A ribonucleoprotein inclusion body in Entamoeba invadens

Summary Cytochemical tests have shown that the chromatoid bodies contain mainly ribonucleic acid and protein. There is no evidence to suggest the presence of any desoxyribonucleic acid, lipid, glycogen, neutral fat, or metaphosphate. The cyclic changes in the chromatoid body have been studied by means of light and electron microscopy and estimations of the total RNA present. The results indicate that the chromatoid bodies arise by a process of aggregation of small groups of 250–300 A units form polycrystalline masses in precystic and early cysts. At the same time there is a steady rise in the amount of RNA present. In the maturing cysts, the crystalline masses fragment into separate particles, but there is no corresponding fall in the amount of RNA. The significance of these results in relation to the previous literature on chromatoid bodies and similarly named cellular inclusions in spermatogenic and plant cells, as well as in the protozoa, is discussed. Attention is drawn to the striking similarity of composition, formation, development and ultimate fate of these various inclusion bodies but as the terminology is still confused it is proposed that the term chromatoid body be retained solely for particulate or lamellar ribonucleo-protein accumulations. Acknowledgements. Most of this study was presented in partial fulfillment of the requirements for the degree of Doctor of Philosophy at the University of Edinburgh. I should like to express my deep appreciation to Prof. M. M. Swann F. R. S. for his interest and helpful guidance in this work. I am grateful to the Melville Trust for Cancer Research who provided the electron microscope.     

Calcitonin-like immunoreactivity in the blue crab: tissue distribution, variations in the molt cycle, and partial characterization.

Calcitonin-like immunoreactivity has been found in blood and tissues of the marine blue crab, Callinectes sapidus Rathbun. The activity in blood rises significantly during the immediate premolt (D4) stage, but tissue immunoreactivity does not vary significantly with molt stage. The immunoreactivity to antisalmon calcitonin serum is due to a 27.2 kDa protein that shows strong similarities in amino acid composition to a similar protein in a lobster and to human calcitonin precursor. The protein is most abundant in the midgut glands (hepatopancreas), but is also found in significant quantities in several other tissues. Immunoreactivity in the blood appears to be primarily due to the same molecular weight fraction, with secondary contributions from smaller molecules, closer in size to vertebrate calcitonin. A physiological function of the calcitonin-like substance in calcium transport during or after molt has yet to be demonstrated.     

Proposal for adoption of the base of the Undulograptus austrodentatus Biozone as a global Ordovician stage and series boundary level

The base of the Undulograptus austrodentatus Biozone appears to be a synchronous event that is widely recognizable within graptolitic facies around the world. It occurs within an interval in which graptolite species ranges are now well known and in which there is a rapid turnover in the composition of graptolite faunas. This turnover reflects the rapid evolutionary radiation of the Diplograptacea simultaneously with the appearance of several distinctive pseudisograptid and glossograptid species. These events provide the basis for the recognition of two thin but widely applicable subzones; a lower Arienigraptus zhejiangensis Subzone and an upper U. sinicus Subzone. The occurrence of the lower boundary of the U. austrodentatus Biozone within a succession of first appearances also permits accurate and reliable identification of the boundary as well as assessment of stratigraphic completeness across the boundary interval in correlated sections. Diverse graptolite faunas of late Yapeenian and early Darriwilian age occur in association with the Histiodella altifrons Biozone of the North American midcontinent conodont zonation and the Paroistodus originalis and Microzarkodina parva biozones of the North Atlantic conodont zonation. They also occur in association with the shelly-fossil zonations developed for several different continents. These features of the base of the U. austrodentatus Biozone make it a suitable level for use as the boundary level for a global stage. Its stratigraphic position within the Ordovician System relative to other likely global stages as well as its coincidence with one of the major events in graptolite evolutionary history suggest that this level also may be a suitable level for the base of a global Middle Ordovician Series.Ordovician System, Ordovician stages, graptolite zonation, chronostratigraphy, international correlation. Charles E. Mitchell and Jörg Maletz, Department of Geology, State University of New York at Buffalo, Buffalo, New York 14260-1550, USA; 13th July, 1994; revised 22nd May, 1995.     

Modulation of cisPlatin cytotoxicity by interleukin-1α and resident tumor macrophages

