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Schistosoma mansoni eggs as a target for praziquantel: efficacy of oral application in mice.

A study was undertaken to investigate the effect of administering a double oral dose of praziquantel (PZQ), given 9 days apart, on the egg viability in Schistosoma mansoni infected mice. The viability of eggs trapped in the tissues and those excreted in the faeces was determined using morphological (oogram) and function (egg hatching) criteria. Nine days after the first dose of PZQ, 90.5% of eggs excreted in the faeces were judged to be viable by the egg hatch method, as were 98.0% of eggs excreted from the control mice. However, only 44.0% of the eggs isolated from the intestine of PZQ treated mice were viable compared with 82.5% in the pretreatment group or 93.0% in the control group. Similar results were obtained for viability of eggs isolated from the intestine, by both the hatching and oogram methods. There was a dramatic reduction in the viability of eggs in liver tissue 9 days after the first dose of PZQ (13.0%) when compared with either the untreated controls (85.3%) or eggs in the livers of mice prior to PZQ treatment (80.0%). Following a second dose of PZQ (69 DPI), no eggs were detected in the faeces by 78 DPI, and eggs in the tissues were no longer viable. These data confirm the susceptibility of mature eggs of S. mansoni to a single dose of PZQ, but also demonstrate that the remaining immature eggs are undamaged and mature in the tissues of infected mice. Administration of a second dose of PZQ, 9 days after the first is suggested as an alternative treatment strategy assuring killing of all the schistosome eggs.

Administration, Oral↗

Labeling hen's egg with 74Se for use in human metabolic experiments.

Experiments were conducted with laying hens to explore quantitative aspects of incorporation of 75Se ( radioselenium ) at various dosing levels and for different chemical forms of an orally administered tracer. Quantitative distribution of the incorporated isotope in egg white and egg yolk was strongly influenced by both the chemical form of the label and the dosing level. The ratio of egg yolk:egg white selenium decreased with increased level of administered dose of selenite. In addition, the rate of incorporation and the amount of selenium in whole egg were higher when [75Se]selenomethionine was given as compared to [75Se]selenite. Characterization of the chemical form of selenium in egg white and egg yolk labeled biologically by giving hens radioactive selenite or selenomethionine was performed by classification as: selenite, selenoprotein and fat-bound selenium. Studies were then undertaken to achieve intrinsic labeling of egg white and egg yolk with stable isotope 74Se for purposes of exploring selenium bioavailability in humans. Enrichments of 74Se in egg white and egg yolk of hens given high dose selenite (54.4 micrograms 74Se ) were 20- and 28-fold, whereas in egg white and egg yolk of hens given low dose (10.9 micrograms 74Se ) they were 4- and 10-fold the level of natural abundance, respectively. The stable isotope-labeling studies indicated that a 7-day sequential dosing protocol with 20-100 micrograms Se per dose permitted sufficient enrichment of egg white (only high dose) and of yolk with the stable isotope 74Se for use in human metabolic studies.

Animals↗

Failure of hamster eggs fertilized in vitro to extrude the second polar body correlates with high levels of polyspermy.

The occurrence of second polar body (PB2) retention in in-vitro fertilized hamster eggs is reported. Of 2,872 eggs examined, 6.9% (199 eggs) failed to extrude PB2. Eggs that failed to extrude PB2 were more likely to be polyspermic than eggs with PB2 (63% and 14.6%, respectively), and the number of fusing sperm per egg was higher in the abnormal eggs. These data indicate that eggs which fail to extrude PB2 have an impaired block to polyspermy. The level of PB2 retention varied between females and ranged from all eggs extruding PB2 to 20% of the eggs failing to extrude PB2 (41% and 4% of the females, respectively). There was no correlation within a female between the percentage of eggs that failed to extrude PB2 and the level of polyspermy in the sister eggs with PB2. Therefore, regardless of the condition of their sister eggs, eggs that fail to extrude PB2 have an impaired block to polyspermy and eggs that extrude PB2 have a normal block.

