Radiation Quality Effects on Transcriptome Profiles in 3-d Cultures After Particle Irradiation

In this work, we evaluate the differential effects of low- and high-LET radiation on 3-D organotypic cultures in order to investigate radiation quality impacts on gene expression and cellular responses. Reducing uncertainties in current risk models requires new knowledge on the fundamental differences in biological responses (the so-called radiation quality effects) triggered by heavy ion particle radiation versus low-LET radiation associated with Earth-based exposures. We are utilizing novel 3-D organotypic human tissue models that provide a format for study of human cells within a realistic tissue framework, thereby bridging the gap between 2-D monolayer culture and animal models for risk extrapolation to humans. To identify biological pathway signatures unique to heavy ion particle exposure, functional gene set enrichment analysis (GSEA) was used with whole transcriptome profiling. GSEA has been used extensively as a method to garner biological information in a variety of model systems but has not been commonly used to analyze radiation effects. It is a powerful approach for assessing the functional significance of radiation quality-dependent changes from datasets where the changes are subtle but broad, and where single gene based analysis using rankings of fold-change may not reveal important biological information. We identified 45 statistically significant gene sets at 0.05 q-value cutoff, including 14 gene sets common to gamma and titanium irradiation, 19 gene sets specific to gamma irradiation, and 12 titanium-specific gene sets. Common gene sets largely align with DNA damage, cell cycle, early immune response, and inflammatory cytokine pathway activation. The top gene set enriched for the gamma- and titanium-irradiated samples involved KRAS pathway activation and genes activated in TNF-treated cells, respectively. Another difference noted for the high-LET samples was an apparent enrichment in gene sets involved in cycle cycle/mitotic control. It is plausible that the enrichment in these particular pathways results from the complex DNA damage resulting from high-LET exposure where repair processes are not completed during the same time scale as the less complex damage resulting from low-LET radiation

Bacillus subtilis SbcC protein plays an important role in DNA inter-strand cross-link repair

BACKGROUND: Several distinct pathways for the repair of damaged DNA exist in all cells. DNA modifications are repaired by base excision or nucleotide excision repair, while DNA double strand breaks (DSBs) can be repaired through direct joining of broken ends (non homologous end joining, NHEJ) or through recombination with the non broken sister chromosome (homologous recombination, HR). Rad50 protein plays an important role in repair of DNA damage in eukaryotic cells, and forms a complex with the Mre11 nuclease. The prokaryotic ortholog of Rad50, SbcC, also forms a complex with a nuclease, SbcD, in Escherichia coli, and has been implicated in the removal of hairpin structures that can arise during DNA replication. Ku protein is a component of the NHEJ pathway in pro- and eukaryotic cells. RESULTS: A deletion of the sbcC gene rendered Bacillus subtilis cells sensitive to DNA damage caused by Mitomycin C (MMC) or by gamma irradiation. The deletion of the sbcC gene in a recN mutant background increased the sensitivity of the single recN mutant strain. SbcC was also non-epistatic with AddAB (analog of Escherichia coli RecBCD), but epistatic with RecA. A deletion of the ykoV gene encoding the B. subtilis Ku protein in a sbcC mutant strain did not resulted in an increase in sensitivity towards MMC and gamma irradiation, but exacerbated the phenotype of a recN or a recA mutant strain. In exponentially growing cells, SbcC-GFP was present throughout the cells, or as a central focus in rare cases. Upon induction of DNA damage, SbcC formed 1, rarely 2, foci on the nucleoids. Different to RecN protein, which forms repair centers at any location on the nucleoids, SbcC foci mostly co-localized with the DNA polymerase complex. In contrast to this, AddA-GFP or AddB-GFP did not form detectable foci upon addition of MMC. CONCLUSION: Our experiments show that SbcC plays an important role in the repair of DNA inter-strand cross-links (induced by MMC), most likely through HR, and suggest that NHEJ via Ku serves as a backup DNA repair system. The cell biological experiments show that SbcC functions in close proximity to the replication machinery, suggesting that SbcC may act on stalled or collapsed replication forks. Our results show that different patterns of localization exist for DNA repair proteins, and that the B. subtilis SMC proteins RecN and SbcC play distinct roles in the repair of DNA damage

Factors Associated with Time to Cease Exclusive Breastfeeding Among Infants Aged 6–9 Months, Kirkos Sub-City, Addis Ababa, Ethiopia: Survival Analysis

