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Influence of betaine and salinomycin on intestinal absorption of methionine and glucose and on the ultrastructure of intestinal cells and parasite developmental stages in chicks infected with Eimeria acervulina.

The effect of betaine and salinomycin on absorption of methionine and glucose in tissue from the duodenal loops of Eimeria acervulina-infected chicks was determined. Differences in the ultrastructure of the intestinal cells and parasite developmental stages were also examined. With a drug-resistant isolate of E. acervulina, methionine absorption was significantly higher in chicks fed a basal diet supplemented with 0.15% betaine as compared with absorption in chicks fed the unsupplemented basal diet. Addition of 66 ppm salinomycin to the diet containing betaine did not further enhance absorption. Conversely, with a drug-sensitive isolate, methionine absorption was significantly higher in chicks fed a diet supplemented with both betaine and salinomycin than in chicks fed the unsupplemented basal diet. Tissue from chicks fed any of the supplemented diets was usually significantly heavier than that from chicks fed the unsupplemented diet, even when weight gains of the birds were similar. Glucose absorption was similar in all diet groups. Epithelial cells in coccidia-infected and uninfected chicks fed diets supplemented with betaine or betaine plus salinomycin were less electron dense than cells from chicks fed diets that were not supplemented with betaine. Merozoites of E. acervulina in chicks fed diets supplemented with salinomycin had extensive membrane disruption and vacuolization, but the damage was prevented when betaine was added to the diet. Numerous merozoites and intact schizonts were seen in the intestinal lumen of chicks fed the diet containing betaine plus salinomycin.

Animals↗

Intermediate filament protein expression in early developmental stages of the mouse. A confocal scanning laser microscopy study of in vitro fertilized and in vitro cultured pre-implantation mouse embryos.

Expression patterns of intermediate filament proteins have been studied during early mouse embryo development. For this purpose, pre-implantation embryos at different stages of development after in vitro fertilization were studied using antibodies to cytokeratins, vimentin and lamins, using the indirect immunofluorescence assay. The levels of expression were quantitated and localization of the protein constituents was assessed by means of confocal scanning laser microscopy. Our studies showed that, although the embryos grew in culture, vimentin could not be detected in a filamentous organization. Immunofluorescence for cytokeratins was only positive from the 8-cell stage onwards. In the morula stage an increased level of cytokeratin expression was observed with a transitional staining pattern, combining a filamentous and a diffuse occurrence. In the blastocyst stages profound cytokeratin filaments were seen in trophoblast cells but not in the inner cell mass. When the cytokeratin subtypes were analysed separately, it became apparent that expression levels of cytokeratins 8 and 18 increased gradually up to a filamentous pattern in the blastocyst stage. Cytokeratins 7 and 19, although elevated in the latter stage and showing a filamentous distribution, were not found as prominently as cytokeratins 8 and 18. A-type as well as B-type lamins could be detected in all developmental stages examined, as a faintly reactive nuclear lamina. In blastocysts both lamin types were detected in trophoblast as well as in inner cell mass.

Animals↗

Early developmental stages of a protozoan parasite, Marteilioides chungmuensis (Paramyxea), the causative agent of the ovary enlargement disease in the Pacific oyster, Crassostrea gigas.

A paramyxea, Marteilioides chungmuensis, causes the irregular enlargement of the ovary in the Pacific oyster, Crassostrea gigas in Korea and Japan. The knowledge about the life cycle of the parasite has been limited to the sporulation stages within the oocyte of oysters. In this study, we used the parasite-specific DNA probes and electron microscopy to experimentally infected oysters in a field and successfully clarified early developmental stages of the parasite. The parasite invaded the oysters through the epithelial tissues of the gills, mantle and labial palps. Extrasporogony repeatedly occurred in the connective tissues by binary fusion. The inner cell of the extrasporogonic stage migrated into the gonadal epithelium, invaded the oocyte to start sporulation.

Animals↗

Developmental stage dependent toxicity of aminoguanidine on chick embryos.

