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PubMed2000

Full-length sequence of a Canadian porcine reproductive and respiratory syndrome virus (PRRSV) isolate.

Presently, one of the most economically important pathogens affecting swine is the porcine reproductive and respiratory syndrome virus (PRRSV). This virus is prevalent in herds throughout the world and continues to pose a significant threat as newer and more virulent disease phenotypes emerge. In this report we describe the full-length nucleotide sequence of a Canadian PRRSV isolate, designated PA8. A consecutive sequence of 15,411 nucleotides was obtained from a set of overlapping cDNA clones. In order to determine the extent of genetic variation among isolates recovered from swine in Canada and the US, as well as to understand the molecular mechanisms governing the evolution of PRRSV, the full-length sequence of PA8 was compared with that of two US isolates, VR2332 and 16244B. The genomic sequence of PA8 shared 98.2% and 99.2% identity with 16244B and VR2332, respectively. The untranslated regions (UTR) at the 5' and 3' ends of the genome were very well conserved. Notable exceptions include an eight nucleotide difference at the 5' end of the 5' UTR of VR2332 relative to PA8 and 16244B and a two nucleotide difference in the 3' UTR of PA8 relative to VR2332 and 16244B. In contrast to PA8 and VR2332, 16244B possessed two nucleotide differences within the RNA pseudoknot structure of the ribosomal frameshift region between open reading frame (ORF)1a and ORF1b. Amino acid differences were distributed throughout the genome, however they appeared to be most extensive in Nsp1beta and ORF5 of the nonstructural and structural coding regions, respectively, suggesting that the evolutionary pressure to conserve these viral genes is somewhat lower.

3' Untranslated Regions↗
PubMed2006

The history of simulation in medical education and possible future directions.

INTRODUCTION: Clinical simulation is on the point of having a significant impact on health care education across professional boundaries and in both the undergraduate and postgraduate arenas. SCOPE OF SIMULATION: The use of simulation spans a spectrum of sophistication, from the simple reproduction of isolated body parts through to complex human interactions portrayed by simulated patients or high-fidelity human patient simulators replicating whole body appearance and variable physiological parameters. GROWTH OF SIMULATION: After a prolonged gestation, recent advances have made available affordable technologies that permit the reproduction of clinical events with sufficient fidelity to permit the engagement of learners in a realistic and meaningful way. At the same time, reforms in undergraduate and postgraduate education, combined with political and societal pressures, have promoted a safety-conscious culture where simulation provides a means of risk-free learning in complex, critical or rare situations. Furthermore, the importance of team-based and interprofessional approaches to learning and health care can be promoted. CONCLUSION: However, at the present time the quantity and quality of research in this area of medical education is limited. Such research is needed to enable educators to justify the cost and effort involved in simulation and to confirm the benefit of this mode of learning in terms of the outcomes achieved through this process.

Education, Medical↗
PubMed1994

Porcine reproductive and respiratory syndrome virus: morphological, biochemical and serological characteristics of Quebec isolates associated with acute and chronic outbreaks of porcine reproductive and respiratory syndrome.

Cytolytic and noncytolytic strains of the porcine reproductive and respiratory syndrome virus (PRRSV) were isolated in primary cultures of porcine alveolar macrophages (PAM) from lung homogenates of stillborn fetuses or blood samples of dyspneic piglets collected from Quebec pig farms having experienced acute or chronic outbreaks of PRRS. Serological identification of the virus was confirmed by indirect immunofluorescence and indirect protein A-gold immunoelectron microscopy using reference antiserum prepared from experimentally-infected specific pathogen free (SPF) piglets and monoclonal antibodies (MoAbs) directed against the p15 nucleocapsid (N) protein of the reference ATCC-VR2332 isolate. Intracytoplasmic enveloped viral particles that tended to accumulate into cytoplasmic vesicles were observed in the infected PAM; no budding was demonstrated at the level of the cytoplasmic membrane. The extracellular virions appeared as pleomorphic but mostly spherical enveloped particles, 50-72 nm in diameter (averaged diameter of 50 particles was 58.3 nm), with an isometric core about 25-30 nm. Buoyant density of the virus in CsCL density gradients was estimated to 1.18-1.20 g/mL. No hemagglutinating activity was demonstrated. Analysis of semipurified virions of isolate IAF-exp91 by radioimmunoprecipitation (RIPA) and Western immunoblotting experiments, using reference rabbit and porcine hyperimmune sera, revealed four major viral proteins, a predominant 15 kD N protein and three other proteins with predicted M(r_ of 19, 26 and 42 kD. Progeny viral particles produced in PRRSV-infected PAM in the presence of tunicamycin lacked the 42 kD protein, thus confirming its N-glycosylated nature. Immunoprecipitation experiments using the anti-ATCC-VR2332 MoAbs confirmed the close antigenic relationships between Quebec and American reference isolates of PRRSV.

