F-1 Food Assistance Program State Summary, August 2016

The Food Assistance Monthly Participation Report is a monthly summary of Food Assistance program participation, statewide and for each Iowa county. Breakouts are reported for participants also in the FIP program, those only receiving Food Assistance, and those that are receiving economic assistance under other programs (primarily Medicaid). This report may also be known as the F-1 Report.

F-1 Food Assistance Program State Summary, July 2016

The Food Assistance Monthly Participation Report is a monthly summary of Food Assistance program participation, statewide and for each Iowa county. Breakouts are reported for participants also in the FIP program, those only receiving Food Assistance, and those that are receiving economic assistance under other programs (primarily Medicaid). This report may also be known as the F-1 Report.

F-1 Food Assistance Program State Summary, October 2016

The Food Assistance Monthly Participation Report is a monthly summary of Food Assistance program participation, statewide and for each Iowa county. Breakouts are reported for participants also in the FIP program, those only receiving Food Assistance, and those that are receiving economic assistance under other programs (primarily Medicaid). This report may also be known as the F-1 Report.

F-1 Food Assistance Program State Summary, November 2016

The Food Assistance Monthly Participation Report is a monthly summary of Food Assistance program participation, statewide and for each Iowa county. Breakouts are reported for participants also in the FIP program, those only receiving Food Assistance, and those that are receiving economic assistance under other programs (primarily Medicaid). This report may also be known as the F-1 Report.

F-1 Food Assistance Program State Summary, December 2016

The Food Assistance Monthly Participation Report is a monthly summary of Food Assistance program participation, statewide and for each Iowa county. Breakouts are reported for participants also in the FIP program, those only receiving Food Assistance, and those that are receiving economic assistance under other programs (primarily Medicaid). This report may also be known as the F-1 Report.

Low fat loss response after medium-term supervised exercise in obese is associated with exercise-induced increase in food reward

Objective: To examine exercise-induced changes in the reward value of food during medium-term supervised exercise in obese individuals. ---------- Subjects/Methods: The study was a 12-week supervised exercise intervention prescribed to expend 500 kcal/day, 5 d/week. 34 sedentary obese males and females were identified as responders (R) or non-responders (NR) to the intervention according to changes in body composition relative to measured energy expended during exercise. Food reward (ratings of liking and wanting, and relative preference by forced choice pairs) for an array of food images was assessed before and after an acute exercise bout. ---------- Results. 20 responders and 14 non-responders were identified. R lost 5.2 kg±2.4 of total fat mass and NR lost 1.7 kg±1.4. After acute exercise, liking for all foods increased in NR compared to no change in R. Furthermore, NR showed an increase in wanting and relative preference for high-fat sweet foods. These differences were independent of 12-weeks regular exercise and weight loss. ---------- Conclusion. Individuals who showed an immediate post-exercise increase in liking and increased wanting and preference for high-fat sweet foods displayed a smaller reduction in fat mass with exercise. For some individuals, exercise increases the reward value of food and diminishes the impact of exercise on fat loss.

Appetite, satiety, and food reward in obese individuals : a behavioral phenotype approach

Why do some people remain lean while others are susceptible to obesity, and why do obese individuals vary in their successes in losing weight? Despite physiological processes that promote satiety and satiation, some individuals are more susceptible to overeating. While the phenomena of susceptibility to weight gain, resistance to treatment or weight loss, and individual variability are not novel, they have yet to be exploited and systematically examined to better understand how to characterise phenotypes of obesity. The identification and characterisation of distinct phenotypes not only highlight the heterogeneous nature of obesity but may also help to inform the development of more tailored strategies for the treatment and prevention of obesity. This review examines the evidence for different susceptible phenotypes of obesity that are characterised by risk factors associated with the hedonic and homeostatic systems of appetite control.

