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Characterisation of complementary DNAs from the expressed sequence tag analysis of life cycle stages of Laminaria digitata (Phaeophyceae).

Laminariales (Phaeophyceae, Heterokonta) are characterised by a heteromorphic digenetic life cycle with a filamentous, microscopic gametophyte and a highly evolved, macroscopic sporophyte. With the ultimate goal of comparing gene expression in each life cycle stage, complementary DNA libraries were constructed from sporophytes and gametophytes of Laminaria digitata. A set of ca. 500 expressed sequence tags (EST) was generated from each life history phase, by single-run partial sequencing of randomly picked cDNA clones. Comparison of the EST deduced amino acid sequences with database protein sequences assigned a putative identity for 39% of the 412 gametophyte clones and 48% of the 493 sporophyte clones sequenced thus far. These data represent more than 152 different proteins now probably identified in L. digitata. Several of those newly identified proteins are of interest to our understanding of the molecular physiology of kelps, for example their carbon-concentrating mechanisms, cell wall biosynthesis and halogen metabolism. EST analysis also confirmed that genes with long 3'-UTRs are widespread in Laminariales and the study of 5'-UTRs allowed the identification of a Kozak consensus sequence, c(A/C)A(A/C)CAUGGc(G/T). Several potential developmentally regulated differences in gene expression are discussed.

Algal Proteins↗

Inequalities in women's health in England and Wales: mortality among married women according to social circumstances, employment characteristics and life-cycle stage.

"Data obtained from follow-up of the Office of Population Censuses and Surveys Longitudinal Study 1971 Census sample have been used to look at women's mortality differentials at ages 15-59 in England and Wales....In this paper we focus on married women and use age of youngest child as a measure of life-cycle stage. We relate this to whether the woman was a housewife, or was in full or part-time paid employment, so as to examine how these affect differences in mortality by social class. We find that socio-economic mortality differences persist irrespective of life-cycle stage.... Housewives married to men in manual occupations experienced death rates over one and a half times as high as those married to men in non-manual occupations. For women in employment the differences by husbands' social class are of a lesser magnitude.... Differences in the mortality of those in full and part-time work depend on the woman's own social class and are greater for non-manual than manual classes." (SUMMARY IN FRE AND ITA)

Demography↗

[Antigenic differentiation among 3 life-cycle stages of Toxoplasma gondii using fluorescent antibodies].

Antigenic differentiation between three stages of the life cycle of Toxoplasma gondii by fluorescent antibodies. Antisera were prepared in three groups of twenty mice each with three different antigens of Toxoplasma gondii: the first group was inoculated with tachyzoites of RH strain and received sulfadiazine treatment; the second with tissue cysts of T-100-cat-6751 strain and the third with oocysts of the same strain, both without treatment. In the indirect immunofluorescent antibody technique each antigen was tested with its homologous and heterologous antisera, determining qualitative and quantitative antigenic differences according to the fluorescence patterns. Some stages of Toxoplasma when reacting with their heterologous antibodies showed a central, partial posterior or total fluorescence during a certain period of development. The difference in fluorescence was sufficient to distinguish whether the origin of the infection was via cyst, oocysts and/or via tachyzoite when observation was made before day 65 post-infection.

Animals↗

Identification of life cycle stages of the nematode Echinocephalus overstreeti by allozyme electrophoresis.

Data presented in this study highlight the potential of allozyme electrophoresis in providing unequivocal genetic evidence for the identification of life cycle stages, particularly where species have complex life cycles. Adults of the nematode Echinocephalus overstreeti parasitize the elasmobranch Heterodontus portusjacksoni. The putative larval form which is morphologically dissimilar is found in two species of marine molluscs, Chlamys bifrons and Pecten albus. Electrophoretic analysis indicated that the adult and larval forms shared alleles at all of the 34 enzyme loci established. Furthermore, there were no fixed allelic differences between larval forms from different mollusc species.

Alleles↗

Life cycle stages of the amphibian chytrid Batrachochytrium dendrobatidis.