Animals↗
PubMed2026

Digest: Speciation involves both barriers and bridges in the tropical Andes.

Mountains are important centers of biodiversity and studies of speciation. In particular, the north-south linear orientation of the Andes allows examination of how geographic isolation, elevation, and latitude influence divergence in closely related species such as Myioborus warblers (Parulidae), the focal taxa of this study. Traditional models of Andean speciation have emphasized allopatric divergence due to geographic barriers, but thanks to the limited sampling of hybrid zones and lack of genome-wide datasets in tropical taxa, we may have underestimated of the role of gene flow in shaping patterns of divergence in the region. Using ddRAD-seq genomic data, Céspedes Arias et al. (2026) demonstrated that geographic isolation and hybridization both contribute to the generation and maintenance of Andean warbler lineages. Their results revealed how complex patterns of genetic divergence, introgression, and isolation-by-distance contribute to the historical buildup of biodiversity in the Andes, challenging strictly isolationist models of mountain evolution.

Animals↗
PubMed1996

Abortion and placentitis in pregnant bison (Bison bison) induced by the vaccine candidate, Brucella abortus strain RB51.

OBJECTIVE: To determine the ability of Brucella abortus strain RB51 to induce placentitis and abortion in bison after SC vaccination. ANIMALS: 10 pregnant bison cows, 3 to 10 years old and at 3 to 8 months' gestation. PROCEDURE: Pregnant bison cows on a Montana ranch were vaccinated SC with 10(9) colony-forming units of B abortus strain RB51. Two cows, identified prior to the study, were euthanatized and examined 5 weeks after vaccination to obtain optimal histologic samples of placenta. Other cows were euthanatized and examined after abortion. After euthanasia, tissue specimens were collected for histologic and immunohistochemical evaluation. Tissue and fluid specimens for bacteriologic culture were also collected during necropsy. RESULTS: Of 8 cows, 2 aborted at 68 and 107 days after vaccination. Aborting cows had endometritis. Strain RB51 was isolated from reproductive tissues and supramammary lymph nodes. Fetal lesions were not seen; however, fetal bronchial lymph nodes and amniotic fluid contained strain RB51. Cows examined 5 weeks after vaccination had placentitis and endometritis, with numerous bacteria within trophoblastic epithelial cells that were immunoreactive for strain RB51 antigen. Strain RB51 was isolated from placentomes and numerous lymph nodes. Fetal lesions were not seen 5 weeks after vaccination; however, strain RB51 was isolated from numerous lymph nodes and lung, allantoic fluid, and rectal swab specimens. CONCLUSIONS: The vaccine candidate B abortus RB51 has tropism for the bison placenta, and can cause placentitis, which induces abortion in pregnant bison. The vaccine dose used was similar to that being tested in cattle, but may not be appropriate for pregnant bison.

Abortion, Veterinary↗
PubMed1980

The effect of arprinocid on the reproductive potential of a field isolate of Eimeria tenella.

Oocyst production by an isolate of Eimeria tenella propagated from birds medicated with 50 ppm arprinocid was compared with that of the same isolate propagated from unmedicated birds. Statistically significant (P less than 0.05) reductions in oocyst production were found for the drug-exposed isolate for inocula levels of 64 and 250 sporulated oocysts per bird. Numerical reductions were observed for inocula levels of 1,000 oocysts per bird, but with higher levels of inocula, no differences were seen. Inhibitory drug effects for the inocula levels of 64 and 250 oocysts per bird appear to be meaningful because they coincide with the maximum reproductive potential (oocysts produced per oocyst inoculated). The results indicated that significant information can be masked by excessive inoculum with consequent crowding and reduction in reproductive potential.

Adenine↗
PubMed2025

Are We Witnessing a Speciation Continuum? Evidence From Current and Past Gene Flow in the Genus Oritrophium s.s. (Asteraceae) From the Tropical High Andes.

