Argyrophile and argentaffin reactions in individual granules of enterochromaffin cells of reserpine treated guinea pigs

Summary The individual granules of enterochromaffin cells of normal and reserpine treated guinea pigs have been studied by staining slides of the duodenum first by an argentaffin method and subsequently by an argyrophile method. Some argentaffin cells can be shown to contain not only argentaffin granules, but also granules that are purely argyrophile. The relative number of such argentaffin cells is greatly increased following administration of reserpine, as depletion of their 5-hydroxytryptamine content converts argentaffin granules into purely argyrophile ones. On the basis of this finding it is confirmed that the argyrophile granule is merely an argentaffin granule depleted of its 5-HT content, and that the argyrophile (nonargentaffin) and the argentaffin cells represent different phases of a secretory cycle.This investigation was supported by a grant from the Indian Council of Medical Research. I am grateful to Ms. Ciba of India Ltd. for making available reserpine (Serpasil) and solvent for reserpine. It is a pleasure to thank Mr. Anand Parkash for technical assistance and Mr. M. L. Sharma for the photographs.     

Bionomics ofTrioxys indicus [Hym.: Aphidiidae], a parasitoid ofAphis craccivora [Hem.: Aphididae]: 31. Effect of host haemolymph on the numerical response of the parasitoid

The influence of kairomones on the numerical response of the parasitoidTrioxys indicus against its hostAphis craccivora at its varying density was studied. The kairomones (applied as aqueous extract of the host) significantly enhanced the rate of parasitisation and multiplication and the area of discovery of the parasitoid and also the K-values of mortality of the host at all parasitoid densities introduced (1, 2, 4, 8, 12 and 16 parasitoids) into troughs having about 200 hosts. The sex-ratio of F₁ offspring decreased at lower parasitoid densities and remained more or less unchanged at higher parasitoid densities after the application of kairomones. The present findings indicate that if kairomones are applied properly, the number of hosts destroyed by a stimulated parasitoid will be about 200, twice the number reported earlier, thus fewer parasitoids will be needed to regulate an estimated population of the hosts.      

Blood rheology in acute mountain sickness and high-altitude pulmonary edema.

The role of blood rheology in the pathogenesis of acute mountain sickness and high-altitude pulmonary edema was investigated. Twenty-three volunteers, 12 with a history of high-altitude pulmonary edema, were studied at low altitude (490 m) and at 2 h and 18 h after arrival at 4,559 m. Eight subjects remained healthy, seven developed acute mountain sickness, and eight developed high-altitude pulmonary edema. Hematocrit, whole blood viscosity, plasma viscosity, erythrocyte aggregation, and erythrocyte deformability (filtration) were measured. Plasma viscosity and erythrocyte deformability remained unaffected. The hematocrit level was lower 2 h after the arrival at high altitude and higher after 18 h compared with low altitude. The whole blood viscosity changed accordingly. The erythrocyte aggregation was about doubled 18 h after the arrival compared with low-altitude values, which reflects the acute phase reaction. There were, however, no significant differences in any rheological parameters between healthy individuals and subjects with acute mountain sickness or high-altitude pulmonary edema, either before or during the illness. We conclude that rheological abnormalities can be excluded as an initiating event in the development of acute mountain sickness and high-altitude pulmonary edema.     

Chromosomal mapping and nucleotide sequence of a human DNA autonomously replicating sequence

A 1.1-kb human DNA fragment (ARSH1) capable of functioning as a putative origin of replication in yeast cells has been characterized both by in situ hybridization to human metaphase chromosomes and by DNA sequencing. Our hybridization studies show a preferential localization of ARSH1 in chromosome regions 1p34-36 and 2q34-37. DNA sequence analysis indicates that in addition to the consensus sequence required for ARS function in yeast cells, nuclear matrix-associated DNA motifs are also present in the 1.1-kb fragment. These results suggest that ARSH1 sequences may serve as points of anchorage to the nuclear matrix for chromosomes 1 and 2.     