Food reward sensitivity predicts overconsumption of high-fat snack food

Overconsumption of snack foods has been linked to rising rates of obesity, with our ‘obesogenic’ environment and its abundance of palatable, high-calorie foods and associated cues especially implicated. However, it is clear that some individuals are particularly susceptible to overconsumption and weight gain. It was hypothesised that individuals sensitive to the rewarding properties of palatable foods, and associated stimuli, would show elevated consumption. Snack food intake was measured in 50 adults (mean age 34.5 years, BMI 23.9 kg/m2, 56% female) in a repeated measures design, both with and without a ‘food cue’. Trait (BIS/BAS scales), behavioural (computerised CARROT) and food reward were assessed. Sensitivity to food reward, but not generalised reward, was positively associated with snack food intake. This relationship was not affected by the presence of a food cue. Findings are discussed in the context of implications for weight management.

Neural effects of cannabinoid CB1 neutral antagonist tetrahydrocannabivarin (THCv) on food reward and aversion in healthy volunteers

Disturbances in the regulation of reward and aversion in the brain may underlie disorders such as obesity and eating disorders. We previously showed that the cannabis receptor subtype (CB1) inverse agonist rimonabant, an antiobesity drug withdrawn due to depressogenic side effects, diminished neural reward responses yet increased aversive responses (Horder et al., 2010). Unlike rimonabant, tetrahydrocannabivarin is a neutral CB1 receptor antagonist (Pertwee, 2005) and may therefore produce different modulations of the neural reward system. We hypothesized that tetrahydrocannabivarin would, unlike rimonabant, leave intact neural reward responses but augment aversive responses. Methods: We used a within-subject, double-blind design. Twenty healthy volunteers received a single dose of tetrahydrocannabivarin (10mg) and placebo in randomized order on 2 separate occasions. We measured the neural response to rewarding (sight and/or flavor of chocolate) and aversive stimuli (picture of moldy strawberries and/or a less pleasant strawberry taste) using functional magnetic resonance imaging. Volunteers rated pleasantness, intensity, and wanting for each stimulus. Results: There were no significant differences between groups in subjective ratings. However, tetrahydrocannabivarin increased responses to chocolate stimuli in the midbrain, anterior cingulate cortex, caudate, and putamen. Tetrahydrocannabivarin also increased responses to aversive stimuli in the amygdala, insula, mid orbitofrontal cortex, caudate, and putamen. Conclusions: Our findings are the first to show that treatment with the CB1 neutral antagonist tetrahydrocannabivarin increases neural responding to rewarding and aversive stimuli. This effect profile suggests therapeutic activity in obesity, perhaps with a lowered risk of depressive side effects. Keywords: reward, THCv, obesity, fMRI, cannabinoid

Delta EEG Activity in Left Orbitofrontal Cortex in Rats Related to Food Reward and Craving

眶额叶皮质与中脑边缘多巴胺奖赏系统有着复杂的相互纤维联系.先前的研究探讨了药物成瘾过程中眶额叶皮质的脑电活动.在本实验中,将探讨食物奖赏和渴求过程中该皮质的脑电活动.实验采用了两个环境:对照环境和食物刺激相关的环境.首先,训练大鼠在食物刺激相关的环境中吃巧克力花生豆,而后在该环境中设置两种不同的刺激方式:能看到和闻到但不能吃到(渴求实验),或者仍旧可以吃到巧克力花生豆(奖赏实验):同时进行左侧眶额叶皮质的脑电记录.结果发现,在食物刺激相关的环境中大鼠Delta频段(2-4Hz)的脑电活动与食物刺激显著相关,此外,与在对照环境中相比,其相对功率在食物渴求时下降而在食物奖赏时升高.本实验表明,食物相关的奖励可以改变大鼠眶额叶皮质的脑电活动,而且,Delta频段的脑电活动能够作为监测该奖励的一个指标.

Opposing neural effects of naltrexone on food reward and aversion: implications for the treatment of obesity

Rationale: Opioid antagonism reduces the consumption of palatable foods in humans but the neural substrates implicated in these effects are less well understood. Objectives: The aim of the present study was to examine the effects of the opioid antagonist, naltrexone, on neural response to rewarding and aversive sight and taste stimuli. Methods: We used functional magnetic resonance imaging (fMRI) to examine the neural responses to the sight and taste of pleasant (chocolate) and aversive (mouldy strawberry) stimuli in 20 healthy volunteers who received a single oral dose of naltrexone (50 mg) and placebo in a double-blind, repeated-measures cross-over, design. Results: Relative to placebo, naltrexone decreased reward activation to chocolate in the dorsal anterior cingulate cortex and caudate, and increased aversive-related activation to unpleasant strawberry in the amygdala and anterior insula. Conclusions: These findings suggest that modulation of key brain areas involved in reward processing, cognitive control and habit formation such as the dorsal anterior cingulate cortex (dACC) and caudate might underlie reduction in food intake with opioid antagonism. Furthermore we show for the first time that naltrexone can increase activations related to aversive food stimuli. These results support further investigation of opioid treatments in obesity.