Animals↗

The fertilization potential provides a fast block to polyspermy in lamprey eggs.

At fertilization, the membrane potential of the egg of the lamprey, Lampetra japonica, shifted rapidly from its resting value of -12 to +36 mV and gradually returned to about the same resting level (fertilization potential). The amplitude of depolarization was influenced by the external Cl- concentration and by an anion channel blocker, DIDS, indicating that the positive shift of membrane potential resulted from Cl- efflux. A similar change in membrane potential (activation potential) was observed when the unfertilized egg was pricked with a fine needle or treated with A23187 to induce parthenogenetic activation. Pricking at the animal pole region (predetermined site for sperm entry) resulted in the occurrence of an immediate activation potential and the initiation of cortical granule exocytosis. A time lag between the pricking and the occurrence of the activation potential was observed when the egg was pricked at a distance from the animal pole. In this instance, the activation potential was produced immediately before the propagating cortical granule exocytosis initiated at the pricked site reached the animal pole region. Sperm-egg fusion was blocked in eggs voltage-clamped at +20 to +40 mV and inseminated, whereas it took place in eggs clamped at -60 to 0 mV. However, most eggs clamped at +20 to +40 mV did activate, indicating that the voltage dependence of egg activation differs from that of sperm-egg fusion. Although eggs voltage-clamped at negative membrane potentials permitted multiple sperm to fuse with the egg plasma membrane, the nucleus of the fused sperm did not necessarily enter the ooplasm. We conclude that: (1) A fast electrical block against polyspermy operates in this species and is effective for about 160 sec of the onset of the positive shift; (2) the opening of Cl- channels is responsible for the potential change; (3) the channels are largely localized in the animal pole region; (4) during voltage clamp at positive potentials, eggs can be activated without sperm-egg fusion; and (5) during voltage clamp at negative potentials, sperm-egg fusion occurs, but sperm entry into the egg cytoplasm does not always proceed.

Animals↗

Schistosoma mansoni: in vitro adhesion of parasite eggs to the vascular endothelium. Subsequent inhibition by a monoclonal antibody directed to a carbohydrate epitope.

Schistosoma mansoni eggs come into direct contact with the vascular endothelium, particularly in the postcapillary venules of the mesenteric tract (oviposition site). We investigated the adhesion of eggs to endothelial cells in a static in vitro assay and in a flow-based in vitro assay. Live S. mansoni eggs rapidly attached, in a time-dependent manner, to the human endothelial cell line ECV 304, but not KOH-treated eggs. Activation of ECV monolayers with interleukin-1 promoted live S. mansoni eggs adhesion. An in vitro flow-based assay of human umbilical vein endothelial cells (HUVEC) showed the influence of wall shear stresses on the attachment of eggs to endothelial cells, particularly under postcapillary venule shear stress conditions. Interleukin-1 activation of HUVEC promoted adhesion between live eggs and endothelial cells. Higher wall shear stresses were needed to obtain the detachment of eggs from activated endothelial cells than control cells. Preincubation of interleukin-1-activated HUVEC, in a static in vitro assay, with monoclonal antibodies specific for intercellular adhesion molecule-1, E-selectin, and vascular cell adhesion molecule-1 significantly decreased adhesion of live eggs. Previous studies have shown that a monoclonal antibody specific for a schistosome carbohydrate epitope abundant in eggs is related to the Lewis X antigen. In this study, the anti-Lewis X-specific monoclonal antibody was used for adhesion-inhibition assays. Preincubation of eggs with this monoclonal antibody significantly decreased adhesion of live eggs to interleukin-1-activated HUVEC cultured in vitro. These results suggest that surface adhesion molecules, expressed by endothelial cells under conditions of interleukin-1 activation, directly participate in egg adhesion and that egg carbohydrate antigens play an important role in live S. mansoni egg adhesion to the vascular endothelium.

Animals↗

Loss of functional sperm entry into Xenopus eggs after activation correlates with a reduction in surface adhesivity.

