O comportamento gregário é comum em algumas espécies de Opiliones. No período noturno indivíduos gregários saem do abrigo para forragear e ao amanhecer retornam ao ambiente de repouso. Químicos de coespecíficos têm sido discutidos como mediadores do retorno ao abrigo nos indivíduos da subordem Eupnoi, mas até o momento nada se sabe sobre as pistas que os indivíduos da subordem Laniatores utilizam para retornar ao abrigo. Recentemente, uma grande diversidade de receptores olfativos foi descrita nessa subordem e o uso do olfato à curta distância (< 2 cm) já foi documentado em agregados. Testamos as hipóteses do uso do olfato à longa distância (50 cm) na atração por (1) químicos voláteis emitidos diretamente de coespecíficos e (2) químicos voláteis próprios e de coespecíficos deixados previamente nos abrigos. Utilizamos um olfatômetro em formato de Y. Usamos como estímulos fêmeas e machos coespecíficos para (1) e abrigos dos sujeitos experimentais, de coespecífico do mesmo sexo e controle para (2). Para (1) machos de M. cuspidatus escolheram significativamente o lado do olfatômetro com machos coespecíficos. Fêmeas de M. cuspidatus não mostraram preferência por nenhum dos estímulos. Para (2) machos e fêmeas de M. cuspidatus não escolheram significativamente nenhum estímulo do olfatômetro. Esses resultados nos mostram que talvez machos de M. cuspidatus tenham maior sensibilidade olfativa comparada ao das fêmeas coespecíficas. Os machos de M. cuspidatus não mostraram preferência por fêmeas coespecíficas talvez porque no momento do experimento as fêmeas não estivessem liberando voláteis. Nossos resultados também mostram que indivíduos de M. cuspidatus não são atraídos à distância por químicos deixados no abrigo, talvez porque esses químicos sejam pouco voláteis ou porque os indivíduos detectaram o odor, mas optaram por não escolher o braço com odor próprio e nem de coespecífico. O olfato parece ser mais importante do que anteriormente mencionado em Opiliones / Gregarious behavior is common in some species of Opiliones. At night, gregarious individuals leave their shelter to forage and at dawn return to the original spot. Chemicals of conspecifics have been suggested to be mediators of homing in individuals of the suborder Eupnoi, but so far nothing is known about the cues used in the suborder Laniatores. Recently, a large diversity of olfactory receptors has been described in this suborder and the use of olfaction at close range (<2 cm) has already been documented in aggregations. We tested the hypotheses of the use of olfaction at long range (50 cm) in the attraction by (1) volatile chemicals emitted directly from conspecifics and (2) volatiles (own chemicals and conspecific chemicals) previously left in shelters. We used a Y-shaped olfactometer. We used conspecific females and males as stimuli for (1) and shelters of the experimental subjects and of conspecifics of the same sex for (2). For (1) males of M. cuspidatus chose significantly the side of the olfactometer with conspecifics males. Females did not choose any stimuli. For (2) males and females of M. cuspidatus did not choose any of the stimuli. Males may therefore be more sensitive than females. Males of M. cuspidatus did not show preference for conspecific females, perhaps because at the time of the experiment females were not releasing volatiles. Individuals of M. cuspidatus are not attracted at a distance by chemicals left in the shelter, perhaps because these chemicals are not volatile or because the individuals detected the odor, but chose not to walk towards it. Olfaction seems to be more important than previously mentioned in Opiliones