Aminoguanidine (AG) sulfate was injected into the thin albumen of fertile eggs on the different days of incubation. The lethality of AG sulfate on the developing chick embryos became lower when the treatment was done at later developmental stage. When AG was injected on the 5th day of incubation, the frequency of retardation of development of the body and liver, aplasia of gallbladder and enlarged spleen was highest without high mortality compared with the other experiments. Embryos with liver damage could not hatch. 14C-AG hydrochloride injected on the 5th day of incubation easily distributed into the embryo through the yolk from the thin albumen of the injection site, but when injected on the 9th day, the agent transferred very little into the yolk and embryo. The change of transport in the eggs with development may elucidate the difference of lethality of AG. Additionally, it was confirmed using thin layer chromatography that a large part of the radioactivity in the embryo might be an unknown metabolite of AG and the substance was accumulated with extremely high concentration in the liver at 24-48 hr after the injection of AG on the 5th day of incubation. This is probably the main cause of the peculier abnormalities in the liver of chick embryos.

Abnormalities, Drug-Induced↗

Acute CO2 tolerance during the early developmental stages of four marine teleosts.

Ocean sequestration of CO2 is proposed as a possible measure to mitigate climate changes caused by increasing atmospheric concentrations of the gas, but its impact on the marine ecosystem is unknown. We investigated the acute lethal effect of CO2 during the early developmental stages of four marine teleosts: red sea bream (Pagrus major), Japanese whiting (Sillago japonica), Japanese flounder (Paralichthys olivaceus), and eastern little tuna (Euthynnus affinis). The percentages of larvae that hatched and survived were not affected by exposure to water with a PCO2 of 1.0 kPa (= 7.5 mmHg) within 24 h. Median lethal PCO2 values for a 360-min exposure were 1.4 kPa (cleavage), 5.1 kPa (embryo), 7.3 kPa (preflexion), 4.2 kPa (flexion), 4.6 kPa (postflexion), and 2.5 kPa (juvenile) for red sea bream; 2.4 kPa (cleavage), 4.9 kPa (embryo), 5.9 kPa (preflexion), 6.1 kPa (flexion), 4.1 kPa (postflexion), and 2.7 kPa (juvenile) for Japanese whiting; 2.8 kPa (cleavage) and > 7.0 kPa (young) for Japanese flounder; and 11.8 kPa (cleavage) for eastern little tuna. Red sea bream and Japanese whiting of all ontogenetic stages had similar susceptibilities to CO2: the most susceptible stages were cleavage and juvenile, whereas the most tolerant stages were preflexion and flexion.

Animals↗

[Investigations into the fire structure of the asexual developmental stages of Frenkelia in the liver of the bank vole (author's transl)].

Bank voles (Clethrionomys glareolus) were infected by stomach tube with Frenkelia sporocysts from the faeces of buzzards (Buteo buteo). The voles were sacrificed at regular intervals and their livers examined electronmicroscopically. Seven days p.i. developmental stages of Frenkelia could be detected in liver parenchymal cells. The youngest schizonts detected are enveloped by a pellicle consisting of two membranes. This pellicle, which is in direct contact with the host cell mitochondria, shows marked invaginations which increase with the development of the schizont. A parasitophorous vacuole is not detectable. In developing schizonts numerous sections through nuclei with nucleic spindles and merozoite anlagen (dome-shaped) structures) are visible. It is not clear whether there are several nuclei or a section through one large and lobed nucleus. Within the merozoite anlagen the conoid and the subpellicular microtubules are formed first. By the prolongation of the dome-shaped structures towards the posterior pole, the nucleus and the other newly formed cell organelles are incorporated into the forming merozoite. The posterior pole of the merozoite still remains open at this stage of development. With increasing differentiation the merozoites become lancet-shaped, their apical poles bing always directed towards the periphery of the schizont. The outer membrane of the pellicle of the schizont forms the outer part of the pellicle of the merozoites by invaginating around them. At this stage of development the inner membrane of the pellicle of the schizont is no longer detectable. Thus the typical pellicle of the motile stages of sporozoaonsisting of three membranes is formed. In the centre of the merozoites which lie freely in the liver cell a residual body is present. The host cell reacts against the parasites by forming a thick border of mitochondria and distinct endoplasmic reticulum.

Animals↗

Identification of volatile compounds in soybean at various developmental stages using solid phase microextraction.