Determining species boundaries is key for appropriately assessing biodiversity. However, the continuity of the speciation process makes delimiting species a difficult task, especially for recently diverged taxa. Furthermore, past introgression may leave traces that result in reticulate evolutionary patterns, challenging the estimation of species relationships. The fastest-evolving biodiversity hotspot on Earth is the Páramo. Its flora in the tropical high Andes is known for extraordinarily high species richness and endemism. However, the recent origin, fast diversification and complex taxonomy of many genera challenge species delimitation and phylogenetic reconstruction. In this study, we reconstructed phylogenetic relationships and addressed the role of introgression in the diversification of Oritrophium s.s. (Asteraceae) based on phylogenomic data. We combined genomic, phenotypic and ecological data to test species boundaries and compared trajectories across the speciation continuum within the taxonomically complex 'O. peruvianum group'. We found that historical introgression played an important role in the evolution of Oritrophium s.s., and many of the taxa within the 'O. peruvianum group' are at various stages of speciation. These results highlight the importance of testing for introgression to understand the diversification of recently evolved groups. Likewise, they suggest that heterogeneous speciation trajectories associated with geographic isolation and secondary contact, possibly during the Pleistocene, contributed to plant diversity in the tropical high Andes.

Genetic Speciation↗
PubMed1999

Genetic structure, reproductive biology and ecology of isolated populations of asplenium csikii (Aspleniaceae, pteridophyta)

The potential for environmental heterogeneity to generate spatial structuring of genotypes in seed-plant populations that occupy patchy habitats has been demonstrated by several studies, but little is known about the population structure of pteridophytes occupying patchy environments. In this study we have examined the genetic structure of isolated populations of the rock fern Asplenium csikii, an ecological specialist, growing almost exclusively on perpendicular walls of natural rock outcrops. All genetic variation observed in this taxon was partitioned between localities; no allozyme variation was found within a site and each site was colonized by a single multilocus phenotype (MLP). In total, five different MLPs were recorded from the nine localities, with two MLPs present at more than one site. Previous examination of population structure and genetic diversity in another rock fern, A. ruta-muraria, showed that the genetic diversity increases through multiple colonization over time. However, we cannot find any such correlation for A. csikii. All populations are genetically uniform, despite the probably considerable age of the populations and sites. Earlier studies concluded that the ample production of wind-borne propagules would lead to multiple colonization of sites and that reproductive features, such as single-spore colonization and subsequent intragametophytic selfing, would lead to very little genetic structuring of fern populations. In contrast to this prediction, it appears that ecological specialization and the scarcity of the narrowly defined niche contribute strongly to the pronounced partitioning of genetic variability observed in populations of A. csikii.

Journal Article↗
PubMed1996

Comparative pathogenicity of nine US porcine reproductive and respiratory syndrome virus (PRRSV) isolates in a five-week-old cesarean-derived, colostrum-deprived pig model.

One hundred forty-six 5-week- old cesarean-derived, colostrum-deprived (CDCD) pigs were inoculated intranasally with 1 of 9 US porcine reproductive and respiratory syndrome virus (PRRSV) isolates. Differences were found in severity of clinical respiratory disease, rectal temperatures (P < or = 0.001), gross lung lesions (P < or = 0.001), and microscopic lung lesions (P < or = 0.05). Gross lung lesions were generally most severe 10 days postinoculation and were distributed primarily in the cranial, middle, and accessory lobes and ventromedial portion of the caudal lung lobes. Mean gross lung lesion scores estimating the percentage of lung affected by pneumonia at 10 days postinoculation ranged from 16.7% +/- 2.8% (mean +/- SEM, n = 10) for isolate ISU-51 to 62.4% +/- 5.7% (n = 10) for isolate ISU-28. Microscopic lung lesions were characterized by hyperplastic and hypertrophied type 2 pneumocytes, septal infiltration by mononuclear cells, and accumulation of necrotic alveolar exudate. Lymph node follicular hyperplasia and focal necrosis was seen with all 9 isolates. This CDCD pig model was useful for demonstration of significant differences in pathogenicity among US PRRSV isolates. This difference in pathogenicity may help explain the variation of severity of clinical disease observed in field outbreaks of porcine reproductive and respiratory syndrome and should provide for meaningful comparison of PRRSV genotypes.

Animals↗
PubMed1982

Prevalence and distribution of Haemophilus somnus in the male bovine reproductive tract.

