Bursts in discontinuous Aeolian saltation.

Close to the onset of Aeolian particle transport through saltation we find in wind tunnel experiments a regime of discontinuous flux characterized by bursts of activity. Scaling laws are observed in the time delay between each burst and in the measurements of the wind fluctuations at the fluid threshold Shields number θc. The time delay between each burst decreases on average with the increase of the Shields number until sand flux becomes continuous. A numerical model for saltation including the wind-entrainment from the turbulent fluctuations can reproduce these observations and gives insight about their origin. We present here also for the first time measurements showing that with feeding it becomes possible to sustain discontinuous flux even below the fluid threshold.

Disease associated balanced chromosome rearrangements: a resource for large scale genotype-phenotype delineation in man.

Disease associated balanced chromosomal rearrangements (DBCRs), which truncate, delete, or otherwise inactivate specific genes, have been instrumental for positional cloning of many disease genes. A network of cytogenetic laboratories, Mendelian Cytogenetics Network (MCN), has been established to facilitate the identification and mapping of DBCRs. To get an estimate of the potential of this approach, we surveyed all cytogenetic archives in Denmark and southern Sweden, with a population of approximately 6.6 million. The nine laboratories have performed 71 739 postnatal cytogenetic tests. Excluding Robertsonian translocations and chromosome 9 inversions, we identified 216 DBCRs ( approximately 0.3%), including a minimum estimate of 114 de novo reciprocal translocations (0.16%) and eight de novo inversions (0.01%). Altogether, this is six times more frequent than in the general population, suggesting a causal relationship with the traits involved in most of these cases. Of the identified cases, only 25 (12%) have been published, including 12 cases with known syndromes and 13 cases with unspecified mental retardation/congenital malformations. The remaining DBCRs were associated with a plethora of traits including mental retardation, dysmorphic features, major congenital malformations, autism, and male and female infertility. Several of the unpublished DBCRs defined candidate breakpoints for nail-patella, Prader-Willi, and Schmidt syndromes, ataxia, and ulna aplasia. The implication of the survey is apparent when compared with MCN; altogether, the 292 participating laboratories have performed >2.5 million postnatal analyses, with an estimated approximately 7500 DBCRs stored in their archives, of which more than half might be causative mutations. In addition, an estimated 450-500 novel cases should be detected each year. Our data illustrate that DBCRs and MCN are resources for large scale establishment of phenotype-genotype relationships in man.

Primary and secondary infections with Cryptosporidium parvum in immunosuppressed adult mice.

The present study was undertaken to determine if infection of immunocompetent adult C57BL/6N mice with Cryptosporidium parvum would render them more resistant to a challenge infection following immunosuppression with dexamethasone (DEX). Fecal oocyst shedding and parasite colonization of the terminal ilea were greater in immunosuppressed mice than in nonimmunosuppressed mice. Secondary infections with C. parvum resulted in decreased oocyst shedding and reduced parasite colonization compared with primary infections. Flow cytometry revealed fewer splenic B cells but more splenic total T, CD4+ T, CD8+ T cells and macrophages in immunosuppressed mice than in nonimmunosuppressed mice. The CD4+ to CD8+ T cell ratios and blastogenic responses to lipopolysaccharide (LPS), but not to concanavalin A, were decreased in immunosuppressed mice compared with nonimmunosuppressed mice. Blastogenic responses to LPS and percentages of splenic total B cells and macrophages were increased in secondary infections compared to primary infections. Enhanced susceptibility to C. parvum infection in immunosuppressed mice revealed DEX-mediated effects on both cell-mediated and humoral immunity. Our results suggest that increased resistance in immunosuppressed mice to secondary infections with C. parvum may involve increases in B cells and macrophages.

Effects of dehydroepiandrosterone in immunosuppressed adult mice infected with Cryptosporidium parvum.