Soybean (Glycine max) seed volatiles were analyzed using a solid phase microextraction (SPME) method combined with gas chromatography-mass spectrometry (GC-MS). Thirty volatile compounds already reported for soybean were recovered, and an additional 19 compounds not previously reported were identified or tentatively identified. The SPME method was utilized to compare the volatile profile of soybean seed at three distinct stages of development. Most of the newly reported compounds in soybean seed were aldehydes and ketones. During early periods of development at maturity stage R6, several volatiles were present at relatively high concentrations, including 3-hexanone, (E)-2-hexenal, 1-hexanol, and 3-octanone. At maturity stage R7 and R8, decreased amounts of 3-hexanone, (E)-2-hexenal, 1-hexanol, and 3-octanone were observed. At maturity stage R8 hexanal, (E)-2-heptenal, (E)-2-octenal, ethanol, 1-hexanol, and 1-octen-3-ol were detected at relatively high concentrations. SPME offers the ability to differentiate between the three soybean developmental stages that yield both fundamental and practical information.

Aldehydes↗

Effects of praziquantel on different developmental stages of Schistosoma mansoni in vitro and in vivo.

To discern whether stage-specific resistance of Schistosoma mansoni to praziquantel occurs in vitro, we determined minimal effective concentrations (MECs) of drug needed to increase motor activity, produce contraction and/or paralysis, and cause tegumental vesiculation of developmental stages from day 0 to day 42 of S. mansoni. Recovery of these stages from exposure to praziquantel in vitro was also evaluated. MECs of praziquantel inducing increased motor activity and muscular contraction or paralysis or both were 0.005-0.01 micrograms/ml, irrespective of the stage examined. However, day-3 lung forms were more resistant than other stages when either drug-induced tegumental vesiculation (MEC, 1 microgram/ml) or recovery from drug exposure was tested. Three-day infections with S. mansoni in CF1 mice were also less responsive to praziquantel treatment than were infections of shorter or longer duration. The concentrations of praziquantel and periods of drug exposure causing gross tegumental damage to S. mansoni in vitro correlated with the peak serum levels and time course of unchanged praziquantel associated with reduction of worm burden in vivo. Thus, stage-specific resistance of S. mansoni to praziquantel does occur in vitro and correlates better with the tegumental than the muscular action of the drug.

Animals↗

Differential expression of the 27 kDa cathepsin L-like cysteine protease in developmental stages of Spirometra erinacei.

The 27 kDa cathepsin L-like cysteine protease of Spirometra erinacei plerocercoid is known to play an important function in tissue penetration, nutrient uptake and immune modulation in human sparganosis. In the present study, the expression of this enzyme was examined at different developmental stages of S. erinacei including immature egg, coracidium, plerocercoid in tadpole and rat, and adult. Proteolytic activity against carboxybenzoyl-phenylalanyl-arginyl-7-amino-4-methylcoumarin was detected in the extracts of coracidia and plerocercoid while no activity was observed in those of immature egg and adult. The specific activity in coracidial extracts was lower than that in the plerocercoid. Reverse transcription-polymerase chain reaction and Northern blot analysis demonstrated that the gene was expressed in the coracidium and plerocercoid but not in immature egg and adult. These results suggest that the 27 kDa cysteine protease is only expressed in the stages involving active migration of the parasite in the host tissue.

Animals↗

In vitro estradiol synthesis and secretion by tadpole ovaries of different developmental stages.

The tadpole ovaries at TK stage XIX can synthesize and secrete estradiol (E2), yet it is unknown when this ovarian function starts and how it develops. To this end, the present work has been carried out. The ovaries of different developmental stages of tadpoles and young frogs of Rana catesbeiana were taken, cut into small pieces, and incubated for 6 hr at 20 degrees in Krebs-Ringer bicarbonate buffer. The media were then analyzed for E2 by radioimmunoassay. The results indicate that the tadpoles synthesize and release E2 as early as stage X, that E2 increases slowly and gradually through stage XXV, and that E2 increases rapidly in juvenile frogs. This trend of growth of the ovarian function in estradiol secretion in vitro is in accordance with that reported for female chick embryos. There exists the possibility that the growth of E2 secretion could be biphasic, one with slow increment of autonomous E2 secretion at early stages and the other with quick increase due to pituitary stimulation at the stage of metamorphic climax.

Analysis of Variance↗

Developmental stages of cortical Lewy bodies and their relation to axonal transport blockage in brains of patients with dementia with Lewy bodies.

