Under-carboxylated osteocalcin regulates glucose and lipid metabolism during pregnancy and lactation in rats.

PURPOSE: Under-carboxylated osteocalcin (UcOC), a bone-released hormone is suggested to regulate energy metabolism. Pregnancy and lactation physiological conditions that require high levels of energy. The current study attempts to examine whether UcOC is involved in regulating energy metabolism during these conditions using adult Wistar rats. METHODS AND RESULTS: Insulin tolerance tests indicated insulin resistance during late pregnancy (day 19 of pregnancy; P19) and insulin sensitivity during early lactation (day 6 of lactation; L6). Gene expression analyses suggested that muscle glucose metabolism was downregulated during P19 and enhanced during L6. Concomitantly, circulatory UcOC levels were lower during pregnancy but higher during early lactation; the rise in UcOC levels was tightly linked to the lactation process. Altering endogenous UcOC levels pharmacologically with warfarin and alendronate in P19 and L6 rats changed whole-body insulin response and muscle glucose transporter (Glut4) expression. Glut4 expression can be increased by either UcOC or estrogen receptors (ERs), both of which act independent of each other. A high fat diet decreased UcOC levels and insulin sensitivity in lactating rats, suggesting that diet can compromise UcOC-established energy homeostasis. Gene expression of lipid metabolism markers and triglyceride levels suggested that UcOC suppression during early pregnancy is an essential step in maternal lipid storage. CONCLUSION: Taken together, we found that UcOC plays an important role in energy homeostasis via regulation of glucose and lipid metabolism during pregnancy and lactation.

Staying Safe from Radiation Exposure in Cath Lab: A Review.

A successful cardiac catheterization Laboratory radiation safety program must manage patient and staff safety by reducing exposure to ionizing radiation to a level that is as low as reasonably achievable (ALARA). The first step in radiation safety is to avoid unnecessary use of ionizing radiation by justification of exposure, one of the basic principles of radiation protection. All individuals involved in fluoroscopic imaging procedures should familiarize themselves to the measures to protect themselves from exposure to unsafe doses of ionizing radiation.

Motility of the oocyte of Helisoma (Mollusca).

Mature oocytes isolated from the ovotestis of Helisoma showed amoeboid movement when treated in vitro with a homogenate of the cerebral ganglia (containing dorsal bodies). This movement ceased when either cytochalasin B or colchicine was added to the preparation. Actin filaments were only visualized in in vitro and in vivo preparation of mature oocytes. We suggest tht in order to ovulate, the oocyte uses its own contractile system to become amoeboid and is thereby separated from the envelope of the follicle cells. We also suggest that the ovulation factor(s) is present in the dorsal bodies and that this factor could be responsible for the induction of microfilaments.

FMRFamide-related peptides, partial serotonin depletion, and osmoregulation in Helisoma duryi (Mollusca: Pulmonata).

Serotonergic neurons were studied by specific histological methods, and neurons containing Phe-Met-Arg-Phe-NH2 (FMRFamide)-related heptapeptides were identified with an antiserum specific for these substances in the central nervous system of the freshwater snail Helisoma duryi. Serotonergic neurons and their axons are present in all of the ganglia (paired buccal, cerebral, pedal, pleural, parietal, and single visceral) and major nerves of the central nervous system. Large neurons containing FMRFamide-related peptide immunoreactivity are located in the left parietal and visceral ganglia, whereas a few small neurons are located in the cerebral and pedal ganglia. Both serotonergic and FMRFamide-related peptide-immunoreactive dendrites and varicosities were observed in the kidney. A second antiserum with high affinity for FMRFamide-related heptapeptides was used to measure the levels of the immunoreactive material in various tissues, and such material was found in every tissue analyzed. When snails were exposed to a medium isosmotic to their hemolymph, the levels of immunoreactive FMRFamide-related peptides increased in the hemolymph, central nervous system, mantle, and kidney. Injection of dihydroxytryptamine, which is known to deplete serotonin content in the snail, also reduced the levels of FMRFamide-related-immunoreactive material in the above tissues. Therefore, serotonin may influence the levels of FMRFamide-related peptides in tissues by regulating the rate of their synthesis, axonal transport, or release. Both serotonin and FMRFamide-related peptides could be involved in osmoregulation.

FMRFamide-related peptides from the kidney of the snail, Helisoma trivolvis.

Three FMRFamide-related peptides (FaRPs) were purified and characterized from the kidney of the snail, Helisoma trivolvis, by HPLC and detected using two radioimmunoassays (RIA) for FaRPs. Automated sequencing and mass spectrometry of the isolated peptides suggest the following sequences: Phe-Met-Arg-Phe-NH2 (FMRFamide), Phe-Leu-Arg-Phe-NH2 (FMRFamide), and Gly-Asp-Pro-Phe-Leu-Arg-Phe-NH2 (GDPFLRFamide). The FaRPs, predominantly the heptapeptides, were also detected by HPLC and RIA in other osmoregulatory tissues such as the skin, mantle, and the hemolymph. The level of FaRPs, detected by radioimmunoassay, appears to be lower in snails kept under hyposmotic stress than in snails kept under isosmotic stress. The FaRPs appear to be involved in osmoregulation in H. trivolvis.

