The development of an online neonatal intensive care unit family education

Background: British Columbia’s Fraser Health Authority (FHA) neonatal intensive care units (NICUs) value family centered care (FCC). Nevertheless, there is limited evidence that FCC is actually incorporated into practice, as well as some concern that FHA NICU education is inaccessible, inconsistent, or disorganized. Purpose: The mission of this project is to support the principles of FCC throughout the development of an FHA online NICU family education guide by reflecting upon the needs of families throughout their NICU journey. Methods: A needs assessment was initially completed and included literature reviews, consultations, and an environmental scan. This data informed development of an online NICU family education guide which plots current education materials along key stages of the NICU journey: prenatal, admission, early days, growing and developing, discharge and at home. For the purposes of this practicum, only the prenatal stage was fully developed and will serve as a template for other stages following a formative evaluation. A pamphlet and revised FHA Neonatal Checkpoint will also be developed to augment teaching by health care professionals. Implementation and evaluation plans were adapted from the Center for Disease Control Framework for Program Evaluation in Public Health. Results: The needs assessment validates and directs the development, implementation, and evaluation of the online guide illustrating an FCC approach. The online guide centralizes and organizes education by selecting education topics that relate to each stage of the NICU journey. This family-directed design enables families’ access to consistent and reliable information and offers them an opportunity to learn at their own pace. Conclusion: The process of creating, implementing, and evaluating an online family education program for FHA NICUs elucidates the intricacies and the advantages of integrating FCC into NICU practice.

Biological and Physical Factors Affecting the Natural History and Evolution of Encapsulated Development

The evolution of reproductive strategies involves a complex calculus of costs and benefits to both parents and offspring. Many marine animals produce embryos packaged in tough egg capsules or gelatinous egg masses attached to benthic surfaces. While these egg structures can protect against environmental stresses, the packaging is energetically costly for parents to produce. In this series of studies, I examined a variety of ecological factors affecting the evolution of benthic development as a life history strategy. I used marine gastropods as my model system because they are incredibly diverse and abundant worldwide, and they exhibit a variety of reproductive and developmental strategies.

The first study examines predation on benthic egg masses. I investigated: 1) behavioral mechanisms of predation when embryos are targeted (rather than the whole egg mass); 2) the specific role of gelatinous matrix in predation. I hypothesized that gelatinous matrix does not facilitate predation. One study system was the sea slug Olea hansineensis, an obligate egg mass predator, feeding on the sea slug Haminoea vesicula. Olea fed intensely and efficiently on individual Haminoea embryos inside egg masses but showed no response to live embryos removed from gel, suggesting that gelatinous matrix enables predation. This may be due to mechanical support of the feeding predator by the matrix. However, Haminoea egg masses outnumber Olea by two orders of magnitude in the field, and each egg mass can contain many tens of thousands of embryos, so predation pressure on individuals is likely not strong. The second system involved the snail Nassarius vibex, a non-obligate egg mass predator, feeding on the polychaete worm Clymenella mucosa. Gel neither inhibits nor promotes embryo predation for Nassarius, but because it cannot target individual embryos inside an egg mass, its feeding is slow and inefficient, and feeding rates in the field are quite low. However, snails that compete with Nassarius for scavenged food have not been seen to eat egg masses in the field, leaving Nassarius free to exploit the resource. Overall, egg mass predation in these two systems likely benefits the predators much more than it negatively affects the prey. Thus, selection for environmentally protective aspects of egg mass production may be much stronger than selection for defense against predation.

