Peptides (v.72, #C)

Playing a critical role in the metabolic homeostasis of living systems, the circulating concentrations of peptides/proteins are influenced by a variety of patho-physiological events. These peptide/protein concentrations in biological fluids are measured using various methods, the most common of which is enzymatic immunoassay EIA/ELISA and which guide the clinicians in diagnosing and monitoring diseases that inflict biological systems. All the techniques where enzymes are employed to show antigen–antibody reactions are generally referred to as enzymatic immunoassay EIA/ELISA method. Since the basic principles of EIA and ELISA are the same. The main objective of this review is to present an overview of the historical journey that had led to the invention of EIA/ELISA, an indispensible method for medical and research laboratories, types of ELISA developed after its invention [direct (the first ELISA method invented), indirect, sandwich and competitive methods], problems encountered during peptide/protein analyses (pre-analytical, analytical and post-analytical), rules to be followed to prevent these problems, and our laboratory experience of more than 15 years.
Keywords: Analytical error; ELISA; General protocol; Peptide analysis;

Peptides and the blood–brain barrier by William A. Banks (16-19).
The demonstration that peptides and regulatory proteins can cross the blood–brain barrier (BBB) is one of the major contributions of Dr. Abba J. Kastin. He was the first to propose that peptides could cross the BBB, the first to show that an endogenous peptide did so, and the first to describe a saturable transport system at the BBB for peptides. His work shows that in crossing the BBB, peptides and regulatory proteins act as informational molecules, informing the brain of peripheral events. Brain-to-blood passage helps to control levels of peptides with the brain and can deliver information in the brain-to-blood direction. He showed that the transporters for peptides and proteins are not static, but respond to developmental and physiological changes and are affected by disease states. As such, the BBB is adaptive to the needs of the CNS, but when that adaption goes awry, the BBB can be a cause of disease. The mechanisms by which peptides and proteins cross the BBB offer opportunities for drug delivery of these substances or their analogs to the brain in the treatment of diseases of the central nervous system.
Keywords: Peptides; Blood–brain barrier; AJ Kastin; Delta sleep-inducing peptide; Regulatory proteins; Cytokines;

Functional elucidation of the endogenous opioid system temporally paralleled the creation and growth of the journal, Peptides, under the leadership of its founding editor, Dr. Abba Kastin. He was prescient in publishing annual and uninterrupted reviews on Endogenous Opiates and Behavior that served as a microcosm for the journal under his stewardship. This author published a 2004 review, “Endogenous opioids and feeding behavior: a thirty-year historical perspective”, summarizing research in this field between 1974 and 2003. The present review “closes the circle” by reviewing the last 10 years (2004–2014) of research examining the role of endogenous opioids and feeding behavior. The review summarizes effects upon ingestive behavior following administration of opioid receptor agonists, in opioid receptor knockout animals, following administration of general opioid receptor antagonists, following administration of selective mu, delta, kappa and ORL-1 receptor antagonists, and evaluating opioid peptide and opioid receptor changes in different food intake models.
Keywords: Food intake; Mu opioid receptor; Delta opioid receptor; Kappa opioid receptor; ORL-1 opioid receptor;

Peptides: Basic determinants of reproductive functions by Onder Celik; Suleyman Aydin; Nilufer Celik; Musa Yilmaz (34-43).
Mammalian reproduction is a costly process in terms of energy consumption. The critical information regarding metabolic status is signaled to the hypothalamus mainly through peripheral peptides from the adipose tissue and gastrointestinal tract. Changes in energy stores produce fluctuations in leptin, insulin, ghrelin and glucose signals that feedback mainly to the hypothalamus to regulate metabolism and fertility. In near future, possible effects of the nutritional status on GnRH regulation can be evaluated by measuring serum or tissue levels of leptin and ghrelin in patiens suffering from infertility. The fact that leptin and ghrelin are antagonistic in their effects on GnRH neurons, their respective agonistic and antagonistic roles make them ideal candidates to use instead of GnRH agonist and antagonist. Similarly, kisspeptin expressing neurons are likely to mediate the well-established link between energy balance and reproductive functions. Exogenous kisspeptin can be used for physiological ovarian hyperstimulation for in-vitro fertilization. Moreover, kisspeptin antagonist therapy can be used for the treatment of postmenapousal women, precocious puberty, PCOS, endometriosis and uterine fibroids. In this review, we will analyze the central mechanisms involved in the integration of metabolic information and their contribution to the control of the reproductive function. Particular attention will be paid to summarize the participation of leptin, kisspeptin, ghrelin, NPY, orexin, urocortin, VIP, insulin, galanin, galanin like peptide, oxytocin, agouti gene-related peptide, and POMC neurons in this process and their possible interactions to contribute to the metabolic control of reproduction.
Keywords: Peptides; Reproduction; Infertility;

