Blastocystis hominis: pathogenic potential in human patients and in gnotobiotes.

Germfree guinea pigs were inoculated orally, in some experiments, and intracecally, in others, with Blastocystis hominis and the enteric flora from symptomatic patients. Other germfree guinea pigs received the parasite from axenic culture and still others from monoxenic culture with Proteus vulgaris. Fourteen of 43 animals inoculated orally with B. hominis and patient's enteric flora developed B. hominis infections and those with particularly heavy infections developed watery diarrhea of more than 1 week's duration immediately prior to sacrifice. Similar results were obtained from intracecal inoculations in that 13 of 28 animals developed infections and those with the greatest numbers of B. hominis had watery diarrhea for more than 1 week prior to sacrifice. Gross pathologic changes in these animals were mostly unremarkable, with only a slight hyperemia observed in several of the symptomatic animals. Microscopic examination, however, revealed frequent penetration of intestinal epithelium by B. hominis and the parasites in significant numbers were observed within the epithelium. There was a slight increase in cellularity in the lamina propria but parasites were not observed therein and their presence in the epithelium did not provoke an inflammatory response. Only one of eight animals inoculated from monoxenic cultures developed B. hominis infection (asymptomatic), and infections were not produced in animals inoculated from axenic culture. As a result of our observations of diarrhea in patients with particularly heavy infections with B. hominis together with the demonstration of similar symptoms in animals heavily infected with this parasite, we believe B. hominis may occasionally be related causally to the production of such symptoms by a mechanism not completely understandable.     

In-Vitro and In-Vivo Effects of Aldicarb on Survival and Development of Heterodera schachtii

Aqueous solutions of 5-500 μg/ml aldicarb inhibited hatching of Heterodera schachtii. Addition of hatching agents, zinc chloride, or sugarbeet root diffusate, to the aldicarb solutions did not decrease the inhibition of hatching. When cysts were removed from the aldicarb solufions and then treated for 4 wk in sugarbeet root diffusate, larvae hatched and emerged. Treatments of newly hatched larvae of H. schachtii with 5-100 μg/ml aldicarb depressed later development of larvae on sugarbeet (Beta vulgaris). Similar treatments with aldicarb sulfoxide had less effect on larval development, and aldicarb sulfone had no effect. Numbers of treated larvae that survived and developed were inversely proportional to concentration (0.1-5.0 μg/ml) and duration (0-14 days) of aldicarb treatments. Development of H. schachtii on sugarbeet grown in aldicarb-treated soil was inversely proportional to the concentration of aldicarb in the tested range of 0.75 - 3.0 μg aldicarb/g of soil. Transfer of nematode-infected plants to soil with aldicarb retarded nematode development, whereas transfer of plants first grownin treated soil to nematode-infested soil only slightly suppressed nematode development. Development of H. schachtii was inhibited in slices of storage roots of table beet (B. vulgaris), sugarbeet and turnip, (Brassica rapa), that had grown in soil treated with aldicarb.     

Behaviour of phenylalanine ammonia-lyase in carrot cells in suspension cultures

The high PAL activity in carrot cells in suspension culture was found at the linear and early stationary phases, with concomitant increases in phenylal     

Isoperoxidases from tobacco tissue cultures

Two anodic isoperoxidases (A₁ and A₂) from tobacco tissue culture W-38 and two cathodic isoperoxidases (C₃ and C₄) from tobacco suspension culture WR-132 have been separated and characterized. Molecular weights for each of the isoperoxidases have been determined by two different methods. Only C₄ contained a carbohydrate component. The substrate specificity and the pH optima for the four enzymes with each of five substrates were determined.     