In the second study, I examined desiccation resistance in intertidal egg masses made by Haminoea vesicula, which preferentially attaches its flat, ribbon-shaped egg masses to submerged substrata. Egg masses occasionally detach and become stranded on exposed sand at low tide. Unlike adults, the encased embryos cannot avoid desiccation by selectively moving about the habitat, and the egg mass shape has high surface-area-to-volume ratio that should make it prone to drying out. Thus, I hypothesized that the embryos would not survive stranding. I tested this by deploying individual egg masses of two age classes on exposed sand bars for the duration of low tide. After rehydration, embryos midway through development showed higher rates of survival than newly-laid embryos, though for both stages survival rates over 25% were frequently observed. Laboratory desiccation trials showed that >75% survival is possible in an egg mass that has lost 65% of its water weight, and some survival (<25%) was observed even after 83% water weight lost. Although many surviving embryos in both experiments showed damage, these data demonstrate that egg mass stranding is not necessarily fatal to embryos. They may be able to survive a far greater range of conditions than they normally encounter, compensating for their lack of ability to move. Also, desiccation tolerance of embryos may reduce pressure on parents to find optimal laying substrata.

The third study takes a big-picture approach to investigating the evolution of different developmental strategies in cone snails, the largest genus of marine invertebrates. Cone snail species hatch out of their capsules as either swimming larvae or non-dispersing forms, and their developmental mode has direct consequences for biogeographic patterns. Variability in life history strategies among taxa may be influenced by biological, environmental, or phylogenetic factors, or a combination of these. While most prior research has examined these factors singularly, my aim was to investigate the effects of a host of intrinsic, extrinsic, and historical factors on two fundamental aspects of life history: egg size and egg number. I used phylogenetic generalized least-squares regression models to examine relationships between these two egg traits and a variety of hypothesized intrinsic and extrinsic variables. Adult shell morphology and spatial variability in productivity and salinity across a species geographic range had the strongest effects on egg diameter and number of eggs per capsule. Phylogeny had no significant influence. Developmental mode in Conus appears to be influenced mostly by species-level adaptations and niche specificity rather than phylogenetic conservatism. Patterns of egg size and egg number appear to reflect energetic tradeoffs with body size and specific morphologies as well as adaptations to variable environments. Overall, this series of studies highlights the importance of organism-scale biotic and abiotic interactions in evolutionary patterns.

Roles of CTCF and YY1 in T Cell Receptor Gene Rearrangement And T Cell Development

Diversity of T cell receptors (TCR) and immunoglobulins (Ig) is generated by V(D)J recombination of antigen receptor (AgR) loci. The Tcra-Tcrd locus is of particular interest because it displays a nested organization of Tcrd and Tcra gene segments and V(D)J recombination follows an intricate developmental program to assemble both TCRδ and TCRα repertoires. However, the mechanisms that dictate the developmental regulation of V(D)J recombination of the Tcra-Tcrd locus remain unclear.

We have previously shown that CCCTC-binding factor (CTCF) regulates Tcra gene transcription and rearrangement through organizing chromatin looping between CTCF- binding elements (CBEs). This study is one of many showing that CTCF functions as a chromatin organizer and transcriptional regulator genome-wide. However, detailed understanding of the impact of specific CBEs is needed to fully comprehend the biological function of CTCF and how CTCF influences the generation of the TCR repertoire during thymocyte development. Thus, we generated several mouse models with genetically modified CBEs to gain insight into the CTCF-dependent regulation of the Tcra-Tcrd locus. We revealed a CTCF-dependent chromatin interaction network at the Tcra-Tcrd locus in double-negative thymocytes. Disruption of a discrete chromatin loop encompassing Dδ, Jδ and Cδ gene segments allowed a single Vδ segment to frequently contact and rearrange to diversity and joining gene segments and dominate the adult TCRδ repertoire. Disruption of this loop also narrowed the TCRα repertoire, which, we believe, followed as a consequence of the restricted TCRδ repertoire. Hence, a single CTCF-mediated chromatin loop directly regulates TCRδ diversity and indirectly regulates TCRα diversity. In addition, we showed that insertion of an ectopic CBE can modify chromatin interactions and disrupt the rearrangement of particular Vδ gene segments. Finally, we investigated the role of YY1 in early T cell development by conditionally deleting YY1 in developing thymocytes. We found that early ablation of YY1 caused severe developmental defects in the DN compartment due to a dramatic increase in DN thymocyte apoptosis. Furthermore, late ablation of YY1 resulted in increased apoptosis of DP thymocytes and a restricted TCRα repertoire. Mechanistically, we showed that p53 was upregulated in both DN and DP YY1-deficient thymocytes. Eliminating p53 in YY1-deficient thymocytes rescued the survival and developmental defects, indicating that these YY1-dependent defects were p53-mediated. We conclude that YY1 is required to maintain cell viability during thymocyte development by thwarting the accumulation of p53.