Le rôle des acides gras oméga-3 sur la balance énergétique, la régulation de l’appétit, l’état émotionnel et l’implication du récepteur GPR120

L’obésité est un facteur de risque lié à des problèmes physiques, émotionnels et comportementaux. Aujourd’hui, l’alimentation est composée d’un régime typiquement occidental «Western diet» qui est riche en acides gras saturés (AGS) et pauvre en acides gras polyinsaturés (AGPI) tel que les oméga-3 (N-3) et occasionnant un déséquilibre du ratio alimentaire N-6/N-3. Ce déséquilibre est une des causes de la prévalence des maladies mentales y compris celles des troubles de l'humeur et de l’anxiété. L’acide docosahexaénoïque (ADH, 22: 6 n-3) est l’acide gras (AG) le plus abondant dans le cerveau et son accumulation est particulièrement élevée pendant la période périnatale. Il joue un rôle important dans le développement neuronal et d'autres fonctions du cerveau tel l'apprentissage et la mémoire. Des perturbations de l’environnement périnatal peuvent influencer à très long terme l’avenir de la descendance en la rendant plus susceptible de développer des problèmes d’obésité dans un contexte nutritionnel riche. On ignore cependant si le déficit alimentaire chez la mère et particulièrement en ADH aura un impact sur la motivation alimentaire de la progéniture. L’objectif principal de cette thèse est d’étudier le rôle potentiel des N-3 sur la balance énergétique, la motivation alimentaire, la dépression et le niveau d’anxiété des descendants de souris mâles adultes assujetties à une alimentation riche en gras. Nos données ont démontré qu‘un régime maternel déficitaire en ADH durant la période périnatale incitait la descendance à fournir plus d’effort afin d’obtenir un aliment palatable. Ceci entraînerait un dérèglement de l’homéostasie énergétique en augmentant le gain de poids et en diminuant l’activité locomotrice tout en exacerbant le comportement de type anxieux dès que les souris sont exposées à un milieu obésogène. Les acides gras libres (AGL) sont des nutriments essentiels fonctionnant comme des molécules de signalisation dans le cerveau en ayant des récepteurs qui jouent un rôle important dans le contrôle du métabolisme énergétique. Parmi eux, on distingue un récepteur couplé à la protéine G (GPCR), le GPR120. Ce récepteur activé par les AGPI ω-3 intervient dans les mécanismes anti-inflammatoires et insulino-résistants via les N-3. Une mutation dans le gène GPR120 occasionnée par une réduction de l’activité de signalisation du gène est liée à l’obésité humaine. L'objectif premier de cette deuxième étude était d’évaluer l'impact de la stimulation pharmacologique de GPR120 dans le système nerveux central (SNC) sur l'alimentation, les dépenses d'énergie, le comportement de type anxieux et la récompense alimentaire. Nos résultats démontrent qu’une injection centrale aiguë d'agoniste GPR120 III réduit la prise alimentaire ad libitum et la motivation alimentaire pour un aliment riche en gras et en sucre; ainsi que les comportements de type anxieux. L’injection centrale chronique (21 jours) de ce même agoniste GPR120 III transmis par une pompe osmotique a démontré que les souris placées sous diète hypercalorique (HFD n’ont présenté aucune modification lors de la prise alimentaire ni de gain de poids mais qu’il y avait comparativement au groupe de véhicule, une réduction du comportement de type anxieux, que ce soit dans le labyrinthe en croix surélevé (LCS) ou dans le test à champ ouvert (OFT). L’ADH est reconnu pour ses propriétés anorexigènes au niveau central. De plus, la stimulation des récepteurs de GPR120 au niveau du cerveau avec un agoniste synthétique peut produire un effet intense intervenir sur le comportement lié à l'alimentation des rongeurs. Trouver une approche visant à contrôler à la fois la neuroinflammation, la récompense alimentaire et les troubles émotionnels aiderait assurément au traitement de l'obésité et du diabète de type 2.