"Haemophilus somnus' was isolated from 77% of 31 reproductive tracts of bulls from an Ontario slaughterhouse. Identification of H somnus was based on morphologic and cultural characteristics and on fluorescent antibody and immunodiffusion tests, using antisera prepared against a known pathogenic encephalitic isolate of H somnus. The infection rate and distribution of H somnus within the tract were as follows: preputial orifice--71% preputial cavity--71%; urinary bladder--26%; accessory sex glands--19% and ampulla of ductus deferens--10%. Isolates were not obtained from the testes or epididymides. On 2 occasions, H somnus was isolated in pure culture from the preputial cavity. A higher prevalence of infection was found in young bulls. There were no differences found in infection rates between breeds. Differences in hemolytic activity and minor antigenic variation between isolates indicated that a series of biotypes within the species H somnus may exist. The study indicates that organisms presently identified as H somnus may normally form part of the flora of the bovine prepuce and that dissemination from the male bovine reproductive tract is one possible means of infection in H somnus-associated diseases. The pathogenic significance of genital isolates of H somnus awaits further study.

Animals↗
PubMed1983

Hemophilus somnus in the reproductive tracts of slaughtered cows: location and frequency of isolations and lesions.

One hundred bovine female reproductive tracts were examined for the presence of Hemophilus somnus, and fifty were examined for gross and microscopic lesions. H. somnus was isolated from eight of 100 tracts. The isolation was made from the vagina in seven cows. Of 24 tracts in which the major vestibular gland was sampled, three were positive at this site, and two of these three also had the organism isolated from the vagina. Further investigations into the role of the major vestibular gland as a source for continuing infections of the reproductive tract of cattle were indicated. In this investigation, H. somnus was as likely to be found in a region without lesions as one with lesions.

Abattoirs↗
PubMed2006

Expression of mRNAs encoding receptors that mediate stress signals in gonadotropin-releasing hormone neurons of the mouse.

Neurons that synthesize and secrete gonadotropin-releasing hormone (GnRH) represent the neural control point for fertility modulation in vertebrates. As such GnRH neurons are ideally situated to integrate stress responses on reproduction. By isolating individual GnRH neurons from acute brain slices of adult female GnRH-EGFP transgenic mice and using microarray analyses, we have identified a range of transcripts encoding receptors known to be involved in stress responses in GnRH neurons. Prominent among these were receptors for corticotropin-releasing hormone (CRH), vasopressin, interleukins, prostaglandins, tumor necrosis factor alpha and other inflammatory mediators. We selected 4 of these targets [interleukin 1 receptor accessory protein (IL-1Racc), prostaglandin E(2) receptor subtype EP2 (PGER2), CRH receptor type 1 (CRH-R1), and arginine-vasopressin receptor type 1b (AVP-R1b)] for validation using single-cell RT-PCR from individual GnRH neurons. In total, 54% of GnRH neurons (n = 26) were found to express at least 1 of these transcripts. The IL-1Racc, PGER2 and CRH-R1 mRNAs were each detected in approximately 25% of the GnRH neurons tested, but no evidence was found for AVP-R1b transcripts. Overlap was found between the expression of CRH-R1 and PGER2, and IL-1Racc and PGER2 in individual GnRH neurons. Dual immunofluorescence experiments confirmed the expression of CRH-R1/2 in a subpopulation ( approximately 30%) of GnRH neurons. These observations indicate that a variety of different stressors and stress pathways have the capacity to have an impact directly upon a subpopulation of GnRH neurons to influence the reproductive axis.

Animals↗
PubMed1996

Isolation and developmental expression of male reproductive organ-specific genes in a dioecious campion, Melandrium album (Silene latifolia).

The dioecious campion Melandrium album (Silene latifolia) has sex chromosomes, and male and female plants form different unisexual flowers. The very youngest buds have both sex organs but the development of either a gynoecium in the male bud or stamens in the female bud is arrested as the bud matures. To examine the molecular mechanism of male-specific differentiation, a cDNA library from male buds was differentially screened with cDNA from male and female buds. Four male reproductive organspecific cDNAs (corresponding to genes designated MROS1, MROS2, MROS3 and MROS4, respectively) were isolated. Northern blot analysis showed that the transcripts of MROS genes were expressed only in male reproductive organs. The transcripts of MROS3 and MROS4 accumulated in the early buds, while transcripts of MROS1 and MROS2 accumulated in the late buds and in the anthers of open flowers. The putative MROS2 protein has glycine-rich regions, while the putative MROS3 protein exhibits no similarity to known proteins. Genomic Southern blot analysis showed that male and female genomes included MROS2 and MROS3 genes. Non-radioactive in situ hybridization revealed the localization of the transcripts of MROS2 and MROS3 in male buds. MROS3 mRNA was distributed throughout the tapetum. After MROS2 mRNA had accumulated in the epidermis and the endothecium near the stomium, the accumulation of the mRNA increased significantly in the mature pollen. MROS2 seems to be a novel anther-specific gene that functions in dehiscence of anthers and the maturation of pollen.

