Blow-up behavior of collocation solutions to Hammerstein-type volterra integral equations

We analyze the blow-up behavior of one-parameter collocation solutions for Hammerstein-type Volterra integral equations (VIEs) whose solutions may blow up in finite time. To approximate such solutions (and the corresponding blow-up time), we will introduce an adaptive stepsize strategy that guarantees the existence of collocation solutions whose blow-up behavior is the same as the one for the exact solution. Based on the local convergence of the collocation methods for VIEs, we present the convergence analysis for the numerical blow-up time. Numerical experiments illustrate the analysis

SNP genotyping using alkali cleavage of RNA/DNA chimeras and MALDI time-of-flight mass spectrometry

Single nucleotide polymorphisms (SNPs) are now widely used for many DNA analysis applications such as linkage disequilibrium mapping, pharmacogenomics and traceability. Many methods for SNP genotyping exist with diverse strategies for allele-distinction. Mass spectrometers are used most commonly in conjunction with primer extension procedures with allele-specific termination. Here we present a novel concept for allele-preparation for SNP genotyping. Primer extension is carried out with an extension primer positioned immediately upstream of the SNP that is to be genotyped, a complete set of four ribonucleotides and a ribonucleotide incorporating DNA polymerase. The allele-extension products are then treated with alkali, which results in the cleavage immediately after the first added ribonucleotide. In addition, to obtain fragments easily detectable by mass spectrometry, we have included a ribonucleotide in the primer usually at the fourth nucleotide from the 3′ terminus. The method was tested on four SNPs each with a different combination of nucleotides. The advantage over other mass spectrometry-based SNP genotyping assays is that this one only requires a PCR, a primer extension reaction with a universal extension mix and an inexpensive facile cleavage reaction, which makes it overall very cost effective and easy in handling

Information recovery from low coverage whole-genome bisulfite sequencing.

The cost of whole-genome bisulfite sequencing (WGBS) remains a bottleneck for many studies and it is therefore imperative to extract as much information as possible from a given dataset. This is particularly important because even at the recommend 30X coverage for reference methylomes, up to 50% of high-resolution features such as differentially methylated positions (DMPs) cannot be called with current methods as determined by saturation analysis. To address this limitation, we have developed a tool that dynamically segments WGBS methylomes into blocks of comethylation (COMETs) from which lost information can be recovered in the form of differentially methylated COMETs (DMCs). Using this tool, we demonstrate recovery of ∼30% of the lost DMP information content as DMCs even at very low (5X) coverage. This constitutes twice the amount that can be recovered using an existing method based on differentially methylated regions (DMRs). In addition, we explored the relationship between COMETs and haplotypes in lymphoblastoid cell lines of African and European origin. Using best fit analysis, we show COMETs to be correlated in a population-specific manner, suggesting that this type of dynamic segmentation may be useful for integrated (epi)genome-wide association studies in the future

TRANSFER TEKNOLOGI PEMBENAH TANAH PADA GAPOKTAN SINAR HARAPAN DI DESA KRAWANGSARI, LAMPUNG SELATAN, SEBAGAI UPAYA PERBAIKAN KESUBURAN TANAH

Saat ini, petani di Desa Krawang Sari yang tergabung dalam Gapoktan Sinar Harapan dihadapkan dengan permasalahan kondisi lahan yang menurun status kesuburannya (pH tanah rendah). Kegiatan ini bertujuan untuk meningkatkan pengetahuan dan keterampilan anggota Gapoktan Sinar Harapan dalam penggunaan teknologi pembenah tanah sebagai upaya untuk menjaga kesuburan lahan pertanian. Kegiatan ini dilaksanakan selama empat bulan, yaitu pada bulan Mei—Agustus 2023. Proses transfer teknologi dilakukan dengan cara penyuluhan pertanian menggunakan teknik ceramah/presentasi, diskusi, demonstrasi cara, dan demonstrasi hasil. Evaluasi dilakukan dengan memberikan kuisioner yang berisi pertanyaan terkait materi penyuluhan yang telah diberikan. Perubahan kemampuan dalam menjawab pertanyaan menjadi indikator adanya perubahan pengetahuan yang dimiliki oleh petani. Sementara untuk peningkatan keterampilan petani dapat dilihat pada saat dilakukannya kegiatan praktik, baik dalam bentuk demonstrasi cara dan hasil. Dari kegiatan demonstrasi cara dan hasil yang telah dilakukan pada demplot tanah percobaan didapati adanya peningkatan pH tanah pada tanah yang telah diaplikasikan pembenah tanah, baik zeolite maupun dolomit. Pada demplot pertama, diperoleh pH tanah awal dengan nilai rata-rata 5,7 kemudian setelah diaplikasikan zeolite nilai rata-rata pH tanahnya naik menjadi 5,9. Sementara itu, pada demplot kedua diperoleh pH tanah awal dengan nilai rata-rata 5,6 kemudian setelah diaplikasikan dolomit nilai rata-rata pH tanahnya menjadi 6. &nbsp