In Xenopus, the plasma membrane of the unactivated egg is receptive to sperm only in the animal hemisphere (R. Grey, M. Bastiani, D. Webb, and E. Schertel, 1982, Dev. Biol. 89, 475-487). The reinsemination experiments of investment-free eggs reported in this paper demonstrate that functional sperm entry is lost after activation. Supernumerary sperm were excluded even though the fertilization envelope was absent and the membrane potential had returned to the level found in the unfertilized egg. Even when the electrical block to polyspermy was suppressed by 40 mM NaI (which reduces the membrane potential), polyspermy could be induced only if denuded eggs were initially inseminated in this medium. We estimate that the loss of functional sperm entry, independent of the electrical block, occurs during the first 10 min following fertilization. Sperm readily adhere to the surface of the animal hemisphere of unactivated eggs divested of their extracellular coats, but they do not adhere to the surface of activated eggs. Denuded eggs also adhere to each other, with the surface of the animal hemisphere of unactivated eggs exhibiting the greatest degree of adhesivity. We used electric field-induced fusion (EFIF), without prior dielectrophoresis, to quantify the regional and temporal adhesiveness of eggs. At electric field strengths greater than 8 V/cm, the probability of fusion during EFIF is highest with the animal hemisphere of unactivated eggs, moderate with both the vegetal hemisphere of unactivated eggs and the animal hemisphere of activated eggs, and lowest with the vegetal hemisphere of activated eggs. When pairs of eggs are constructed with different hemispheres in contact, the fusion characteristics of the pair are similar to the more adhesive member of the pair. The regional and temporal differences in the adhesiveness of the Xenopus egg surface correlate with its receptivity to sperm and could possibly account for the plasma membrane's activation-induced loss of functional sperm entry.

Animals↗

Co-culture of ovine eggs with oviductal cells and trophoblastic vesicles.

Three experiments were conducted to determine whether coculture of early sheep eggs with oviductal cells would improve the ability of eggs to survive in culture. Eggs recovered from superovulated ewes were cultured in Ham's F-10 medium supplemented with 10% fetal calf serum (F10FCS) at 37.5 degrees C in 95% air:5% CO(2). In Experiment 1, eggs with one to eight cells were either transferred into recipient ewes immediately after collection or were cultured for 24 h in 5 ml Ham's F10 medium supplemented with 10% fetal calf serum (F10FCS), 5 ml F10FCS on a confluent monolayer of oviductal cells; in 25 ml of fresh F10FCS; or in 25 ml of F10FCS removed cultures of oviductal cells, 25 mul of fresh F10FCS or 25 mul of F10FCS removed from cultures of oviductal cells. After 24 h, the cultured eggs were transferred to recipient ewes synchronous with donors and subsequently recovered at necropsy on Day 8 post estrus. Coculture of sheep eggs with oviductal cells improved (P < 0.05) the development of transferred eggs compared to culture in F10FCS alone. In Experiment 2, eggs recovered from superovulated ewes on Days 3 to 6 after estrus had undergone 1.8 cleavages by Day 3 and 4.1 cleavages by Day 6. In Experiment 3, single-cell eggs were cultured for 3 d in 5 ml F10FCS, cocultured with ovine trophoblastic vesicles or cultured on a confluent monolayer of oviductal cells. Coculture of eggs in F10FCS on a monolayer of oviductal cells supported in vitro egg cleavage to a greater degree than did F10FCS alone or F10FCS with trophoblastic vesicles (P < 0.05). In Experiment 4, single-cell eggs were cultured for 3 d then transferred to recipients. Eggs were cultured in 5 ml F10FCS on confluent monolayers of oviductal cells from luteal or estrous ewes or on cells that had been frozen after recovery from a culture of oviductal cells. After culture, the eggs were transferred to oviducts of recipients and recovered 3 d later at necropsy. Coculture of eggs for 72 h with oviductal cell monolayers did not increase the in vitro, or subsequent in vivo, cleavage rate regardless of the type of oviductal cells.

Journal Article↗

Studies on ticks of veterinary importance in Nigeria. VI. Comparisons of oviposition and the hatching of eggs of Hyalomma species.