Cloning and expression of ovine placental lactogen

Ovine placental lactogen (oPL) is active in a wide range of GH and PRL assays, a property that it shares with human GH (hGH). In addition, oPL is one of a small number of hormones that bind the human GH receptor with high affinity. In order to compare the sequence of oPL to the sequences of other members of the GH family, full-length cDNA clones have been isolated. These clones predict that the full sequence of oPL contains 198 amino acids preceded by a 38 amino acid signal sequence. The mature oPL sequence includes six cysteine and two tryptophan residues and shows substantially more identity to bovine PL (67%) and oPL (49%) than to mouse (31%) or human (25%) PL or to oGH (28%) or (26%) hGH. Like the natural hormone, oPL expressed in mammalian tissue cells binds with high affinity to a soluble form of the recombinant hGH receptor. Thus, oPL binds to the human receptor in spite of having a sequence that is considerably divergent from hGH. Interestingly, the sequence of oPL differs from hGH at most of the amino acids recently found by mutagenesis studies to be important residues in the binding of hGH to the human receptor.     

Effect of GA3 on the Phytotoxicity of Aldicarb and Carbofuran on Seedling Growth in Capsicum frutescens var. California Wonder and Rate of Root Knot Nematode Infestation

The present investigation dealt with the effect of the two granular systemic nematicides aldicarb and carbofuran, individually and in combination with 100 mg · 1^-1 GA₃, on plant growth and root knot formation in C. frutescens var. California Wonder. Both nematicides and GA₃ were applied as a seed-dip treatment. GA₃ increased seed germination and seedling growth. All concentrations (2 to 20 g a.i. · 1^-1) of aldicarb and carbofuran, when applied independently, reduced seed germination and length of epicotyl and hypocotyl. However, application of these nematicides in combination with 100 mg · 1^-1 GA₃ showed antagonistic behaviour. All the treated seedlings were inoculated with 1000 nematode larvae of Meloidogyne incognita. Vegetative parameters of the plant were studied 15 days after transplantation. A combination of GA₃ (100 mg · 1^-1) with different concentrations of nematicides showed a synergistic effect on treated plants. Root knot index in plants treated with nematicides was found to be reduced with increasing concentrations of both nematicides. Maximum reduction in root knot index was observed at highest dose (20 g a.i. · 1^-1) of both nematicides, when applied in combination with 100 mg · 1^-1 GA₃. Proteolytic activities were decreased in plants treated with both aldicarb and carbofuran. Supplementation of the nematicides with GA₃ resulted in an increase in enzyme activities of the treated plants. Toxic residues increased with increasing concentration of both nematicides, whereas it was reduced at highest concentration of both nematicides when applied in combination with GA₃.     

A study on large radial motion of arteries in vivo

This study analyses the radial periodic motion of an artery which is modelled as a thin cylinder of uniform cross-section subjected to dynamic inner pressure using the theory of finite deformation of elastic materials. The arterial tissue properties (anisotropy, homogeneity and incompressibility) are taken into account in an analysis based on the use of the strain energy function. The validity of the mathematical analysis is illustrated through numerical computation applying the available in vivo data for elastic constants of the canine middle descending thoracic aorta to the expressions for the intramural pressure and circumferential stresses obtained by solving the necessary equation of motion together with the boundary conditions. Results obtained in this study indicate very low stresses which suggest that the arteriosclerosis resulting from high stress gradients is effectively ruled out in this model.     

A comparative study of the epidermis of the common carp and the three Indian major carp

A comparative study has been made of the mucogenic epidermis of the common carp, Cyprinus carpio var. communis, and the three Indian major carps, Catla catla, Labeo rohita and Cirrhina mrigala: on the basis of epidermis structural organization, these species are easily differentiated. The epithelial cells in the superficial layer, as in most fishes, show secretory activity, evidenced by positive histochemical reactions, which is high in C. carpio var. communis, moderate in C. catla and low in L. rohita and C. mrigala. The epithelial cells in the underlying two or three layers also give positive reactions, though their intensity is relatively weak. The mucous cells in C. carpio var. communis are distributed in large numbers arranged in several superimposed layers in the outer regions of the epidermis, whereas in C. catla they are fewer in number and are widely separated in the surface layers as well as in the deeper layers of the epidermis; in both species the mucous cells appear rounded, large, and open on the surface by wide pores. In contrast, in L. rohita and C. mrigala the mucous cells are smaller, restricted mainly to the superficial layer, close together in a single row, and open on the surface by narrow pores. The overall density of mucous cells in L. rohita and C. mrigala, as in C. catla, is much lower than in C. carpio var. communis. In the epidermis of C. carpio var. communis there are a large number of mucous cells, and the few club cells are restricted to the deeper layers. In contrast, in the epidermis of the three Indian major carp the overall density of the mucous cells is much lower and the club cells are very numerous. It is suggested that the high density of club cells compensates an overall low density of mucous cells as an adaptation for an effective defence mechanism. Increased mucus production in the epidermis of C. carpio var. communis, as evidenced by a large number of mucous cells in outer regions and high secretory activity of superficial layer epithelial cells, is associated with increased precipitation of mud held in suspension, needed as an adaptation to the species’peculiar bottom-scooping habits. The varied density of the taste buds in the epidermis of the four carp is associated with their feeding habits.     