Cryptosporidiosis is a diarrheal disease in humans and other animals caused by the coccidian parasite, Cryptosporidium parvum. This study was undertaken to determine the effectiveness of dehydroepiandrosterone (DHEA) in reducing C. parvum infections in immunosuppressed adult C57BL/6N mice and to identify the immunomodulatory effects of DHEA that result in increased resistance to cryptosporidiosis. Dexamethasone-immunosuppressed mice were readily infected with C. parvum following orogastric intubation with 10(6) oocysts/mouse. DHEA treatment of these mice significantly reduced (P < 0.01) both fecal oocyst shedding and parasite colonization of the ilea. Immunosuppressed mice treated with DHEA had more splenic total T cells, CD4+ T cells, and CD8+ T cells than immunosuppressed mice that were not treated, but the differences were not always significant. Moreover, nonimmunosuppressed mice treated with DHEA had significantly more (P < 0.05) splenic total T cells, CD4+ T cells, and total B cells than nonimmunosuppressed mice that did not receive DHEA. Of particular interest was the significantly larger (P < 0.05) number of CD8+ T cells in immunosuppressed, C. parvum-infected, DHEA-treated mice compared with the same mice that were not treated. Up-regulation of the immune system by exogenous DHEA may be useful in the treatment and palliation of cryptosporidiosis.

Therapeutic efficacy of paromomycin in immunosuppressed adult mice infected with Cryptosporidium parvum.

The intent of this study was to evaluate the therapeutic efficacy of paromomycin in immunosuppressed adult C57BL/6N mice infected with Cryptosporidium parvum. Seven groups of 10 mice/group were used. Groups 1, 2, and 7 served as normal, toxicity, and placebo controls, respectively. Groups 2-7 were immunosuppressed with dexamethasone phosphate administered ad libitum in drinking water. Groups 3-7 were infected with C. parvum on day 7 postimmunosuppression. Groups 3-6 were treated by administering paromomycin per os for 10 consecutive days, beginning on day 10 postinfection, at dosage levels of 0.25, 0.5, 1, and 2 g/kg/day, respectively. Paromomycin was judged to be nontoxic at the dosage levels used. Groups 1 and 2 remained uninfected while groups 3-7 began shedding oocysts by day 3 postinfection. Paromomycin was therapeutically effective against C. parvum at 1 and 2 g/kg/day as determined by significant reductions in fecal oocyst shedding (P < 0.01), parasite colonization (P < 0.05), and villus atrophy (P < 0.05) in the ilea and terminal ilea of infected mice. We conclude that paromomycin may be useful in the treatment and palliation of cryptosporidiosis.

Effects of dehydroepiandrosterone in immunosuppressed rats infected with Cryptosporidium parvum.

Cryptosporidium parvum is a coccidian parasite that causes diarrheal disease in animals and humans. Severe cryptosporidial infections were noted in young adult rats immunosuppressed with the glucocorticosteroid dexamethasone (DEX). B-cell and T-cell responses to the mitogens lipopolysaccharide and concanavalin A, respectively, were depressed in the DEX-treated rats. In addition, DEX treatment suppressed serum IgG levels, in vitro IgG production, and natural killer cell activities. Previous results have shown that DEX-immunosuppressed rats treated with dehydroepiandrosterone (DHEA) exhibit significant reductions in cryptosporidiosis as determined by monitoring oocyst shedding in the feces and parasite colonization of the small intestine. Results from this study indicated that B- and T-cell responses to their respective mitogens, serum IgG levels, and in vitro IgG production were greater in DHEA-treated immunosuppressed rats than in untreated DEX-immunosuppressed rats infected with C. parvum. Similar results were demonstrated in DHEA-treated versus normal control rats infected with C. parvum. These results suggest that the effects of DHEA in reducing cryptosporidiosis are the result of a potentiation of the immune system in the immunosuppressed rats.

Complete development of Cryptosporidium parvum in a human endometrial carcinoma cell line.