Fully engorged Hyalomma spp. in Nigeria oviposited greater numbers of eggs than those partially engorged. Hyalomma impressum was a more prolific egg layer than H. rufipes, H. impeltatum and H. truncatum. The variations in the egg output as well as the recognizable peaks in the number of eggs during oviposition were described for each species. No species of Hyalomma below the engorged weight of 0.2 g oviposited; oviposition started with ticks of weight 0.3 g. Eggs produced by ticks weighing below 0.3 g did not hatch; the highest percent egg eclosion occurred with ticks of weight 0.6 g (H. rufipes) and 0.7 g (other Hyalomma species). The pre-oviposition, oviposition and eclosion periods were shortened when eggs were laid and incubated at high temperatures, although the number of oviposited eggs did not increase significantly. At the standard temperature of 24 degrees C, the longest eclosion period was seen in the eggs of H. rufipes (41 days) while those of H. truncatum, H. impressum and H. impeltatum were similar to each other (29 days). Only eggs of H. rufipes hatched at an incubation temperature of 15 degrees C. Eggs of Hyalomma species laid at the same time hatched over a 2--4 day period, except at 15 degrees C when the hatching period of H. rufipes lasted 10 days. The eclosion period was longest in the earlier ovipositions and shorter in the later ones. It is suggested that some intrauterine larval development might have started in the eggs before they were released at a later oviposition period. The percentage mortality of eggs at various temperatures showed that eggs of H. rufipes were more tolerant of low temperatures than those of H. impressum, H. truncatum and H. impeltatum, while the eggs of the latter 3 species were more tolerant of high temperatures than those of H. rufipes. The relevance of these results of the distribution and abundance of the Hyalomma species in Nigeria was discussed.

Animals↗

Partial characterization of the carbohydrates of the eggs (oncospheres) of the tapeworm, Hymenolepis diminuta.

The carbohydrate constituents of whole eggs and fractions derived from the eggs of Hymenolepis diminuta were partly characterized. Whole eggs contained 9.6 ng of carbohydrate (phenol-sulphuric acid-positive material) per egg, and 3.6 ng of glucose. Analysis of hydrolysed eggs by HPLC demonstrated the presence of glucose, galactose, glycerol, N-acetylgalactosamine, and 2-deoxyribose. Isolated egg shells and the KOH-stable, ethanol-precipitable fraction (putative glycogen) of eggs contained 26 and 76%, respectively, of the total carbohydrate associated with eggs. Glucose and galactose were present in both of these samples, but only glucose was present in the KOH-stable, ethanol-precipitable fraction isolated from chemically deshelled eggs; thus, the source of the galactose in the KOH-stable, ethanol-precipitable fraction isolated from whole eggs was the egg-shells. These data confirm earlier reports that the shells contain carbohydrate; the data demonstrate further that only two monosaccharides (glucose and galactose) are present in the shells, and that they are present as high molecular weight polymers. The ethanol-soluble fraction of eggs contained 2.5% of the total carbohydrate in eggs, and glucose accounted for < 1% of the ethanol-soluble carbohydrate. Analysis of the ethanol-soluble fraction by HPLC demonstrated that the predominant monosaccharides were glycerol and mannose, with smaller quantities of 2-deoxyribose and 2-deoxyglucose. The absence of a 'free pool' of glucose in eggs, and the presence of large amounts of glycerol and mannose, suggest that the pathways of intermediary carbohydrate metabolism in eggs might be very different from those in adult tapeworms.

Acetylgalactosamine↗

Tetrodotoxin levels in eggs of the rough-skin newt, Taricha granulosa, are correlated with female toxicity.