Getachew Sale Mezgebu,1 Luladey Getachew,2 Fentaw Wassie Feleke,1,3 Rediet Kidane2 1Department of Human Nutrition, School of Nutrition, Food Science and Technology, Academic Center of Excellence in Human Nutrition, College of Agriculture, Hawassa University, Hawassa, Ethiopia; 2Department of Nutrition and Dietetics, Faculty of Public Health, Institute of Health, Jimma University, Jimma, Ethiopia; 3Department of Public Health, School of Public Health, College of Medicine and Health Sciences, Woldia University, Woldia, EthiopiaCorrespondence: Fentaw Wassie Feleke, Tel +251917008650, Fax +251335400643, Email [email protected]: Notwithstanding the World Health Organization’s (WHO) guideline for exclusive breastfeeding (EBF) for the first six months of an infant’s life, its prevalence remains low in both developed and developing countries. Although breastfeeding is widely practiced in Ethiopia, only 59% of the children under the age of six months were exclusively breastfed.Objective: This study aimed to assess the time to cessation of EBF and associated factors among infants aged 6– 9 months in health facilities under Kirkos sub-city, Addis Ababa, Ethiopia.Methods: An institutional-based cross-sectional study was conducted among 287 mothers attending Kirkos sub-city health facilities from September 1 to 30, 2020. Bivariable and multivariable Cox regression models were fitted to identify factors associated with the time to cessation of EBF.Results: According to this finding, the median period for EBF discontinuation was 6 months. Early cessation of EBF was 130 (45.8%) days. Mothers with average household monthly income < 15 USD [AHR, 5.31 (95% CI: 2.82, 9.99)] and 15– 30 USD [AHR, 2.59 (95% CI: 1.41, 4.77)], EBF counseling at post-natal care [AHR, 0.18 (95% CI: 0.10, 0.31)], media access [AHR, 1.67 (95% CI: 1.06, 2.63)], child comorbidity [AHR, 0.51 (95% CI: 0.29, 0.88)], bottle feeding [AHR, 3.28 (95% CI: 2.14 5.01)], unfavorable attitude towards EBF [AHR, 3.59 (95% CI: 2.02, 6.40)] were significant factors associated with the hazard of cessation of EBF.Conclusion: In this investigation, the cumulative survival probability of EBF to 6 months was 45.8%, with a median duration of 6 months (95% CI 5.83– 6.18) months. Unfavorable attitudes, child comorbidity, and bottle feeding were factors associated with earlier cessation of EBF. Media access, household monthly higher income, and EBF counseling at postnatal care were factors associated with prolonged EBF. As a result, supporting behavioral change and media access for effective communication on EBF duration practice following the established standard should be targeted during both ante-natal and post-natal care visits.Keywords: Addis Ababa, hazard ratio, postnatal care, survival analysis, time to cessation of EB

Intra- and interspecific polymorphisms ofLeishmania donovani andL. tropica minicircle DNA

A pair of degenerate polymerase chain reaction (PCR) primers (LEI-1, TCG GAT CC[C,T] [G,C]TG GGT AGG GGC GT; LEI-2, ACG GAT CC[G,C] [G,C][A,C]C TAT [A,T]TT ACA CC) defining a 0.15-kb segment ofLeishmania minicircle DNA was constructed. These primers amplified not only inter- but also intraspecifically polymorphic sequences. Individual sequences revealed a higher intraspecific than interspecific divergence. It is concluded that individual sequences are of limited relevance for species determination. In contrast, when a data base of 19 different sequences was analyzed in a dendrographic plot, an accurate species differentiation was feasible

A novel SMC-like protein, SbcE (YhaN), is involved in DNA double-strand break repair and competence in Bacillus subtilis

Bacillus subtilis and most Gram positive bacteria possess four SMC like proteins: SMC, SbcC, RecN and the product of the yhaN gene, termed SbcE. SbcE is most similar to SbcC but contains a unique central domain. We show that SbcE plays a role during transformation in competent cells and in DNA double-strand break (DSB) repair. The phenotypes were strongly exacerbated by the additional deletion of recN or of sbcC, suggesting that all three proteins act upstream of RecA and provide distinct avenues for presynapsis. SbcE accumulated at the cell poles in competent cells, and localized as a discrete focus on the nucleoids in 10% of growing cells. This number moderately increased after treatment with DNA damaging agents and in the absence of RecN or of SbcC. Damage-induced foci of SbcE arose early after induction of DNA damage and rarely colocalized with the replication machinery. Our work shows that SMC-like proteins in B. subtilis play roles at different subcellular sites during DNA repair. SbcC operates at breaks occurring at the replication machinery, whereas RecN and SbcE function mainly, but not exclusively, at DSBs arising elsewhere on the chromosome. In agreement with this idea, we found that RecN-YFP damage-induced assemblies also arise in the absence of ongoing replication