An overview of the morphology and life cycle of Batrachochytrium dendrobatidis, the cause of chytridiomycosis of amphibians, is presented. We used a range of methods to examine stages of the life cycle in culture and in frog skin, and to assess ultrastructural pathology in the skin of 2 frogs. Methods included light microscopy, transmission electron microscopy with conventional methods as well as high pressure freezing and freeze substitution, and scanning electron microscopy with critical point drying as well as examination of bulk-frozen and freeze-fractured material. Although chytridiomycosis is an emerging disease, B. dendrobatidis has adaptations that suggest it has long been evolved to live within cells in the dynamic tissue of the stratified epidermis. Sporangia developed at a rate that coincided with the maturation of the cell, and fungal discharge tubes usually opened onto the distal surface of epidermal cells of the stratum corneum. A zone of condensed, fibrillar, host cytoplasm surrounded some sporangia. Hyperkeratosis may be due to (1) a hyperplastic response that leads to an increased turnover of epidermal cells, and (2) premature keratinization and death of infected cells.

Animals↗

Experimental infections of farmed eels with different Trypanosoma granulosum life-cycle stages and investigation of pleomorphism.

Trypanosoma granulosum, a flagellate protozoon commonly found in the blood of the European eel Anguilla anguilla, was injected experimentally into uninfected eels purchased from a local farm. In order to investigate the infectivity of different stages in the life cycle, trypanosomes from various sources were used for inoculation. Infectivity was greatly reduced in in vitro culture stages inoculated at 20 C. Isolated bloodstream stages injected into groups of animals held at 12 and 20 C could be detected for over 70 days but did not appear to multiply. Naturally infected Hemiclepsis marginata, a piscivorous leech known to serve as vector, produced detectable, single-peak infections in eels held at 20 C. Infections were characterized by a prepatent stage and a phase of rising parasitemia. Peak infection intensities ranged between 1 and 7 x 10(4) trypanosomes/ml. Trypanosomes in the bloodstream of eels experimentally infected with leeches, divided at a very low rate during the early stages of infection. Small morphs present during the early phase of rising parasitemia were gradually replaced by larger trypanosomes. The overall length frequency distribution of trypanosomes was unimodal.

Anguilla↗

[Differentiation of closely related species Hyalomma anatolicum and H. excavatum (Acari: Ixodidae) based on a study of all life cycle stages, throughout entire geographical range].