Ultrastructure of the Invasion of Eimeria magna Sporozoites into Cultured Cells*†‡

SYNOPSIS. Monolayers of bovine kidney cells were overlaid with Eimeria magna sporozoites and observed with phase-contrast optics until penetration of the cells by the parasites had begun. Cells and penetrating parasites were fixed with glutaraldehyde and OsO₄-containing ruthenium red, dehydrated, and embedded in situ. Cells being penetrated were selected for study in the electron microscope. The lack of intracellular staining with ruthenium red and intact plasmalemmas of cells being penetrated, was accepted as evidence that the sporozoites did not disrupt the plasma membranes. The sporozoite caused invagination of the host cell plasmalemma until the parasite was entirely within the cell, after which the invagination was sealed off by short pseudopodia enclosing the sporozoite within a membrane-lined vacuole inside the cell. Often myelin-forms, apparently of host cell origin, were seen in the space between the sporozoite and the cell.     

The effect of different media and culture techniques on plating efficiency of strawberry mesophyll cells in culture

Mesophyll cells of strawberry were isolated mechanically by a hand homogenizer, One g healthy, fully-expanded leaves yielded 10⁷ cells. The agar culture method was the only culture technique found suitable to obtain any quantitative data.     

Neural Tube Closure in Mouse Whole Embryo Culture

Genetic mouse models are an important tool in the study of mammalian neural tube closure (Gray & Ross, 2009; Ross, 2010). However, the study of mouse embryos in utero is limited by our inability to directly pharmacologically manipulate the embryos in isolation from the effects of maternal metabolism on the reagent of interest. Whether using a small molecule, recombinant protein, or siRNA, delivery of these substances to the mother, through the diet or by injection will subject these unstable compounds to a variety of bodily defenses that could prevent them from reaching the embryo. Investigations in cultures of whole embryos can be used to separate maternal from intrinsic fetal effects on development.Here, we present a method for culturing mouse embryos using highly enriched media in a roller incubator apparatus that allows for normal neural tube closure after dissection (Crockett, 1990). Once in culture, embryos can be manipulated using conventional in vitro techniques that would not otherwise be possible if the embryos were still in utero. Embryo siblings can be collected at various time points to study different aspects of neurulation, occurring from E7-7.5 (neural plate formation, just prior to the initiation of neurulation) to E9.5-10 (at the conclusion of cranial fold and caudal neuropore closure, Kaufman, 1992). In this protocol, we demonstrate our method for dissecting embryos at timepoints that are optimal for the study of cranial neurulation. Embryos will be dissected at E8.5 (approx. 10-12 somities), after the initiation of neural tube closure but prior to embryo turning and cranial neural fold closure, and maintained in culture till E10 (26-28 somities), when cranial neurulation should be complete.     

Site‐specific interactions of neurotrophin‐3 and fibroblast growth factor (FGF2) in the embryonic development of the mouse cochlear nucleus

Neurotrophins and FGF2 contribute to formation of the cochlea, but their roles in cochlear nucleus development are unknown. The effects of these factors may differ in the cochlea and cochlear nucleus, which may influence each other's development. It is important to analyze the effects of these factors on cellular structures at well‐defined steps in the normal morphogenetic sequence. The present study used immunohistochemistry to localize factors in situ and to test hypotheses about their roles in an in vitro model. Specific antibody staining revealed that TrkC, the NT3 receptor, is present in neural precursors prior to embryonic day E11 until after birth. NT3 appeared in precursor cells during migration (E13–E15) and disappeared at birth. TrkC and NT3 occurred in the same structures, including growing axons, terminals, and their synaptic targets. Thus, NT3 tracks the migration routes and the morphogenetic sequences within a window defined by TrkC. In vitro, the cochlear nucleus anlage was explanted from E11 embryos. Cultures were divided into groups fed with defined medium, with or without FGF2, BDNF, and NT3 supplements, alone or in combinations, for 7 days. When neuroblasts migrated and differentiated, immunostaining was used for locating NT3 and TrkC in the morphogenetic sequence, bromodeoxyuridine for proliferation, and synaptic vesicle protein for synaptogenesis. By time‐lapse imaging and quantitative measures, the results support the hypothesis that FGF2 promotes proliferation and migration. NT3 interacts with FGF2 and BDNF to promote neurite outgrowth, fasciculation, and synapse formation. Factors and receptors localize to the structural sites undergoing critical changes. © 2006 Wiley Periodicals, Inc. J Neurobiol, 2006