Expression of Glial and Vimentin Type Intermediate Filaments in Cultures Derived from Human Glial Material

Several cultures established from biopsies of apparently normal adult human glial material showed no cells positive for glial fibrillary acidic protein (GFA) when examined after seven or more cumulative population doublings (CPD), although the established glioma line U251 MG showed ∼3% GFA-positive cells, and U333 CG/343 MG clone 3 showed >98% GFA-positive cells. Both the human glia derived cultures and the glioma lines were positive when assayed with sera specific for vimentin. We therefore investigated the expression of GFA as a function of cumulative population doublings after the establishment of primary cultures. Under our experimental conditions, although GFA-positive cells were clearly present in the primary cultures accounting for some 3%–14% of the cells present, the GFA marker was subsequently lost, and the proliferating cultures expressed only the vimentin type of intermediate filament. Those cells that were GFA-positive also appeared to be vimentin-positive. GFA expression was not reinduced in cultures that had lost the GFA marker by treatment with dibutyryl cyclic AMP. We discuss two alternate hypotheses for the origin of the GFA-negative cells: (1) the cultures are of astrocyte origin but lose the ability to express GFA on culturing; (2) the cultures originate from cells of nonastrocyte origin present in the primary material.     

Non-axenic and Axenic Growth of Vorticella microstoma

SYNOPSIS. Vorticella microstoma was grown non-axenically and axenically at pH 6.4. Vorticellas were maintained indefinitely on Bacillus cereus in a medium composed of Proteose-Peptone, Cerophyl, and the filtrate from boiled wheat kernels. Prolific growth occurred in 2-membered cultures. A medium containing hydrolyzed gelatin, aqueous liver extract, yeast nucleic acid hydrolysate, glucose, and penicillin is recommended for axenic growth.
The potential value of vorticellids as research tools is discussed together with metabolic implications of supplementing sterile Proteose-Peptone broth with natural substances in particle form. The ineffectiveness of adding tryptophan, thiamine, glycine, and chelating agents to the axenic medium was considered. Refinements of the axenic medium are on trial.     

Studies on the morphology and ultrastructure of the Malpighian tubules of Halobiotus crispae Kristensen, 1982 (Eutardigrada)

The excretory and osmoregulatory system of Halobiotus crispae consists of two lateral and one smaller dorsal Malpighian tubules, which empty into the digestive tract in the transition zone of the midgut and rectum. The tubules are identical at the ultrastructural level, and consist of an initial segment with three large cells, a thin transitional distal part lacking a nucleus, and a proximal part with 9–12 nuclei. The initial segment possesses deep basal infoldings and interdigitating, finger-shaped processes of the plasma membrane, large mitochondria and giant nuclei. The distal part is a short section which supports the initial segment. Cellular offshoots from the succeeding proximal part constitute the distal part. The distal and proximal parts contain intercellular canals with concretions of variable size. The exit of the proximal part into the digestive tract is characterized by the presence of microvilli. Correlated with the different stages in the cyclomorphosis of H. crispae , we observed size variation of the Malpighian tubules; thus, pseudosimplex stages have the largest tubules. We present suggestions concerning the physiology of the tubules and compare the Malpighian tubules of Tardigrada with the Malpighian papillae of Protura.     

Eutherian phylogeny from a primate perspective

Determinant formulas for up to 22 serum proteins from nineteen species in seven Eutherian orders (Chiroptera, Carnivora, Artiodactyla, Perrissodactyl, Rodentia, Proboscidea, Xenarthra) and Metatheria were derived by comparative determinant analysis from a primate (human) perspective. Plesiomorphic character states for taxon-pairs were inferred, including epitope patterns of the putative last common ancestor of Primates and non-primate Eutheria. Our approach, comparative determinant analysis, is exceptional among immunological methods in being able to define molecular character states for cladistic systematics. Average degrees of phylogenetic conservation of protein determinants varied between loci by 6.5-fold. Against the background of possible lineage-specific rates of molecular substitution, the clear immunological differentiation of the Asiatic elephant from (other) ungulates is discussed as to its phylogenetic relevance, as are the well-marked distances between guinea-pig and myomorph rodents, fissipeds and harbour seal, Mega- and Microchiroptera, or pig and ruminants. It is concluded that genetic drift influenced the rate of molecular evolution of mammalian orders to a different extent, e.g. more in the Proboscidean lineage than in the clade leading to the horse. It is concluded that the substitution rate of antigenic determinants of serum proteins correlates positively with speciation frequency in a phyletic lineage.     