Overall, this thesis work has shown that CTCF-dependent looping provides a central framework for lineage- and developmental stage-specific regulation of Tcra-Tcrd gene expression and rearrangements. In addition, we identified YY1 as a novel regulator of thymocyte viability.

Development and Application of Covalent-Labeling Strategies for the Large-Scale Thermodynamic Analysis of Protein Folding and Ligand Binding

Thermodynamic stability measurements on proteins and protein-ligand complexes can offer insights not only into the fundamental properties of protein folding reactions and protein functions, but also into the development of protein-directed therapeutic agents to combat disease. Conventional calorimetric or spectroscopic approaches for measuring protein stability typically require large amounts of purified protein. This requirement has precluded their use in proteomic applications. Stability of Proteins from Rates of Oxidation (SPROX) is a recently developed mass spectrometry-based approach for proteome-wide thermodynamic stability analysis. Since the proteomic coverage of SPROX is fundamentally limited by the detection of methionine-containing peptides, the use of tryptophan-containing peptides was investigated in this dissertation. A new SPROX-like protocol was developed that measured protein folding free energies using the denaturant dependence of the rate at which globally protected tryptophan and methionine residues are modified with dimethyl (2-hydroxyl-5-nitrobenzyl) sulfonium bromide and hydrogen peroxide, respectively. This so-called Hybrid protocol was applied to proteins in yeast and MCF-7 cell lysates and achieved a ~50% increase in proteomic coverage compared to probing only methionine-containing peptides. Subsequently, the Hybrid protocol was successfully utilized to identify and quantify both known and novel protein-ligand interactions in cell lysates. The ligands under study included the well-known Hsp90 inhibitor geldanamycin and the less well-understood omeprazole sulfide that inhibits liver-stage malaria. In addition to protein-small molecule interactions, protein-protein interactions involving Puf6 were investigated using the SPROX technique in comparative thermodynamic analyses performed on wild-type and Puf6-deletion yeast strains. A total of 39 proteins were detected as Puf6 targets and 36 of these targets were previously unknown to interact with Puf6. Finally, to facilitate the SPROX/Hybrid data analysis process and minimize human errors, a Bayesian algorithm was developed for transition midpoint assignment. In summary, the work in this dissertation expanded the scope of SPROX and evaluated the use of SPROX/Hybrid protocols for characterizing protein-ligand interactions in complex biological mixtures.

Hispanic Orientalism: The Literary Development of a Cultural Paradigm, from Medieval Spain to Modern Latin America