Host-defense and trefoil factor family peptides in skin secretions of the Mawa clawed frog Xenopus boumbaensis (Pipidae) by J. Michael Conlon; Milena Mechkarska; Jolanta Kolodziejek; Jérôme Leprince; Laurent Coquet; Thierry Jouenne; Hubert Vaudry; Norbert Nowotny; Jay D. King (44-49).
Peptidomic analysis of norepinephrine-stimulated skin secretions from the octoploid Mawa clawed frog Xenopus boumbaensis Loumont, 1983 led to the identification and characterization of 15 host-defense peptides belonging to the magainin (two peptides), peptide glycine-leucine-amide (PGLa; three peptides), xenopsin precursor fragment (XPF; three peptides), caerulein precursor fragment (CPF; two peptides), and caerulein precursor fragment-related peptide (CPF-RP; five peptides) families. In addition, caerulein and three peptides with structural similarity to the trefoil factor family (TFF) peptides, xP2 and xP4 from Xenopus laevis were also present in the secretions. Consistent with data from comparisons of the nucleotides sequence of mitochondrial and nuclear genes, the primary structures of the peptides suggest a close phylogenetic relationship between X. boumbaensis and the octoploid frogs Xenopus amieti and Xenopus andrei. As the three species occupy disjunct ranges within Cameroon, it is suggested that they diverged from a common ancestor by allopatric speciation.
Keywords: Host-defense; Frog skin; Pipidae; Polyploidy; Xenopus; Trefoil factor family;

Peptides at the blood brain barrier: Knowing me knowing you by Thomas P. Davis; Thomas J. Abbruscato; Richard D. Egleton (50-56).
When the Davis Lab was first asked to contribute to this special edition of Peptides to celebrate the career and influence of Abba Kastin on peptide research, it felt like a daunting task. It is difficult to really understand and appreciate the influence that Abba has had, not only on a generation of peptide researchers, but also on the field of blood brain barrier (BBB) research, unless you lived it as we did. When we look back at our careers and those of our former students, one can truly see that several of Abba's papers played an influential role in the development of our personal research programs.
Keywords: Blood brain barrier; Neuropeptides; Drug delivery; Tight junction proteins; Transporter proteins; P-glycoprotein;

The influence of intracellular angiotensin II (Ang II) on the process of chemical communication and metabolic cooperation between cardiac cells is discussed. Emphasis is given to the influence of pathological conditions like heart failure, myocardial ischemia or hyperglycemia on the activation of the intracrine renin angiotensin aldosterone system (RAAS) and its consequence for the metabolic cooperation between heart cells. Furthermore, the influence of high glucose on the process of chemical communication was described as well as its implication for the failing and diabetic heart. The major conclusion is that the activation of the intracrine renin angiotensin induced by heart failure, hyperglycemia, aldosterone or myocardial ischemia generates metabolic imbalance in the heart with serious consequences for the cardiac function.
Keywords: Intracellular angiotensin; Chemical communication; Heart cells;