Sporozoites of Cryptosporidium parvum, excysted from oocysts isolated from calves, were applied to monolayers of the human endometrial carcinoma cell line RL95-2. Cells were grown as monolayers in 24-well plates at concentrations ranging from 5 x 10(4) to 1 x 10(5) RL95-2 cells per well. At 1 or 7 days postculturing, C. parvum sporozoites (ranging from 1 x 10(5) to 2 x 10(5) were added to the monolayers of RL95-2 cells. The cells were fixed and stained to estimate the extent of parasite colonization. Light microscopy and electron microscopy confirmed the development and replication of C. parvum within the RL95-2 cells. A standardized and reproducible in vitro culture system for C. parvum is necessary to evaluate therapies against cryptosporidiosis.

Dehydroepiandrosterone-induced reduction of Cryptosporidium parvum infections in aged Syrian golden hamsters.

Cryptosporidiosis, a parasitic disorder caused by Cryptosporidium parvum, is frequently a fulminating and life-threatening disease in immunocompromised hosts. The immune status of the host plays a critical role in determining the length and severity of the disease. Dehydroepiandrosterone (DHEA) is an immunomodulator that has been demonstrated to upregulate immune parameters. Ten aged (20-24 mo) Syrian golden hamsters were treated with DHEA for 7 days prior to intragastric inoculation with 1 x 10(6) C. parvum oocysts. An additional 10 aged hamsters were infected similarly but retained as untreated controls. The untreated hamsters presented with generalized infections as determined by oocyst shedding in the feces and parasite colonization of the small intestine. Hamsters treated with DHEA exhibited a significant reduction in cryptosporidial infection when compared to untreated hamsters. These results suggest that DHEA may be an effective prophylactic agent for cryptosporidiosis in immunocompromised patients.

Experimental Cryptosporidium parvum infections in immunosuppressed adult mice.

Five strains of adult mice were immunosuppressed with the synthetic glucocorticosteroid dexamethasone (DEX), administered either orally or intraperitoneally. The strains of mice used were C57BL/6N, DBA/2N, CBA, C3H/HeN, and BALB/cAnN. All mice were evaluated for susceptibility to Cryptosporidium parvum after intragastric inoculation with 10(6) oocysts per mouse. The DBA/2N, CBA, C3H/HeN, and BALB/cAnN mice given 0.25 micrograms of DEX per g per day orally (the dose and route previously used to infect rats with C. parvum) failed to develop chronic infections. However, the C57BL/6N mice sustained light infections during the entire 28-day experiment. The five strains of mice were also administered DEX intraperitoneally at concentrations ranging from 62.5 to 500 micrograms/day. Only the C57BL/6N mice given DEX at 125 micrograms/day developed chronic infections which persisted over 10 weeks, suggesting that the genetic background of the mouse plays a role in determining susceptibility to cryptosporidosis following immunosuppression with DEX. We believe that the C57BL/6N mouse model will prove to be superior to other animal models for evaluating potential anticryptosporidial agents, as well as for elucidating the immunological defects that allow C. parvum to establish chronic infections, because of cost effectiveness and ease in maintenance, breeding, and handling. We also evaluated the C3H/HeJ/beige mouse (lacks natural killer cell activity) and the C57BL/6N mouse maintained on a low-protein diet to induce immunosuppression. Neither of these mice exhibited heavy cryptosporidial infections.

Cryptosporidium parvum: experimental infections in aged Syrian golden hamsters.

Young (8-12 weeks) and aged (20-24 months) Syrian golden hamsters were intragastrically inoculated with Cryptosporidium parvum oocysts to compare the susceptibility to cryptosporidiosis between these two age groups. Oocyst shedding was significantly more intense in the aged than in the young hamsters. Moreover, parasite colonization of the small intestine was observed in the aged but not the young hamsters. Splenocytes from aged hamsters exhibited significantly lower T, B, and natural killer cell activities than did those from young hamsters. These studies suggest that susceptibility to cryptosporidial infections may be greater in aged individuals than has been previously realized.

