynthesis, photoinhibition, early light-induced proteins (ELIPs) gene expression, and accumulation of antioxidant metabolites and enzymes during seasonal CA were investigated. During seasonal CA, maximum photosynthetic rate (P_max) and maximum quantum efficiency of PSII (F_v/F_m) were significantly down-regulated. Compared with R. catawbiense, R. ponticum showed less photoinhibition and higher overall accumulation (in magnitude) of antioxidant systems while R. catawbiense exhibited more efficient up-regulation of ELIP expression and antioxidant system (i.e., greater efficiency of increasing this pool in winter months relative to the summer levels). The two species respond differently to winter conditions and have evolved strategies to avoid, reduce and/or tolerate photooxidative stress in winter. These include down-regulation of photosynthesis and up-regulation of ELIPs and antioxidant systems, together with specialized leaf anatomy and thermonasty behaviour.     

Assessment of genetic diversity among sorghum landraces and their wild/weedy relatives in western Kenya using simple sequence repeat (SSR) markers

Understanding the extent of gene exchange between cultivated sorghum and its wild/weedy relatives and the evolutionary processes (including farmers’ practices) that act to shape the structure of genetic diversity within and between them is an important aspect for germplasm conservation strategies, biosafety risk assessment, and crop improvement programs. In this study, molecular characterization and genetic diversity analyses were conducted on wild, weedy and cultivated sorghums collected at a local-scale in a traditional farming system in the Lambwe Valley of western Kenya. Nine simple sequence repeat (SSR) markers were used to genotype 294 cultivated sorghum and 200 wild sorghum individuals. The nine SSR markers were highly polymorphic with a number of alleles that varied from 2 to 19. Overall, wild sorghums had higher genetic diversity, observed heterozygosity, total number of alleles, polymorphic information content and more genotypes per locus than the cultivated types. A Mantel test demonstrated that there was significant isolation-by-distance for wilds and cultivated materials. STRUCTURE, cluster and principal coordinate analyses consistently assigned wild and cultivated individuals to different groups but failed to place hybrids/weedy types as a single separate group from wilds. Our results provide strong evidence of significant genetic diversity retained within wilds, larger divergence between wild and cultivated materials and reduced gene flow than those previously reported in Kenya. These results demonstrate the value of the Lambwe Valley region as a genetic reservoir and the importance to conduct genetic diversity studies at the local scale to design and execute appropriate in situ conservation programs and policies.     

Adiponectin Agonist ADP355 Attenuates CCl4-Induced Liver Fibrosis in Mice

Liver fibrosis is a growing global health problem characterized by excess deposition of fibrillar collagen, and activation of hepatic stellate cells (HSCs). Adiponectin is known to possess anti-fibrotic properties; however a high physiological concentration and multiple forms circulating in blood prohibit clinical use. Recently, an adiponectin-like small synthetic peptide agonist (ADP355: H-DAsn-Ile-Pro-Nva-Leu-Tyr-DSer-Phe-Ala-DSer-NH2) was synthesized for the treatment of murine breast cancer. The present study was designed to evaluate the efficacy of ADP355 as an anti-fibrotic agent in the in vivo carbon tetrachloride (CCl₄)-induced liver fibrosis model. Liver fibrosis was induced in eight-week old male C57BL/6J mice by CCl₄-gavage every other day for four weeks before injection of a nanoparticle-conjugated with ADP355 (nano-ADP355). Control gold nanoparticles and nano-ADP355 were administered by intraperitoneal injection for two weeks along with CCl₄-gavage. All mice were sacrificed after 6 weeks, and serum and liver tissue were collected for biochemical, histopathologic and molecular analyses. Biochemical studies suggested ADP355 treatment attenuates liver fibrosis, determined by reduction of serum aspartate aminotransferase (AST), alanine aminotransferase ALT) and hydroxyproline. Histopathology revealed chronic CCl₄-treatment results in significant fibrosis, while ADP355 treatment induced significantly reversed fibrosis. Key markers for fibrogenesis–α-smooth muscle actin (α-SMA), transforming growth factor-beta1 (TGF-β1), connective tissue growth factor (CTGF), and the tissue inhibitor of metalloproteinase I (TIMP1) were also markedly attenuated. Conversely, liver lysates from ADP355 treated mice increased phosphorylation of both endothelial nitric oxide synthase (eNOS) and AMPK while AKT phosphorylation was diminished. These findings suggest ADP355 is a potent anti-fibrotic agent that can be an effective intervention against liver fibrosis.