Le rôle des acides gras oméga-3 sur la balance énergétique, la régulation de l’appétit, l’état émotionnel et l’implication du récepteur GPR120

L’obésité est un facteur de risque lié à des problèmes physiques, émotionnels et comportementaux. Aujourd’hui, l’alimentation est composée d’un régime typiquement occidental «Western diet» qui est riche en acides gras saturés (AGS) et pauvre en acides gras polyinsaturés (AGPI) tel que les oméga-3 (N-3) et occasionnant un déséquilibre du ratio alimentaire N-6/N-3. Ce déséquilibre est une des causes de la prévalence des maladies mentales y compris celles des troubles de l'humeur et de l’anxiété. L’acide docosahexaénoïque (ADH, 22: 6 n-3) est l’acide gras (AG) le plus abondant dans le cerveau et son accumulation est particulièrement élevée pendant la période périnatale. Il joue un rôle important dans le développement neuronal et d'autres fonctions du cerveau tel l'apprentissage et la mémoire. Des perturbations de l’environnement périnatal peuvent influencer à très long terme l’avenir de la descendance en la rendant plus susceptible de développer des problèmes d’obésité dans un contexte nutritionnel riche. On ignore cependant si le déficit alimentaire chez la mère et particulièrement en ADH aura un impact sur la motivation alimentaire de la progéniture. L’objectif principal de cette thèse est d’étudier le rôle potentiel des N-3 sur la balance énergétique, la motivation alimentaire, la dépression et le niveau d’anxiété des descendants de souris mâles adultes assujetties à une alimentation riche en gras. Nos données ont démontré qu‘un régime maternel déficitaire en ADH durant la période périnatale incitait la descendance à fournir plus d’effort afin d’obtenir un aliment palatable. Ceci entraînerait un dérèglement de l’homéostasie énergétique en augmentant le gain de poids et en diminuant l’activité locomotrice tout en exacerbant le comportement de type anxieux dès que les souris sont exposées à un milieu obésogène. Les acides gras libres (AGL) sont des nutriments essentiels fonctionnant comme des molécules de signalisation dans le cerveau en ayant des récepteurs qui jouent un rôle important dans le contrôle du métabolisme énergétique. Parmi eux, on distingue un récepteur couplé à la protéine G (GPCR), le GPR120. Ce récepteur activé par les AGPI ω-3 intervient dans les mécanismes anti-inflammatoires et insulino-résistants via les N-3. Une mutation dans le gène GPR120 occasionnée par une réduction de l’activité de signalisation du gène est liée à l’obésité humaine. L'objectif premier de cette deuxième étude était d’évaluer l'impact de la stimulation pharmacologique de GPR120 dans le système nerveux central (SNC) sur l'alimentation, les dépenses d'énergie, le comportement de type anxieux et la récompense alimentaire. Nos résultats démontrent qu’une injection centrale aiguë d'agoniste GPR120 III réduit la prise alimentaire ad libitum et la motivation alimentaire pour un aliment riche en gras et en sucre; ainsi que les comportements de type anxieux. L’injection centrale chronique (21 jours) de ce même agoniste GPR120 III transmis par une pompe osmotique a démontré que les souris placées sous diète hypercalorique (HFD n’ont présenté aucune modification lors de la prise alimentaire ni de gain de poids mais qu’il y avait comparativement au groupe de véhicule, une réduction du comportement de type anxieux, que ce soit dans le labyrinthe en croix surélevé (LCS) ou dans le test à champ ouvert (OFT). L’ADH est reconnu pour ses propriétés anorexigènes au niveau central. De plus, la stimulation des récepteurs de GPR120 au niveau du cerveau avec un agoniste synthétique peut produire un effet intense intervenir sur le comportement lié à l'alimentation des rongeurs. Trouver une approche visant à contrôler à la fois la neuroinflammation, la récompense alimentaire et les troubles émotionnels aiderait assurément au traitement de l'obésité et du diabète de type 2.