Species status is stated for Hyalomma anatolicum and H. excavatum, based on numerous material from the territory of their distribution. The differentiation of species is recovered for all stages of the life cycle. Diagnosis of Hyalomma anatolicum. Female: small tick--length of scutum commonly less than 2 mm; width of scutum commonly less than 1.9 mm; color of scutum, gnathosoma and coxae light, yellowish- or reddish-brown; whitish enameled pigment on scutum absent; posterolateral ledges of scutum weakly expressed or absent; cervical and lateral grooves more shallow (than in H. excavatum) (fig. 1); setae of scutum, alloscutum, sternal setae and ventromedian setae of second article of palpae more tapering to the apex, narrow rounded or acute (fig. 2, 1, 2, 5, 8). Male: small tick--length of conscutum, as a rule, less than 4 mm; width of scutum, as a rule, less than 2.3 mm; shape of conscutum marrow oval (fig. 3), widest in the midlevel; color of conscutum, anal shields, gnathosoma and coxae light, yellowish- or reddish-brown; whitish enameled pigment on conscutum absent; caudal depression with less dense and more large punctuations (than in H. excavatum); ridges lateral to caudal field less high and sharp (than in H. excavatum); posteromedian groove separated from parma by contiguous punctuations or smooth non high area; setae of conscutum conus-like, more sharpening (fig. 4, 1-3); posterior dorsal margin of gnathosoma straight or slightly concave (fig. 4, 8); segments of leg IV not swallowed. Nymph: large tick (see description). Scutum more narrow. Dorsal tale of spiracular plates clear expressed, marginal perforations distant from margin of spiracular plates at the base of tale (fig. 5, 3); anterolateral side of basis capituli approximately equal to half of gnathosoma width; ventrally lateral projections of gnathosoma situated in posterior half of capitulum base (fig. 5, 4, 5). Hypostome more longer and narrow (fig. 5, 5). Larva: large tick (see description). Diagnosis of Hyalomma excavatum. Female: large tick--length of scutum commonly more than 2 mm; width of scutum commonly more than 1.9 mm; color of scutum, gnathosoma and coxae dark, reddish- or black-brown; whitish enameled pigment commonly present; posterolateral ledges of scutum clearly expressed; cervical and lateral grooves more dipper (than in H. anatolicum) (fig. 7). Setae of scutum, alloscutum, sternal setae and ventromedian setae of second article of palpae more stick-like, more widely obtuse (fig. 8, 1-5). Male: large tick--length of conscutum more than 4 mm; width of scutum more than 2.3 mm; shape of conscutum wide oval (fig. 9), widest in posterior half of length; color of conscutum, anal shields, gnathosoma and coxae dark, reddish- or black-brown; whitish enameled pigment often present; caudal depression with more dense and more small punctuations (than in H. anatolicum); ridges lateral to caudal field more high and sharp (than in H. anatolicum); posteromedian groove separated from parma by strong elevations fused with paraparmal festoons; setae of conscutum more obtuse, often stick-like (fig. 10, 1, 2); posterior dorsal margin of gnathosoma straight or lightly concave. (fig. 10, 3); segments of leg IV swallowed. Nymph: small tick (see description). Scutum more wide. Dorsal tale of spiracular plates weakly expressed, marginal perforations not distant from margin of spiracular plates at the base of tale (fig. 11, 3); anterolateral side of basis capituli clearly shorter than half of gnathosoma width; ventrally lateral projections situated in the middle of or anterior half of capitulum base (fig. 11, 4, 5). Hypostome more shorter and wider (fig. 5, 5). Larva: small tick (see description). Established taxonomical independence of H. anatolicum and H. excavatum is confirmed by several criteria: external morphology (see diagnoses); morphometrical data (see diagnoses and fig. 12, 13); size inversion: larva and nymph of H. anatolicum are large, but females and males are small; while H. excavatum vise versa--larva and nymph are small, but adults--large; host specialization of immature stages: larvae and nymphs of H. anatolicum, as rule, parasitize larger mammals (cattle, camels), while immature stages of H. excavatum parasitize smaller mammals (rodents, hares); presence of sympatry areas: almost entire area of H. excavatum include in area of H. anatolicum and cattle help to mix its populations.

Africa↗

Nippostrongylus brasiliensis: the effect of mitochondrial inhibitors on life-cycle stages.

The effects of mitochondrial inhibitors on the in vitro development of Nippostrongylus brasiliensis have been studied in free-living and parasitic life-cycle stages. Mitochondrial inhibitors were chosen as being representative of established electron transport inhibitors and oxidative phosphorylation inhibitors and uncouplers of the classical mammalian respiratory chain. All mitochondrial inhibitors tested were highly effective in killing or retarding development of free-living stages of N. brasiliensis. Free-living stages were particularly susceptible to such inhibitors upon hatching of embryonated eggs to 1st-stage larvae. Concentrations of inhibitors effective against free-living stages were consistent with their level of inhibition against isolated mitochondria from embryonated eggs and 3rd-stage infective larvae. Results suggest an absolute requirement in the development of free-living stages for the mammalian-like respiratory chain and associated oxidative phosphorylation. Electron transport inhibitors were effective in retarding at least the initial development of 4th-stage larvae to adults, but only antimycin A and azide produced a lasting effect leading to worm death. Oxidative phosphorylation inhibitors and uncouplers were ineffective against developing parasitic stages of N. brasiliensis. Experiments on whole-worm respiration indicated that most electron transport inhibitors were able to penetrate the adult worm, but oxidative phosphorylation inhibitors were without effect on whole-worm respiration. Results suggest that the mammalian-like electron transport chain is a necessary requirement to adult N. brasiliensis, but oxidative phosphorylation in the adult worm may not be required for development and survival in vitro although it could be necessary to support the parasite in vivo.

2,4-Dinitrophenol↗

Expressed sequence tags from life cycle stages of Trichinella spiralis: application to biology and parasite control.

