Binding of [3H]Neurotensin in Human Brain: Properties and Distribution

The binding of [3H]neurotensin to membranes from human brain at 0 degrees C was specific, saturable, and reversible. In the frontal cortex, the equilibrium dissociation constant (KD) for [3H]neurotensin determined from the ratio of rate constants (k-1/k1), saturation isotherms, and inhibition binding experiments was 0.80, 2.0, and 2.0 nM, respectively, and the maximum number of binding sites (Bmax) from the saturation isotherms and the competitive binding experiments was 2.4 and 2.2 pmol/g of tissue, respectively. Hill coefficients for binding were equal to 1, indicating the presence of single, noncooperative binding sites. Inhibition of specific binding of [3H]-neurotensin by several analogs of neurotensin showed that [Gln4]neurotensin and neurotensin(8-13) had the highest affinities for these binding sites in human frontal cortex, with each analog being approximately 13-fold more potent than neurotensin. In addition, these data showed that the carboxy-terminal portion of neurotensin played an important part in the binding of this neuropeptide in human brain, a result described for other species. Regional distribution of binding sites was different from that reported for animal brains. Of the 33 different regions investigated, the uncus and substantia nigra showed the highest specific binding of [3H]neurotensin, whereas such areas as the pineal body, medulla, and corpus callosum had few binding sites.     

Latitudinal clines in the properties of a circadian pacemaker

The circadian rhythm of eclosion activity and its pacemaker were analyzed in a series of latitudinal races of Drosophila auraria ranging from 34.2 degrees to 42.9 degrees N in Japan. The phase of the rhythm (psi EL) to the daily photoperiod (PP) changes as daylength is increased, and the slope of psi EL (PP) changes with latitude. Is is sufficiently greater in the north to cause a phase reversal of northern and southern races on long versus short photoperiods. This reversal is found in assays of the pacemaker's phase (psi PL) as well as that of the rhythm (psi EL). Assay of the pacemaker shows that its period (tau) is longer in northern than in southern races, and that the amplitude of its phase response curve (PRC) is lower in the north. The period of the rhythm in all latitudinal races is longer than 24 hr in short photoperiods (LD 1:23), but is probably less than 24 hr (as an aftereffect of photoperiod) in longer days such as LD 14:10. The observed north-south differences in the phase relation of both pacemaker and rhythm to the light cycle are explained by the latitudinal clines in pacemaker properties and a postulated aftereffect of photoperiod on tau. It is suggested that the latitudinal cline in PRC amplitude has functional significance in conserving the amplitude of the pacemaker's signal to the rest of the system it times. Computer simulation shows that without such a reduction in the perceived light intensity, pacemaker amplitude will be lowered by the increase in duration of the daily light at higher latitudes.     

Feeding behaviour of the Japanese loach,Misgurnus anguillicaudatus (Cobitididae)

The feeding behaviour of a Japanese loach,Misgurnus anguillicaudatus was experimentally analyzed. From observations the behaviour patterns were described. The adult fish responded to a piece of thread soaked with filteredTubifex homogenate (Tubifex solution) and showed feeding behaviour. But they ignored an unsoaked piece of thread. LivingTubifex kept in a glass cell failed to elicit feeding behaviour in the fish. A gauze-covered vessel withTubifex solution buried in the sand substrate released feeding behaviour in fish in its vicinity. It follows that the feeding behaviour of this fish is induced by some chemical stimuli. WhenTubifex solution was introduced at the central area of the plastic vessel with a fish, the fish suddenly started the Feeding-crawl and showed Gulp which was composed of Dig and Twist. Using sand particles, it was shown that intake of materials into the digestive tract occurs only at the moment of Twist and not any other time. The standard feeding patterns ofMisgurnus anguillicaudatus are thus quick inhalation of organic matter together with substrate, occurring at the moment of Twist, which is released at the place where the fish received the appropriate chemical stimuli.Misgurnus anguillicaudatus is regarded as a typical “detritus feeder”.     