Extracellular application of cobalt: a fast and simple method for delineating invertebrate neurosecretory pathways.

The extracellular cobalt backfilling technique was shown to be an excellent method to obtain cobalt backfills of invertebrate neurosecretory cells (NSCs). Aqueous cobalt was placed in an extracellular suction electrode into which a portion of a neurohaemal (NH) region containing the axons and/or terminals of NSCs was drawn. Spontaneously discharging extracellular action potentials were recorded as the cobalt was applied to the NH region, and the greater the electrical activity, the more extensive the cobalt backfilling. The greatest success occurred with 0.5 M cobalt chloride dissolved in physiological saline. No backfills were obtained in the absence of electrical activity. This technique was shown to backfill NSCs terminating in NH regions of the insect, Rhodnius prolixus, the isopod crustacean, Oniscus asellus, and the freshwater pulmonate snail, Helisoma. Combined with a light insensitive silver intensification method, this paper describes a relatively fast and simple method for delineating invertebrate neurosecretory pathways.

Insulin-like peptide(s) in the central nervous system of the snail Helisoma duryi.

The central nervous system of the snail Helisoma duryi contains porcine insulin-immunoreactive cells. Most of these cells are known as neurosecretory mediodorsal cells (MDC), and are involved in regulation of growth. There are about 25-40 large and small MDC in each cerebral ganglion near the commissure. Besides the MDC, 6-8 insulin-immunoreactive cells are also seen in the left parietal ganglion and the visceral ganglion. Insulin-immunoreactive material can be traced from the MDC perikarya into their axons in the median lip nerve. The elementary granules of the MDC are 150-250 nm in diameter, and are insulin-immunoreactive. Insulin-like material is released by the CNS in vitro, and can be measured by radioimmunoassay using an antiserum against human insulin. High potassium and 4-aminopyridine increase such release in vitro. It is likely that insulin-like peptide is a growth hormone in H. duryi.

Effect of vanadium(IV) compounds in the treatment of diabetes: in vivo and in vitro studies with vanadyl sulfate and bis(maltolato)oxovandium(IV).

Vanadyl sulfate (VOSO(4)) was given orally to 16 subjects with type 2 diabetes mellitus for 6 weeks at a dose of 25, 50, or 100 mg vanadium (V) daily [Goldfine et al., Metabolism 49 (2000) 1-12]. Elemental V was determined by graphite furnace atomic absorption spectrometry (GFAAS). There was no correlation of V in serum with clinical response, determined by reduction of mean fasting blood glucose or increased insulin sensitivity during euglycemic clamp. To investigate the effect of administering a coordinated V, plasma glucose levels were determined in streptozotocin (STZ)-induced diabetic rats treated with the salt (VOSO(4)) or the coordinated V compound bis(maltolato)oxovandium(IV) (abbreviated as VO(malto)(2)) administered by intraperitoneal (i.p.) injection. There was no relationship of blood V concentration with plasma glucose levels in the animals treated with VOSO(4), similar to our human diabetic patients. However, with VO(malto)(2) treatment, animals with low plasma glucose tended to have high blood V. To determine if V binding to serum proteins could diminish biologically active serum V, binding of both VOSO(4) and VO(malto)(2) to human serum albumin (HSA), human apoTransferrin (apoHTf) and pig immunoglobulin (IgG) was studied with EPR spectroscopy. Both VOSO(4) and VO(malto)(2) bound to HSA and apoHTf forming different V-protein complexes, while neither V compound bound to the IgG. VOSO(4) and VO(malto)(2) showed differences when levels of plasma glucose and blood V in diabetic rodents were compared, and in the formation of V-protein complexes with abundant serum proteins. These data suggest that binding of V compounds to ligands in blood, such as proteins, may affect the available pool of V for biological effects.

Immunocytochemical localization of FMRFamide in the central nervous system and the kidney of Helisoma duryi (Mollusca): its possible antidiuretic role.

The distribution of FMRFamide-immunoreactive neurons in the central nervous system of the freshwater pulmonate, Helisoma duryi is described. All parts of the central nervous system except the two pleural and the right parietal ganglia, contain immunoreactive neurons. By immunogold techniques, only one kind of neurosecretory FMRFamide-immunoreactive cell (previously identified as the type-3 cell) was localized in the visceral and left parietal ganglia. This cell type has been previously implicated in an antidiuretic role. FMRFamide-immunoreactive material is found in the whole mount of the kidney as well as in kidney sections. Electron microscopic examination shows that the axons innervating either the smooth muscles of the kidney or the kidney itself contain neurosecretory granules morphologically similar to type-3 cells of the visceral and left parietal ganglia. When incubated in saline containing nanogram quantities of FMRFamide, the wet weight of the kidney increased. It is suggested that FMRFamide-like substance may function as an antidiuretic factor and that the kidney is a target organ of this peptide for osmoregulation.

