Effects of fatty acid chain length and saturation on gastric inhibitory polypeptide release in obese hyperglycaemic (ob/ob) mice

Plasma gastric inhibitory polypeptide (GIP) responses to equimolar intragastrically administered emulsions of fatty acids (2.62 mmol/7.5 ml/kg) were examined in 18 h fasted obese hyperglycaemic (ob/ob) mice. Propionic acid (C3:0), a saturated short-chain fatty acid, and capric acid (C10:0), a saturated medium chain fatty acid, did not signilicantly stimulate GIP release. However, the saturated long-chain fatty acid stearic acid (C18:0), and especially the unsaturated long-chain fatty acids oleic (C18:1), linoleic (C18:2) and linolenic (C18:3) acids produced a marked GIP response. The results show that chain length and to a lesser extent the degree of saturation are important determinants of fatty acid-stimulated GIP release. The GIP-release action of long-chain, but not short-chain, fatty acids may be r e l a t e d to differences in their intracellular handling.     

The ins and outs of reticulocyte maturation revisited: The role of autophagy in sickle cell disease

Autophagy plays an important role in the removal of membrane bound organelles during the last stage of erythropoiesis as the enucleate reticulocyte matures into the erythrocyte. Autophagic vesicles are expelled from the reticulocyte as intact, inside-out, phosphatidylserine (PS) decorated vesicles and are subsequently removed during splenic passage. Failure to remove these vesicles causes the elevation in PS exposed red cells in Sickle Cell Disease.     

How NOT to Approach the Obesity Problem

FLATT, JP. How NOT to approach the obesity problem. The emphasis given to the energy balance equation has fostered the widespread belief that obesity is a problem of energy balance. This mistaken view has led to many unjustified and unfortunate interpretations, because obesity is, rather, a problem of the interaction between body composition and food intake regulation.     

Role of ketone bodies in the diabetes-induced changes in hepatic mixed-function oxidase activities

In order to assess the role of ketone bodies in the diabetes-induced changes in hepatic mixed-function oxidase activity, rats rendered hyperketonaemic by dietary administration of medium chain triacylglycerols were compared with streptozotocin treated rats. Both groups of animals became hyperketonaemic but only the latter were hyperglycaemic. Treatment with streptozotocin or medium chain triacylglycerols gave rise to marked increases in the O-dealkylations of ethoxyresorufin, ethoxycoumarin and pentoxyresorufin, the p-hydroxylation of aniline and the N-demethylation of dimethylnitrosamine. It is concluded that the streptozotocin-induced changes in hepatic mixed-function oxidases are mediated, at least partly, by the high levels of ketone bodies.     

Glucose-mediated Ca(2+) signalling in single clonal insulin-secreting cells: evidence for a mixed model of cellular activation

Using clonal insulin-secreting BRIN-BD11 cells, we have assessed whether the graded response of the whole cell population to glucose can be accounted for by a dose-dependent recruitment of individual cells, an amplification of the response of the recruited cells or both. Cytosolic free Ca(2+) concentration ([Ca(2+)](i)) is an established index of beta-cell function. We used fura-2 microfluorescence techniques to assess the [Ca(2+)](i) responsiveness of single BRIN-BD11 cells to glucose and other secretagogues. Glucose (1-16.7 mM) evoked oscillatory [Ca(2+)](i) rises in these cells resembling those found in parental rat pancreatic beta-cells. The percentage of glucose-responsive cells was 11% at 1 mM and increased to 40-70% at 3-16.7 mM glucose, as assessed by a single-stimulation protocol. This profile was unrelated to possible differences in the cell cycle, as inferred from experiments where the cultured cells were synchronized by a double thymidine block protocol. Individual cells exhibited variable sensitivities to glucose (threshold range: 1-5 mM) and a variable dose-dependent amplification of the [Ca(2+)](i) responses (EC(50) range: 2-10 mM), as assessed by a multiple-stimulation protocol. Glyceraldehyde and alpha-ketoisocaproic acid had glucose-like effects on [Ca(2+)](i). The data support a mixed model for the activation of insulin-secreting cells. Specifically, the graded secretory response of the whole cell population is likely to reflect both a recruitment of individual cells with different sensitivities to glucose and a dose-dependent amplification of the response of the recruited cells.     