Genetic Recombination in Mycobacteria

Evidence for genetic recombination between Mycobacterium smegmatis strain Rabinowitchi (Rab) and strain Jucho or PM5 is presented. Backcrosses of recombinants by either parental strain indicated four different types of mating behavior, suggesting that the mycobacterial compatibilities are controlled by at least two different factors. No sex factor that transfers at a high frequency or that is sensitive to acridine dyes was detected. Analysis of segregation of unselected markers revealed that strain Jucho, or PM5, contributes the majority of alleles in almost all recombinants obtained from different selective media. Efforts to construct linkage maps for the markers employed failed because of ordering ambiguities. Mating medium containing streptomycin prevented genetic recombination when strain Rab was resistant to the antibiotic and Jucho, or PM5, was sensitive, but it did not prevent recombination when Rab was sensitive to streptomycin and Jucho, or PM5, was resistant. Very low frequency of recombinant formation was observed when Jucho, or PM5, had been treated with streptomycin, whereas recombinants were formed at fairly high frequencies when Rab had been treated with the antibiotic, suggesting that the roles of parental strains in zygote formation were not identical. The results suggest a polar transfer of genetic material from Rab to Jucho, or PM5, although an alternative possibility of cell fusion followed by exclusion could not be excluded.     

Synthesis of copper(II) and nickel(II) complexes of N-(hydroxyalkylaminoalkyl)salicylamides

Copper(II) and nickel(II) complexes are prepared of potentially quadridentate ligands (LH₃), N-{2-(2- hydroxyethylamino)ethyl}- (seeH₃), N-{3-(2-hydroxy- ethylamino)propyl}- (steH₃), and N-{2-(3-hydroxypropylamino)ethyl}-salicylamide (setH₃). The nickel complexes Na[Ni(see)] and Na[Ni(set)]·1/2H₂O are diamagnetic and square-planar, in which the ligands act as a quadridentate one coordinated through secondary amino-N, and deprotonated phenolic-O, alcoholic-O, and amido-N atoms. The three copper complexes Na [CuL]·H₂O (L = see, ste, set) with a normal magnetic moment have a similar square-planar structure. In another type complexes Cu(LH)·H₂O (LH = seeH, setH) an alcohol group is not deprotonated. Two isomers are present in Cu(seeH)·H₂O: one has a normal and the other a subnormal magnetic moment. The difficulty of complex formation of steH₃ may be attributed to an unfavourably fused 6-6-5 membered chelate ring with strain.     

Characterization of a C-5,13-Cleaving Enzyme of 13(S)-Hydroperoxide of Linolenic Acid by Soybean Seed

An activity was found in mature soybean seeds (Glycine max L. cv Century) that cleaved 13(S)-hydroperoxy-9(Z),11(E),15(Z)-octadecatrienoic acid (13S-HPOT) into 13-oxo-9(Z),11(E)-tridecadienoic acid and two isomeric pentenols, 2(Z)-penten-1-ol and 1-penten-3-ol. Isomeric pentene dimers were also produced and were presumably derived from the combination of two pentene radicals. 13(S)-Hydroperoxy-9(Z),11(E)-octadecadienoic acid (13S-HPOD) was, by contrast, a poor substrate. Activity with 13S-HPOT increased 24-fold under anaerobic conditions reminiscent of a similar anaerobic promoted reaction of 13S-HPOD catalyzed by lipoxygenase (LOX) in the presence of linoleic acid. However, prior to ion-exchange chromatography, cleavage activity did not require linoleic acid. After separation by gel filtration followed by ion-exchange chromatography, cleavage activity was lost but reappeared in the presence of either linoleic acid or dithiothreitol. Under these conditions cleavage activity was coincident with the activity of types 1 and 2 LOX. LOX inhibitors suppressed the cleavage reaction in a manner similar to inhibition of LOX activity. Heat-generated alkoxyl radicals derived from either 13S-HPOT or 13S-HPOD afforded similar products and yields of 13-oxo-9(Z),11(E)-tridecadienoic acid compared to the enzymic reaction. The product 1-penten-3-ol may be the precursor of the "raw-bean" volatile ethylvinylketone.     