This dissertation offers a novel approach to Hispanic Orientalism, developing a dynamic paradigm from its origins in medieval and Renaissance Iberia during the process of the Christian Reconquest, to its transatlantic migration and establishment in the early years of the Colony, from where it changed in late colonial and post-Independence Latin America, and onto modernity. The study argues that Hispanic Orientalism does not necessarily imply a negative depiction of the Other, a quality associated with the traditional critique of Saidian Orientalism. Neither, does it entirely comply with the positivist approach suggested in the theoretical research of Said’s opponents, like Julia Kushigian. This dissertation also argues that sociopolitical changes and the shift in the discourse of powers, from imperial to non-imperial, had a significant impact of the development of Hispanic Orientalism, shaping the relationship with the Other. The methodology involves close reading of representative texts depicting the interactions of the dominant and dominated societies from each of the four historic periods that coincided with significant sociopolitical transformations in Hispanic society. Through an intercultural approach to literary studies, social history, and religious studies, this project develops an original paradigm of Hispanic Orientalism, derived from the image of the reinvented Semitic Other portrayed in the literary works depicting the relationship between the hegemonic and the subaltern cultures during the Reconquest period in Spain. Then, it traces the turn of the original paradigm towards reinterpretation during its transatlantic migration to Latin America through the analysis of the chronicles and travelogs of the first colonizers and explorers. During the transitional late colonial and early Independence periods Latin America sees a significant change in the discourse of powers, and Hispanic Orientalism reflects this oscillation between the past and the present therough the works of the Latin American authors from the seventeenth to the nineteenth centuries. Finally, once the non-imperial discourse of power established itself in the former Colony, a new modern stage in the development of Hispanic Orientalist paradigm takes place. It is marked by the desire to differentiate itself from the O(o)thers, as manifested in the works of the representatives of Modernism and the Boom.

Establishing a Baseline Measurement of Intercultural Competence of Ukrainian Faculty: Setting the Stage for Domestic and Global Intercultural Understanding

Post-Soviet Ukraine is in a time of upheaval and transition. Internal relations between pro-Western and pro-Russian supporters have deteriorated in the light of recent political events of Euro Revolution, Russia’s occupation of the Crimean peninsula, and the militant confrontations in the southeastern regions of the country. In the light of these developments, intercultural competence is greatly needed to alleviate domestic tensions and enable effective intercultural communication with the representatives of different cultures within the country and beyond its borders. This study established a baseline of psychometric estimates of intercultural competence of Ukrainian higher education faculty. A sample of 276 professors of different academic majors from one university in Western Ukraine participated in the research. The Global Perspective Inventory (GPI; Merrill, Braskamp, & Braskamp, 2012) was chosen as a research instrument to measure intercultural competence of the faculty members. The GPI takes into account cognitive, intrapersonal, and interpersonal domains, each of which contains two scales reflective of theories of cultural development and intercultural communication – Cognitive-Knowing, Cognitive-Knowledge, Intrapersonal-Identity, Intrapersonal-Affect, Interpersonal-Social Responsibility, and Interpersonal-Social Interaction. Because the research instrument has neither been previously used as a measure of intercultural competence, nor administered in Ukraine, it was cross-validated using a Table of Specification (Newman, Lim, & Pineda, 2013) and two sets of factor analyses. As a result, a modified version of the GPI was created for use in Ukraine. Multiple linear regression analyses were used to test relationships between the participants’ GPI scores on intercultural competence, and several independent variables that consisted of academic discipline, intercultural experience, and how long the participants taught at the university. The analyses determined a positive relationship between the scores on three out of six scales of the original version and two out of five scales of the modified version of the GPI and all the independent variables simultaneously. The relationship between the faculty responses on the six scales of both GPI versions and the independent variables controlling for each other produced mixed results. A unique role of intercultural professional development in predicting intercultural competence was discussed.

Analyses of benthic and planktonic foraminifera during Marine Isotope Stage 5 of ODP Hole 172-1058C

Subtropical Gyres are an important constituent of the ocean-atmosphere system due to their capacity to store vast amounts of warm and saline waters. Here we decipher the sensitivity of the (sub)surface North Atlantic Subtropical Gyre with respect to orbital and millennial scale climate variability between ~140 and 70 ka, Marine Isotope Stage (MIS) 5. Using (isotope)geochemical proxy data from surface and thermocline dwelling foraminifers from Blake Ridge off the west coast of North America (ODP Site 1058) we show that the oceanographic development at subsurface (thermocline) level is substantially different from the surface ocean. Most notably, surface temperatures and salinities peak during the penultimate deglaciation (Termination II) and early MIS 5e, implying that subtropical surface ocean heat and salt accumulation might have resulted from a sluggish northward heat transport. In contrast, maximum thermocline temperatures are reached during late MIS 5e when surface temperatures are already declining. We argue that the subsurface warming originated from intensified Ekman downwelling in the Subtropical Gyre due to enhanced wind stress. During MIS 5a-d a tight interplay of the subtropical upper ocean hydrography to high latitude millennial-scale cold events can be observed. At Blake Ridge, the most pronounced of these high latitude cold events are related to surface warming and salt accumulation in the (sub)surface. Similar to Termination II, heat accumulated in the Subtropical Gyre probably due to a reduced Atlantic Meridional Overturning Circulation. Additionally, a southward shift and intensification of the subtropical wind belts lead to a decrease of on-site precipitation and enhanced evaporation, coupled to intensified gyre circulation. Subsequently, the northward advection of these warm and saline water likely contributed to the fast resumption of the overturning circulation at the end of these high latitude cold events.