Effectiveness of dehydroepiandrosterone in reduction of cryptosporidial activity in immunosuppressed rats.

The aim of this study was to evaluate the efficacy of the immunomodulator dehydroepiandrosterone in the treatment of cryptosporidiosis in glucocorticoid-induced immunosuppressed rats. A significant reduction in cryptosporidial activity occurred in treated versus untreated rats as assessed by oocyst shedding intensities and examination of parasite colonization in the small intestine.

Production and partial characterization of monoclonal antibodies specific for the gamonts of Eimeria tenella.

Thirteen hybridomas secreting monoclonal antibodies (Mabs) specific for the sexual stages (gamonts) of Eimeria tenella were produced by fusing spleen cells of gamont-immunized RBF/Dn mice with FOX-NY myeloma cells. A Mab subisotype profile revealed 1 IgG2a and 12 IgG1. All Mabs demonstrated a similar binding pattern when incubated with parasitic gamonts as determined by the indirect fluorescent antibody test. Ascitic fluid containing Mab (GD9 (IgG1) was produced and used to immunize chicks passively per os. There was a 34% decrease (P less than 0.05) in oocyst output from immunized chicks when compared to control chicks. Passively immunized chicks also had reduced cecal lesion scores when compared to control chicks. These results suggest that Mab GD9 partially inhibited the fertilization process of E. tenella.

Effects of dexamethasone and dehydroepiandrosterone in immunosuppressed rats infected with Cryptosporidium parvum.

Treatment of rats immunosuppressed with dexamethasone and inoculated with Cryptosporidium parvum oocysts were treated with dehydroepiandrosterone. A significant reduction in cryptosporidial activity was observed as determined by oocyst shedding and colonization of host tissue by parasites. Dexamethasone treatment alone resulted in decreases in T-, B- and natural killer (NK) cell responses and antibody production that, with the exception of NK-cell activity, were all reversed after administration of dehydroepiandrosterone.

Isolation and characterization of an estrogen-regulated ribosome-associated inactivator of tRNA aminoacylation in the uterus.

Estradiol (E2) induces an increase in the peptide elongation rate of isolated uterine ribosomes assayed in a cell-free protein synthesis system. An inhibitory factor, extracted from ribosomes of E2-deprived rats, was found to inhibit the peptide elongation reaction by acting on certain tRNAs to render them incapable of binding to aminoacyl-tRNA synthetases, thus reducing the availability of specific aminoacylated tRNAs required for the sequential translation of the codons in mRNA. The uterine ribosome-associated tRNA inactivator (RATI) has been partially purified and monoclonal antibodies (MABs) to RATI have been prepared. Specificity of the MABs for RATI was indicated by the inactivation of RATI in vitro by the anti-RATI MABs. RATI selectively inactivates deacylated, but not acylated, tRNAs and the inactivation does not appear to involve nuclease cleavage of the tRNA. Within 1 h after E2 treatment 50% of both RATI activity and immunoreactivity were lost from the uterine ribosome extracts, suggesting that E2 regulation of tRNA reutilization may occur through dissociation of RATI from the ribosomal site of tRNA deacylation or alteration in the structure of RATI resulting in inactivation both biologically and immunologically. We propose that RATI may function as an E2-regulatable 'switch' mechanism which inactivates, delays or defers the aminoacylation of certain tRNAs in the absence of E2 and which participates in the regulation of protein synthesis at the translational level by creating rate-limiting levels of certain tRNAs in the E2-deprived uterus.

Estradiol regulation of the synthesis of uterine proteins with clusters of proline- and glycine-rich peptide sequences.