We investigated 10 dementia with Lewy bodies (DLB) brains showing various degrees of Lewy pathology using alpha-synuclein-immunohistochemistry, and morphologically divided cortical Lewy bodies (LB) into six developmental stages. Further, we demonstrated the ultrastructure of each stage of cortical LB using alpha-synuclein-immunoelectron microscopy. In the initial stage, alpha-synuclein accumulated in part of the neuronal cytoplasm without filamentous components, then formed LB and LB-related neurites composed of granulo-filamentous components. Finally, LB degraded to extracellular LB composed of loose filamentous components with involved astroglial processes. In addition, we immunohistochemically investigated the accumulation of axonal transported substances in cortical LB, and showed that APP, chromogranin-A, synphilin-1 and synaptophysin accumulated in cortical LB from stages 1, 2, 3 and 4, respectively. These findings suggest that chronic axonal transport blockage is implicated in the development of cortical LB in DLB brains.

Amyloid beta-Protein Precursor↗

Effect of lipid polarization by centrifugation at different developmental stages on post-thaw survival of bovine in vitro produced 16-cell embryos.

The developmental rate to the blastocyst stage of frozen-thawed bovine in vitro produced embryos at stages earlier than Day 6 morula is not sufficiently high for practical utilization. The present study was undertaken to determine the effect of polarization of lipid droplets in the cytoplasm of bovine in vitro produced embryos from zygotes to the 8-cell stage, by centrifugation without following micromanipulation, on the survival rate of Day 4 16-cell embryos. After centrifugation at 15,500 x g in medium containing cytochalasin D, embryos were cultured to the 16-cell stage, classified as either mostly or partially delipidated by degree of lipid droplet removal, and then frozen. Embryos centrifuged at the 2-cell stage developed to the 16-cell stage similarly to those centrifuged at the 8-cell stage. The developmental rate to blastocysts after freezing of the mostly delipidated 16-cell embryos centrifuged at the 2-cell stage was higher than that of those centrifuged at the zygote stage, those that were partially delipidated at the 2-cell stage, and those that were not centrifuged. The results demonstrate that polarization of lipid droplets at the 2-cell stage by centrifugation without micromanipulation improved the survival rate of mostly delipidated 16-cell embryos after freezing.

Animals↗

The Streptomyces aureofaciens homologue of the whiB gene is essential for sporulation; its expression correlates with the developmental stage.

In previous experiments, a Streptomyces aureofaciens gene highly similar to the sporulation-specific whiB gene of Streptomyces coelicolor was identified. By integrative transformation, via double cross-over, a stable null mutant of the whiB-homologous gene of S. aureofaciens was obtained. The disruption blocked differentiation at a stage between the formation of aerial mycelium and the development of mature spores, producing white aerial hyphae without septation. Expression of the whiB gene was investigated during differentiation by S1 nuclease mapping, using RNA prepared from S. aureofaciens in various developmental stages. Two putative promoters were identified upstream of the whiB coding region. The stronger promoter, whiB-P2, was induced at the beginning of aerial mycelium formation, and the weaker promoter, whiB-P1, was expressed fairly constantly during differentiation. No differences in the expression of the whiB promoters were detected in an rpoZ-disrupted S. aureofaciens strain. The promoter bearing DNA fragment was inserted into the promoter-probe vector pARC1 to produce an expression pattern consistent with the results of direct RNA analysis.

Bacterial Proteins↗

[Developmental stages of Taenia krabbei Moniez, 1879 (Cysticercus tarandi) in deer (Capreolus capreolus L.) in Czechoslovakia].

The breast and shoulder-blade muscles of a roe (Caprelous capreolus L.), shot in a hunting district in western Bohemia, were found to contain cysticerci with morphological characters and size corresponding to those of Cysticercus tarandi, which is a developmental stage of the tapeworm Taenia krabbei Moniez, 1879. The surface as well as deeper layers of muscle contained one to two cysticerci per 100 cm2 of area. The authors describe the larvocysts and histological changes in the site where the micro-organisms were found. The cysticerci are discussed as to the species to which they belong.

Animals↗

Genetic analysis of developmental stages of the cellular slime mold Dictyostelium purpureum.