15 years on the Editorial Board of PEPTIDES by Luciano Debeljuk (61-63).
In this article, part of an issue of PEPTIDES celebrating the many years of dedication of Dr. A.J. Kastin as Editor-in-Chief of the journal, I describe how I first met him and how our friendship and collaboration developed. In 1970 I joined Dr. A.V. Schally's team at the Endocrine and Polypeptide Laboratories of Tulane University and the V.A. Hospital and it was at that time that I met Dr. Kastin for the first time. In 1984 I joined Dr. Schally's team for the second time and I began publishing part of my research findings in PEPTIDES. Afterward many more results from the research work were also published in PEPTIDES. I became a member of the Editorial Board of the same journal in 1999. It was going to be a very interesting experience. Some of my observations as a reviewer for PEPTIDES are described in the present article. Finally, I expressed my warmest congratulations to Dr. A.J. Kastin for the outstanding job that he carried out as Editor-in-Chief of PEPTIDES.
Keywords: PEPTIDES; Dr. A.J. Kastin; Tachykinins; GnRH;

Cysteine-stabilized αβ defensins: From a common fold to antibacterial activity by Renata de Oliveira Dias; Octavio Luiz Franco (64-72).
Antimicrobial peptides (AMPs) seem to be promising alternatives to common antibiotics, which are facing increasing bacterial resistance. Among them are the cysteine-stabilized αβ defensins. These peptides are small, with a length ranging from 34 to 54 amino acid residues, cysteine-rich and extremely stable, normally composed of an α-helix and three β-strands stabilized by three or four disulfide bonds and commonly found in several organisms. Moreover, animal and plant CSαβ defensins present different specificities, the first being mainly active against bacteria and the second against fungi. The role of the CSαβ-motif remains unknown, but a common antibacterial mechanism of action, based on the inhibition of the cell-wall formation, has already been observed in some fungal and invertebrate defensins. In this context, the present work aims to group the data about CSαβ defensins, highlighting their evolution, conservation, structural characteristics, antibacterial activity and biotechnological perspectives.
Keywords: Defensin; Antimicrobial peptide; Antibacterial; Plant; Insect;

In commemoration of Abba J. Kastin's exceptional service as the founding editor for the international journal Peptides, I review our collaborative work on how neuropeptides are involved in depression and other neuropsychiatric behavior. A special focus is on MIF-1 (prolyl-leucyl-glycinamide) that was discovered in the Kastin laboratory and shown effective to treat human depression with greater efficacy and faster onset of action than traditional antidepressants at the time of clinical trial. My personal reflection of the evolving changes of translational research on neuropeptides will hopefully provide some insight to young investigators.
Keywords: AJ Kastin; Peptides; Depression; MIF-1; TRH; MSH/ACTH4-10;

ADNP is a protein necessary for brain development, important for brain plasticity, cognitive and social functioning, characteristics that are all impaired in autism and in the Adnp +/− mouse model, in a sex-dependent manner. ADNP was originally discovered as a protein that is secreted from glial cells in response to vasoactive intestinal peptide (VIP). VIP is a major neuroprotective peptide in the CNS and PNS and was also associated with social recognition in rodents and aggression, pair-bonding and parental behaviors in birds. Comparative sequence alignment revealed high evolutionary conservation of ADNP in Chordata. Despite its importance in brain function, ADNP has never been studied in birds. Zebra finches (Taeniopygia guttata) are highly social songbirds that have a sexually dichotomous anatomical brain structure, with males demonstrating a developed song system, presenting a model to study behavior and potential sexually dependent fundamental differences. Here, using quantitative real time polymerase chain reaction (qRT-PCR), we discovered sexually dichotomous and age related differences in ADNP mRNA expression in three different regions of the song bird brain–cerebellum, cerebrum, and brain stem. Higher levels of ADNP mRNA were specifically found in young male compared to the female cerebrum, while aging caused a significant 2 and 3-fold decrease in the female and male cerebrum, respectively. Furthermore, a comparison between the three tested brain regions revealed unique sex-dependent ADNP mRNA distribution patterns, affected by aging. Future studies are aimed at deciphering the function of ADNP in birds, toward a better molecular understanding of sexual dichotomy in singing behavior in birds.
Keywords: Activity-dependent neuroprotective protein (ADNP); Zebra finch; Songbird brain; qRT-PCR; Sexual dimorphism; Aging;

