Effect of Media and Estrogen on Morphological Change in Candida albicans

Introduction: Candida albicans (C. albicans), an opportunistic pathogen, lives symbiotically within the intestine of its human host. Temperature and chemical factors have been shown to induce a morphological change in C. albicans from yeast to filamentous form turning C. albicans pathogenic. In this study, we investigated the intestinal cues that might be responsible for the change. We found that different solid media impact the morphological phenotype so we focused on characterizing these before further testing. We tested Estradiol (E2) because of its known linkage to sepsis and higher levels during infections. Experiments were conducted to compare solid agar plates of YEPD, Minimal Media (MM), and Spider Media (SP) for C. albicans growth to choose the best one for further testing with E2 and other factors that could be prone to causing morphological changes. Methods: C. albicans was inoculated through streak method on different solid media (YEPD, MM, SP) and incubated at 30℃. The effect of 0.1nM E2 on C. albicans morphology was also tested. Morphological changes were assayed through bright-field microscopy. Results: Using the three different medias, we found three distinctive phenotypes: A, B, and C. Out of 6 experiments of 14 MM plates, the expressed phenotype was 86% A and 14% inconclusive of the time. 8 experiments of 17 SP plates showed 100% of phenotype B. 6 experiments of 14 YEPD plates presented phenotype C 92% of the time and 8% inconclusive. For E2 trials, 2 experiments, 6 MM plates showed 50% phenotype A and 50% inconclusive. 4 experiments, 10 SP plates had phenotype B 100%. YEPD 2 experiments, 2 plates had phenotype C at 100%. Conclusion: We have established experimental conditions of media controls for further testing whether E2 and other cues, such as inflammatory cytokines, have inhibitory or positive effects on the growth of C. albicans

Testing Environmental Cues on Candida albicans Morphology

Introduction C. albicans is a commensal fungus which under certain environmental cues (e.g., pH, oxidative stress) shifts morphology from spores to filamentous and becomes invasive within the human body. This work aims to identify the environmental gut cues responsible for this morphological shift. Estrogen (E2) becomes elevated during sepsis, thus the guiding hypothesis states that E2 may represent a factor responsible for the morphological change in C. albicans. Methods A calibration curve of growth of C. albicans in liquid minimal media (MM) was established using a spectrophotometer and correlating optical density with cell counts measured with a hematocyter. Liquid MM was inoculated in quadruplets of three different amounts of C. albicans. To test the effect of estrogen at 1nM concentration, E2 was added at the time of inoculation to one of each tube set, and fetal bovine serum was the positive control in another tube. All tubes were anaerobically grown over 3 nights in a shaking incubator at 30℃. Morphological changes were assayed using bright-field microscopy. Results C. albicans was inoculated in amounts of 1, 2, and 4 million cells into sets of 4 tubes each based on the established growth curve. The MM relationship between OD and number of cells is described by the following equation: 1.06×106 + 1.83×10 7x + 1.68×10 7x 2 , R 2= 0.867. Adding E2 at 1 nM to the liquid media appeared to induce filamentous growth and budding, as with positive control 10% FBS. Conclusion Our preliminary experiments indicate that regardless of initial cell amount, tubes containing E2 seem to induce more filamentous growth in MM, as observed with FBS (positive control). Further experiments to determine effects of E2 at other concentrations would bring more insight, as well as trials combining E2 and FBS to explore if there is an additive or inhibitory effect on filamentation