We quantified the amount of the neurotoxin tetrodotoxin (TTX) present in females and newly deposited eggs of the rough-skin newt, Taricha granulosa, to examine the relationship between the toxicity of an individual female and the toxicity of her eggs. We found high levels of TTX in individual eggs as well as substantial variation among clutches. Variation in the amount of TTX per egg within individual clutches was extremely low. Female skin toxicity was positively correlated with the mean egg toxicity of her clutch. Neither egg volume nor female size was significantly correlated with egg TTX levels. Tetrodotoxin stereoisomer-analog profiles were identical for females and their eggs. The presence of high levels of TTX in individual eggs coupled with the relationship between levels of TTX in female skin and levels of TTX in her eggs suggests that the TTX present in eggs of T. granulosa is maternally derived. The lack of correlation between egg size and TTX levels in individual eggs, as well as the low levels of within clutch variation, may indicate that deposition of TTX in eggs of T. granulosa is not linked to the deposition of other egg resources (e.g.. lipids or other yolk components).

Animals↗

Attempts to modify lung granulomatous responses to Schistosoma japonicum eggs in low and high responder mouse strains.

A radioisotopic assay for acute granulomatous hypersensitivity (AGH) to lyophilized eggs of Schistosoma japonicum has been used to further examine responses to egg antigens in various inbred strains of mice. The ranking of responsiveness in mice from high (C57BL/6), intermediate (BALB/c) to low (CBA/H) was not influenced by high or low egg-sensitization regimens. However, the low responsiveness of responder mice sensitized with eggs by the intraperitoneal compared with the subcutaneous route of injection appears to be an egg dose-related phenomenon. The high AGH responsiveness of C57BL/6 mice can be increased further by sensitization with eggs in the presence of purified pertussigen from Bordetella pertussis but CBA/H mice treated identically remain low responders. The monoclonal anti-egg antibody, P.41, which produces a prominent bleb-type circumoval precipitate with eggs, has been shown to be directed against major 'immunopathologic antigen(s)' of S. japonicum eggs. Thus, C57BL/6 mice were sensitized for AGH by injection of soluble extracted egg antigen (SEA) bound to an immunoabsorbent of P.41 antibody on Sepharose. No success has been achieved in modulating AGH in C57BL/6 mice by injection of hyperimmune antisera raised against lyophilized eggs in either high or low responder mouse strains. This failure is in line with previous results using antisera as well as monoclonal anti-egg antibodies. The consistent failure to demonstrate a modulating effect of antibodies in this compared with other laboratories may be related to the use of lyophilized rather than viable eggs. The data suggest that activities of antisera in granuloma modulation in murine schistosomiasis japonica result from egg destruction or inhibition of production of immunopathologic antigens by eggs rather than through effects on immunopathologic immune responses.

Animals↗

A comparison of the effects of feeding treatments and lighting on age at first egg and subsequent laying performance and carcase composition of broiler breeder hens.

The effects of the growth curve from 15 to 20 weeks, age at photostimulation and pattern of photostimulus on sexual maturity, egg production and egg weight were evaluated in two trials with broiler breeder females to 56 and 34 weeks of age (housed in litter pens and individually caged, respectively). Carcase composition and reproductive morphology of hens varying in laying efficiency were measured in the second trial. Trial 1: Four growth curves were applied from 15 to 20 weeks to pullets housed in litter pens. Birds were transferred from 8- to 10-h photoperiods at 20 or 24 weeks of age, followed by weekly increments of one hour to reach a 16-h final photoperiod. Lower-than-recommended body weights at 20 weeks significantly delayed sexual maturity, reduced peak rate of lay, total eggs and mean egg weight. However, double-yolked egg production was lower, resulting in non-significant differences in settable egg numbers between body weight treatments. Birds reared to the heaviest body weight exhibited a significantly advanced sexual maturity, but total egg numbers, peak rate of lay and mean egg weight were not significantly affected. However, the laying of more double-yolked eggs resulted in a decrease in the number of settable eggs. Delaying photostimulation to 24 weeks significantly retarded sexual maturity, reduced total and settable egg numbers, and increased mean egg weight. A tendency for fewer double-yolked eggs was observed. Trial 2: At 19 weeks, birds were selected from the 4 body weight categories in Trial 1 and moved to individual cages in 8 rooms. Five lighting programmes were applied. The pattern of photostimulation applied did not affect any of the production traits measured. At 34 weeks, 24 birds were selected for the analysis of reproductive morphology, presence of internal or multiple ovulations, and carcase composition. Eight hens showing an early age at first egg and regular egg production records, 8 birds showing erratic laying performance and 8 non-layers were killed for this purpose. There were no differences in carcase composition or reproductive morphology between these groups. No internal ovulations or double or multiple ovarian hierarchies were observed. The results presented confirm that broiler breeders do not require a lighting stimulus in order to initiate ovarian activity and that, where no lighting stimulus is given, body weight or feeding level plays a critical role in stimulating the birds to attain sexual maturity. However, when a lighting stimulus is given, factors such as body weight and body composition become relatively less important in regulating the age at sexual maturity.

