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Fusion-positive alveolar rhabdomyosarcoma (FP-RMS) is an aggressive pediatric sarcoma driven primarily by the PAX3-FOXO1 fusion oncogene, for which therapies targeting PAX3-FOXO1 are lacking. We screened 62,643 compounds using an engineered cell line that monitors PAX3-FOXO1 transcriptional activity identifying a hitherto uncharacterized compound, PFI-63. RNA-seq, ATAC-seq, and docking analyses implicated histone lysine demethylases (KDMs) as its targets. Enzymatic assays confirmed the inhibition of multiple KDMs with highest selectivity for KDM3B. Structural similarity search of PFI-63 identified PFI-90 with improved solubility and potency. Biophysical binding of PFI-90 to KDM3B was demonstrated using NMR and SPR. PFI-90 suppressed the growth of FP-RMS in vitro and in vivo through downregulating PAX3-FOXO1 activity, and combined knockdown of KDM3B and KDM1A phenocopied PFI-90 effects. Thus, we report novel KDM inhibitors with highest specificity for KDM3B. Its potent suppression of PAX3-FOXO1 activity can be exploited as a new therapeutic approach for FP-RMS and other transcriptionally driven cancers.

Background: Rural households in Ethiopia are dependent on biomass for their energy needs and this exacerbates deforestation. Therefore, building small-scale biogas is one of the abating means of these challenges. The objective of this study was to assess the practices and opportunities of small-scale biogas technology in selected Districts’ Hadiya zone, southern Ethiopia. Methods: To achieve this objective, fifty-six informants were selected from the seven districts of Hadiya Zone, semi-structured questionnaires and field observations were used as the instruments to collect the data. Result: This survey revealed that small-scale biogas technology in the study area is recent and it needs certain requirements for its construction and due to the stopped feeding, most of the biogas plants were non-functional. Despite this, the presence of many cattle and optimum climate are among the essential opportunities to expand biogas distribution in the study area. Therefore, farmers and the concerned stakeholders should use the available opportunity to maximize the distribution of biogas technology in the study area.

BACKGROUND: The PAX3/7-FOXO1 (P3F) fusion transcription factor is the oncogenic driver in fusion-positive rhabdomyosarcoma (FP-RMS). P3F drives oncogenesis in FP-RMS through transcriptional modulation of downstream target genes. Thus, P3F represents a unique vulnerability in FP-RMS. A screen for inhibitors of P3F action identified novel histone lysine demethylase (KDM) inhibitors characterized in this study. MATERIALS/METHODS: A specific luciferase assay which monitors P3F activity was utilized to screen 62,643 compounds. The top candidate with unknown mechanism of action, PFI-63, and its analog PFI-90, were characterized. Western blotting was used to detect MYOG, PARP, and PAX3-FOXO1. KDM enzyme inhibition assays were conducted. Ligand-observed NMR analysis was used to determine binding of PFI-90 to KDM3B. KDM knockdown using CRISPRi was carried out. ChIP-seq analysis on H3K4me3, H3K9me2, H3K27me3, H3K27ac, and PAX3-FOXO1 was performed. Mouse xenograft models of FP-RMS were used to determine in vivo efficacy. RESULTS: 64 compounds that inhibited P3F activity without general inhibition of transcription or induction of cell death were further characterized. PFI-63 and a more water-soluble analog, PFI-90, were identified. GSEA of RNA-seq showed activation of apoptosis and myogenesis while P3F targets were repressed. Activation of apoptosis and myogenesis were validated by Western blotting showing PARP cleavage and increased MYOG levels, respectively. RNA-seq suggested that PFI-63 and PFI-90 were KDM inhibitors. In vitro enzymatic inhibition assays confirmed activity against multiple KDMs with most potent inhibition of KDM3B. Western analysis for methylation at H3K4 and H3K9 showed increases after PFI-90 treatment. NMR techniques confirmed biophysical binding of PFI-90 to KDM3B. RNA-seq of KDM knockdowns demonstrated that KDM3B knockdown most closely recapitulated PFI-90’s downregulation of P3F targets. Knockdown of KDM1A recapitulated PFI-90’s upregulation of myogenesis and apoptosis. ChIP-seq analysis showed increased levels of H3K9me2 at P3F sites while H3K4me3 was increased in muscle differentiation and apoptosis. In two different in vivo xenograft models of FP-RMS, PFI-90 treatment delayed tumor progression vs DMSO control. CONCLUSION: We identified novel multi-KDM inhibitors with highest potency for KDM3B. Downregulation of P3F by KDM3B inhibition is associated with increased H3K9me2 at P3F sites. PFI-90 also inhibits KDM1A which increases H3K4me3 at myogenesis and apoptosis genes. Thus, PFI-90 is a novel multi-KDM inhibitor whose biological effect on FP-RMS is by inhibition of KDM3B and KDM1A. Pre-clinical validation via FP-RMS xenograft models showed that PFI-90 delayed tumor progression. PFI-90 thus represents a promising novel compound for the treatment of FP-RMS, and potentially, other transcriptionally driven cancers. Citation Format: Yong Yean Kim, Girma Woldemichael, Berkley Gryder, Silvia Pomella, Ranuka Sinniah, Josh Kowalczyk, Young Song, Mehal Churiwal, Joseph Barchi, John Schneekloth, Xinyu Wen, Hsein-Chao Chou, Barry Okeefe, John Shern, Robert Hawley, Javed Khan. Novel histone lysine demethylase inhibitors disrupt PAX3-FOXO1-driven transcriptional output in fusion-positive rhabdomyosarcoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 703.