Effect of Media and Estrogen on Morphological Change in Candida albicans

Introduction: Candida albicans (C. albicans), an opportunistic pathogen, lives symbiotically within the intestine of its human host. Temperature and chemical factors have been shown to induce a morphological change in C. albicans from yeast to filamentous form turning C. albicans pathogenic. In this study, we investigated the intestinal cues that might be responsible for the change. We found that different solid media impact the morphological phenotype so we focused on characterizing these before further testing. We tested Estradiol (E2) because of its known linkage to sepsis and higher levels during infections. Experiments were conducted to compare solid agar plates of YEPD, Minimal Media (MM), and Spider Media (SP) for C. albicans growth to choose the best one for further testing with E2 and other factors that could be prone to causing morphological changes. Methods: C. albicans was inoculated through streak method on different solid media (YEPD, MM, SP) and incubated at 30℃. The effect of 0.1nM E2 on C. albicans morphology was also tested. Morphological changes were assayed through bright-field microscopy. Results: Using the three different medias, we found three distinctive phenotypes: A, B, and C. Out of 6 experiments of 14 MM plates, the expressed phenotype was 86% A and 14% inconclusive of the time. 8 experiments of 17 SP plates showed 100% of phenotype B. 6 experiments of 14 YEPD plates presented phenotype C 92% of the time and 8% inconclusive. For E2 trials, 2 experiments, 6 MM plates showed 50% phenotype A and 50% inconclusive. 4 experiments, 10 SP plates had phenotype B 100%. YEPD 2 experiments, 2 plates had phenotype C at 100%. Conclusion: We have established experimental conditions of media controls for further testing whether E2 and other cues, such as inflammatory cytokines, have inhibitory or positive effects on the growth of C. albicans

WWOX expression in colorectal cancer—a real-time quantitative RT-PCR study

The WWOX gene is a tumour suppressor gene affected in various types of malignancies. Numerous studies showed either loss or reduction of the WWOX expression in variety of tumours, including breast, ovary, liver, stomach and pancreas. Recent study demonstrated that breast cancer patients exhibiting higher WWOX expression showed significantly longer disease-free survival in contrast to the group with lower relative WWOX level. This work was undertaken to show whether similar phenomena take place in colon tumours and cell lines. To assess the correlation of WWOX gene expression with prognosis and cancer recurrence in 99 colorectal cancer patients, we performed qRT-PCR analysis. We also performed analysis of WWOX promoter methylation status using MethylScreen method and analysis of loss of heterozygosity (LOH) status at two WWOX-related loci, previously shown to be frequently deleted in various types of tumours. A significantly better disease-free survival was observed among patients with tumours exhibiting high level of WWOX (hazard ratio = 0.39; p = 0.0452; Mantel–Cox log-rank test), but in multivariate analysis it was not an independent prognostic factor. We also found that although in colorectal cancer WWOX expression varies among patients and correlates with DFS, the exact mode of decrease in this type of tumour was not found. We failed to find the evidence of LOH in WWOX region, or hypermethylation in promoter regions of this gene. Although we provide the evidence for tumour-suppressive role of WWOX gene expression in colon, we were unable to identify the molecular mechanism responsible for this

Genetically Determined Height and Risk of Non-hodgkin Lymphoma

Although the evidence is not consistent, epidemiologic studies have suggested that taller adult height may be associated with an increased risk of some non-Hodgkin lymphoma (NHL) subtypes. Height is largely determined by genetic factors, but how these genetic factors may contribute to NHL risk is unknown. We investigated the relationship between genetic determinants of height and NHL risk using data from eight genome-wide association studies (GWAS) comprising 10,629 NHL cases, including 3,857 diffuse large B-cell lymphoma (DLBCL), 2,847 follicular lymphoma (FL), 3,100 chronic lymphocytic leukemia (CLL), and 825 marginal zone lymphoma (MZL) cases, and 9,505 controls of European ancestry. We evaluated genetically predicted height by constructing polygenic risk scores using 833 height-associated SNPs. We used logistic regression to estimate odds ratios (OR) and 95% confidence intervals (CI) for association between genetically determined height and the risk of four NHL subtypes in each GWAS and then used fixed-effect meta-analysis to combine subtype results across studies. We found suggestive evidence between taller genetically determined height and increased CLL risk (OR = 1.08, 95% CI = 1.00–1.17, p = 0.049), which was slightly stronger among women (OR = 1.15, 95% CI: 1.01–1.31, p = 0.036). No significant associations were observed with DLBCL, FL, or MZL. Our findings suggest that there may be some shared genetic factors between CLL and height, but other endogenous or environmental factors may underlie reported epidemiologic height associations with other subtypes