This work presents a survey of blue-stain fungi found in Northwestern Ontario, characterization of a homing endonuclease gene within Grosmannia piceiperda and finally an examination of the introns and homing endonuclease genes found in the large ribosomal subunit gene in species of Ceratocystis; using molecular techniques and phylogenetic analysis, we studied the molecular evolution of these mobile genetic elements. The blue-stain fungi of Northwestern Ontario were identified based on phylogenic analysis of rDNA internal transcribed spacer region sequences. This data was supplemented with morphological characteristics of the fungal cultures. The second project was an examination of a LAGLIDADG homing endonuclease and its IC2 group I intron. This intron is uniquely positioned within the group I intron-encoded rps3 gene of the large subunit ribosomal RNA gene. The final chapter is an investigation of the large subunit ribosomal RNA gene in species of Ceratocystis. The 3’ segment of this gene contains several novel introns and homing endonuclease genes. There is also much diversity between strains despite their close relation on the rDNA internal transcribed spacer region phylogenetic tree. Further, our data also suggest that the single motif LAGLIDADG homing endonuclease of the rDNA mL1923 intron is likely to be an ancestor to other homing endonucleases in the area. The results of these studies demonstrate the role that these elements play in the genetic diversity observed in the blue-stain fungi.

With the deployment of 3G networks and gradual implementation of wireless networks, seamless handover between these wireless networks is becoming an increasingly desirable. mSCTP (Mobile Stream Control Transmission Protocol) is a new protocol developed from SCTP (Stream Control Transmission Protocol) to provide seamless handover based on IP networks. This thesis studies how to use this new protocol to handle handovers on transport level between CDMA2000 and WLAN networks. A survey of recently proposed and used mobility protocols is presented, comparing three common handover protocols operating on different layers: MIP (mobile IP) for the network layer, mSCTP for the transport layer and SIP (Session Initial Protocol) for the session layer. The results show mSCTP is the future for mobility support. Lastly, I will present a detailed procedure on how to set up handover testbed between CDMA2000 network and 802.11 WLAN based on mSCTP and the results show that the handover performed between these two networks is fast and smooth but it is affected by the signal strength of the CDMA2000.

This work presents a survey of blue-stain fungi found in Northwestern Ontario, characterization of a homing endonuclease gene within Grosmannia piceiperda and finally an examination of the introns and homing endonuclease genes found in the large ribosomal subunit gene in species of Ceratocystis; using molecular techniques and phylogenetic analysis, we studied the molecular evolution of these mobile genetic elements. The blue-stain fungi of Northwestern Ontario were identified based on phylogenic analysis of rDNA internal transcribed spacer region sequences. This data was supplemented with morphological characteristics of the fungal cultures. The second project was an examination of a LAGLIDADG homing endonuclease and its IC2 group I intron. This intron is uniquely positioned within the group I intron-encoded rps3 gene of the large subunit ribosomal RNA gene. The final chapter is an investigation of the large subunit ribosomal RNA gene in species of Ceratocystis. The 3’ segment of this gene contains several novel introns and homing endonuclease genes. There is also much diversity between strains despite their close relation on the rDNA internal transcribed spacer region phylogenetic tree. Further, our data also suggest that the single motif LAGLIDADG homing endonuclease of the rDNA mL1923 intron is likely to be an ancestor to other homing endonucleases in the area. The results of these studies demonstrate the role that these elements play in the genetic diversity observed in the blue-stain fungi.

Thesis (Ph. D.)--University of Washington, 2007. / Vita. Includes bibliographical references (leaves 122-140).

For most of the animals, if not all, finding their way to a particular place is crucial for survival. To address this challenge of way-finding, different animals have evolved with different homing strategies. Social hymenopterans like honey bees, ants and wasps are of special interest – foragers of these insects show excellent homing capabilities while having simple neural resources. In this study field, honey bees and ants (desert ants, in particular) are among the most studied animals. Compared to these insects, our understanding on the homing mechanisms of social wasp is rather poor. For my thesis, I have studied homing behaviour of the tropical social wasp Ropalidia marginata, a predator in their foraging habit. To begin with, first I had to know their typical foraging range, which I found to be within about 500 m from their nest. Forager wasps possess a surprisingly well-developed familiarity with their foraging landscape, apparently more intricate than honey bees and desert ants. They acquire this spatial familiarity through flying around the landscape before starting foraging for food. Compared to honey bees and desert ants, this learning period in wasps appears to be much longer – this can be attributed to the much higher density of the tropical landscape in which they have evolved. I have also found that, if needed, they can fly to a distance of about 1.5 km for foraging and can return to their nest even if passively displaced to familiar and unfamiliar places. To return from unfamiliar places, they probably use some sort of searching mechanisms – a skill that they improve with their age. Such searching behaviour is prevalent throughout other hymenopteran insects. I conclude that capability and mechanisms of spatial orientation, navigation and homing in animals are much influenced by their evolutionary origin and the environment in which they have evolved.