While the poisonous effects of botulinum neurotoxin (BoNT) have been recognized since antiquity, the overall actions and mechanisms of effects of BoNT have been elucidated primarily over the past several decades. The general utility of BoNT is described in the paper, but the focus is mainly on the approaches towards negating the toxic effects of BoNT, and on the projection of an engineered BoNT molecule serving as a Trojan Horse to deliver a therapeutic load for treatment of a host of medical disorders. The BoNT molecule is configured with a binding domain, a zinc-dependent protease with specificity primarily for vesicular proteins, and a translocation domain for delivery of the metalloprotease into the cytoplasm. The anti-toxin approaches for BoNT include the use of vaccines, antibodies, block of BoNT binding or translocation, inhibition of metalloprotease activity, impeded translocation of the protease/catalytic domain, and inhibition of the downstream Src signaling pathway. Projections of BoNT as a therapeutic include its targeting to non-cholinergic nerves, also targeting to non-neuronal cells for treatment of hypersecretory disorders (e.g., cystic fibrosis), and treatment of hormonal disorders (e.g., acromegaly). Still in the exploratory phase, there is the expectation of major advances in BoNT neuroprotective strategies and burgeoning utility of engineered BoNTs as therapeutics.
Keywords: Botulinum; Neurotoxin; Trojan horse; Therapeutic;

Antimicrobial peptides: Possible anti-infective agents by Jayaram Lakshmaiah Narayana; Jyh-Yih Chen (88-94).
Multidrug-resistant bacterial, fungal, viral, and parasitic infections are major health threats. The Infectious Diseases Society of America has expressed concern on the decrease of pharmaceutical companies working on antibiotic research and development. However, small companies, along with academic research institutes, are stepping forward to develop novel therapeutic methods to overcome the present healthcare situation. Among the leading alternatives to current drugs are antimicrobial peptides (AMPs), which are abundantly distributed in nature. AMPs exhibit broad-spectrum activity against a wide variety of bacteria, fungi, viruses, and parasites, and even cancerous cells. They also show potential immunomodulatory properties, and are highly responsive to infectious agents and innate immuno-stimulatory molecules. In recent years, many AMPs have undergone or are undergoing clinical development, and a few are commercially available for topical and other applications. In this review, we outline selected anion and cationic AMPs which are at various stages of development, from preliminary analysis to clinical drug development. Moreover, we also consider current production methods and delivery tools for AMPs, which must be improved for the effective use of these agents.
Keywords: Antimicrobial peptides; Antibiotics; Multidrug resistance; Bacteria; Fungi; Virus; Parasites;

Blockade of nociceptin/orphanin FQ receptor signaling reverses LPS-induced depressive-like behavior in mice by Iris U. Medeiros; Chiara Ruzza; Laila Asth; Remo Guerrini; Pedro R.T. Romão; Elaine C. Gavioli; Girolamo Calo (95-103).
Nociceptin/orphanin FQ is the natural ligand of a Gi-protein coupled receptor named NOP. This peptidergic system is involved in the regulation of mood states and inflammatory responses. The present study aimed to investigate the consequences of blocking NOP signaling in lipopolysaccharide (LPS)-induced sickness and depressive-like behaviors in mice. LPS 0.8 mg/kg, ip, significantly induced sickness signs such as weight loss, decrease of water and food intake and depressive-like behavior in the tail suspension test. Nortriptyline (ip, 60 min prior the test) reversed the LPS-induced depressive states. The NOP receptor antagonist SB-612111, 30 min prior LPS, did not modify LPS-induced sickness signs and depressive-like behavior. However, when injected 24 h after LPS, NOP antagonists (UFP-101, icv, and SB-612111, ip) significantly reversed the mood effects of LPS. LPS evoked similar sickness signs and significantly increased tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) plasma levels 6 h post-injection in wild-type ((NOP(+/+)) and NOP knockout ((NOP(−/−)) mice. However, LPS treatment elicited depressive-like effects in NOP(+/+) but not in NOP(−/−) mice. In conclusion, the pharmacological and genetic blockade of NOP signaling does not affect LPS evoked sickness signs while reversing depressive-like behavior.
Keywords: Nociceptin/orphanin FQ; NOP receptor; Lipopolysaccharide-induced depressive; Inflammation; Depression; Tail suspension test;