While the approach taken to date to study Trichinella spp., involves mainly characterization of individual genes of interest, we initiated a genomics approach as an antecedent to more complete genome sequencing. Our approach involves use of expressed sequence tags (ESTs) obtained from three life cycle stages of Trichinella spiralis; adult worms (AD), mature muscle larvae (ML) and immature L1 larvae (immL1, also known as newborn larvae) () to improve the technical capacity for research on Trichinella spp. and to generate information that will aid prospective development of relevant hypotheses. In this review, we will summarize findings of our EST analysis and discuss how they relate to topics mentioned above. The foundation laid by this data will also contribute toward development of a more substantial genomic database and technical capacity to dissect molecular interactions between vertebrate hosts and Trichinella spp.

Animals↗

Pathologic and clinical findings in patients with cyclosporiasis and a description of intracellular parasite life-cycle stages.

Cyclospora cayetanensis has been observed in the feces of persons with prolonged diarrhea. A description of the symptoms and histopathologic findings for patients with cyclosporiasis is presented. The intracellular life-cycle stages of these parasites in the enterocytes of patients will also be described. Seventeen Peruvian patients positive for Cyclospora organisms were surveyed and underwent endoscopy, and their symptoms were recorded. Patients presented with gastrointestinal symptoms, including diarrhea, flatulence, weight loss, abdominal discomfort, and nausea. Jejunal biopsies showed an altered mucosal architecture with shortening and widening of the intestinal villi due to diffuse edema and infiltration by a mixed inflammatory cell infiltrate. There was reactive hyperemia with vascular dilatation and congestion of villous capillaries. Parasitophorous vacuoles contained sexual and asexual forms. Type I and II meronts, with 8-12 and 4 fully differentiated merozoites, respectively, were found at the luminal end of epithelial cells. These findings demonstrate the complete developmental cycle associated with host changes due to Cyclospora organisms.

Animals↗

Differentiation of species and life cycle stages of Brugia spp. by isoenzyme analysis.

The isoenzyme patterns of glucose phosphate isomerase and phosphoglucomutase of 3 species of Brugia, B. pahangi, subperiodic B. malayi, and B. patei, and 3 life cycle stages, adult, third-stage larva, and microfilaria were compared using the technique of isoelectricfocusing on polyacrylamide gels. The results demonstrated that the adults of all 3 species could be identified from one another and that differences existed between the sexes of any one species. Hybridization between B. pahangi and B. patei could be detected in the progeny of the cross. Both the third-stage larvae and microfilariae of B. malayi and B. pahangi were differentiated and the epidemiological significance and the application of these findings to arthropod-borne filarial infections were discussed.

Animals↗

Brachylaima cribbi (Digenea: Brachylaimidae): scanning electron microscopical observations of the life-cycle stages.

Brachylaima cribbi is a recently described species of terrestrial trematode that infects mammals and birds with helicid land snails as its first and second intermediate hosts. The adult worm is 2.5-6.0 mm long by 0.5-0.8 mm wide being a long slender cylindrical worm with oral and ventral suckers in the anterior quarter and genital pore in the posterior quarter. Scanning electron microscopy shows that there is a dense covering of tegumental spines at the anterior end which diminishes towards the posterior extremities of the worm. Development of spines was observed in juvenile and mature adult worms. In young worms 1-3 weeks post infection (wpi) spines appear as buds with a serrated edge each having 1-4 spikes per spine. As the worm ages the spines broaden and by 5 wpi the number of spikes per spine increases to an average of 8.1. The serial development of oral sucker papillae in the cercaria, metacercaria and adult worm was observed with the finding of an elongated papilla with a bifurcated tip on the cercaria becoming a shorter and thicker elongated papilla with a large central stoma on the metacercaria. In the adult worm, this papilla becomes dome-shaped with a small central stoma. For some of these papillae a cilium could be seen extended from the central stoma. Other life-cycle stages illustrated were the hatched egg with an extruded egg membrane minus an operculum and a portion of the branched sporocyst dissected from the digestive gland of the land snail Theba pisana showing a terminal birth pore. Scanning electron microscopy morphological features of the adult worm observed for the first time in a Brachylaima were the unarmed cirrus extended from the genital pore with released sperm present and the Laurer's canal opening visible in tegumental folds on the dorsal surface approximately 300 microm posterior to the genital pore.

Animals↗

Description of the life-cycle stages of Brachylaima cribbi n. sp. (Digenea: Brachylaimidae) derived from eggs recovered from human faeces in Australia.