Implementing the Physical Literacy Component of the 2015 Ontario Health and Physical Education Curriculum: Is the Stage Set?

Introduction: Current physical activity levels among children and youth are alarmingly low; a mere 7% of children and youth are meeting the Canadian Physical Activity Guidelines (Colley et al., 2011), which means that the vast majority of this population is at risk of developing major health problems in adulthood (Janssen & Leblanc, 2010). These high inactivity rates may be related to suboptimal experiences in sport and physical activity stemming from a lack of competence and confidence (Lubans, Morgan, Cliff, Barnett, & Okely, 2010). Developing a foundation of physical literacy can encourage and maintain lifelong physical activity, yet this does not always occur naturally as a part of human growth (Hardman, 2011). An ideal setting to foster the growth and development of physical literacy is physical education class. Physical education class can offer all children and youth an equal opportunity to learn and practice the skills needed to be active for life (Hardman, 2011). Elementary school teachers are responsible for delivering the physical education curriculum, and it is important to understand their will and capacity as the implementing agents of physical literacy development curriculum (McLaughlin, 1987). Purpose: The purpose of this study was to explore the physical literacy component of the 2015 Ontario Health and Physical Education curriculum policy through the eyes of key informants, and to explore the resources available for the implementation of this new policy. Methods: Qualitative interviews were conducted with seven key informants of the curriculum policy development, including two teachers. In tandem with the interviews, a resource inventory and curriculum review were conducted to assess the content and availability of physical literacy resources. All data were analyzed through the lens of Hogwood and Gunn’s (1984) 10 preconditions for policy implementation. Results: Participants discussed how implementation is affected by: accountability, external capacity, internal capacity, awareness and understanding of physical literacy, implementation expertise, and policy climate. Discussion: Participants voiced similar opinions on most issues, and the overall lack of attention given to physical education programs in schools will continue to be a major dilemma when trying to combat such high physical inactivity levels.

Expertise Development in Volleyball: the Role of Early Sport Activities and Players’ Age and Height

The purpose of this study was to analyse the developmental pathway of skilled and less skilled volleyball players by focusing on the quantity and type of sporting activities, as well as their age and height in comparison to peers in those experiences. Retrospective interviews were conducted to provide a longitudinal and detailed account of sport involvement of 30 skilled and 30 less skilled volleyball players (15 male and 15 female players per group) throughout different developmental stages (stage 1: 8-12 years; stage 2: 13-16 years; stage 3: 17-20 years). Results indicated that the developmental pathway of these volleyball players (i.e. skilled and less skilled) was characterized by an early diversified sport involvement with a greater participation in sport activities during stages 1 and 2. However, skilled players specialized later in volleyball (between age 14 and 15) and performed more hours of volleyball at stage 3 (from 17 years of age onwards). Also, skilled players (male and female) were younger in both the diversified sport activities and volleyball at the later stages of development (i.e. stages 2 and 3), and skilled female players were taller than peers in those activities in the early stages of development (i.e. stages 1 and 2). The present findings suggest early diversification as a feasible pathway to reach expertise in volleyball and highlight the importance of practicing with older peers once specialization in the main sport has occurred. The findings highlight the need for coaches and sport programs to consider different stimuli existing within the training environment (i.e. characteristics of athletes, such as age and height) that influence the quality of practice and contribute to players’ expertise development.