PICOPHYTOPLANKTON BIOMASS IN RELATION TO LAKE TROPHIC STATE AND THE TN:TP RATIO OF LAKE WATER IN JAPAN1

Picophytoplankton biomass and its contribution to total phytoplankton biomass were investigated in relation to the nutrient concentration and total N: total P ratio of the epilimnetic waters of 42 Japanese lakes during the warm season in 1991 (April–October). Picophytoplankton biomass (as chlorophyll a) in meso-, eu-, and hypertrophic lakes was significantly higher than those observed in oligotrophic lakes. However, picophytoplankton biomass increased significantly with increased total P concentrations in all systems excluding hypertrophic lakes. Picophytoplankton contribution to total chlorophyll a content was significantly higher in oligo- and mesotrophic lakes than in eu- and hypertrophic lakes and was inversely correlated with total P concentrations in lake water. Picophytoplankton contribution to the total phytoplankton biomass was positively (r = 0.54, n = 42, P = 0.0003) correlated with the total N: total P ratio of lake waters. Each lake trophic type, with the exception of hypertrophic lakes, showed this trend, although the correlation was not significant. We suggest that picophytoplankton contribution is influenced by the total N: total P ratio rather than lake trophic state; however, picophytoplankton were of little importance in hypertrophic lakes.     

Structural changes of immunoglobulin G oligosaccharides with age in healthy human serum

Age-related changes of IgG N-linked oligosaccharides isolated from normal human serum are reported for 403 individuals (male 227 and female 176), varying in age from 0 to 85 years. The IgG N-linked oligosaccharides were released from the protein by digestion with a glycoamidase and reductively aminated with the fluorescent reagent, 2-aminopyridine. The mixture of pyridylaminated oligosaccharides was separated at high resolution by HPLC using a reverse-phase column. From the results of neutral oligosaccharide analysis, agalactosyl glycoform and bisecting GlcNAc-containing glycoform were shown to increase with increasing age. Spearman's correlation coefficients were 0.503 and 0.473, respectively. Thus, in healthy people, an increase of both types of glycoforms correlates weakly with age. In addition, differences were demonstrated between male and female groups in their twenties. The quantity of agalactosyl glycoform was found to be lower in females than in males. No significant differences, however, were observed in the quantity of bisecting GlcNAc-containing glycoforms between males and females. Abbreviations: Gal, D-galactose: GlcNAc, N-acetyl-D-glucosamine; Man, D-mannose; Fc, C-terminal half of the heavy chain dimers of IgG; HPLC, high-performance liquid chromatography; IgG, immunoglobulin G; ODS, octadecylsilyl; PA, pyridylamino This revised version was published online in November 2006 with corrections to the Cover Date.     

Cyclosporine A inhibits the secretion of certain anterior pituitary hormones in patients with nephrotic syndrome.

To clarify the effects of cyclosporine A (CsA) on the secretion of serum thyrotropin (TSH), prolactin (PRL), luteinizing hormone (LH) and follicular stimulating hormone (FSH), we performed TRH and LH-RH testing in 4 patients with the nephrotic syndrome before and after the administration of CsA, 6 mg/kg/day for 4 to 12 weeks. Prior to CsA all patients responded normally to TRH with respect to TSH and PRL secretion. Two patients showed normal response of LH and FSH to LH-RH stimulation while the response in 2 other patients, who were both menopausal, was exaggerated. By the third or fourth week of CsA administration the basal and peak TSH and PRL values declined significantly in all patients in response to TRH stimulation while those of LH and FSH showed only a modest decrease in response to LH-RH stimulation. Two to 4 weeks after the cessation of CsA the response of TSH, PRL and FSH returned to the pretreatment level. These observations suggest that: 1) CsA exerts an inhibitory effect on the secretion of at least TSH and PRL in humans, and 2) the effect of CsA on the pituitary may be partially reversible after the cessation of the therapy.     

Antitumor action of an acidic glycoprotein (SAGP) fromStreptococcus pyogenes in mice

We have shown that an acidic glycoprotein (SAGP) isolated from a cell-free extract ofStreptococcus pyogenes (Su strain) prolonged the life-span of Ehrlich ascites carcinoma (EAC)bearing mice. The present study shows that the life-span prolonging effect of SAGP in EAC-bearing mice was reduced by whole body X-irradiation before EAC inoculation. SAGP (500g protein/mouse/day X 4, i.p.) also showed a life-span prolonging effect (T/C (%) = 169) on Meth A fibrosarcoma (Meth A)-bearing mice, but the effect of SAGP was abrogated by an i.p. pretreatment of the host with carrageenan, an antimacrophage agent. The spleen cells from the Meth A-inoculated and SAGP-treated mice were found to have a considerable cytostatic activity by a³H-thymidine incorporation assay. But the activity disappeared in the presence of carrageenan. These results suggest that thein vivo antitumor effects of SAGP are mediated through its immunomodulating action.