Biological activity and antidiabetic potential of synthetic fragment peptides of glucose-dependent insulinotropic polypeptide, GIP(1-16) and (Pro3)GIP(1-16)

Glucose-dependent insulinotropic polypeptide (GIP) is a key physiological insulin releasing peptide and potential antidiabetic agent. The present study was undertaken in an attempt to develop small molecular weight GIP agonist and antagonist molecules. The bioactivity of two modified C-terminally truncated fragment GIP peptides, GIP(1-16) and (Pro3)GIP(1-16), was examined in terms of insulin secretion and glucose homeostasis using BRIN-BD11 cells and type 2 diabetic mice. In vitro insulin release studies demonstrated that GIP(1-16) and (Pro3)GIP(1-16) possessed weak GIP-receptor agonist and antagonistic properties, respectively. Intraperitoneal administration of GIP(1-16) in combination with glucose to obese diabetic (ob/ob) mice did not effect the glycaemic excursion and had a marginal effect on insulin release. GIP(1-16) was substantially less effective than the native GIP(1-42). (Pro3)GIP(1-16) administration significantly curtailed (P < 0.05) the insulinotropic and glucose lowering effects of native GIP, but was significantly less effective than (Pro3)GIP. Based on the established concept of a therapeutic benefit of GIP receptor antagonism in obesity-diabetes, ob/ob mice received once daily injection of (Pro3)GIP(1-16) for 14 days. No significant effects were observed on food intake, body weight, HbA1c, glucose tolerance, metabolic response to feeding and either insulin secretion or insulin sensitivity following prolonged (Pro3)GIP(1-16) treatment. These data demonstrate that C-terminal truncation of GIP or (Pro3)GIP yields small molecular weight GIP molecules with significantly reduced biological activity that precludes therapeutic utility.     

Histamine-releasing and antimicrobial peptides from the skin secretions of the dusky gopher frog, Rana sevosa

Seven novel peptides were isolated from the skin secretions of the North American dusky gopher frog, Rana sevosa, on the basis of antimicrobial activity and histamine release from rat peritoneal mast cells. The peptides were purified to homogeneity using HPLC and characterized by electrospray ion-trap mass spectrometry, MALDI-TOF mass spectrometry and Edman sequencing. Bioinformatic analysis of primary structures revealed that the novel peptides could be assigned to four established families of ranid frog antimicrobial peptides, namely esculentin-1, esculentin-2, brevinin-1 and ranatuerin-2. The peptides were named in accordance with accepted terminology as ranatuerin 2SEa, etc., reflecting the peptide family name, the species of origin (SE for sevosa) and the isotype (a). Of major interest was the fact that brevinin 1SE displayed significant structural similarity to ponericin W5, an antibacterial venom peptide from the ant, Pachyconyla goeldii. This is a further example of amphibian skin defensive peptides showing striking structural similarities to peptides from insects. These data may shed some light on the functional biological relevance of defensive peptides that possess both antimicrobial and histamine-releasing activities.     

NMR and alanine scan studies of glucose-dependent insulinotropic polypeptide in water

Glucose-dependent insulinotropic polypeptide (GIP) is an incretin hormone that stimulates the secretion of insulin after ingestion of food. GIP also promotes the synthesis of fatty acids in adipose tissue. Therefore, it is not surprising that numerous literature reports have shown that GIP is linked to diabetes and obesity-related diseases. In this study, we present the solution structure of GIP in water determined by NMR spectroscopy. The calculated structure is characterized by the presence of an alpha-helical motif between residues Ser(11) and Gln(29). The helical conformation of GIP is further supported by CD spectroscopic studies. Six GIP-(1-42)Ala(1-7) analogues were synthesized by replacing individual N-terminal residues with alanine. Alanine scan studies of these N-terminal residues showed that the GIP-(1-42)Ala(6) was the only analogue to show insulin-secreting activity similar to that of the native GIP. However, when compared with glucose, its insulinotropic ability was reduced. For the first time, these NMR and modeling results contribute to the understanding of the structural requirements for the biological activity of GIP.     