The modulation of cisPlatin cytotoxicity by interleukin-1 (IL-1α) was studied in cultures of SCC-7 tumor cells with and without tumor macrophages to examine potential mechanisms for the synergistic antitumor activity of cisPlatin and IL-1α in SCC-7 solid tumors. Neither IL-1α nor tumor macrophages affected the survival of clonogenic tumor cells and IL-1α had no direct effect on tumor cell growthin vitro. Macrophages had no direct effect on cisPlatin sensitivity (IC₉₀=6.0 μM), but, the addition of IL-1α (500–2000U/ml) to co-cultures of cisPlatin pretreated tumor cells and resident tumor macrophages increased cell killing (IC₉₀=3.1 μM). Similar responses were seen in primary cultures treated with cisPlatin before IL-1α. The modulation of cisPlatin cytotoxicity by IL-1α exhibited a biphasic dose response that paralleled the IL-1α dose dependent release of H₂O₂by resident tumor macrophages. Further, IL-1α modification of cisPlatin cytotoxicity was prompt and inhibited by catalase. CisPlatin and exogenous H₂O₂ (50 μM) produced more than additive SCC-7 clonogenic cell kill and hydroxyl radicals played an important role in the response. Interleukin-1 modulation of cisPlatin cytotoxicity was schedule dependent. IL-1α treatment for 24 hrs, before cisPlatin, produced drug resistance (IC₉₀=11.1 μM). Our study shows that IL-1α can stimulate tumor macrophages to release pro-oxidants that modify cellular chemosensitivity in a schedule and dose dependent fashion. Our findings may also provide a mechanistic explanation for the synergistic antitumor activity of cisPlatin and IL-1αin vivo.     

Deletion of the Leader Peptide of the Mitochondrially Encoded Precursor of Saccharomyces Cerevisiae Cytochrome C Oxidase Subunit II

Cytochrome c oxidase subunit II (Cox2p) of Saccharomyces cerevisiae is synthesized within mitochondria as a precursor, pre-Cox2p. The 15-amino acid leader peptide is processed after export to the intermembrane space. Leader peptides are relatively unusual in mitochondrially coded proteins: indeed mammalian Cox2p lacks a leader peptide. We generated two deletions in the S. cerevisiae COX2 gene, removing either the leader peptide (cox2-20) or the leader peptide and processing site (cox2-21) without altering either the promoter or the mRNA-specific translational activation site. When inserted into mtDNA, both deletions substantially reduced the steady-state levels of Cox2p and caused a tight nonrespiratory phenotype. A respiring pseudorevertant of the cox2-20 mutant was heteroplasmic for the original mutant mtDNA and a ρ(-) mtDNA whose deletion fused the first 251 codons of the mitochondrial gene encoding cytochrome b to the cox2-20 sequence. The resulting fusion protein was processed to yield functional Cox2p. Thus, the presence of amino-terminal cytochrome b sequence bypassed the need for the pre-Cox2p leader peptide. We propose that the pre-Cox2p leader peptide contains a targeting signal necessary for membrane insertion, without which it remains in the matrix and is rapidly degraded.     

Shell variation in sympatric freshwater Lymnaea peregra and L. ovata (Gastropoda: Lymnaeidae)

Snails of the genus Lymnaea are morphologically variable and their taxonomy is unclear. In particular, the forms peregra and ovata , distinguished by shell shape, are often considered variants of the same species, L. peregra. We studied a rare situation in a Swiss mountain lake where both forms are sympatric. First, we found that the forms shows complete separation for a number of allozymes. Second, we examined the response of the two snail forms to infection by the trematode Diplostomum phoxini . For the ovata form, there was a transitory, 10% increase in the growth of infected snails compared to uninfected controls. The peregra form showed no gigantism and had higher parasite-induced mortality. Third, we assessed differences between the forms in reproduction under different environmental conditions. Density negatively influenced egg production to the same degree in both forms. However, decreasing water levels, characteristic of part of the lake studied, led to a 30% decrease in egg production for the ovata form, but only a 10% decrease for the peregra form. These differences are discussed in relation to the microdistributions of the snails in the lake and we conclude that the two forms are almost certainly separate species.     

Molecular evidence that diploid Stylosanthes humilis and diploid Stylosanthes hamata are progenitors of allotetraploid Stylosanthes hamata cv. Verano.

Restriction fragment length polymorphism analysis, using peroxidase, O-methyltransferase, phenylalanine ammonia-lyase, and coniferyl alcohol dehydrogenase cDNAs isolated from Stylosanthes humilis, as probes, provided molecular evidence for the genetic origin of the naturally occuring allotetraploid genotype Stylosanthes hamata cv. Verano (2n = 4x = 40). Hybridization patterns strongly suggest that the likely progenitors of S. hamata cv. Verano were a diploid S. humilis (2n = 2x = 20) and a diploid S. hamata (2n = 2x = 20) species.     

Randomised double blind controlled clinical trial of intranasal budesonide in treatment of hay fever.

The effectiveness of budesonide, a new non-halogenated glucocorticoid administered by nasal inhaler, was evaluated in a double blind comparison with placebo in patients presenting with hay fever. Patients were supplied with antihistamine tablets and eye drops for use when they considered that their symptoms were inadequately controlled by their inhaler. Patients recorded the severity of their symptoms in a daily diary card and visited their general practitioner for assessment weekly for four weeks. All nasal symptoms of hay fever were appreciably reduced in the group taking budesonide and, although their eye symptoms were more severe than in the group taking placebo, they did not use appreciably more eye drops than the latter. The placebo group used appreciably more antihistamine tablets than those in the budesonide group. No patients were withdrawn from the budesonide group because of treatment failure or unacceptable side effects. The results suggest that budesonide is an acceptable and effective treatment for the nasal symptoms of hay fever.