Some Factors Influencing the in vitro Infectivity and Replication of Encephalitozoon cuniculi*

SYNOPSIS. Rabbit Encephalitozoon cuniculi were propagated in vitro using rabbit choroid plexus (RCP) cells. The organisms reached maximum titer and numbers by 15 days. The source and in vitro passage level of RCP cells moderately influenced the sensitivity of the cells to infection. Cells less than 1 week old were significantly less sensitive than older cells. A moderate increase in infectivity for RCP cells was demonstrated with increasing organism passage level in vitro. Rabbit E. cuniculi were not affected by penicillin-streptomycin or gentamicin in the culture medium. The organism survived more than 9 days in buffer at 37 C and least 24 days at 4 and 20 C. Storage at -70 C or in liquid nitrogen was successful for at least 6 months. Encephalitozoon cuniculi survived 60 but not 120 min at 56 C. They were killed after 10 min of autoclaving and by 2% (v/v) Lysol, 10% (v/v) formalin and 70% (v/v) ethyl alcohol. The organisms survived at least 24 h at pH 9 or pH 4 and were not affected by sonication, freezing and thawing, or distilled water but lost significant infectivity after 24 h in CsCl or 40% (w/v) sucrose.     

Heterogeneity of Intermediate Filament Expression in Vascular Smooth Muscle: A Gradient in Desmin Positive Cells from the Rat Aortic Arch to the Level of the Arteria Iliaca Communis

The display of the two distinct intermediate filament proteins, desmin and vimentin, in rat vascular smooth muscle tissue was studied by immunofluorescence microscopy on frozen sections of aorta and other blood vessels. Vascular smooth muscle cells present in these vessels always appeared rich in vimentin. However, staining of sections covering six distinct but contiguous parts of the aorta showed that the number of desmin containing cells was low distal to the truncus brachiocephalicus, but increases until in distal parts of the aorta and in the arteria iliaca communis almost all cells appear positive for desmin. Thus blood vessels show heterogeneity of intermediate filament expression not only in cross-section but can also display heterogeneity along their length. Muscular arteries such as the renal artery and the arteria femoralis, as well as arterioles and veins including the vena jugularis and the vena cava also contain desmin. Thus it may be that low numbers of desmin-positive cells are typical of elastic arteries, while muscular arteries and other blood vessels are characterized by large numbers of desmin-positive cells. We discuss whether desmin-positive and desmin-negative vascular smooth muscle cells may perform different functions and raise the possibility that desmin expression may coincide with the turn on of a specially regulated contractility program.     

The significance of alkaline phosphatase activity in Lough Neagh

SUMMARY. A study of the eutrophic Lough Neagh was undertaken to ascertain the use of alkaline phosphatase activity as an indicator of phosphorus-limited algal growth. Lough Neagh and its surrounding rivers were monitored during 1974 and 1975. Alkaline phosphatase activity increased in the lough when the algal crop increased and depleted orthophosphate to a growth-limiting concentration. Enzyme activity was appreciable, however, at all other times of the year and may on occasions have been associated with zooplankton. The enzyme activity in the inflowing rivers was much greater than in the lough itself. Depending on enzyme stability the enzyme inputs from rivers could contribute significantly to the lough activity. Sewage disposal works effluent appeared to be a major source of enzyme activity in river water. As the alkaline phosphate activity in Lough Neagh resulted from several sources, increased activity could not be used as a reliable indicator of phosphorus limited algal growth.     

Anatomical variation in some perennial Triticeae

JARVIE, J. K. & BARKWORTH, M. E., 1992. Anatomical variation in some perennial Triticeae. Cross-sectional anatomy of glumes and leaf blades was examined in 22 taxa of the perennial Triticeae. The taxa included diploids and polyploids based largely on various combinations of the E, J and S genomes. The objective was to determine how much correlation exists between anatomical characteristics and genomic constitution. The data were analysed by principal co-ordinate and cluster analyses. The results showed that monogenomic species based on different genomes were readily distinguishable on glume and leaf blade features. Most heterogenomic taxa exhibited some aspects of each genome present except in the case of JE allotetraploids, which exhibited no E genome characteristics.     