O comportamento gregário é comum em algumas espécies de Opiliones. No período noturno indivíduos gregários saem do abrigo para forragear e ao amanhecer retornam ao ambiente de repouso. Químicos de coespecíficos têm sido discutidos como mediadores do retorno ao abrigo nos indivíduos da subordem Eupnoi, mas até o momento nada se sabe sobre as pistas que os indivíduos da subordem Laniatores utilizam para retornar ao abrigo. Recentemente, uma grande diversidade de receptores olfativos foi descrita nessa subordem e o uso do olfato à curta distância (< 2 cm) já foi documentado em agregados. Testamos as hipóteses do uso do olfato à longa distância (50 cm) na atração por (1) químicos voláteis emitidos diretamente de coespecíficos e (2) químicos voláteis próprios e de coespecíficos deixados previamente nos abrigos. Utilizamos um olfatômetro em formato de Y. Usamos como estímulos fêmeas e machos coespecíficos para (1) e abrigos dos sujeitos experimentais, de coespecífico do mesmo sexo e controle para (2). Para (1) machos de M. cuspidatus escolheram significativamente o lado do olfatômetro com machos coespecíficos. Fêmeas de M. cuspidatus não mostraram preferência por nenhum dos estímulos. Para (2) machos e fêmeas de M. cuspidatus não escolheram significativamente nenhum estímulo do olfatômetro. Esses resultados nos mostram que talvez machos de M. cuspidatus tenham maior sensibilidade olfativa comparada ao das fêmeas coespecíficas. Os machos de M. cuspidatus não mostraram preferência por fêmeas coespecíficas talvez porque no momento do experimento as fêmeas não estivessem liberando voláteis. Nossos resultados também mostram que indivíduos de M. cuspidatus não são atraídos à distância por químicos deixados no abrigo, talvez porque esses químicos sejam pouco voláteis ou porque os indivíduos detectaram o odor, mas optaram por não escolher o braço com odor próprio e nem de coespecífico. O olfato parece ser mais importante do que anteriormente mencionado em Opiliones / Gregarious behavior is common in some species of Opiliones. At night, gregarious individuals leave their shelter to forage and at dawn return to the original spot. Chemicals of conspecifics have been suggested to be mediators of homing in individuals of the suborder Eupnoi, but so far nothing is known about the cues used in the suborder Laniatores. Recently, a large diversity of olfactory receptors has been described in this suborder and the use of olfaction at close range (<2 cm) has already been documented in aggregations. We tested the hypotheses of the use of olfaction at long range (50 cm) in the attraction by (1) volatile chemicals emitted directly from conspecifics and (2) volatiles (own chemicals and conspecific chemicals) previously left in shelters. We used a Y-shaped olfactometer. We used conspecific females and males as stimuli for (1) and shelters of the experimental subjects and of conspecifics of the same sex for (2). For (1) males of M. cuspidatus chose significantly the side of the olfactometer with conspecifics males. Females did not choose any stimuli. For (2) males and females of M. cuspidatus did not choose any of the stimuli. Males may therefore be more sensitive than females. Males of M. cuspidatus did not show preference for conspecific females, perhaps because at the time of the experiment females were not releasing volatiles. Individuals of M. cuspidatus are not attracted at a distance by chemicals left in the shelter, perhaps because these chemicals are not volatile or because the individuals detected the odor, but chose not to walk towards it. Olfaction seems to be more important than previously mentioned in Opiliones

No description available.