Single cell analyses reveal contrasting life strategies of the two main nitrifiers in the ocean

Nitrification, the oxidation of ammonia via nitrite to nitrate, is a key process in marine nitrogen (N) cycling. Although oceanic ammonia and nitrite oxidation are balanced, ammonia-oxidizing archaea (AOA) vastly outnumber the main nitrite oxidizers, the bacterial Nitrospinae. The ecophysiological reasons for this discrepancy in abundance are unclear. Here, we compare substrate utilization and growth of Nitrospinae to AOA in the Gulf of Mexico. Based on our results, more than half of the Nitrospinae cellular N-demand is met by the organic-N compounds urea and cyanate, while AOA mainly assimilate ammonium. Nitrospinae have, under in situ conditions, around four-times higher biomass yield and five-times higher growth rates than AOA, despite their ten-fold lower abundance. Our combined results indicate that differences in mortality between Nitrospinae and AOA, rather than thermodynamics, biomass yield and cell size, determine the abundances of these main marine nitrifiers. Furthermore, there is no need to invoke yet undiscovered, abundant nitrite oxidizers to explain nitrification rates in the ocean

Purple sulfur bacteria fix N-2 via molybdenum-nitrogenase in a low molybdenum Proterozoic ocean analogue

Biological N-2 fixation was key to the expansion of life on early Earth. The N-2-fixing microorganisms and the nitrogenase type used in the Proterozoic are unknown, although it has been proposed that the canonical molybdenum-nitrogenase was not used due to low molybdenum availability. We investigate N-2 fixation in Lake Cadagno, an analogue system to the sulfidic Proterozoic continental margins, using a combination of biogeochemical, molecular and single cell techniques. In Lake Cadagno, purple sulfur bacteria (PSB) are responsible for high N-2 fixation rates, to our knowledge providing the first direct evidence for PSB in situ N-2 fixation. Surprisingly, no alternative nitrogenases are detectable, and N-2 fixation is exclusively catalyzed by molybdenum-nitrogenase. Our results show that molybdenum-nitrogenase is functional at low molybdenum conditions in situ and that in contrast to previous beliefs, PSB may have driven N-2 fixation in the Proterozoic ocean. N-2 fixation was key to the expansion of life on Earth, but which organisms fixed N-2 and if Mo-nitrogenase was functional in the low Mo early ocean is unknown. Here, the authors show that purple sulfur bacteria fix N-2 using Mo-nitrogenase in a Proterozoic ocean analogue, despite low Mo conditions

A Study of Reconfigurable Accelerators for Cloud Computing

Due to the exponential increase in network traffic in the data centers, thousands of servers interconnected with high bandwidth switches are required. Field Programmable Gate Arrays (FPGAs) with Cloud ecosystem offer high performance in efficiency and energy, making them active resources, easy to program and reconfigure. This paper looks at FPGAs as reconfigurable accelerators for the cloud computing presents the main hardware accelerators that have been presented in various widely used cloud computing applications such as: MapReduce, Spark, Memcached, Databases

First experiences with a novel farmer citizen science approach: crowdsourcing participatory variety selection through on-farm triadic comparisons of technologies (TRICOT)

Rapid climatic and socio-economic changes challenge current agricultural R&D capacity. The necessary quantum leap in knowledge generation should build on the innovation capacity of farmers themselves. A novel citizen science methodology, triadic comparisons of technologies or tricot, was implemented in pilot studies in India, East Africa, and Central America. The methodology involves distributing a pool of agricultural technologies in different combinations of three to individual farmers who observe these technologies under farm conditions and compare their performance. Since the combinations of three technologies overlap, statistical methods can piece together the overall performance ranking of the complete pool of technologies. The tricot approach affords wide scaling, as the distribution of trial packages and instruction sessions is relatively easy to execute, farmers do not need to be organized in collaborative groups, and feedback is easy to collect, even by phone. The tricot approach provides interpretable, meaningful results and was widely accepted by farmers. The methodology underwent improvement in data input formats. A number of methodological issues remain: integrating environmental analysis, capturing gender-specific differences, stimulating farmers' motivation, and supporting implementation with an integrated digital platform. Future studies should apply the tricot approach to a wider range of technologies, quantify its potential contribution to climate adaptation, and embed the approach in appropriate institutions and business models, empowering participants and democratizing science