Major contributions of comparative endocrinology to the development and exploitation of the incretin concept

An incretin is a factor released by the gut in response to nutrients that facilitates uptake of glucose by peripheral tissues. The incretin concept predates the discovery of insulin but it is now clear that incretins act by stimulating secretion of this hormone. As glucagon has insulin-releasing activity, it was speculated that intestinal glucagon-like immunoreactivity (enteroglucagon) was involved in the incretin effect but it was an achievement in the field of comparative endocrinology that led to the demonstration that the preproglucagon gene encodes the most potent incretin in the human. Characterization of cloned cDNAs encoding two preproglucagons from the Brockmann body of the anglerfish Lophius americanus demonstrated that the glucagon sequence is flanked by a 34 amino-acid-residue sequence with appreciable structural similarity to glucagon that was termed glucagon-like peptide (GLP). A 36 amino-acid-residue ortholog of anglerfish GLP was subsequently identified in human preproglucagon but this peptide had only weak insulin-releasing activity. However, alignment of GLP sequences from human and teleost fish showed that the human ortholog is extended from its N-terminus by a hexapeptide. Removal of this extension by an endogenous protease generates GLP-1-(7-36)amide, the potent and effective form of the incretin. More recently, comparative endocrinology has contributed to the exploitation of incretins as antidiabetic drugs. Exendin-4, a GLP-1 receptor agonist first isolated from the venom of the Gila monster Heloderma suspectum, is a clinically valuable, long-acting incretin and the skins of several species of frogs synthesize potent insulin-releasing peptides with therapeutic potential.     

SIMPLE: implementation of recommendations from international evidence-based guidelines on caesarean sections in the Netherlands. Protocol for a controlled before and after study

Background

Caesarean section (CS) rates are rising worldwide. In the Netherlands, the most significant rise is observed in healthy women with a singleton in vertex position between 37 and 42 weeks gestation, whereas it is doubtful whether an improved outcome for the mother or her child was obtained. It can be hypothesized that evidence-based guidelines on CS are not implemented sufficiently. Therefore, the present study has the following objectives: to develop quality indicators on the decision to perform a CS based on key recommendations from national and international guidelines; to use the quality indicators in order to gain insight into actual adherence of Dutch gynaecologists to guideline recommendations on the performance of a CS; to explore barriers and facilitators that have a direct effect on guideline application regarding CS; and to develop, execute, and evaluate a strategy in order to reduce the CS incidence for a similar neonatal outcome (based on the information gathered in the second and third objectives).

Methods

An independent expert panel of Dutch gynaecologists and midwives will develop a set of quality indicators on the decision to perform a CS. These indicators will be used to measure current care in 20 hospitals with a population of 1,000 women who delivered by CS, and a random selection of 1,000 women who delivered vaginally in the same period. Furthermore, by interviewing healthcare professionals and patients, the barriers and facilitators that may influence the decision to perform a CS will be measured. Based on the results, a tailor-made implementation strategy will be developed and tested in a controlled before-and-after study in 12 hospitals (six intervention, six control hospitals) with regard to effectiveness, experiences, and costs.

Discussion

This study will offer insight into the current CS care and into the hindering and facilitating factors influencing obstetrical policy on CS. Furthermore, it will allow definition of patient categories or situations in which a tailor-made implementation strategy will most likely be meaningful and cost effective, without negatively affecting the outcome for mother and child.

Trial registration

http://www.clinicaltrials.gov: NCT01261676