DEVELOPMENT OF A MECHANISM AND PROCESS TO CONTINUOUSLY PRODUCE SPHERICAL, TORREFIED BIOMASS PELLETS

A recently developed novel biomass fuel pellet, the Q’ Pellet, offers significant improvements over conventional white pellets, with characteristics comparable to those of coal. The Q’ Pellet was initially created at bench scale using a proprietary die and punch design, in which the biomass was torrefied in-situ¬ and then compressed. To bring the benefits of the Q’ Pellet to a commercial level, it must be capable of being produced in a continuous process at a competitive cost. A prototype machine was previously constructed in a first effort to assess continuous processing of the Q’ Pellet. The prototype torrefied biomass in a separate, ex-situ reactor and transported it into a rotary compression stage. Upon evaluation, parts of the prototype were found to be unsuccessful and required a redesign of the material transport method as well as the compression mechanism. A process was developed in which material was torrefied ex-situ and extruded in a pre-compression stage. The extruded biomass overcame multiple handling issues that had been experienced with un-densified biomass, facilitating efficient material transport. Biomass was extruded directly into a novel re-designed pelletizing die, which incorporated a removable cap, ejection pin and a die spring to accommodate a repeatable continuous process. Although after several uses the die required manual intervention due to minor design and manufacturing quality limitations, the system clearly demonstrated the capability of producing the Q’ Pellet in a continuous process. Q’ Pellets produced by the pre-compression method and pelletized in the re-designed die had an average dry basis gross calorific value of 22.04 MJ/kg, pellet durability index of 99.86% and dried to 6.2% of its initial mass following 24 hours submerged in water. This compares well with literature results of 21.29 MJ/kg, 100% pellet durability index and <5% mass increase in a water submersion test. These results indicate that the methods developed herein are capable of producing Q’ Pellets in a continuous process with fuel properties competitive with coal.

A Global Review on Peri-Urban Development and Planning

Urban regions worldwide are increasingly facing the challenge of dealing with highly dynamic metropolitan growth and, at the same time, institutional changes like decentralisation and globalisation. These kinds of changes express themselves most evidently in peri-urban areas, where urban and rural life meets. These peri-urban areas in particular have been the stage for rapid physical, social and economic transformations, both in developed and developing countries. Peri-urbanization takes place here. Based on literature review, this paper presents an effort to identify generic attributes of peri-urbanisation and the way in which development planning tends to reply. Three major attributes are identified: peri-urban space (the spatial expression of peri-urban development), peri-urban life (the functional appearance of land uses, activities and peri-urban innovation), and peri-urban change (a causal and temporal perspective featuring flows and drivers of change). It is also shown that prevalent institutional replies in planning and development generally fail to acknowledge the dynamic and increasingly fragmented attributes of global peri-urbanisation.

Importance of risk factor management in diabetic patients and reduction in Stage B heart failure

BACKGROUND: A number of studies have demonstrated the presence of a diabetic cardiomyopathy, increasing the risk of heart failure development in this population. Improvements in present-day risk factor control may have modified the risk of diabetes-associated cardiomyopathy.

AIM: We sought to determine the contemporary impact of diabetes mellitus (DM) on the prevalence of cardiomyopathy in at-risk patients with and without adjustment for risk factor control.

DESIGN: A cross-sectional study in a population at risk for heart failure.

METHODS: Those with diabetes were compared to those with other cardiovascular risk factors, unmatched, matched for age and gender and then matched for age, gender, body mass index, systolic blood pressure and low density lipoprotein cholesterol.