The life-cycle of Brachylaima cribbi n. sp. was established in the laboratory. Asymmetrical brachylaimid eggs, measuring 26-32 microm (29.1 microm) long and 16 -17.5 microm (16.6 microm) wide, were recovered from human faeces and fed to the helicid land snail Theba pisana as the first intermediate host. Sporocysts and cercariae were recovered from the T. pisana eight weeks after infection. The cercariae were used to infect the helicid land snails Cernuella virgata and Helix aspersa as second intermediate hosts. Metacercariae were recovered from the kidneys of these snails and used to infect mice. Adults of Brachylaima cribbi n. sp. were recovered from the small intestine of the mice. The differential features of B. cribbi n. sp. are the specificity for helicid snails as first and second intermediate hosts; characteristic ventral sucker and body cercarial chaetotaxy; and a long slender adult worm with equal size suckers in the first quarter of the worm, the ventral sucker occupying 41% of the body width, the uterus extending anterior to the ventral sucker and the vitelline follicles falling short of the posterior margin of the ventral sucker. No other known Brachylaima species exhibits all of these features. B. cribbi n. sp. is the first brachylaimid known to have infected humans and is probably of European origin, as the intermediate host snails were all introduced into Australia from Europe.

Aged↗

De novo ribosome biosynthesis is transcriptionally regulated in Eimeria tenella, dependent on its life cycle stage.

Protozoan parasites go through various developmental stages during their parasitic life, which requires the expression of different genes. To identify stage specific gene products in Eimeria tenella, a differential screening was performed comparing the intracellular schizont stage with the extracellular oocyst stage. De novo transcripts of 18S-5.8S-26S rRNA transcription units and of two ribosomal proteins (RPL5 and RPL23) were specifically identified in schizonts and were undetectable in oocysts. The stage specific transcription of pre-rRNAs (prior to processing) was confirmed with Northern blot analysis. Since the E. tenella genome contains a repeated gene cluster with an estimated 140 large rRNA transcription units, they all might be similarly regulated. Specific expression of RPL5 and RPL23 in E. tenella schizonts was also confirmed by Northern blotting. Furthermore, an analysis of the E. tenella EST database with 26,705 ESTs showed that 9.5% of all merozoite ESTs and only 0.2% of the sporozoite ESTs encoded ribosomal proteins (RPs). These ESTs encoded 69 different RPs, suggesting that most and possibly all RPs are differentially transcribed in E. tenella. Analysis of EST data from other Coccidia, such as Toxoplasma gondii, indicated a similar stage dependent transcription of RP genes. We conclude that ribosome biosynthesis is transcriptionally regulated in E. tenella and other Coccidia, such that rapidly growing parasite stages utilize much of their resources to de novo biosynthesis of ribosomes, and that "dormant" oocyst stages do not synthesize new ribosomes. The 50- to 100-fold reduction in transcription of RPs together with the reduced rRNA transcription prevents that unnecessary new ribosomes are synthesized in oocysts.

Animals↗

Distribution of life cycle stages of Sarcoptes scabiei var wombati and effects of severe mange on common wombats in Victoria.

Seven female and three male common wombats (Vombatus ursinus) collected from forested areas of Victoria (Australia) over a 10 mo period, 10 April 1997 to 22 February 1998 had at least 30% of their skin affected by severe hyperkeratotic sarcoptic mange. Mangy wombats were grazing during the day, could be readily approached, were in poor body condition, and lacked subcutaneous fat. The anterolateral surface of the body was most heavily parasitised with Sarcoptes scabiei var wombati followed by the posterolateral surface, the dorsal region between the ears, the ears, ventral abdomen, medial aspect of the legs, axillary and inguinal areas, and the dorsal midline. Larvae were the most prevalent life-cycle stage followed by eggs, nymphs, females, and males. Mite numbers and the severity of clinical signs, namely thickness of scale crust and the degree of alopecia, were correlated and were symmetrical on each side of the body. Fissuring of crust and skin only occurred when scale crust was present. Bacterial infections occurred in three of 10 wombats within lymph nodes or the pleural cavity. Lymphoid depletion did not occur in lymph nodes or spleens and prescapular lymph nodes contained a greater amount of nuclear debris in germinal centres than non-mangy wombats. Seven wombats had fatty change in their livers. Gonads of mature wombats were not active or had minimal activity. Significant histopathological changes were not seen in the gastrointestinal tract, kidney, brain, myocardium, spleen, thyroid, reproductive tract, and gonads. Hematocrit, mean corpuscular volume, mean corpuscular hemoglobin, and concentrations of hemoglobin, lymphocytes, calcium, glucose, creatinine, total solids, total protein, albumin determined both colormetrically and electrophoretically, and globulins were significantly lower and concentrations of neutrophils, monocytes, phosphorus, urea, glutamate dehydrogenase, aspartate aminotransferase and creatine kinase were significantly higher in mangy versus captive wombats. Concentrations of erythrocytes, mean corpuscular hemoglobin, leucocytes, band neutrophils, eosinophils, nucleated erythrocytes, sodium, potassium, chloride, total bilirubin, alkaline phosphatase, and gamma glutamyltransferase for mangy wombats were not significantly different from that reported for captive wombats. Hematological and pathological changes in mangy wombats were consistent with anemia, inflammation, and changes seen with starvation.