A study on the cytoplasmic granules of the pericardial gland cells of some bivalve molluscs.

The pericardial glands of three bivalve molluscs are composed of convoluted epithelium that appears as pouches on the auricles of Mytilus and as tubules in the connective tissue at the anterior-lateral sides of the pericardial cavity of Mercenaria and Anodonta. The pericardial gland cells are attached to each other by many randomly placed desmosome-like cell junctions and gap junctions. Belt-desmosomes that are characteristic of epithelial cells were not observed. The basal membrane of these cells is invaginated producing complex interdigitating cytoplasmic processes and filtration slits. The pericardial gland cells stain for the presence of iron with Prussian blue stain. Electron-dense and electron-lucent granules of various diameters are present in the cytoplasm. Many electron-dense granules contain ferritin-like particles in which the presence of iron has been demonstrated by microanalysis. It is suggested that these particles are the iron storage protein ferritin since they contain iron, and are water soluble, heat stable, and morphologically similar to mammalian ferritin. Ferritin particles are probably both synthesized and broken down by the pericardial gland cells; thus the pericardial gland cells may be involved in iron homeostasis in these molluscs.

Cell junctions in the endocrine dorsal body cells of Helisoma duryi (Mollusca: Pulmonata): an in vitro study.

The endocrine dorsal bodies of gastropod molluscs regulate reproduction and are closely associated with the central nervous system. Previous studies on Helisoma duryi have shown that the dorsal body cells of reproducing snails contain more gap junctions than those of non-reproducing virgin snails. More dorsal body cells were isolated from virgin snails than reproducing snails in Ca(2+) and Mg(2+) free experimental saline. The isolated spherical cells attached to the culture dish, spread and formed filopodium-like processes within a few hours in culture medium containing Ca(2+) and Mg(2+). Many isolated cells reaggregated after 4-6 h in culture forming septate junction-like and gap junction-like cell contacts, as revealed by thin section and freeze-fracture studies. Following 10 min incubation in carboxyfluorescein diacetate the isolated cells fluoresced and, after aggregation, these cells transferred fluorescent dye to unlabelled cells. Cell aggregation was inhibited by cytochalasin D. Staining by NBD-phallacidin revealed the presence of actin in the filopodium-like processes of spread cells and in the perinuclear cytoplasm. It is likely that the septate junction-like contacts provide sites of cell attachment between aggregating cells; gap junctions are involved in intercellular communication, and actin is required in this process.

An electron microscopic study of the endocrine dorsal bodies in reproductively active and inactive Siphonaria pectinata (Pulmonata: Mollusca).

The fine structure of the dorsal bodies of the pulmonate limpet Siphonaria pectinata is described in the context of female reproduction involving egg production. In reproductively-active (egg-laying) animals, the ciliated dorsal body cells are filled with lipid droplets and mitochondria. Gap junctions are commonly seen between the cells. The Golgi complexes and the smooth endoplasmic reticulum constitute the other prominent cell organelles. In reproductively-inactive (non-egg-laying) animals, there is a significant reduction in the number of lipid droplets and evidence of reduced synthetic activity in the dorsal bodies. About 12 dorsal body cells are present immediately underneath the perineurium of each cerebral ganglion of the central nervous system. These internal cells are structurally similar to those outside the central nervous system. Cell processes of some of these cells exit the central nervous system at a minimum of three locations on each side and they come in close proximity to the dorsal body cells outside the cerebral ganglia. Like the external cells, the internal cells also communicate via gap junctions and exhibit structural differences according to whether or not the animals are reproductively active. The dorsal body cells, inside and outside the central nervous system, appear to be innervated by neurosecretory axons suggesting neuronal control of dorsal body activity.

Osmoregulation and FMRFamide-related peptides in the salt marsh snail Melampus bidentatus (Say) (Mollusca: Pulmonata).

The pulmonate snail Melampus bidentatus occupies the high intertidal zone of salt marshes in a nearly terrestrial environment. The hemolymph osmolarity of the snails collected in the field paralleled that of the adjacent water and was affected by the tides and precipitation. The snails initially gained or lost weight when submerged in hypo- or hyperosmotic media, respectively, but returned to their original weight after 24 h. The content of their immunoreactive (IR)-FMRFamide-Related Peptides (FaRPs) was measured in various tissues by radioimmunoassay, and IR-FaRPs were found in every tissue analyzed. The subesophageal part of the central nervous system (CNS) contained more IR-FaRPs than the supraesophageal part, and the kidney and the tissues of the reproductive tract contained more than other peripheral tissues. The levels of IR-FaRPs in the CNS, kidney, and hemolymph were higher in snails that were immersed in higher concentrations of seawater. Many IR neurons are present in all ganglia of the CNS except the pleural ganglia, and IR neurites are extensively distributed within the CNS and its connective tissue sheath. The visceral nerve from the visceral ganglion is immunoreactive and could be seen to innervate the kidney, which contains IR-varicosities. An osmoregulatory role for the FaRPs is suggested.