Animal Feed↗

Stimulation of repetitive calcium transients in mouse eggs.

1. We have combined cell membrane electroporation by electrical field (EF) stimulation with a rapid perfusion system in order to stimulate repetitive increases in cytoplasmic free [Ca2+] ([Ca2+]i) in mouse eggs. [Ca2+]i was monitored by ratio fluorescent measurements of intracellular indo-1 on individual eggs. The conditions required to cause different types of [Ca2+]i increases were established and the effects of these [Ca2+]i changes upon egg activation examined. 2. The rapid perfusion of non-ionic medium caused a single [Ca2+]i increase. However, to generate repetitive [Ca2+]i increases, eggs were exposed to EF pulses in the presence of Ca2+ and then washed rapidly with culture medium. Sequential EF pulse application led to prolonged elevation of [Ca2+]i levels and eventual cell lysis unless rapid reperfusion with culture medium was achieved. Transient increases in [Ca2+]i in eggs could also be generated by EF pulses in the presence of inositol 1,4,5-trisphosphate (InsP3). 3. In response to EF stimulation fertilized eggs showed [Ca2+]i increases that were enhanced relative to unfertilized eggs. The responses in these fertilized eggs were often followed by repetitive [Ca2+]i oscillations, despite the fact that the [Ca2+]i oscillations associated with sperm penetration had ceased by this stage. 4. In unfertilized mouse eggs the [Ca2+]i increases appeared to be due to direct cation influx since repeated EF pulses caused repeated influx of Mn2+ as monitored by quenching of fluorescence of fura-2 loaded eggs. 5. Under conditions that stimulated reproducible patterns of [Ca2+]i transients we found that a single large [Ca2+]i transient did not cause significant egg activation, but that inducing repetitive [Ca2+]i transients was effective in activating eggs. The speed of activation as judged by the rate of pronuclear formation was also dependent upon the frequency of pulse application. 6. These data show that combining EF pulses with a rapid and precise sequential perfusion system can be used to manipulate [Ca2+]i levels in mammalian eggs. This provides a means of artificial mimicry of the [Ca2+]i transients seen after fertilization. It appears that Ca2+ influx during EF pulses does not cause significant Ca2+ release from internal stores in unfertilized eggs, but after fertilization Ca2+ influx does induce Ca2+ release. It is also apparent that mouse eggs are more successfully activated by repetitive [Ca2+]i increases than by single large [Ca2+]i rises. We suggest that our data provide direct evidence for the hypothesis that a cellular response to oscillations of intracellular [Ca2+]i can be distinct from that to monotonic rises in [Ca2+]i.

Animals↗

Circadian clock controlling egg hatching in the cricket (Gryllus bimaculatus).