Alopecia↗

Characterization of a life-cycle-stage-regulated membrane protein tyrosine phosphatase in Trypanosoma brucei.

We report the first characterization of plasma-membrane-bound tyrosine phosphatase activity in the haemoprotozoan. Trypanosoma brucei. Several enzymic properties of the membrane fraction were identical to other protein tyrosine phosphatases (PTPases), such as (a) insensitivity to inhibitors of other protein phosphatases, including tetramisole, sodium tartrate and okadaic acid, (b) inhibition by sodium vanadate, and (c) activation by spermidine. Additionally, T. brucei PTPase activity presented two novel features, an acidic pH optimum at pH 4.0-5.0 and a very low Km value (2.5 nM) for the specific synthetic substrate, Tyr(P)Raytide. Higher Km values of 170 nM for Tyr(P)-RCML (RCML, reduced, carboxamidomethylated and maleylated lysozyme) and of 3 mM for the non-specific inorganic substrate p-nitrophenyl phosphate, suggested that the PTPase activity of T. brucei was substrate specific. Reconstitution experiments on bloodstream-stage membrane proteins revealed that three polypeptides of 148, 115 and 72 kDa contained vanadate-inhibitable PTPase activity. Modulator assays revealed that the 72-kDa protein was responsible for the observed spermidine stimulation, but indicated that the modulator profile of the 148-kDa protein was most similar to the whole membrane fraction. Furthermore, the PTPase activity of T. brucei was life-cycle-stage regulated. Neither the whole membrane fraction nor the reconstituted proteins of the procyclic insect stage dephosphorylated tyrosine residues.

Animals↗

Endocytosis by African trypanosomes. II. Occurrence in different life-cycle stages and intracellular sorting.

Horseradish peroxidase (HRP) and colloidal gold-labeled proteins enter many of the endocytic organelles of bloodstream forms of Trypanosoma brucei and T. congolense. However, the colloidal gold markers were excluded from substantial parts of the pathway that contained HRP. Morphometric studies revealed that HRP entered organelles that accounted for approximately 5% of the total cell volume while transferrin-gold entered organelles that comprised approximately 2% of the total cell volume. In addition, large colloidal gold particles were excluded from organelles that contained smaller gold particles. Antibodies, raised against the variable surface glycoprotein, when applied to thawed cryosections were found to label structures from which endocytosed colloidal gold coupled to bovine serum albumin (BSA) was excluded. Endocytosis was shown to occur in two in vitro propagated forms of trypanosomes, similar to those found in the insect vector (Glossina spp.). The mammal-infective metacyclic forms were similar to bloodstream forms in that they endocytosed HRP and colloidal gold markers but excluded colloidal gold from approximately 3% of the endocytic organelles. Estimation of the flagellar pocket volumes of bloodstream form T. brucei showed that this organelle occupied 0.5% to 1.4% of the total cell volume. The flagellar pocket volume of T. congolense varied between life-cycle stages, with a fractional volume of 4.4% for bloodstream forms, 2.3% for metacyclic forms and 1.4% for procyclic forms. Endocytosis of HRP, but not of protein-gold markers, occurred in procyclic (uncoated) forms. Endocytosis by procyclic forms has heretofore not been reported.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