The roles of the preproghrelin-derived peptides - ghrelin, desacyl ghrelin and obestatin - in prostate cancer

Prostate cancer is the second most common cause of cancer related deaths in Western men. Despite the significant improvements in current treatment techniques, there is no cure for advanced metastatic, castrate-resistant disease. Early detection and prevention of progression to a castrate-resistant state may provide new strategies to improve survival. A number of growth factors have been shown to act in an autocrine/paracrine manner to modulate prostate cancer tumour growth. Our laboratory has previously shown that ghrelin and its receptors (the functional GHS-R1a and the non-functional GHS-R1b) are expressed in prostate cancer specimens and cell lines. We have shown that ghrelin increases cell proliferation in the PC3 and LNCaP prostate cancer cell lines through activation of ERK1/2, suggesting that ghrelin could regulate prostate cancer cell growth and play a role in the progression of the disease. Ghrelin is a 28 amino-acid peptide hormone, identified to be the natural ligand of the growth hormone secretagogue receptor (GHS-R1a). It is well characterised as a growth hormone releasing and as an orexigenic peptide that stimulates appetite and feeding and regulates energy expenditure and bodyweight. In addition to its orexigenic properties, ghrelin has been shown to play a regulatory role in a number of systems, including the reproductive, immune and cardiovascular systems and may play a role in a number of pathological conditions such as chronic heart failure, anorexia, cachexia, obesity, diabetes and cancer. In cancer, ghrelin and its receptor are expressed in a range of tumours and cancer cell lines and ghrelin has been demonstrated to modulate cell proliferation, apoptosis, migration and invasion in some cell types. The ghrelin gene (GHRL) encodes preproghrelin peptide, which is processed to produce three currently known functional peptides - ghrelin, desacyl ghrelin and obestatin. Prohormone convertases (PCs) have been shown to cleave the preproghrelin peptide into two primary products - the 28 amino acid peptide, ghrelin, and the remaining 117 amino acid C-terminal peptide, C-ghrelin. C-ghrelin can then be further processed to produce the 23 amino acid peptide, obestatin. Ghrelin circulates in two different forms - an octanoylated form (known as ghrelin) and a non-octanoylated form, desacyl ghrelin. The unique post-translational addition of octanoic acid to the serine 3 residue of the propeptide chain to form acylated ghrelin is catalysed by ghrelin O-acyltransferase (GOAT). This modification is necessary for binding of ghrelin to its only known functional receptor, the GHS-R1a. As desacyl ghrelin cannot bind and activate the GHS-R1a, it was initially thought to be an inactive peptide, despite the fact that it circulates at much higher levels than ghrelin. Further research has demonstrated that desacyl ghrelin is biologically active and shares some of the actions of ghrelin, as well as having some opposing and distinct roles. Interestingly, both ghrelin and desacyl ghrelin have been shown to modulate apoptosis, cell differentiation and proliferation in some cell types, and to stimulate cell proliferation through activation of ERK1/2 and PI3K/Akt pathways. The third known peptide product of the ghrelin preprohormone, obestatin, was initially thought to oppose the actions of ghrelin in appetite regulation and food intake and to mediate its effects through the G protein-coupled receptor 39 (GPR39). Subsequent research failed to reproduce the initial findings, however, and the possible anorexigenic effects of obestatin, as well as the identity of its receptor, remain unclear. Obestatin plays some important physiological roles, including roles in improving memory, the inhibition of thirst and anxiety, increased secretion of pancreatic juice, and regulation of cell proliferation, survival, apoptosis and differentiation. Preliminary studies have also shown that obestatin stimulates cell proliferation in some cell types through activation of ERK1/2, Akt and PKC pathways. Overall, however, at the commencement of this PhD project, relatively little was known regarding the functions and mechanisms of action of the preproghrelin-derived functional peptides in modulating prostate cancer cell proliferation. The roles of obestatin, and desacyl ghrelin as potential growth factors had not previously been investigated, and the potential expression and regulation of the preproghrelin processing enzymes, GOAT and prohormone convertases was unknown in prostate cancer cell lines. Therefore, the overall objectives of this study were to: 1. investigate the effects of obestatin on cell proliferation and signaling in prostate cancer cell lines 2. compare the effects of desacyl ghrelin and ghrelin on cell proliferation and signaling in prostate cancer cell lines 3. investigate whether prostate cancer cell lines possess the necessary enzymatic machinery to produce ghrelin and desacyl ghrelin and if these peptides can regulate GOAT expression Our laboratory has previously shown that ghrelin stimulates cell proliferation in the PC3 and LNCaP prostate cancer cell line through activation of the ERK1/2 pathway. In this study it has been demonstrated that treatments with either ghrelin, desacyl ghrelin or obestatin over 72 hours significantly increased cell proliferation in the PC3 prostate cancer cell line but had no significant effect in the RWPE-1 transformed normal prostate cell line. Ghrelin (1000nM) stimulated cell proliferation in the PC3 prostate cancer cell line by 31.66 6.68% (p<0.01) with the WST-1 method, and 13.55 