Adult crickets (Gryllus bimaculatus) were maintained under a 12-h light:12-h dark cycle (LD 12:12). After oviposition, their eggs were incubated under different lighting regimens at 23 degrees C, and temporal profiles of egg hatching were examined. When the eggs were incubated in LD 12:12 or in DL 12:12 with a phase difference of 12h from LD 12:12, throughout embryogenesis, 88% to 97% of hatching occurred within 3 h of the dark-light transition on days 17 and 18 of embryogenesis; the phases of the egg-hatching rhythms in the LD 12:12 and DL 12:12 groups differed by about 12 h. In eggs incubated in constant darkness (DD) throughout embryogenesis, a circadian (about 24 h) rhythm of hatching was found, and the phase of the rhythm was similar to that seen in eggs incubated in LD 12:12, but not DL 12:12, throughout embryogenesis. When eggs that had been incubated in DD after oviposition were transferred to DL 12:12 in the middle or later stages of embryogenesis and were returned to DD after three cycles of DL 12:12, the rhythm of hatching synchronized (entrained) to DL 12:12. However, when eggs in the earlier stages of embryogenesis were transferred from DD to DL 12:12 and returned to DD after three cycles, 52% to 94% of hatching did not entrain to DL 12:12. To determine whether photoperiodic conditions to which the parents had been exposed influenced the timing of egg hatching, adult crickets were maintained in DL 12:12, and their eggs were incubated in LD 12:12, DL 12:12, or DD throughout embryogenesis. The egg-hatching rhythm was also found in the eggs incubated under these three lighting regimens. In DD, the phase of the rhythm was similar to that seen in eggs incubated in DL 12:12, not LD 12:12, throughout embryogenesis. The results indicate that in the cricket, the timing of egg hatching is under circadian control and that the circadian rhythm of hatching entrains to 24-h light:dark cycles, but only if the light:dark cycles are imposed midway through embryogenesis. Therefore, by midembryogenesis, a circadian clock has been formed in the cricket, and this is entrainable to light:dark cycles. In addition, the photoperiodic conditions to which the parents (probably the mothers) have been exposed influence the timing of hatching, suggesting that maternal factors may regulate the timing of egg hatching.

Animals↗

OXYGEN TRANSPORT IN EGG MASSES OF THE AMPHIBIANS RANA SYLVATICA AND AMBYSTOMA MACULATUM: CONVECTION, DIFFUSION AND OXYGEN PRODUCTION BY ALGAE

Many amphibians lay their eggs in gelatinous masses up to 10&shy;20 cm in diameter, posing problems for diffusive oxygen delivery. Oxygen may also be provided by water convection between eggs or by oxygen production by endogenous algae. We studied egg masses of two local amphibians, Rana sylvatica and Ambystoma maculatum, to estimate the importance of each of these processes. We injected dye to check for water channels, measured oxygen partial pressures within egg masses to determine the influence of external water convection and lighting, measured oxygen consumption and production in darkness and light and calculated expected gradients through egg masses with a cylindrical, homogeneous egg mass model. Rana sylvatica had relatively loose egg masses with water channels between the eggs; water convection was important for oxygen delivery. Ambystoma maculatum had firm egg masses with no spaces in the jelly between eggs; thus, there was no opportunity for convective oxygen delivery. The egg masses were cohabited by Oophila ambystomatis, a green alga found specifically in association with amphibian egg masses. Oxygen delivery in A. maculatum was by diffusion and by local production by the algal symbiont. Analysis of a cylindrical egg mass model and measurement of oxygen gradients through egg masses indicated that diffusion alone was not adequate to deliver sufficient O2 to the innermost embryos at late developmental stages. In the light, however, egg masses had a net oxygen production and became hyperoxic. Over the course of a day with a 14 h:10 h light:dark cycle, the innermost embryos were alternately exposed to hyperoxia and near anoxia.

Journal Article↗

Tolerization of mice to Schistosoma mansoni egg antigens causes elevated type 1 and diminished type 2 cytokine responses and increased mortality in acute infection.