Genome-wide association analysis implicates dysregulation of immunity genes in chronic lymphocytic leukaemia

Several chronic lymphocytic leukaemia (CLL) susceptibility loci have been reported; however, much of the heritable risk remains unidentified. Here we perform a meta-analysis of six genome-wide association studies, imputed using a merged reference panel of 1,000 Genomes and UK10K data, totalling 6,200 cases and 17,598 controls after replication. We identify nine risk loci at 1p36.11 (rs34676223, P=5.04 × 10−13), 1q42.13 (rs41271473, P=1.06 × 10−10), 4q24 (rs71597109, P=1.37 × 10−10), 4q35.1 (rs57214277, P=3.69 × 10−8), 6p21.31 (rs3800461, P=1.97 × 10−8), 11q23.2 (rs61904987, P=2.64 × 10−11), 18q21.1 (rs1036935, P=3.27 × 10−8), 19p13.3 (rs7254272, P=4.67 × 10−8) and 22q13.33 (rs140522, P=2.70 × 10−9). These new and established risk loci map to areas of active chromatin and show an over-representation of transcription factor binding for the key determinants of B-cell development and immune response

Genetic overlap between autoimmune diseases and non-Hodgkin lymphoma subtypes

Epidemiologic studies show an increased risk of non-Hodgkin lymphoma (NHL) in patients with autoimmune disease (AD), due to a combination of shared environmental factors and/or genetic factors, or a causative cascade: chronic inflammation/antigen-stimulation in one disease leads to another. Here we assess shared genetic risk in genome-wide-association-studies (GWAS). Secondary analysis of GWAS of NHL subtypes (chronic lymphocytic leukemia, diffuse large B-cell lymphoma, follicular lymphoma, and marginal zone lymphoma) and ADs (rheumatoid arthritis, systemic lupus erythematosus, and multiple sclerosis). Shared genetic risk was assessed by (a) description of regional genetic of overlap, (b) polygenic risk score (PRS), (c)"diseasome", (d)meta-analysis. Descriptive analysis revealed few shared genetic factors between each AD and each NHL subtype. The PRS of ADs were not increased in NHL patients (nor vice versa). In the diseasome, NHLs shared more genetic etiology with ADs than solid cancers (p = .0041). A meta-analysis (combing AD with NHL) implicated genes of apoptosis and telomere length. This GWAS-based analysis four NHL subtypes and three ADs revealed few weakly-associated shared loci, explaining little total risk. This suggests common genetic variation, as assessed by GWAS in these sample sizes, may not be the primary explanation for the link between these ADs and NHLs

Genome-wide association analysis implicates dysregulation of immunity genes in chronic lymphocytic leukaemia

Several chronic lymphocytic leukaemia (CLL) susceptibility loci have been reported; however, much of the heritable risk remains unidentified. Here we perform a meta-analysis of six genome-wide association studies, imputed using a merged reference panel of 1,000 Genomes and UK10K data, totalling 6,200 cases and 17,598 controls after replication. We identify nine risk loci at 1p36.11 (rs34676223, P=5.04 × 10−13), 1q42.13 (rs41271473, P=1.06 × 10−10), 4q24 (rs71597109, P=1.37 × 10−10), 4q35.1 (rs57214277, P=3.69 × 10−8), 6p21.31 (rs3800461, P=1.97 × 10−8), 11q23.2 (rs61904987, P=2.64 × 10−11), 18q21.1 (rs1036935, P=3.27 × 10−8), 19p13.3 (rs7254272, P=4.67 × 10−8) and 22q13.33 (rs140522, P=2.70 × 10−9). These new and established risk loci map to areas of active chromatin and show an over-representation of transcription factor binding for the key determinants of B-cell development and immune response