5.68% (p<0.05) with the CyQUANT assay. Desacyl ghrelin (1000nM) increased cell proliferation in PC3 cells by 21.73 2.62% (p<0.01) (WST-1), and 15.46 7.05% (p<0.05) (CyQUANT) above untreated control. Obestatin (1000nM) induced a 28.37 7.47% (p<0.01) (WST-1) and 12.14 7.47% (p<0.05) (CyQUANT) significant increase in cell proliferation in the PC3 prostate cancer cell line. Ghrelin and desacyl ghrelin treatments stimulated Akt and ERK phosphorylation across a range of concentrations (p<0.01). Obestatin treatment significantly stimulated Akt, ERK and PKC phosphorylation (p<0.05). Through the use of specific inhibitors, the MAPK inhibitor U0126 and the Akt1/2 kinase inhibitor, it was demonstrated that ghrelin- and obestatin-induced cell proliferation in the PC3 prostate cancer cell line is mediated through activation of ERK1/2 and Akt pathways. Although desacyl ghrelin significantly stimulated Akt and ERK phosphorylation, U0126 failed to prevent desacyl ghrelin-induced cell proliferation suggesting ghrelin and desacyl ghrelin might act through different mechanisms to increase cell proliferation. Ghrelin and desacyl ghrelin have shown a proliferative effect in osteoblasts, pancreatic -cells and cardiomyocytes through activation of ERK1/2 and PI3K/Akt pathways. Here it has been shown that ghrelin and its non-acylated form exert the same function and stimulate cell proliferation in the PC3 prostate cancer cell line through activation of the Akt pathway. Ghrelin-induced proliferation was also mediated through activation of the ERK1/2 pathway, however, desacyl ghrelin seems to stimulate cell proliferation in an ERK1/2-independent manner. As desacyl ghrelin does not bind and activate GHSR1a, the only known functional ghrelin receptor, the finding that both ghrelin and desacyl ghrelin stimulate cell proliferation in the PC3 cell line suggests that these peptides could be acting through the yet unidentified alternative ghrelin receptor in this cell type. Obestatin treatment also stimulated PKC phosphorylation, however, a direct role for this pathway in stimulating cell proliferation could not be proven using available PKC pathway inhibitors, as they caused significant cell death over the extended timeframe of the cell proliferation assays. Obestatin has been shown to stimulate cell proliferation through activation of PKC isoforms in human retinal epithelial cells and in the human gastric cancer cell line KATO-III. We have demonstrated that all of the prostate-derived cell lines examined (PC3, LNCaP, DU145, 22Rv1, RWPE-1 and RWPE-2) expressed GOAT and at least one of the prohormone convertases, which are known to cleave the proghrelin peptide, PC1/3, PC2 and furin, at the mRNA level. These cells, therefore, are likely to possess the necessary machinery to cleave the preproghrelin protein and to produce the mature ghrelin and desacyl ghrelin peptides. In addition to prohormone convertases, the presence of octanoic acid is essential for acylated ghrelin production. In this study octanoic acid supplementation significantly increased cell proliferation in the PC3 prostate cancer cell line by over 20% compared to untreated controls (p<0.01), but surprisingly, not in the DU145, LNCaP or 22Rv1 prostate cancer cell lines or in the RWPE-1 and RWPE-2 prostate-derived cell lines. In addition, we demonstrated that exogenous ghrelin induced a statistically significant two-fold decrease in GOAT mRNA expression in the PC3 cell line (p<0.05), suggesting that ghrelin could pontentially downregulate its own acylation and, therefore, regulate the balance between ghrelin and desacyl ghrelin. This was not observed, however, in the DU145 and LNCaP prostate cancer cell lines. The GOAT-ghrelin system represents a direct link between ingested nutrients and regulation of ghrelin production and the ghrelin/desacyl ghrelin ratio. Regulation of ghrelin acylation is a potentially attractive and desirable tool for the development of better therapies for a number of pathological conditions where ghrelin has been shown to play a key role. The finding that desacyl ghrelin stimulates cell proliferation in the PC3 prostate cancer cell line, and responds to ghrelin in the same way, suggests that this cell line expresses an alternative ghrelin receptor. Although all the cell lines examined expressed both GHS-R1a and GHS-R1b mRNA, it remains uncertain whether these cell lines express the unidentified alternative ghrelin receptor. It is possible that the varied responses seen could be due to the expression of different ghrelin receptors in different cell lines. In addition to GOAT, prohormone convertases and octanoic acid availability may regulate the production of different peptides from the ghrelin preprohormone. The studies presented in this thesis provide significant new information regarding the roles and mechanisms of action of the preproghrelin-derived peptides, ghrelin, desacyl ghrelin and obestatin, in modulating prostate cancer cell line proliferation. A number of key questions remain to be resolved, however, including the identification of the alternative ghrelin/desacyl ghrelin receptor, the identification of the obestatin receptor, a clarification of the signaling mechanisms which mediate cell proliferation in response to obestatin treatment and a better understanding of the regulation at both the gene and post-translational levels of functional peptide generation. Further studies investigating the role of the ghrelin axis using in vivo prostate cancer models may be warranted. Until these issues are determined, the potential for the ghrelin axis, to be recognised as a novel useful target for therapy for cancer or other pathologies will be uncertain.