Estradiol (E2) regulates the synthesis of uterine proteins at both the transcriptional and translational levels. E2 induces an increase in the specific amino acid acceptor activity of uterine tRNA, with the largest increases seen for proline, glycine and methionine. The synthesis of three uterine proteins that are rich in proline and glycine, estrogen receptor, progesterone receptor and glucose-6-phosphate dehydrogenase, is induced by E2. E2-induced increases in these proteins were preceded by an correlated with stimulation of tRNA acceptor activity for proline and glycine and these responses were specifically and simultaneously inhibited by prior azaserine treatment, which inhibits the E2-induced repair and synthesis of the 3'-CCA acceptor terminus of tRNAs. The high frequency and clustering of proline and glycine residues in estrogen receptor, progesterone receptor and glucose-6-phosphate dehydrogenase suggests that the translating ribosomes may slow down during synthesis of these proteins due to limiting levels of these tRNAs in E2-deprived uteri.

Schistosoma mansoni: interactions of adult parasites with the complement system.

The interaction of the mouse complement system with adult male Schistosoma mansoni was studied by immunocytochemical localization procedures and in-vitro assays for complement mediated tegument damage. Mouse C3 was demonstrated to be associated with the parasite's tegument, but was localized only in the infoldings of the tegument and not on its free surface. Freshly harvested parasites manifested no detectable tegumental modification when incubated in normal mouse serum or in immune mouse serum. However, parasites which had been allowed to lose their adsorbed host components by elution (incubation in serum free media for 3 h at 37 degrees C) were severely damaged by incubation in normal mouse serum, but not by incubation in immune mouse serum. This damage was shown to be mediated by the alternative complement pathway and appeared to be initially limited to the tubercles of the adult male parasite. Tegument disruption could be blocked by pre-incubation of the eluted worms in either immune mouse serum or an IgG fraction of immune mouse serum. An IgG fraction of normal mouse serum did not protect the parasite, and infected mouse serum (IMS) which had been depleted of IgG produced tegument damage equivalent to that observed with normal mouse serum (NMS). The addition of I-IgG to NMS abrogated tegument damage. These data suggest that while adult schistosomes possess surface molecules bearing alternative pathway complement activation sites, these sites are masked by adsorbed host components in vivo. These results further indicate that in the absence of these masking host molecules anti-schistosome IgG may play a role in protecting the adult worm from alternative pathway activation, perhaps by binding to and blocking the activation sites on the tegument associated molecules.

Biology of tegument associated IgG-Fc and C3 receptors inSchistosoma mansoni.

Mechanisms by which schistosomes escape host immune responses are reviewed, with particular emphasis regarding the possible contributions of host or host-like Fc and C3 receptors on adult parasites.

Isolation and partial characterization of an antigen shared between Schistosoma mansoni, Fasciola hepatica, and Biomphalaria glabrata.

An antigen was isolated and partially characterized from adult Schistosoma mansoni which immunologically cross-reacted with Fasciola hepatica and Biomphalaria glabrata, a schistosome intermediate snail host. The antigen was isolated from a solubilized freeze-thaw preparation containing 0.5% Triton X-100 by passage through a CNBr-activated Sepharose 4B column coupled with rabbit IgG prepared against a homogenate of B. glabrata hepatopancreas. The eluted antigen (designated as SMw-53P) stained with Coomassie Brilliant Blue R250, but not with Nile Blue A or periodic acid-Schiff's stains. The antigen did not bind to a Concanavalin A affinity column. SMw-53P was determined to have a molecular weight of 53,000 daltons by SDS-polyacrylamide gel electrophoresis. Western blotting, using sera from mice infected with S. mansoni, revealed the presence of the antigen in whole worm preparations of both S. mansoni and F. hepatica. The serodiagnostic potential of the antigen was evaluated by ELISA utilizing sera from S. mansoni-infected mice, or rabbits injected with homogenates of F. hepatica or S. mansoni whole worm, or B. glabrata hepatopancreas. SMw-53P was shown to strongly react with all antisera, but not normal mouse or rabbit sera. These data suggest a limited value for the antigen for the specific immunodiagnosis of schistosomiasis mansoni, but do suggest a possible potential as a general screening tool for detecting trematode infections. Further studies regarding the antigen's potential as a vaccine are indicated.