DNA methylation in human gastric epithelial cells defines regional identity without restricting lineage plasticity

BACKGROUND: Epigenetic modifications in mammalian DNA are commonly manifested by DNA methylation. In the stomach, altered DNA methylation patterns have been observed following chronic Helicobacter pylori infections and in gastric cancer. In the context of epigenetic regulation, the regional nature of the stomach has been rarely considered in detail. RESULTS: Here, we establish gastric mucosa derived primary cell cultures as a reliable source of native human epithelium. We describe the DNA methylation landscape across the phenotypically different regions of the healthy human stomach, i.e., antrum, corpus, fundus together with the corresponding transcriptomes. We show that stable regional DNA methylation differences translate to a limited extent into regulation of the transcriptomic phenotype, indicating a largely permissive epigenetic regulation. We identify a small number of transcription factors with novel region-specific activity and likely epigenetic impact in the stomach, including GATA4, IRX5, IRX2, PDX1 and CDX2. Detailed analysis of the Wnt pathway reveals differential regulation along the craniocaudal axis, which involves non-canonical Wnt signaling in determining cell fate in the proximal stomach. By extending our analysis to pre-neoplastic lesions and gastric cancers, we conclude that epigenetic dysregulation characterizes intestinal metaplasia as a founding basis for functional changes in gastric cancer. We present insights into the dynamics of DNA methylation across anatomical regions of the healthy stomach and patterns of its change in disease. Finally, our study provides a well-defined resource of regional stomach transcription and epigenetics. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s13148-022-01406-4

Involvement of a citrus meiotic recombination TTC-repeat motif in the formation of gross deletions generated by ionizing radiation and MULE activation

Background: Transposable-element mediated chromosomal rearrangements require the involvement of two transposons and two double-strand breaks (DSB) located in close proximity. In radiobiology, DSB proximity is also a major factor contributing to rearrangements. However, the whole issue of DSB proximity remains virtually unexplored. Results: Based on DNA sequencing analysis we show that the genomes of 2 derived mutations, Arrufatina (sport) and Nero (irradiation), share a similar 2 Mb deletion of chromosome 3. A 7 kb Mutator like element found in Clemenules was present in Arrufatina in inverted orientation flanking the 5' end of the deletion. The Arrufatina Mule displayed "dissimilar" 9-bp target site duplications separated by 2 Mb. Fine-scale single nucleotide variant analyses of the deleted fragments identified a TTC-repeat sequence motif located in the center of the deletion responsible of a meiotic crossover detected in the citrus reference genome. Conclusions: Taken together, this information is compatible with the proposal that in both mutants, the TTC-repeat motif formed a triplex DNA structure generating a loop that brought in close proximity the originally distinct reactive ends. In Arrufatina, the loop brought the Mule ends nearby the 2 distinct insertion target sites and the inverted insertion of the transposable element between these target sites provoked the release of the in-between fragment. This proposal requires the involvement of a unique transposon and sheds light on the unresolved question of how two distinct sites become located in close proximity. These observations confer a crucial role to the TTC-repeats in fundamental plant processes as meiotic recombination and chromosomal rearrangements

DNA sequencing by MALDI-TOF MS using alkali cleavage of RNA/DNA chimeras

Approaches developed for sequencing DNA with detection by mass spectrometry use strategies that deviate from the Sanger-type methods. Procedures demonstrated so far used the sequence specificity of RNA endonucleases, as unfortunately equivalent enzymes for DNA do not exist and therefore require transcription of DNA into RNA prior to fragmentation