From a personal viewpoint, Abba was always a congenial colleague and a good friend. I recall the hours of unraveling complex, confusing, and many times contradictory data sets with him and the excitement we both felt as what had been puzzling began to make sense. Most of all, I feel privileged to have Abba as a long and valued friend.
Keywords: Abba Kastin; MSH/ACTH 4-10; Proteopathies; Behavior; Attention; Learning;

Neuropeptides as lung cancer growth factors by Terry W. Moody; Paola Moreno; Robert T. Jensen (106-111).
This manuscript is written in honor of the Festschrift for Abba Kastin. I met Abba at a Society for Neuroscience meeting and learned that he was Editor-in-Chief of the Journal Peptides. I submitted manuscripts to the journal on “Neuropeptides as Growth Factors in Cancer” and subsequently was named to the Editorial Advisory Board. Over the past 30 years I have published dozens of manuscripts in Peptides and reviewed hundreds of submitted manuscripts. It was always rewarding to interact with Abba, a consummate professional. When I attended meetings in New Orleans I would sometimes go out to dinner with him at the restaurant “Commanders Palace”. When I chaired the Summer Neuropeptide Conference we were honored to have him receive the Fleur Strand Award one year in Israel. I think that his biggest editorial contribution has been the “Handbook of Biologically Active Peptides.” I served as a Section Editor on “Cancer/Anticancer Peptides” and again found that it was a pleasure working with him. This review focuses on the mechanisms by which bombesin-like peptides, neurotensin and vasoactive intestinal peptide regulate the growth of lung cancer.
Keywords: Lung cancer; Bombesin; Vasoactive intestinal peptide; Neurotensin;

The rapid aging of the world's population has led to a need to increase our understanding of the pathophysiology of the factors leading to frailty and cognitive decline. Peptides have been shown to be involved in the pathophysiology of frailty and cognitive decline. Weight loss is a major component of frailty. In this review, we demonstrate a central role for both peripheral peptides (e.g., cholecystokinin and ghrelin) and neuropeptides (e.g., dynorphin and alpha-MSH) in the pathophysiology of the anorexia of aging. Similarly, peripheral peptides (e.g., ghrelin, glucagon-like peptide 1, and cholecystokinin) are modulators of memory. A number of centrally acting neuropeptides have also been shown to modulate cognitive processes. Amyloid-beta peptide in physiological levels is a memory enhancer, while in high (pathological) levels, it plays a key role in the development of Alzheimer's disease.
Keywords: Peptides; Memory; Frailty; Cognitive decline; Neuropeptides; Anorexia of aging;

Abba J. Kastin, the Zeitgeist, and the inception of Peptides by Gayle A. Olson; Richard D. Olson (119-120).
It is wonderful to be able to record the establishment and growth of a professional journal after thirty-five years, and to celebrate the splendid career of Abba J. Kastin as an editor as well as a scientist and educator. Abba is also an enriched human being who is both sophisticated and simple, and we are proud to be life-long friends of his. This Festschrift reviews how we (the Olsons) started our careers as neuropsychologists, our interactions with Abba, reflection of the job as neuroscientists, and discussion of the growth and future of Peptides with the new publishing fads.
Keywords: Abba J. Kastin; Editor; Peptides; Behavior;

In this Festschrift, I discuss the career and guiding principles to which Abba J. Kastin has adhered during the last 20 years we worked together. I briefly describe the history of our joint laboratory group, the context of studies of peptide permeation across the blood–brain barrier (BBB), and newer developments in the BBB Group as Abba steps down after serving 35 years as the founding Editor-in-Chief for Peptides. Abba's BBB studies on peptides have contributed to concepts in the neuroendocrinology of feeding and developed information on molecular trafficking across BBB cells. The astroglial leptin signaling studies and the interactions of sleep and BBB are two major directions, whereas the long-term MIF-1 project demarcates a tortuous road on translational research.
Keywords: Leptin; TNF; MIF-1; Obesity; Circadian; Sleep;