The small ribosomal subunit gene of the Chaetomium thermophilum DSM 1495 is invaded by a nested intron at position mS1247, which is composed of a group I intron encoding a LAGLIDADG open reading frame interrupted by an internal group II intron. The first objective was to examine if splicing of the internal intron could reconstitute the coding regions and facilitate the expression of an active homing endonuclease. Using in vitro transcription assays, the group II intron was shown to self-splice only under high salt concentration. Both in vitro endonuclease and cleavage mapping assays suggested that the nested intron encodes an active homing endonuclease which cleaves near the intron insertion site. This composite arrangement hinted that the group II intron could be regulatory with regards to the expression of the homing endonuclease. Constructs were generated where the codon-optimized open reading frame was interrupted with group IIA1 or IIB introns. The concentration of the magnesium in the media sufficient for splicing was determined by the Reverse Transcriptase-Polymerase Chain Reaction analyses from the bacterial cells grown under various magnesium concentrations. Further, the in vivo endonuclease assay showed that magnesium chloride stimulated the expression of a functional protein but the addition of cobalt chloride to the growth media antagonized the expression. This study showed that the homing endonuclease expression in Escherichia coli can be regulated by manipulating the splicing efficiency of the group II introns which may have implications in genome engineering as potential ‘on/off switch’ for temporal regulation of homing endonuclease expression . Another objective was to characterize native homing endonucleases, cytb.i3ORF and I-OmiI encoded within fungal mitochondrial DNAs, which were difficult to express and purify. For these, an alternative approach was used where two compatible plasmids, HEase.pET28b (+)-kanamycin and substrate.pUC57-chloramphenicol, based on the antibiotic markers were maintained in Escherichia coli BL21 (DE3). The in vivo endonuclease assays demonstrated that these homing endonucleases were able to cleave the substrate plasmids when expressed, leading to the loss of the antibiotic markers and thereby providing an indirect approach to screen for potential active homing endonucleases before one invests effort into optimizing protein overexpression and purification strategies. / October 2016

Syngeneic graft-versus-host disease (SGVHD) is induced by reconstituting lethally irradiated mice with syngeneic BM cells followed by a 21 day treatment with the immunosuppressive agent cyclosporine A (CsA). Clinical symptoms of the disease appear 2-3 weeks following cessation of CsA therapy and disease-associated inflammation occurs primarily in the colon and liver. The development of SGVHD is a complex process resulting from the cooperative interaction of multiple effector cell populations including NK cells, T cells and macrophages. TH1 cytokines (IL-12, TNF-α, IFN- γ), produced by these effector cells, serve as inflammatory mediators contributing to the pathogenesis of SGVHD. The SGVHD conditioning agents, irradiation and CsA, are both required for the development of disease and contribute to the production of oxidative stress. Time course studies revealed increased reactive oxygen and nitrogen species (ROS/RNS), as well as, increased colon mRNA levels for TNF-α and iNOS in CsA-treated versus control BMT animals. Since ROS/RNS are known to mediate CsA toxicity, studies were undertaken to determine the effect of oxidative stress on the induction of SGVHD. In vivo treatment with the antioxidant MnTBAP caused a reduction in colon mRNA levels for iNOS and TNF-α, as well as delayed disease development, suggesting a role for oxidative stress in the development of SGVHD. In addition, CD4+ T cells have been shown to play an important role in the inflammatory response observed in the gut of SGVHD mice. Time course studies revealed significant increases in the migration of CD4+ T cells as early as day 14 post- BMT into the colon of CsA mice as well as significant elevated mRNA levels of cell adhesion molecules. Homing studies revealed that a labeled CD4+ T cell line, generated from SGVHD mice, migrated in larger numbers into the gut of CsA-treated mice compared to control animals. This study demonstrated that CD4+ T cells responsible for the pathogenesis observed in murine SGVHD are present early after BMT in colons of CsA-treated mice, suggesting that during the 21 days of immunosuppression therapy functional mechanisms are in place that result in increased homing of effector cells to colons of CsA-treated mice.