Rat atrial natriuretic factor: Isolation,structure and biological activities of four major peptides

Four peptides possessing both natriuretic activity and smooth muscle relaxant activity were isolated from rat atrium and their amino acid sequences determined. The four peptides designated ANF-I, ANF-II, ANF-III and ANF-IV containing 35, 31, 30 and 25 amino acid residues, respectively, were obtained in a molar ratio of 4:60:20:16. The predominant species ANF-II, which may represent the native form of ANF, has the following sequence: (H₂N)-G-P-R-S-L-R-R-S-S-C-F-G-G-R-I-D-R-I-G-A-Q-S-G-L-G-C-N-S-F-R-Y-(COOH) in which Cys-10 and Cys-26 are disulfide linked. Cleavage of the aspartyl linkage at position 16 by staphylococcal protease caused complete inactivation, indicating that the ring conformation is essential. The dose-response relationships determined for the four pepdides in relaxing norepinephrine-induced contraction of rabbit thoracic aorta showed half-maximum relaxation at concentrations ranging from 1.5 × 10^?9 to 2.5 × 10^?9 M. Comparable dose-response relationships were observed in relaxation of carbacol-induced contraction of chick rectum strips as tested with ANF-II and ANF-IV.     

Soluble, prolonged-acting insulin derivatives. II. Degree of protraction and crystallizability of insulins substituted in positions A17, B8, B13, B27 and B30

It has previously been found that insulins, to which positive charge has been added by substitutions in position B30, thus raising the isoelectric point towards pH 7, had a prolonged action when injected as slightly acidic solutions because such derivatives crystallize very readily upon neutralization. Positive charge has now been added by substituting the B13 and A17 glutamic acid residues with glutamines and B27 threonine with lysine or arginine. These substitutions were introduced by site-specific mutagenesis in a gene coding for a single-chain insulin precursor. By tryptic transpeptidation the single-chain precursors were transformed to the double-chain insulin structure, concomitantly with incorporation of residue B30. Thus insulins combining B13 glutamine, A17 glutamine and B27 lysine or arginine with B30 threonine, threonine amide or lysine amide were synthesized. The time course of blood glucose lowering effect and the absorption were studied after subcutaneous injection in rabbits and pigs. The prolonged action of B30-substituted insulins was markedly enhanced by B27 lysine or arginine substitutions and by B13 glutamine. The B27 residue is located on the surface of the hexamer, so a basic residue in this position presumably promotes the packing of hexamers at neutral pH. The B13 residues cluster in the centre of the hexamer. When the electrostatic repulsive forces from six glutamic acid residues are abolished by substitution with glutamine, a stabilization of the hexamer can be envisaged.(ABSTRACT TRUNCATED AT 250 WORDS)     

Diazepam tolerance: effect of age,regular sedation,and alcohol.

The dose of intravenous diazepam required for sedation was estimated in a series of 78 patients aged 17-85 years given the drug for dental and endoscopic procedures. Multiple regression analysis showed a significant correlation (r = 0.71; p less than 0.001) between dose and age, body weight, the taking of regular sedation, and the taking of more than 40 g alcohol daily, but there were no differences in the doses required between men and women, smokers and non-smokers, inpatients and outpatients, or dental and endoscopy patients. Patients aged 80 required an average dose of 10 mg and patients aged 20 an average dose of 30 mg, and the dose required was much higher in those receiving regular sedation or having a high alcohol intake. Plasma total and free diazepam concentrations were measured in the second half of the series of patients (n = 37). Plasma concentrations required for sedation fell twofold to threefold between the ages of 20 and 80 and were significantly higher in those taking regular sedation or alcohol. Differences in the acute response to diazepam appeared to be due to differences in the sensitivity of the central nervous system (pharmacodynamic tolerance) rather than to differences in pharmacokinetic factors.     

Nitroimidazole conjugates of bis(thiosemicarbazonato)Cu(II) - Potential combination agents for the PET imaging of hypoxia

Combination agents comprising two different pharmacophores with the same biological target have the potential to show additive or synergistic activity. Bis(thiosemicarbazonato)copper(II) complexes (e.g. ⁶⁴Cu-ATSM) and nitroimidazoles (e.g. ¹⁸F-MISO) are classes of tracer used for the delineation of tumor hypoxia by positron emission tomography (PET). Three nitroimidazole-bis(thiosemicarbazonato)copper(II) conjugates were produced in order to investigate their potential as combination hypoxia imaging agents. Two were derived from the known bifunctional bis(thiosemicarbazone) H₂ATSM/A and the third from the new precursor diacetyl-2-(4-N-methyl-3-thiosemicarbazone)-3-(4-N-ethylamino-3-thiosemicarbazone) - H₂ATSM/en. Oxygen-dependent uptake studies were performed using the ⁶⁴Cu radiolabelled complexes in EMT6 carcinoma cells. All the complexes displayed appreciable hypoxia selectivity, with the nitroimidazole conjugates displaying greater selectivity than a simple propyl derivative used as a control. Participation of the nitroimidazole group in the trapping mechanism is indicated by the increased hypoxic uptake of the 2- vs. the 4-substituted ⁶⁴Cu-ATSM/A derivatives. The 2-nitroimidazole derivative of ⁶⁴Cu-ATSM/en demonstrated superior hypoxia selectivity to ⁶⁴Cu-ATSM over the range of oxygen concentrations tested. Biodistribution of the radiolabelled 2-nitroimidazole conjugates was carried out in EMT6 tumor-bearing mice. The complexes showed significantly different uptake trends in comparison to each other and previously studied Cu-ATSM derivatives. Uptake of the Cu-ATSM/en conjugate in non-target organs was considerably lower than for derivatives based on Cu-ATSM/A.     