Cellular oxidation of lignoceric acid is regulated by the subcellular localization of lignoceroyl-CoA ligases

The acyl-CoA ligases convert free fatty acids to acyl-CoA derivatives, and these enzymes have been shown to be present in mitochondria, peroxisomes, and endoplasmic reticulum. Because their activity is obligatory for fatty acid metabolism, it is important to identify their substrate specificities and subcellular distributions to further understand the cellular regulation of these pathways. To define the role of the enzymes and organelles involved in the metabolism of very long chain (VLC) fatty acids, we studied human genetic cell mutants impaired for the metabolism of these molecules. Fibroblast cell lines were derived from patients with X-linked adrenoleukodystrophy (X-ALD) and Zellweger's cerebro-hepato-renal syndrome (CHRS). While peroxisomes are present and morphologically normal in X-ALD, they are either greatly reduced in number or absent in CHRS. Palmitoyl-CoA ligase is known to be present in mitochondria, peroxisomes, and endoplasmic reticulum (microsomes). We found enzyme-dependent formation of lignoceroyl-CoA in these same organelles (specific activities were 0.32 +/- 0.12, 0.86 +/- 0.12, and 0.78 +/- 0.07 nmol/h per mg protein, respectively). However, lignoceroyl-CoA synthesis was inhibited by an antibody to palmitoyl-CoA ligase in isolated mitochondria while it was not inhibited in peroxisomes or endoplasmic reticulum (ER). This suggests that palmitoyl-CoA ligase and lignoceroyl-CoA are different enzymes and that mitochondria lack lignoceroyl-CoA ligase. This conclusion is further supported by data showing that oxidation of lignoceric acid was found almost exclusively in peroxisomes (0.17 nmol/h per mg protein) but was largely absent from mitochondria and the finding that monolayers of CHRS fibroblasts lacking peroxisomes showed a pronounced deficiency in lignoceric acid oxidation in situ (1.8% of control). In spite of the observation that lignoceroyl-CoA ligase activity is present on the cytoplasmic surface of ER, our data indicate that lignoceroyl-CoA synthesized by ER is not available for oxidation in mitochondria. This organelle plays no physiological role in the beta-oxidation of VLC fatty acids. Furthermore, the normal peroxisomal oxidation of lignoceroyl-CoA but deficient oxidation of lignoceric acid in X-ALD cells indicates that cellular VLC fatty acid oxidation is dependent on peroxisomal lignoceroyl-CoA ligase. These studies allow us to propose a model for the subcellular localization of various acyl-CoA ligases and to describe how these enzymes control cellular fatty acid metabolism.     

SIRIUS. An automated method for the analysis of the preferred packing arrangements between protein groups

Automated methods have been developed to determine the preferred packing arrangement between interacting protein groups. A suite of FORTRAN programs, SIRIUS, is described for calculating and analysing the geometries of interacting protein groups using crystallographically derived atomic co-ordinates. The programs involved in calculating the geometries search for interacting pairs of protein groups using a distance criterion, and then calculate the spatial disposition and orientation of the pair. The second set of programs is devoted to analysis. This involves calculating the observed and expected distributions of the angles and assessing the statistical significance of the difference between the two. A database of the geometries of the 400 combinations of side-chain to side-chain interaction has been created. The approach used in analysing the geometrical information is illustrated here with specific examples of interactions between side-chains, peptide groups and particular types of atom. At the side-chain level, an analysis of aromatic-amino interactions, and the interactions of peptide carbonyl groups with arginine residues is presented. At the atomic level the analyses include the spatial disposition of oxygen atoms around tyrosine residues, and the frequency and type of contact between carbon, nitrogen and oxygen atoms. This information is currently being applied to the modelling of protein interactions.