Comparison of Postembryonic Organization of the Genital Segments in Trichoceridae, Tipulidae, and Anisopodidae (Diptera, Nematocera)

The number of abdominal segments in Tipulomorpha and Bibionomorpha larvae and aduts is discussed. For Nematocera, the most primitive number of abdominal segments in both male and female larvae is nine. Reduction of the IX abdominal segment and its subsequent fusion with the VIII segment occurs in different phyletic lines in Nematocera and might have evolved several times. In Trichocera spp. nine abdominal segments are present. In the genital segments the main interrelationships in the position of the anus and some main innervation areas, especially the ventral lobes, and the derivatives of the genital primordia were followed during postembryonic reorganization by studying variously stained serial sections from all developmental stages from the first larval instar to the adult stage. Homologies between male and female derivatives of the IX segment genital primordium were established for Trichoceridae. The trichoceroid male claspers and female ovipositor were found to be of sternal origin and highly specialized structures. They appear to be unique features of the Tipulomorpha. Postembryonic development in Limonia nubeculosa Meigen, 1804 and Sylvicola cinctu (Fab-ricius, 1787) was studied in the same way. In Limonia males the trichoceroid functional system for grasping is present. In Anisopodidae ( Sylvicola ), another functional system for grasping has been evolved by the male, which only includes primordial derivatives. In the adults, fusion of the VIII and IX segments prevents development of outer clasping organs or special structures for egg guidance.     

Temperature and the metabolic balance of streams

1. It is becoming increasingly clear that fresh waters play a major role in the global C cycle. Stream ecosystem respiration (ER) and gross primary productivity (GPP) exert a significant control on organic carbon fluxes in fluvial networks. However, little is known about how climate change will influence these fluxes. 2. Here, we used a ‘natural experiment’ to demonstrate the role of temperature and nutrient cycling in whole‐system metabolism (ER, GPP and net ecosystem production – NEP), in naturally heated geothermal (5–25 °C) Icelandic streams. 3. We calculated ER and GPP with a new, more accurate method, which enabled us to take into account the additional uncertainties owing to stream spatial heterogeneity in oxygen concentrations within a reach. ER ranged 1–25 g C m^?2 day^?1 and GPP 1–10 g C m^?2 day^?1. The median uncertainties (based on 1 SD) in ER and GPP were 50% and 20%, respectively. 4. Despite extremely low water nutrient concentrations, high metabolic rates in the warm streams were supported by fast cycling rates of nutrients, as revealed from inorganic nutrient (N, P) addition experiments. 5. ER exceeded GPP in all streams (with average GPP/ER = 0.6) and was more strongly related to temperature than GPP, resulting in elevated negative NEP with warming. We show that, as a first approximation based on summer investigations, global stream carbon emission to the atmosphere would nearly double from 0.12 Pg C year^?1 at 13 °C to 0.21 (0.15–0.33) Pg C year^?1 with a 5 °C warming. 6. Compared to previous studies from natural systems (including terrestrial ecosystems), the temperature dependence of stream metabolism was not confounded by latitude or altitude, seasonality, light and nutrient availability, water chemistry, space availability (water transient storage), and water availability. 7. Consequently, stream nutrient processing is likely to increase with warming, protecting downstream ecosystems (rivers, estuaries, coastal marine systems) during the summer low flows from nutrient enrichment, but at the cost of increased CO₂ flux back to the atmosphere.     