Cimetidine-resistant Zollinger-Ellison syndrome: successful management with ranitidine.

In a patient with the Zollinger-Ellison syndrome who had undergone vagotomy, pyloroplasty and, later, gastrojejunostomy, epigastric pain and stomal ulceration recurred despite the use of high doses (2700 mg/d) of cimetidine. Ranitidine, a new histamine H2-receptor antagonist, reversed all symptoms and healed the stomal ulcer without side effects, thus obviating the need for further surgery. Ranitidine may prove to be the drug of choice in the medical management of patients with the Zollinger-Ellison syndrome.     

Hexapeptide analogue of somatostatin, Sandoz 201-456. Conformational study in dimethylsulfoxide by 1D and 2D n.m.r. methods

The conformational properties of the somatostatin analogue 201-456 (1) have been studied by high field n.m.r. in DMSO. This analogue is the base structure of nine derivates synthesized by Bauer et al. and shows a very low biological activity, although derived structures such as SMS 201-995 (2) are very potent. Our study has shown an important difference between the most stable conformation of the two compounds: although the beta turn type II' structure at the Phe3-Trp4-Lys5 level is present in both analogues, an important conformational change appears at the cystine bridge. In SMS 201-995 the beta turn/beta sheet conformation is stabilized by the additional amino-acids D-Phe1 and Thr8 (ol) through intramolecular H-bonds.     

Capillary permeability in the pancreas and colon: restriction of exogenous and endogenous molecules by fenestrated endothelia

The permeability properties of fenestrated capillaries in the colon and exocrine and endocrine pancreas to exogenous and endogenous molecules were examined. The exogenous tracers horseradish peroxidase (Einstein-Stokes radius [ESR], 3.0 nm), hemoglobin (ESR, 3.2 nm), and ferritin (ESR, 6.1 nm) were injected intravenously and allowed to circulate for 5-90 min. Tissues were removed and processed for cytochemical or standard electron microscopic examination. The endogenous plasma proteins albumin (ESR, 3.5 nm) and IgG (ESR, 5.5 nm) were localized by immunocytochemistry using the protein A-gold technique. All vessels examined were permeable to HRP in less than 5 min. In contrast, these vessels were restrictive to the slightly larger hemoglobin molecule (60-min circulation) and to ferritin (90-min circulation). Capillaries in the exocrine and endocrine pancreas were restrictive to albumin and IgG. These results demonstrate the presence of fenestrated capillary beds, in addition to the choriocapillaris, that are restrictive to molecules with ESR greater than or equal to 3.2 nm. Capillaries in the mucosa of the colon were restrictive to hemoglobin and ferritin but did not restrict albumin or IgG. This indicates that these vessels are of the permeable type. However, the rate of transendothelial movement of molecules is slower than that of other permeable vessels, such as in the ileo-jejunum. This study has provided further evidence for the existence of fenestrated endothelia that are restrictive to exogenous and/or endogenous molecules.     

Influence of age,weaning, season and body weight on the levels of progesterone,oestradiol-17β and luteinizing hormone in growing buffalo heifers (Bubalus bubalis)

The influence of age, weaning, season of the year and body weight on the peripheral levels of progesterone, oestradiol-17β and luteinizing hormone (LH) were studied during neonatal, perinatal and peripubertal periods in buffalo heifers. The buffalo heifers exhibited oestrus only after 30 months of age and had higher levels of LH and oestradiol-17β and a lower level of progesterone on the day of oestrus. The progesterone concentration was affected significantly (P < 0.01) by different seasons, by weaning (P < 0.05) and varied between pubertal and neonatal periods (P < 0.01), whereas the oestradiol-17β level was affected significantly (P < 0.01) by weaning and varied at different seasons and with body weight. However, the LH concentration was greater during the neonatal period than the pre- and peripubertal periods and changed significantly (P < 0.01) between groups of ages and body weights. The results suggest that increases in the levels of oestradiol-17β and progesterone after 30 months of age are probably indicative of the onset of puberty in buffalo heifers. However, a further increase in oestradiol-17β, LH, and a decrease in progesterone are essential for oestrus and cyclicity to be exhibited in buffalo heifers.     

Propagation of Zea mays L. by Shoot Tip Culture: A Feasibility Study

A procedure for the stimulation of axillary bud developmentfrom young shoots of maize, their subculture to root-inducingmedia and transfer as rooted plants to soil is described. Axillarybud development was enhanced by the addition of kinetin andauxin to the culture medium. Root initiation on explanted axillarybuds, while successful with some cultivars, was variable. Anumber of mature plants with normal tassels and ears were producedfrom the lowermost buds of an original stem explant. Buds fromhigher positions on the explant exhibited different potentialitieswith some, those normally from cob producing nodes, producingshort-stalked plants with terminal female influorescences. Agradient of bud potentiality along the stem appears to be establishedextremely early after each is initiated. Zea mays., corn, maize, shoot tip culture, clone, vegetative propagation