Translating the Biology of IL-9 into the Pathogenesis of Chronic Rhinosinusitis

Background: Chronic rhinosinusitis (CRS) is an inflammatory disease of the nasal and paranasal sinus mucosa that can have profound effects on patient quality of life and US healthcare costs. Increased IL-9 expression has been identified in CRS patients, particularly those with nasal polyposis (CRSwNP) or eosinophilic CRS (ECRS). Il-9 gene expression has been found to be uniquely regulated by super enhancer RNA (eRNA). Aim 1: Perform an in-depth literature review to summarize the current understanding of IL-9 biology, with an emphasis on IL-9’s contribution to CRS pathology. Aim 2: Use antisense treatment of mouse Th9 cells in vitro to suppress IL-9 as initial validation of a possible immunotherapeutic treatment strategy for CRS patients. Methods: CD4+ T-cells harvested from mouse lymph nodes and spleen were cultured under Th9 polarizing conditions, confirming Th9 differentiation with flow cytometry. Best strategies for robust baseline IL-9 and super enhancer RNA expression were determined by comparing qPCR values with various IL-9 stimulating conditions and RNA extraction methods. Antisense oligonucleotides (ASOs) targeting the Il-9 gene and super enhancer eRNA were designed using the UCSC In-Silico PCR tool and IDT’s OligoAnalyzer tool and were introduced to Th9 cell cultures using electroporation with the Lonza Nucelofector 2b. The fold decrease in gene expression in ASO vs. control conditions were determined using qPCR relative quantification. Results: Baseline super enhancer expression was 8.9, 7.6, and 3.8 times higher for 3 different super enhancer targets using organic extraction compared to a spin column method for RNA extraction, and 6.8, 7.5, and 6.5 times higher in culture conditions utilizing anti-GITR stimulation compared to OX40L stimulation. Preliminary results of unmodified ASO knockdown studies showed ~2-fold decrease in IL-9 expression in ASO conditions, but this was not consistent across all conditions. Conclusions: The addition of anti-GITR to Th9 cultures lead to the greatest baseline super enhancer expression and super enhancer RNA was best isolated using organic extraction methods. Though preliminary ASO studies are promising for IL-9 suppression, additional testing is necessary to determine the ideal concentrations of ASOs and timing of ASO introduction to cell cultures, as well as replicate experiments with modified ASOs

Expressions 1979

Expressions contains selected work from the 1979 Creative Writing Contest winners and honorable mentions along with Commercial Art students at Des Moines Area Community College. Design, typography and layout was done by Journalism students.https://openspace.dmacc.edu/expressions/1001/thumbnail.jp

Patterns of plant phenology in Amazonian seasonally flooded and unflooded forests

Few studies have successfully monitored community-wide phenological patterns in seasonally flooded Amazonian várzea forests, where a prolonged annual flood pulse arguably generates the greatest degree of seasonality of any low-latitude ecosystem on Earth. We monitored the vegetative and reproductive plant phenology of várzea (VZ) floodplain and adjacent terra firme (TF) forests within two contiguous protected areas in western Brazilian Amazonia, using three complementary methods: monthly canopy observations of 1056 individuals (TF: 556, VZ: 500), twice monthly collections from 0.5-m2 litterfall traps within two 100-ha plots (1 TF, 1 VZ; 96 traps per plot), and monthly ground surveys of residual fruit-fall along transect-grids within each 100-ha plot (12 km per plot). Surveys encompassed the entire annual flood cycle and employed a floating trap design to cope with fluctuating water levels. Phenology patterns were generally similar in both forest types. Leaffall peaked during the aquatic phase in várzea forest and the dry season in terra firme. Flowering typically followed leaffall and leaf flush, extending into the onset of the terrestrial phase and rainy season in várzea and terra firme, respectively. Abiotic seed dispersal modes were relatively more prevalent in várzea than terra firme; the main contrast in fruiting seasonality was more likely a result of differences in community composition and relative abundance of seed dispersal modes than differences within individual genera. We emphasize the difficulty in distinguishing the role of the flood pulse from other seasonal environmental variables without multiannual data or spatially replicated studies across the spectrum of Amazonian forest types

Reproductive phenology of Dipteryx odorata (Aubl.) Willd (Fabaceae) in two forest areas in the Central Amazon

Long term phenological studies in tropical forests are scarce. A phenological study of Dipteryx odorata was carried out from 1974 to 2000 in two areas of Amazon Forest: Reserva Florestal Ducke (RFD) and Estação Experimental de Silvicultura Tropical (EEST). The objective was to compare the phenological patterns between the two areas, to verify the regularity of flowering and fruiting over a period of 27 years, and to verify the influence of climatic factors in these events. Five trees were marked in the RFD and five in the EEST; and were observed monthly for the presence of flowers, unripe and ripe fruits. The flowering and unripe fruiting presented an irregular pattern in both areas, and also varied with regard to duration and period of occurrence between years and seasons. Nevertheless, they presented similar frequency of occurrence during the 27 years of observation. On the other hand, the production of ripe fruits presented a different pattern, with three year intervals without ripe fruits in the RFD, and seven year intervals in the EEST. We concluded that the frequency of flowering and unripe fruiting was annual, and the frequency of occurrence of ripe fruiting was supra-annual, with an irregular pattern, and a duration from intermediate to extended. We also observed the occurrence of one to three year intervals between flowering and unripe fruiting episodes, and one to seven year intervals between episodes with ripe fruiting. The influence of climatic factors in the patterns observed was not clear. We suggest the rational use of natural products extracted from Dipteryx odorata, the "cumaru", especially the fruits, and the development of more long term studies, fundamental for understanding the reproductive phenological patterns as well as the resource offering patterns in tropical forests.Estudos fenológicos de longa duração em florestas tropicais são raros. Foi realizado o acompanhamento fenológico de Dipteryx odorata, no período de 1974 à 2000, em duas áreas de floresta amazônicaa Reserva Florestal Ducke (RFD) e Estação Experimental de Silvicultura Tropical (EEST). O objetivo foi observar os padrões fenológicos nas duas áreas, verificar a regularidade da floração e frutificação num período de 27 anos e a influência dos fatores climáticos nestes eventos. Foram marcados cinco indivíduos na RFD e cinco na EEST e observados quanto à produção de flores, frutos imaturos e maduros. A floração e a produção de frutos imaturos apresentaram padrão irregular nas duas áreas observadas, variando quanto a época de ocorrência e a duração entre anos e estações, mas apresentaram freqüência de ocorrência similar nos 27 anos observados. Para a fenofase frutos maduros este padrão foi diferente, com intervalos de até três anos sem ocorrência de frutos maduros, na RFD e de sete anos na EEST. Conclui-se que a freqüência de ocorrência das fenofases de floração e frutos imaturos foi anual e a de frutos maduros supra-anual, todas com padrão irregular e duração de intermediária a prolongada. Ocorreu variação de um a três anos entre episódios de floração e frutos imaturos e de um a sete anos entre episódios de frutos maduros, não ficando evidente, nesta análise, a influência dos fatores climáticos nos padrões observados. Sugere-se o uso racional dos produtos derivados de Dipteryx odorata, o cumaru, especialmente quanto à exploração de seus frutos e o desenvolvimento de mais estudos de longa duração, fundamentais para entender os padrões fenológicos reprodutivos e de oferta de recursos em florestas tropicais