Piscivory and diet overlap between two non‐native fishes in southern Chilean streams

Trophic relations among introduced species may induce highly variable and complex effects in communities and ecosystems. However, studies that identify the potential impacts for invaded systems and illuminate mechanisms of coexistence with native species are scarce. Here, we examined trophic relations between two introduced fishes in streams of NW Patagonia, rainbow trout (Oncorhynchus mykiss) and brown trout (Salmo trutta). These species originate from different regions of the Northern Hemisphere but they now coexist as invading species over the world. We used gastric contents and stable isotopes analysis to compare the diets of two size‐classes of these two invaders in three localities of southern Chile. Both species displayed similar ontogenic diet shifts with smaller trout consuming mostly invertebrates and larger trout being more piscivorous and epibenthic feeders. However, piscivory was more prevalent in brown trout than in rainbow trout and highest at the site with the greatest density of native fishes suggesting that the availability of native fishes as trout prey may limit the occurrence of trout piscivory. We found an elevated dietary overlap between the two trout species at larger sizes while at smaller size a higher intraspecific dietary overlap occurred suggesting a potential interference competition among the two fish invaders especially at larger sizes. Our results highlight that the impacts of invading species on non‐native fishes are context specific (i.e. species and ontogenic stages) and thus, difficult to generalize.     

Enamel ultrastructure and masticatory function in molars of the American opossum, Didelphis virginiana

Maxillary and mandibular molars of the American opossum, Didelphis virginiana L., were viewed in the scanning electron microscope (SEM) after acid-etching or after cutting and acid-etching. Observations were made on enamel prism patterns as they relate to functional properties of the tooth at a particular site. Molars at different stages of wear were also observed under a dissecting microscope; worn surfaces were correlated with function and enamel ultrastructure. Pounding surfaces of molar cusps wear more rapidly than near-vertical shearing surfaces or crushing basins (i.e. the trigon and talonid basin). Pounding surfaces are subjected to abrasion by food and arc not normally involved in tooth-tooth contact. Near-vertical shearing surfaces and basins used for crushing do experience tooth-tooth contact, but are surprisingly more resistant to wear. Prisms at pounding sites approach the occlusal surface at a near 90° angle and are surrounded with very thick interprismatic (IP) enamel parallel to the occlusal surface of the tooth. The pounding pattern is present at tips of cusps and at occlusal surfaces of ridges of the tooth. At near-vertical shearing surfaces, the prisms approach the outer surface obliquely and are surrounded with IP crystals which are perpendicular to the vertical surface. The angle between prismatic and IP enamel in these patterns is 60–90° in a cervical to occlusal direction. In basins of the tooth used principally for crushing and some shearing, IP enamel is perpendicular to the changing slope of the basin and the prisms are usually at a 55–65° angle to the IP enamel. When the pounding and shearing-crushing patterns meet at a ridge, a distinct seam is observed. Pounding forces occur parallel to the long axis of the prisms and perpendicular to the thick IP enamel (i.e. perpendicular to the long axis of the IP crystals) lying on either side of the prisms. Shearing and crushing forces occur at an oblique angle to the prism, and interprismatic enamel is more evenly distributed about the prism. A spiral pattern is found at the bottoms of the trigon and talonid basins, but not at the bottom of the trigonid which is a non-occluding basin. It is concluded that the differential rates of wear of the enamel surfaces are necessary in maintaining the sharp cutting edges and effective crushing basins of the tribosphenic molar, and the ultrastructural arrangements of the enamel prisms are of functional significance.     

Isoenzyme Comparison of Axenic Giardia lamblia Strains

ABSTRACT We obtained isoenzyme patterns by polyacrylamide gradient gel electrophoresis (PGGE) of water-soluble protein fractions prepared from trophozoites of 11 axenic G. lamblia strains. The strains were isolated from animals and humans (both symptomatic and asymptomatic) from various geographic locations. Isoenzymes were also separated by isoelectric focusing. Of 12 enzymes attempted, eight exhibited well-defined and reproducible isoenzyme patterns by PGGE, based on which the strains were grouped into four zymodemes. Although the 11 strains were grouped into four zymodemes based on PGGE, no correlation between zymodeme and the known characteristics of the strains existed. Thus, a high degree of characteristic sharing appears to occur among genetically different G. lamblia strains.