The granuloma that surrounds the Schistosoma mansoni egg is the cause of pathology in murine schistosomiasis, and its formation is driven by egg Ag-stimulated type 1 and type 2 cytokines. To determine the role of egg-driven immune responses during schistosome infection we rendered CBA/Ca mice unresponsive to schistosome eggs by combined cyclophosphamide treatment and thymectomy. In the early acute stages of schistosome infection, egg-tolerized mice suffered high mortalities. Granuloma size and deposition of collagen in the liver were significantly reduced in egg-tolerized mice. Similarly, limited granuloma responses were detected in the intestines of these mice, and this was associated with a >90% reduction in egg excretion. Histologically, egg-tolerized mice had exacerbated hepatocyte damage, with extensive microvesicular steatosis. Elevated plasma transaminase levels confirmed the damage to hepatocytes. Infected egg-tolerized mice had impaired proliferation responses to egg Ag but intact responses to worm Ag. Tolerized mice had diminished Ab responses to egg Ag and had a type 1 cytokine isotype pattern to worm Ag, with elevated IgG2a and diminished IgG1 and IgE. Egg-tolerized mice failed to down-regulate type 1 cytokines that are normally elicited during early schistosome infection. Hepatic granuloma cells from egg-tolerized mice were also type 1 cytokine dominated, with elevated frequencies of Tc1/Th1 and reduced Tc2/Th2 cells. This study demonstrates that mice tolerized to schistosome eggs have elevated type 1 cytokine responses with diminished type 2 responses and reduced anti-egg Ab during schistosome infection, and these effects are detrimental to the host.

Animals↗

[Integrins mediate the increase of concentration of intracellular free calcium in mouse eggs].

For studying the effect of integrin on the [Ca(2+)](i) of mouse eggs and its transmembrane signaling mechanism, zona-free mouse eggs were loaded with calcium probe Fluo-3/AM and the intensity of fluorescence of the eggs treated with different factors was measured through laser confocal microscopy. The results showed that the [Ca(2+)](i) of zona-free mouse eggs was increased when the eggs were treated with RGD peptide, fibronectin (Fn) and anti-mouse integrin subunit alpha(6) and beta(1) monoclonal antibodies, respectively. The [Ca(2+)](i) of the mouse eggs was also increased when the eggs were placed in calcium-free medium and treated with RGD peptide or Fn. The changes in the mouse egg [Ca(2+)](i) caused by RGD and Fn were similar to those caused by sperm. However, the concentration of Ca(2+) of the zona-free mouse eggs pretreated with tyrosine kinase inhibitor was not increased when the eggs were treated in the same way, and, neither was the intracellular calcium increased in those eggs pretreated with PKC inhibitor when the eggs were treated with RGD peptide. It is therefore suggested that the occupancy of integrins on the membrane of mouse eggs by their ligands mediates the release of Ca(2+) and then the increase in the [Ca(2+)](i) of eggs, which is one of the early events of egg activation. The tyrosine kinase signaling pathway and PKC are involved in this process as well.

Animals↗

A 12-wk egg intervention increases serum zeaxanthin and macular pigment optical density in women.

Two carotenoids found in egg yolk, lutein and zeaxanthin, accumulate in the macular retina where they may reduce photostress. Increases in serum lutein and zeaxanthin were observed in previous egg interventions, but no study measured macular carotenoids. The objective of this project was to determine whether increased consumption of eggs would increase retinal lutein and zeaxanthin, or macular pigment. Twenty-four females, between 24 and 59 y, were assigned to a pill treatment (PILL) or 1 of 2 egg treatments for 12 wk. Individuals in the PILL treatment consumed 1 sugar-filled capsule/d. Individuals in the egg treatments consumed 6 eggs/wk, containing either 331 microg (EGG 1) or 964 microg (EGG 2) of lutein and zeaxanthin/yolk. Serum cholesterol, serum carotenoids, and macular pigment OD (MPOD) were measured at baseline and after 4, 8, and 12 wk of intervention. Serum cholesterol concentrations did not change in either egg treatment group, but total cholesterol (P = 0.04) and triglycerides (P = 0.02) increased in the PILL group. Serum zeaxanthin, but not serum lutein, increased in both the EGG 1 (P = 0.04) and EGG 2 (P = 0.01) groups. Likewise, MPOD increased in both the EGG 1 (P = 0.001) and EGG 2 (P = 0.049) groups. Although the aggregate concentration of carotenoid in 1 egg yolk may be modest relative to other sources, such as spinach, their bioavailability to the retina appears to be high. Increasing egg consumption to 6 eggs/wk may be an effective method to increase MPOD.

Adult↗