Soil is one of the natural resource and under high pressure that is increasing from year to year, resulting in poor fertility. The objective of this study was to assess the attitudes of farmer’s perception to soil fertility management practices. In order to achieve these objectives, random sampling methods was used to select respondents in the study area. The data was collected by using field observation, questionnaires and key informant discussion. The collected data were analyzed through descriptive statistics. The survey revealed that the factors that hinder farmers from using improved ways of soil fertility management practices are: labor problem 27.5%, economic problem 20%, lack of awareness and demographic factors 37.5%. In the Kalisha District, there are a number of major indigenous soil fertility management practices (SFMP) that are using by almost all farmers such as using cattle dung, straw, intercropping legumes crops in their farm land and use of enset in homegarden area. In other form, this study showed that, in Kalisha District the attitudes of farmers to soil fertility management is less, due to the awareness gap in society and less interventions of development agents. Therefore the farmers should be aware of soil fertility management practices on both biological and physical measures to restore soil fertility and they have to scale up the indigenous SFMP to maintain the productivity of the soil.

Diabetes mellitus is fourth most high mortality rate diseases in the world and it is also a cause of kidney disease, blindness, and heart diseases. Data mining techniques support a medical decision for a correct diagnosis, treatment of disease in such way it minimizes the workload of specialists. This study proposed to predict diabetes using data mining techniques. Back propagation algorithm is used to predict whether the person has diabetic or not. And also J48, naive bayes and support vector machine were used to predict diabetes. These neural networks were having an input layer with having 8 parameters, one hidden layer having 6 neurons and produce one output layer.5 fold cross-validation technique and large value learning rate was used to improve the performance of the model. PIMA Indian dataset used to conduct this study. The study implemented in RStudio using R programming language. The performance of Back propagation algorithm is used to predict diabetes diseases gave 83.11 % accuracy, 86.53% sensitivity and 76% specificity, the result shows improvement from previous work. The obtained result is also compared with J48, naive bayes and support vector machine algorithm.

Objectives. We examined correlations between survival and race/ethnicity, age, and gender among persons who died from AIDS-related causes. Methods. We estimated survival among 11 022 persons at 12, 36, and 60 months after diagnosis with AIDS in 1993 through 2001 and reported through 2003 to the Chicago Department of Public Health. We estimated hazard ratios (HRs) by demographic and risk characteristics. Results. All demographic groups had higher 5-year survival rates after the introduction of highly active retroviral therapy (1996–2001) than before (1993–1995). The HR for non-Hispanic Blacks to Whites was 1.18 in 1993 to 1995 and 1.51 (P < .01) in 1996 to 2001. The HR for persons 50 years or older to those younger than 30 years was 1.63 in 1993–1995 and 2.28 (P < .01) in 1996–2001. The female-to-male HR was 0.90 in 1993–1995 and 1.20 (P < .02) in 1996–2001. Conclusions. The risk of death was higher for non-Hispanic Blacks and Hispanics than for non-Hispanic Whites. Interventions are needed to increase early access to care for disadvantaged groups.