RESULTS: In total, 1399 patients enrolled in the St Vincent's Screening to Prevent Heart Failure (STOP-HF) cohort were included. About 543 participants had an established history of DM. In the whole sample, Stage B heart failure (asymptomatic cardiomyopathy) was not found more frequently among the diabetic cohort compared to those without diabetes [113 (20.8%) vs. 154 (18.0%), P = 0.22], even when matched for age and gender. When controlling for these risk factors and risk factor control Stage B was found to be more prevalent in those with diabetes [88 (22.2%)] compared to those without diabetes [65 (16.4%), P = 0.048].

CONCLUSION: In this cohort of patients with established risk factors for Stage B heart failure superior risk factor management among the diabetic population appears to dilute the independent diabetic insult to left ventricular structure and function, underlining the importance and benefit of effective risk factor control in this population on cardiovascular outcomes.

Development of release methods for critically endangered freshwater pearl mussels (Margaritifera margaritifera)

Biodiversity loss is a global problem with freshwater bivalves considered amongst the most
endangered biota. The freshwater pearl mussel, Margaritifera margaritifera, is declining
throughout its range owing to habitat degradation and overexploitation. In most of its range,
populations are regarded as reproductively non-functional which has led to the development
of captive breeding programmes. A novel method of releasing M. margaritifera was trialled,
with captive-bred juveniles being released into the rivers caged in ‘mussels silos’ (protective
concrete domes with ventilation creating upwelling to ensure water through flow). We
released 240 juvenile mussels and survival and growth rates were monitored for 18 months
post-release for three size classes: A (13.01-20.00mm); B (10.01-13.00mm); and C (4.01-
10.00mm). We explicitly tested two experimental treatments; one where sediment was added
to each silo (allowing mussels to orientate and burrow) and one without sediment. Survival
by the end of the experiment at month 18 was significantly higher for the largest size class at
97% (though growth was lowest in this cohort), and lowest for the smallest size class at 61%
(though growth was highest in this cohort). Survival and growth were unaffected by the
experimental treatment suggesting that adding sediment offered no advantage. Growth was
positively correlated with both water temperature and the particle size of suspended solids
(both of which were collinear, peaking in summer). There are a large number of ex situ
breeding programmes for freshwater pearl mussels throughout Europe and our finding
suggest that the use of ‘mussel silos’ could be a useful tool to protecting juvenile mussels
allowing them to be released at a relatively early stage of development, minimising the risk of
domestication.

Intraclonal heterogeneity is a critical early event in the development of myeloma and precedes the development of clinical symptoms.

The mechanisms involved in the progression from monoclonal gammopathy of undetermined significance (MGUS) and smoldering myeloma (SMM) to malignant multiple myeloma (MM) and plasma cell leukemia (PCL) are poorly understood but believed to involve the sequential acquisition of genetic hits. We performed exome and whole-genome sequencing on a series of MGUS (n=4), high-risk (HR)SMM (n=4), MM (n=26) and PCL (n=2) samples, including four cases who transformed from HR-SMM to MM, to determine the genetic factors that drive progression of disease. The pattern and number of non-synonymous mutations show that the MGUS disease stage is less genetically complex than MM, and HR-SMM is similar to presenting MM. Intraclonal heterogeneity is present at all stages and using cases of HR-SMM, which transformed to MM, we show that intraclonal heterogeneity is a typical feature of the disease. At the HR-SMM stage of disease, the majority of the genetic changes necessary to give rise to MM are already present. These data suggest that clonal progression is the key feature of transformation of HR-SMM to MM and as such the invasive clinically predominant clone typical of MM is already present at the SMM stage and would be amenable to therapeutic intervention at that stage.

Digestive enzymes during development of Ceratitis capitata (Diptera:Tephritidae) and effects of SBTI on its digestive serine proteinase targets

SILVA, Fatima C. B. L. et al. Digestive enzymes during development of Ceratitis capitata (Diptera:Tephritidae) and effects of SBTI on its digestive serine proteinase targets. Insect Biochemistry and Molecular Biology, v. 36, p. 561-569, 2006.ISSN: 0965-1748.DOI: 10.1016/j.ibmb.2006.04.004.