Insights into bombesin receptors and ligands: Highlighting recent advances by Irene Ramos-Álvarez; Paola Moreno; Samuel A. Mantey; Taichi Nakamura; Bernardo Nuche-Berenguer; Terry W. Moody; David H. Coy; Robert T. Jensen (128-144).
This following article is written for Prof. Abba Kastin's Festschrift, to add to the tribute to his important role in the advancement of the role of peptides in physiological, as well as pathophysiological processes. There have been many advances during the 35 years of his prominent role in the Peptide field, not only as editor of the journal Peptides, but also as a scientific investigator and editor of two volumes of the Handbook of Biological Active Peptides [146,147]. Similar to the advances with many different peptides, during this 35 year period, there have been much progress made in the understanding of the pharmacology, cell biology and the role of (bombesin) Bn receptors and their ligands in various disease states, since the original isolation of bombesin from skin of the European frog Bombina bombina in 1970 [76]. This paper will briefly review some of these advances over the time period of Prof. Kastin 35 years in the peptide field concentrating on the advances since 2007 when many of the results from earlier studies were summarized [128,129]. It is appropriate to do this because there have been 280 articles published in Peptides during this time on bombesin-related peptides and it accounts for almost 5% of all publications. Furthermore, 22 Bn publications we have been involved in have been published in either Peptides [14,39,55,58,81,92,93,119,152,216,225,226,231,280,302,309,355,361,362] or in Prof. Kastin's Handbook of Biological Active Peptides [137,138,331].
Keywords: Bombesin; Gastrin releasing peptide; Neuromedin b; Obesity; Pruritus;

The maternal endocrine stress system is profoundly altered during the course of human pregnancy. The human placenta expresses the genes for CRH as early as the seventh week of gestation and it is the expotential increase in placental CRH (pCRH) over the course of human gestation that is responsible for the greatest modification in the maternal stress system. The bi-directional placental release of hormones into the maternal and fetal compartments has profound influences for both. The influential Fetal Programming model predicted that early or fetal exposures to maternal signals of threat or adverse conditions have lifelong consequences for health outcomes. A basic assumption of this model was that developing organisms play a dynamic role in their own construction. Data are reviewed and new data are presented that elevated pCRH over the course of human gestation plays a fundamental role in the organization of the fetal nervous system, modifies birth phenotype (the timing of the onset of spontaneous labor and delivery), and influences developmental, temperamental and metabolic trajectories. Evidence for sex differences and conserved function across species is presented. Finally, a model is presented that proposes several pathways that pCRH can program risk for health and disease.
Keywords: Corticotropin-releasing-hormone; Fetal programming; Stress; Human pregnancy; Fetal development; Sex differences;

Keywords: AJ Kastin; CNS effects of GHRH analogs; Alzheimer's disease; TBI; PTSD; GWI;

Structure–activity relationship of mastoparan analogs: Effects of the number and positioning of Lys residues on secondary structure, interaction with membrane-mimetic systems and biological activity by Bibiana Monson de Souza; Marcia Perez dos Santos Cabrera; Paulo Cesar Gomes; Nathalia Baptista Dias; Rodrigo Guerino Stabeli; Natalia Bueno Leite; João Ruggiero Neto; Mario Sergio Palma (164-174).
In this study, a series of mastoparan analogs were engineered based on the strategies of Ala and Lys scanning in relation to the sequences of classical mastoparans. Ten analog mastoparans, presenting from zero to six Lys residues in their sequences were synthesized and assayed for some typical biological activities for this group of peptide: mast cell degranulation, hemolysis, and antibiosis. In relation to mast cell degranulation, the apparent structural requirement to optimize this activity was the existence of one or two Lys residues at positions 8 and/or 9. In relation to hemolysis, one structural feature that strongly correlated with the potency of this activity was the number of amino acid residues from the C-terminus of each peptide continuously embedded into the zwitterionic membrane of erythrocytes-mimicking liposomes, probably due to the contribution of this structural feature to the membrane perturbation. The antibiotic activity of mastoparan analogs was directly dependent on the apparent extension of their hydrophilic surface, i.e., their molecules must have from four to six Lys residues between positions 4 and 11 of the peptide chain to achieve activities comparable to or higher than the reference antibiotic compounds. The optimization of the antibacterial activity of the mastoparans must consider Lys residues at the positions 4, 5, 7, 8, 9, and 11 of the tetradecapeptide chain, with the other positions occupied by hydrophobic residues, and with the C-terminal residue in the amidated form. These requirements resulted in highly active AMPs with greatly reduced (or no) hemolytic and mast cell degranulating activities.
Keywords: Mastoparan; Wasp venom; Structure–activity relationship; Antimicrobial activity; Peptide synthesis;