71 Mutants of Dictyostelium purpureum, unable to carry out normal development, were isolated. These mutants fall into three phenotypic classes. The mutants of the first class do not aggregate. Those of the second class develop a little further and form conical aggregates. The mutants of the third class progress still further, and the conical aggregates are transformed into finger-shaped masses, but no fruiting bodies are formed. Mutants of each class were able to form fruiting bodies when mixed with mutants of other classes. Pairs of mutants belonging to the same class, however, never responded synergistically. This result indicates the presence of three distinct developmental stages in the process of the fruiting-body formation. Experiments with actinomycin D suggested that genes controlling the developmental process of D. purpureum were divided into three groups which were identical to those identified by the analysis of synergism.

Dactinomycin↗

In situ hybridization analysis of developmental stages of Pneumocystis carinii that are transcriptionally active for a major surface glycoprotein gene.

An abundant glycoprotein on the surface of Pneumocystis carinii, termed gpA or gp120, is thought to play a role in the interaction of this opportunistic pathogen with its host. Using RNA:RNA hybridization techniques, the in situ expression of gpA mRNA in developmental forms of the organism was investigated in a ferret model of P. carinii pneumonia. The results suggested that the relative abundance of gpA-specific mRNA was variable in different developmental stages of ferret P. carinii. P. carinii localized along the epithelial lining of alveoli were transcriptionally active. Immunocytochemical detection of gpA and Giemsa staining suggested that many of these organisms were trophic forms of P. carinii. While no detectable gpA mRNA signal was found in the majority of P. carinii cysts, a portion of identifiable cysts co-localized with significant levels of gpA mRNA signal. Differential staining of the cyst wall with Gomori's methenamine silver suggested that the transcriptionally active P. carinii cysts were the intermediate or precyst forms of the organism, while the cysts with no detectable mRNA signal were either mature or empty (excysted). Alveolar macrophages were observed surrounded by transcriptionally active organisms; however, no gpA-transcriptional activity was detected within macrophages. Taken together, the results suggest that transcription of gpA occurs in forms of P. carinii that are actively replicating, and in close proximity or contact with, alveolar epithelial cells.

Animals↗

The effect of proctolin on the adenylate and guanylate cyclases in the Locusta brain at various developmental stages.

Proctolin at concentrations 10(-8)-10(-7) M elevated by 40% brain adenylate cyclase activity of adult Locusta migratoria migratoriodes R.F. In moulting individuals, proctolin caused a decrease in brain adenylate cyclase activity, and it proved to be ineffective in the larvae. Proctolin caused only a slight decrease on guanylate cyclase activity of the brain at every developmental stage.

Adenylyl Cyclases↗

Efficacy of entomopathogenic nematode Steinernema feltiae (Rhabditida: Steinernematidae) as influenced by Frankliniella occidentalis (Thysanoptera: Thripidae) developmental stage and host plant stage.

Entomopathogenic nematodes were investigated as an alternative biological control strategy for western flower thrips, Frankliniella occidentalis (Pergande) (Thysanoptera: Thripidae), in ornamental greenhouse crops, by using potted chrysanthemum as a model crop. The susceptibility of various life stages of F. occidentalis to different concentrations of the nematode Steinernema feltiae (Filipjev) (Rhabditida: Steinernematidae) was investigated in petri dish bioassays. This was followed with trials using potted chrysanthemums comparing the efficacy of nematode application to plants in vegetative (exposed habitat) versus flowering (cryptic habitat) stages. In both trials, the effect of the wetting agent Agral 90 (nonylphenoxy polyethoxyethanol), which is used in combination with the nematode spray, on F. occidentalis mortality was assessed. In petri dish trials, the prepupae and pupae were the most susceptible developmental stages of F. occidentalis to infection by S. feltiae. First and second instars were killed by very high rates of nematodes (> or =20,000 infective juveniles per milliliter), but corrected mortality was only approximately 28-37%. No significant mortality was observed for adult thrips. Results from the petri dish trials were confirmed on chrysanthemum plants. Foliar application of S. feltiae did not result in significant mortality in larvae or adults. No significant differences in efficacy were detected by application of nematodes on vegetative versus flowering chrysanthemum. Agral 90 had a significant impact on mortality on the first stage larvae and prepupae in the petri dish trials but not in the plant trials. Thrips control by S. feltiae in greenhouses may be partly or completely due to prepupal and pupal mortality.

Animals↗