Morphological variation and genome constitution in some perennial Triticeae

JARVIE, J. K. & BARKWORTH, M. E., 1992. Morphological variation and genome constitution in some perennial Triticeae. A numerical analysis of species of five genomically defined genera of the Triticeae was undertaken, based on 42 morphological characters and 142 operational taxonomic units (OTUs). The primary goal was to determine the degree of congruence between morphological variation and genomic constitution. The second goal was to determine which existing supraspecific classification, if any, best reflected the morphological variation encountered. The five genera investigated were Thinopyrum (J genome), Lophopyrum (E genome), Pseudoroegneria (S genome), Trichopyrum (EES genome) and Elytrigia (SJE/SSX genome). Both principal co-ordinate and cluster analysis of the data placed the OTUs in supraspecific groups that reflected their genomic constitution. Monogenomic taxa were clearly separated. Allotetraploids between the E and S genomes were situated between E and S monogenomic taxa. Allotetraploids between the J and E genomes were situated closest to J genome taxa. The EES taxa of Trichopyrum were placed closest to Lophopyrum. OTUs of Elytrigia overlapped those of Pseudoroegneria , but not those of Lophopyrum or Thinopyrum.     

Direct Measurements of Turgor Pressure Potentials of Guard Cells: II. THE MECHANICAL ADVANTAGE OF SUBSIDIARY CELLS, THE SPANNUNQSPHASE, AND THE OPTIMUM LEAF WATER DEFICIT

Evidence of the mechanical advantage of subsidiary cells wasobtained by simultaneous measurements of turgor pressure potentialsin adjacent subsidiary and guard cells using injection circuitswith two separate needles. In Tradescantia virginiana the mechanicaladvantage approaches two. Using the same technique evidencewas obtained that the Spannungsphase is, in the first place,a turgor relations phenomenon due to the mechanical advantageof epidermal or subsidiary cells. In addition, the evidenceindicated that the elastic properties of guard cell walls mayundergo changes during the Spannungsphase when potassium iontransport commences. During these measurements it was confirmedthat the optimum leaf water deficit for maximum stomatal openingoccurs when the epidermal turgor is near zero. Under these conditionsthe width of the stomatal pore is a function of the turgor pressureof the guard cells, since at zero turgor of the subsidiary cellstheir mechanical advantage has disappeared.     

Cystolith Development and Structure in Pilea cadierei (Urticaceae)

Cystolith formation, structure and composition have been investigatedin leaves and stem internodes of Pilea cadierei (Urticaceae)using a variety of techniques at the light and electron microscopelevels. The development of lithocysts from epidermal cells hasbeen followed. These cells are cytoplasmically similar to otherepidermal cells but possess a much more active Golgi apparatusand more numerous mitochondria. The cystolith is a spindle-shapedbody composed of concentric layers of longitudinally orientatedcellulose microfibrils associated with pectins and other cellwall polysaccharides. At maturity it is heavily impregnatedwith calcium carbonate. Some cystoliths also contain siliconand are covered in a sheath of siliceous material. Cystolithformation occurs at the tip of a peg that grows in from thelithocyst wall. Evidence from ultrastructure suggests that thelithocytst cytoplasm transports carbohydrates to the cystolithvia Golgi vesicles, and organizes the deposition of cystolithcellulose microfibrils via a system of microtubules lying beneaththe plasma membrane that envelopes the growing cystolith. Thepeg is composed of heavily staining amorphous material likethat of an apoplastically sealed cell wall. It is incapableof supporting the migration of lanthanum ions into the cystolith.We conclude that cystoliths are isolated volumes of apoplastthat act as repositories for inorganic salts, principally calciumcarbonate. We propose that calcium ions move into the lithocystprotoplast from surrounding cells and are then transported acrossthe plasma membrane boundary into the cystolith. This proposalconflicts with previous suggestions that calcium enters by diffusionthrough the peg. Cystolith, lithocyst, cell wall, calcium, silicon, cytochemistry, electron probe analysis, Pilea cadierei