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The existing methods of callose quantification include epifluorescence microscopy and fluorescence spectrophotometry of aniline blue-stained callose particles, immuno-fluorescence microscopy and indirect assessment of both callose synthase and β-(1,3)-glucanase enzyme activities. Some of these methods are laborious, time consuming, not callose-specific, biased and require high technical skills. Here, we describe a method of callose quantification based on Sandwich Enzyme-Linked Immunosorbent Assay (S-ELISA). Tissue culture-derived banana plantlets were inoculated with Xanthomonas campestris pv. musacearum (Xcm) bacteria as a biotic stress factor inducing callose production. Banana leaf, pseudostem and corm tissue samples were collected at 14 days post-inoculation (dpi) for callose quantification. Callose levels were significantly different in banana tissues of Xcm-inoculated and control groups except in the pseudostems of both banana genotypes. The method described here could be applied for the quantification of callose in different plant species with satisfactory level of specificity to callose, and reproducibility. Additionally, the use of 96-well plate makes this method suitable for high throughput callose quantification studies with minimal sampling and analysis biases. We provide step-by-step detailed descriptions of the method.

Background: Genetic polymorphisms within the gene loci of the promoter region of tumor necrosis factor (TNF) alpha have been associated with the pathogenesis of hepatitis B virus (HBV) infection. In Uganda, there is a wide variation in the HBV endemicity, ranging from low endemicity, through moderate endemicity, to hyper-endemicity. However, the underlying reasons for this disparity in HBV burden are not fully elucidated. Thus, we aimed to test the hypothesis that the TNF-α-863C/A and -1031T/C polymorphic sites may have an effect on the difference between the burden of HBV in our country. We screened 384 participants, from which a sample of 134 was drawn, to determine the HBV, TNF-α-863C/A, and TNF-α-863T/C genotypes. The nucleotide BLAST was used to match the unknown targeted sequence obtained from the Sanger sequence against the known deposited sequence. This process unveiled the base substitution mutation and the HBV genotypes. The odds ratio (OR) and Chi-square test of proportions were used for the analysis. All the analyses were performed using SPSS version 26.0 and MedCalc software version 20.010 at 95% CI. A p < 0.05 was considered statistically significant. Results: The prevalence of both the TNF-α-863C/A and the TNF-α-1031T/C genotypes and their alleles did not differ significantly by endemicity (p > 0.05). However, the prevalence of the nucleotide substitution mutations for TNF-α-863C>A and TNF-α-1031T>C was significantly low for all the study groups (p < 0.05). Conclusion: The TNF-α gene promoter at the TNF-α-863C/A and 1031T/C positions is conserved in our population and may not affect the endemicity of HBV infection. However, future research should focus on the use of nationwide samples in order to reach concreate determinations regarding the role of the TNF-α polymorphisms in the risk/resolution of HBV infections in an African or Black population. [ABSTRACT FROM AUTHOR]

Background: Food taboos and cultural beliefs among pregnant and breast-feeding women influence their food consumption patterns and hence the health of women and unborn children. Cognizant of their neglect in programs aimed to ameliorate hidden hunger among pregnant and breast-feeding women in Buyende and other resource-poor communities in sub-Saharan Africa, we opted for a study to unravel them to inform program design., Methods: We documented food taboos and beliefs amongst pregnant and breast-feeding women from six sub-counties of Buyende district in Eastern Uganda. A mixed-methods approach was used, which was comprised of questionnaire interviews with 462 women, eight focus group discussions with 6-10 participants in each and a total of 15 key informant interviews., Results: The present study revealed that 129 (27.9%) of the respondents practice food taboos and adhere to cultural beliefs related to their dietary habits during pregnancy and breast-feeding that are fuelling the prevalence of hidden hunger. The most tabooed foods during pregnancy were sugarcane (17.8%), fishes which included lung fish, catfish and the Lake Victoria sardine (Rastrineobola argentea) (15.2%), oranges (6.6%), pineapples (5.9%), eggs (3.3%), chicken (3.3%) and cassava, mangoes and Cleome gynandra (each at 3%). Most foods were avoided for reasons associated with pregnancy and labour complications and undesirable effects on the baby. Most women learnt of the taboos and beliefs from the elders, their own mother, grandparents or mother-in-law, but there was also knowledge transmission in social groups within the community., Conclusions: The taboos and cultural beliefs in the study area render pregnant and breast-feeding women prone to micronutrient deficiency since they are denied consumption of a diversity of nutritious foods. There is a need to educate such women about consumption of nutrient-rich foods like fish, eggs, fruits and vegetables in order to improve their health, that of the unborn and children being breast fed. Additionally, culturally appropriate nutrition education may be a good strategy to eliminate inappropriate food taboos and beliefs with negative impact on the health of pregnant and breast-feeding women., (© 2024. The Author(s).)