Amino Acid Sequence↗
PubMed1982

Large-scale selection and analysis of temperature-sensitive mutants for cell reproduction from BHK cells.

Temperature-sensitive (ts) mutant cells for cell reproduction were isolated from the Syrian hamster cell line BHK21/13 by multiple culturing in the presence of 5-fluoro-2'-deoxyuridine (FdU) at 37.5 degrees after N-methyl-N'-nitro-N-nitrosoguanidine mutagenesis. A simple method for cell fusion was devised, which enabled us to perform complementation studies with a large number of ts mutants. By using the method we have analyzed 219 ts mutants and classified them into 18 complementation groups. Mutants that belonged to the same complementation groups tended to exhibit similar patterns of inhibition of DNA synthesis at 39.5 degrees; however, some mutants belonging to the same group showed somewhat different patterns, probably due to occurrence of different mutations in the same gene. Distribution of the ts mutants among the 18 complementation groups was uneven; more than 50% of the mutants examined were assigned to complementation group B and G. The mutations belonging to complementation group B and G were found to be linked to the X chromosome.

Animals↗
PubMed1987

Characterization of acid and neutral alpha-mannosidases in bull semen and reproductive organs.

Acid and neutral alpha-mannosidase activities were studied in the bull reproductive tissues, isolated spermatozoa, epididymal and seminal vesicle secretion and seminal plasma. The acid enzyme in the seminal plasma mainly derived from the epididymal secretion, while the neutral one was enriched in the sperm cells. The latter activity in the seminal plasma appears to be due to an enzyme released from the cytoplasmic droplets in the epididymis. The acid enzyme had a molecular weight of 220,000-320,000, pI 7.3-6.0 and an optimum at pH 4.0. It was sensitive to swainsonine but was stimulated by Zn2+. The neutral enzyme had a molecular weight of 360,000-460,000, pI 5.4-4.7 and showed double optima at pH 5.5 and 6.0-7.0. It was resistant to swainsonine but was markedly activated by Co2+ or Fe2+. The neutral enzyme was also more sensitive to thermal inactivation than the acid one.

Animals↗
PubMed1987

Geographic dialects in blind mole rats: role of vocal communication in active speciation.

We compared and contrasted the physical structure of male "courtship" calls of 59 subterranean mole rats belonging to the Spalax ehrenbergi superspecies in Israel, comprising 11 populations of four chromosomal species (2N = 52, 54, 58, and 60). We also conducted behavioral auditory discrimination tests of 144 females of the four species in the laboratory. The results indicate that each chromosomal species has a vocal dialect significantly different from all others, although the call of 2N = 60, the last derivative of speciation, is not yet fully differentiated. Females of 2N = 52, 54, and 58 preferred their homospecific mates' calls, whereas females of 2N = 60 did not. We conclude that call differentiation builds up gradually and provides an efficient ethological reproductive premating isolation mechanism between the emerging species in the active speciation of mole rats in Israel.

Animals↗
PubMed1999

Control of fertilization-independent endosperm development by the MEDEA polycomb gene in Arabidopsis.

Higher plant reproduction is unique because two cells are fertilized in the haploid female gametophyte. Egg and sperm nuclei fuse to form the embryo. A second sperm nucleus fuses with the central cell nucleus that replicates to generate the endosperm, a tissue that supports embryo development. To understand mechanisms that initiate reproduction, we isolated a mutation in Arabidopsis, f644, that allows for replication of the central cell and subsequent endosperm development without fertilization. When mutant f644 egg and central cells are fertilized by wild-type sperm, embryo development is inhibited, and endosperm is overproduced. By using a map-based strategy, we cloned and sequenced the F644 gene and showed that it encodes a SET-domain polycomb protein. Subsequently, we found that F644 is identical to MEDEA (MEA), a gene whose maternal-derived allele is required for embryogenesis [Grossniklaus, U., Vielle-Calzada, J.-P., Hoeppner, M. A. & Gagliano, W. B. (1998) Science 280, 446-450]. Together, these results reveal functions for plant polycomb proteins in the suppression of central cell proliferation and endosperm development. We discuss models to explain how polycomb proteins function to suppress endosperm and promote embryo development.

Arabidopsis↗