Nesfatin-1 was discovered a decade ago and despite the fact that it represents just one of a multitude of food intake-inhibiting factors it received increasing attention. This led to a detailed characterization of NUCB2/nesfatin-1's physiological property to reduce food intake and also gave rise to an involvement in the long term regulation of body weight, especially under conditions of obesity. In addition, studies indicated the involvement of NUCB2/nesfatin-1 in other homeostatic functions as well: glucose homeostasis, water intake, gastrointestinal functions, temperature regulation, cardiovascular functions, puberty onset and sleep. These pleiotropic actions underline the physiological relevance of this peptide. Recently, the involvement of NUCB2/nesfatin-1 in psychiatric disorders such as anxiety has been investigated giving rise to the speculation that NUCB2/nesfatin-1 represents a peptidergic link between eating and anxiety/depression disorders.
Keywords: Anxiety; Brain-gut; Hypothalamus; NUCB2; Nucleobindin2; Stomach;

Ubiquitous expression and multiple functions of biologically active peptides by Kazuhiro Takahashi; Koji Ohba; Kiriko Kaneko (184-191).
Biologically active peptides are widely expressed throughout in human bodies. For example, endothelin-1 and adrenomedullin are expressed in almost all types of cells, including neurons, glial cells, fibroblasts, macrophages, cardiomyocytes, vascular endothelial cells, epithelial cells and cancer cells of various origins. Expression of both these peptides is induced by stimuli, such as hypoxia and inflammatory cytokines. They have a variety of biological functions, such as effects on brain function, hormone secretion, the cardiovascular system and cell proliferation. By contrast, orexins (hypocretins) and melanin-concentrating hormone (MCH) are specifically expressed in the hypothalamus, particularly in the lateral hypothalamus, although very low concentrations of these peptides are found in the peripheral tissues. Orexins and MCH play coordinated, but distinct physiological roles in the regulation of sleep-wake cycle, appetite, emotion and other brain functions. The cardiovascular system is regulated by cardiovascular peptides, such as natriuretic peptides, endothelins and angiotensin II. The renin-angiotensin system (RAS) is one of the most classical regulatory systems on blood pressure, electrolytes and kidney. (Pro)renin receptor is a novel member of the RAS and may be related to the pathophysiology of microvascular complications of hypertension and diabetes mellitus. Moreover, (pro)renin receptor forms a functional complex with vacuolar-type H+-ATPase, which plays an important physiological role in maintaining the acidic environment of intracellular compartments including secretory vesicles. Perhaps, the complex of (pro)renin receptor and vacuolar-type H+-ATPase may be important for the post-translational processing and secretion of many biologically active peptides.
Keywords: Adrenomedullin; Endothelin; Melanin-concentrating hormone; Orexin; Prorenin receptor;

Understanding peptide biology: The discovery and characterization of the novel hormone, neuronostatin by Gina L.C. Yosten; Mollisa M. Elrick; Alison Salvatori; Lauren M. Stein; Grant R. Kolar; Jun Ren; John A. Corbett; Willis K. Samson (192-195).
The Human Genome Project provided the opportunity to use bioinformatic approaches to discover novel, endogenous hormones. Using this approach we have identified two novel peptide hormones and review here our strategy for the identification and characterization of the hormone, neuronostatin. We describe in this mini-review our strategy for determining neuronostatin's actions in brain, heart and pancreas. More importantly, we detail our deductive reasoning strategy for the identification of a neuronostatin receptor and our progress in establishing the physiological relevance of the peptide.
Keywords: Neuropeptides; Hypothalamus; Pituitary; Receptors;