Probing the role of the C-H center dot center dot center dot O hydrogen bond stabilized polypeptide chain reversal at the C-terminus of designed peptide helices. Structural characterization of three decapeptides

The structural characterization in crystals of three designed decapeptides containing a double D-segment at the C-terminus is described. The crystal structures of the peptides Boc-Leu-Aib-Val-Xxx-Leu-Aib-Val- (D)Ala-(D)Leu-Aib-OMe, (Xxx = Gly 2, (D)Ala 3, Aib 4) have been determined and compared with those reported earlier for peptide 1 (Xxx = Ala) and the all L analogue Boc-Leu-Aib-Val-Ala-Leu-Aib-Val-Ala-Leu-Aib-OMe, which yielded a perfect right-handed a-helical structure. Peptides 1 and 2 reveal a right-handed helical segment spanning residues 1 to 7, ending in a Schellman motif with Ala(8) functioning as the terminating residue. Polypeptide chain reversal occurs at residue 9, a novel feature that appears to be the consequence of a C-(HO)-O-... hydrogen bond between residue 4 (CH)-H-alpha and residue 9 CO groups. The structures of peptides 3 and 4, which lack the pro R hydrogen at the C-alpha atom of residue 4, are dramatically different. Peptide 3 adopts a right-handed helical conformation over the 1 to 7 segment. Residues 8 and 9 adopt at conformations forming a C-terminus type I' beta-turn, corresponding to an incipient left-handed twist of the polypeptide chain. In peptide 4, helix termination occurs at Aib(6), with residues 6 to 9 forming a left-handed helix, resulting in a structure that accommodates direct fusion of two helical segments of opposite twist. Peptides 3 and 4 provide examples of chiral residues occurring in the less favored sense of helical twist; (D)Ala(4) in peptide 3 adopts an alpha(R) conformation, while (L)Val(7) in 4 adopts an alpha(L) conformation. The structural comparison of the decapeptides reported here provides evidence for the role of specific C-(HO)-O-... hydrogen bonds in stabilizing chain reversals at helix termini, which may be relevant in aligning contiguous helical and strand segments in polypeptide structures.