This review summarizes recent studies focusing on diapause hormone (DH) in the Helicoverpa/Heliothis complex of agricultural pests. Moths in this complex overwinter in pupal diapause, a form of developmental arrest used to circumvent unfavorable seasons. DH was originally reported in the silkmoth Bombyx mori, a species that relies on DH to induce an embryonic diapause. But, in the case of Helicoverpa/Heliothis, levels of dh transcripts and DH peptides are more abundant in nondiapausing pupae than in diapausing individuals, and DH effectively terminates diapause within a specific temperature range. A structure activity relationship study indicated that the active core of DH is the C-terminal hepta-peptide, LWFGPRLa. We designed and synthesized a first generation of DH agonists and identified two agonists (PK-2Abf and PK-Etz) that were nearly 50- and 13-fold more potent than the native hormone. These studies revealed two structural characteristics of DH and its agonists that are essential for interaction with the receptor: a trans-Pro configuration to form a type I β-turn and a hydrophobic moiety involved in ligand binding. Modification of DH at the active core yielded a potent DH antagonist (DH-Jo, acetyl-GLWA[Jo]RLa) as well as an agonist (DH-2Abf-K). Three compounds (Decyl-1963, Dodecyl-1967, Heptyl-1965) were identified as agents capable of penetrating the cuticle of young pupae and thereby preventing the entry into diapause. DH receptor cDNA was cloned and an effective in vitro high throughput screen system was established for future use. This work sets the stage for further development of DH analogs and antagonists that have the potential to disrupt insect diapause as a tool for pest management.
Keywords: Insect diapause; Diapause hormone; Gene expression; Diapause termination; Structure activity relationship; Analogs development; Diapause prevention; Receptor;

PACAP as a neuroprotective factor in ischemic neuronal injuries by Seiji Shioda; Tomoya Nakamachi (202-207).
Pituitary adenylate cyclase-activating polypeptide (PACAP) is a 27- or 38-amino acid neuropeptide, which belongs to the vasoactive intestinal polypeptide/glucagon/secretin family. PACAP and its three receptor subtypes are expressed in neural tissues, with PACAP known to exert pleiotropic effects on the nervous system. This review provides an overview of current knowledge regarding the neuroprotective effects, mechanisms of action, and therapeutic potential of PACAP in response to ischemic brain injuries.
Keywords: PACAP; PACAP receptors; Ischemia; Neuronal death; CRMP2; DNA microarray analysis;

We have found various bioactive peptides derived from animal and plant proteins, which interact with receptors for endogenous bioactive peptides such as opioids, neurotensin, complements C3a and C5a, oxytocin, and formyl peptides etc. Among them, rubiscolin, a δ opioid peptide derived from plant RuBisCO, showed memory-consolidating, anxiolytic-like, and food intake-modulating effects. Soymorphin, a μ opioid peptide derived from β-conglycinin showed anxiolytic-like, anorexigenic, hypoglycemic, and hypotriglyceridemic effects. β-Lactotensin derived from β-lactoglobulin, the first natural ligand for the NTS2 receptor, showed memory-consolidating, anxiolytic-like, and hypocholesterolemic effects. Weak agonist peptides for the complements C3a and C5a receptors were released from many proteins and exerted various central effects. Peptides showing anxiolytic-like antihypertensive and anti-alopecia effects via different types of receptors such as OT, FPR and AT2 were also obtained. Based on these study, new functions and post-receptor mechanisms of receptor commom to endogenous and exogenous bioactive peptides have been clarified.
Keywords: Angiotensin; Bradykinin; Complement C3a; Complement C5a; Formyl peptide; Neurotensin; Opioid peptide; Oxytocin; Milk; RuBisCO; Soy;